Professional Documents
Culture Documents
Ectomycorrhizal
Ectomycorrhizal
Ectomycorrhizal
Ectomycorrhizal (ECM) symbiosis a mutualistic
and Neotropical
maintenance and evolution of biodiversity and ecosystems
Neotropical
and
ecology of forest trees,trees,
affecting growth, waterwater
and nutrient
ecology of forest affecting growth, and nutrient
absorption and protection against pathogens. It is a research
absorption and protection against pathogens. It is aItresearch
Symbioses
absorption and protection against pathogens. is a research
imperative in tropical and neotropical forest ecosystems
imperative in tropical
imperative and neotropical
in tropical and neotropical forestforest
because they concern an ecologically and economically
ecosystems
ecosystems Neotropical
Neotropical
Neotropical Forests
Forests
Forests
Forests
because they they
concern an ecologically and and economically
because concern an ecologically economically
Forests
important tree species (e.g. Ceasalpinioid subfamily in Africa,
important tree species (e.g. Ceasalpinioid subfamily in Africa,
important tree species (e.g. Ceasalpinioid subfamily in Africa,
in Tropical
Dipterocarpaceae in Asia). The book is an overview of the
Dipterocarpaceae in Asia). The book is an isoverview of theof the
Dipterocarpaceae in Asia). The book an overview
knowledge of ECM symbioses in tropical and neotropical
knowledge of ECM symbioses in tropical and neotropical
knowledge of ECM symbioses in tropical and neotropical
ecosystem forests. The contents address diversity and function
ecosystem forests. The contents address diversity and function
ecosystem forests. The contents address diversity and function
of ectomycorrhiza associated with forest plants, impacts of
of ectomycorrhiza associated with with
forestforest
plants, impacts of of
of ectomycorrhiza associated plants, impacts
ectomycorrhiza on plant diversity and composition,
ectomycorrhiza on plant diversity and and composition,
ectomycorrhiza on plant diversity composition,
regeneration and dynamics of ecosystems, and biomass
regeneration and anddynamics of ecosystems, and andbiomass
regeneration dynamics of ecosystems, biomass
Editors Krista
Editors
production in forestry, adaptation of ectomycorrhiza to
EditorsAmadou
production in forestry, adaptation of ectomycorrhiza to to
production in forestry, adaptation of ectomycorrhiza
nutrient deficient, salt and ultramafic soils.
Abdala G.
nutrient deficient, salt andsaltultramafic soils. soils.
Krista
nutrient deficient, and ultramafic Abdala G. Diédhiou
Amadou
Krista M.
Abdala
Amadou M. Bâ
Editors
Editors
L. McGuire Editors
L. Diédhiou
L. McGuire
AmadouM. M.Bâ
Bâ
G. Diédhiou
Amadou
M.
McGuire
Amadou M. Bâ
Bâ
Bâ
KristaL.L.McGuire
Krista McGuire
Krista L. McGuire
AbdalaG.G.Diédhiou
Abdala Diédhiou
K20685
Abdala G. Diédhiou
6000 Broken Sound Parkway, NW
Suite 300, Boca Raton, FL 33487
711 Third Avenue
New York, NY 10017 9 781 466 59 468 5
an informa business 2 Park Square, Milton Park 9 781 466 59 468 5
Abingdon, Oxon OX14 4RN, UK 9 781 466 59 468 5 A SCIENCE PUBLISHERS BOOK
w w w. c rc p r e s s . c o m
Ectomycorrhizal Symbioses
in
Tropical and Neotropical Forests
Ectomycorrhizal Symbioses
in
Tropical and Neotropical Forests
Editors
Amadou M. Bâ
Université Antilles-Guyane
Guadeloupe (French West Indies)
France
Krista L. McGuire
Barnard College
Columbia University
New York
USA
Abdala G. Diédhiou
Université Cheikh Anta Diop
Dakar
Sénégal
p,
A SCIENCE PUBLISHERS BOOK
CRC Press
Taylor & Francis Group
6000 Broken Sound Parkway NW, Suite 300
Boca Raton, FL 33487-2742
© 2014 by Taylor & Francis Group, LLC
CRC Press is an imprint of Taylor & Francis Group, an Informa business
This book contains information obtained from authentic and highly regarded sources. Reasonable
efforts have been made to publish reliable data and information, but the author and publisher cannot
assume responsibility for the validity of all materials or the consequences of their use. The authors and
publishers have attempted to trace the copyright holders of all material reproduced in this publication
and apologize to copyright holders if permission to publish in this form has not been obtained. If any
copyright material has not been acknowledged please write and let us know so we may rectify in any
future reprint.
Except as permitted under U.S. Copyright Law, no part of this book may be reprinted, reproduced,
transmitted, or utilized in any form by any electronic, mechanical, or other means, now known or
hereafter invented, including photocopying, microfilming, and recording, or in any information stor-
age or retrieval system, without written permission from the publishers.
For permission to photocopy or use material electronically from this work, please access www.copy-
right.com (http://www.copyright.com/) or contact the Copyright Clearance Center, Inc. (CCC), 222
Rosewood Drive, Danvers, MA 01923, 978-750-8400. CCC is a not-for-profit organization that pro-
vides licenses and registration for a variety of users. For organizations that have been granted a pho-
tocopy license by the CCC, a separate system of payment has been arranged.
Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are
used only for identification and explanation without intent to infringe.
Visit the Taylor & Francis Web site at
http://www.taylorandfrancis.com
and the CRC Press Web site at
http://www.crcpress.com
Foreword
Marc-André SELOSSE
Professor at Muséum national d’Histoire naturelle and
President of the Société botanique de France
Observatoire de la Structure et de l’Evolution de la Biodiversité
(UMR7205 CNRS – Muséum), Département Systématique et Evolution,
45 rue Buffon, 75005 Paris, France
Preface
Amadou M. Bâ
Krista L. McGuire
Abdala G. Diédhiou
Contents
Foreword v
Preface ix
1. Diversity and Community Structure of Ectomycorrhizal 1
Fungi in Mixed and Monodominant African Tropical
Rainforests
Abdala Gamby Diedhiou, Helvyne Christelle Michaella Ebenye,
Marc-André Selosse, Nérée Onguene Awana and Amadou Mustapha Bâ
2. Ectomycorrhizas of Three Species of Nyctaginaceae in the 19
Tropical Mountain Rain Forest of South Ecuador
Ingeborg Haug, Ingrid Kottke and Juan Pablo Suárez
3. Diversity and Abundance of Ectomycorrhizal Associations 29
in Rain Forests of Cameroon under Different Disturbance
Regimes
Onguene Awana Nérée, Judith Marthiale Tsamo, Christelle Michaella
Ebenye, Amadou Mustapha Bâ and Thomas Kuyper
4. Mycorrhizal Fungi Diversity and their Importance on the 51
Establishment of Native Species Seedlings within
Madagascarian Degraded Sclerophyllous Forest
Rondro Harinisainana Baohanta, Herizo Andrianantoandro
Randriambanona, Marc Ducousso, Christophe Nirina Rakotoarimanga,
Yves Prin, Heriniaina Ramanankierana and Robin Duponnois
5. Morpho-anatomical Characterization of Three 79
Sebacinales Ectomycorrhizal Species from a Pakaraimaea
dipterocarpacea ssp. nitida (Dipterocarpaceae) Forest in
Southern Venezuela
Bernard Moyersoen
xiv Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
1. Introduction
Mycorrhizal symbioses play a prominent role in the biology and ecology
of forest trees. They involve soil fungi and roots of trees, which together as
a symbiosis provide the fungi with carbohydrates and enhance the uptake
of water and nutrients for the trees, and also have a major protective role
for the roots (Smith and Read 2008). Forest trees are primarily associated
with two types of mycorrhizas: arbuscular mycorrhizas which include fungi
from the phylum of Glomeromycota and ectomycorrhizas (EcMs) mainly
formed by members of Ascomycota and Basidiomycota. It is currently
estimated that 6,000–10,000 plant species (Smith and Read 2008, Brundrett
2009) and 20,000–25,000 fungal species (Rinaldi et al. 2008) are involved in
Fig. 1. Distribution of EcM trees in tropical Africa: (1) rainforests in the Guinea-Congo region; (2)
open forests in the Sudanian and Zambezian regions; (3) savanna woodlands in the Sudanian
and Zambezian regions (Bâ et al. 2012).
Color image of this figure appears in the color plate section at the end of the book.
Ectomycorrhizal Fungi in African Tropical Rainforests 3
2. Site Description
The mixed forest was located in Southern Guinea, one of the last regions
of West Africa retaining a primary tropical rainforest. Above- and below-
ground EcM fungal diversity surveys were conducted in typical evergreen
rainforests covering hills and mountains ranging in altitude from 500
m in the Ziama forest (8°51′N, 9°31'W) to 1,752 m on the Mount Nimba
forest (7°60'N, 8°49'W). The evergreen rainforests are characterized by a
mean annual rainfall of 2,500–3,000 mm and a dry season (mean rainfall
<15 mm) from January to March. Temperatures are generally above 24°C
with relative humidity up to 80%. The soils are generally poor, lateritic
and prone to heavy leaching (McGinley 2008). Canopy trees are at least 30
m tall, with some emergent individuals reaching 50–60 m in height. The
canopy is dominated by Cryptosepalum tetraphyllum (EcM Caesalpinioideae,
Fabales), as well as the AM trees Erythrophleum ivorense (Fabaceae) and
Heritiera utilis (Malvaceae, Malvales). The other EcM trees are represented
by Caesalpinioideae (Afzelia bella, Anthonotha fragans, A. macrophylla,
Gilbertiodendron limba, Paramacrolobium coeruleum and Pelligriniodendron
diphyllum), and Phyllanthaceae (a family formerly classified within
Euphorbiaceae: Uapaca esculenta, U. chevalieri, U. guineensis and Uapaca
heudelotii). Some of these Caesalpinioideae and Phyllanthaceae tree species
grow in mixed patches where they dominate (Rivière et al. 2007, Diédhiou et
al. 2010). Sporocarps (fruit-bodies) and ECMs of mature trees and seedlings
were collected within 3 plots of 0.5–1 ha each in the mixed forest. For the
mature trees, root tips were sampled around trees by tracing roots from the
base of trunk whenever possible.
The monodominant forest of G. dewevrei is located in the Dja Faunal
Reserve (2°49’ to 3°23’N; 12°25’ to 13°35’E) in the Southeastern region of
Cameroon, which extends from Mouloundou to south of Bertoua. The
climate is characterized by two wet seasons from March to June and from
August to November with a mean annual rainfall varying from 1600 to 1700
mm. The two dry seasons are December–February (18–59 mm) and July (54
mm). Mean annual temperature varies from 23.7° to 24.3°C. Soils are clayey
ferralitic, acidic, and poor in nutrients (Peh et al. 2011). The vegetation has a
main canopy of 30–40 m with highest trees rising to 60 m (Letouzey 1985).
The monodominant forests are surrounded by mixed forests including
many tree and shrub species, which form mainly arbuscular mycorrhiza
(e.g., Afrostyrax lepidophyllus, Afzelia bipendensis, Anthonotha ferruginea,
Baphia pubescens, Beilschmiedia louisii, Cryptosepalum congolum, Drypetes paxii,
Ectomycorrhizal Fungi in African Tropical Rainforests 5
Table 1. contd....
8
Table 1. contd.
Type of forest: Mixed Monodominant
Fungi:
Phylum Class/EcM lineage Family/Genus sporocarps EcMs sporocarps EcMs
/ramaria-gautieria Ramaria +
/suillus-rhizopogon Truncocolumella +
Agaricomycetes Chalciporus + +
Geastrum +
Polyporaceae +
Tremellomycetes Trichosporon +
Tremellaceae +
Ascomycota /marcelleina-peziza Marcelleina +
gerardii
/tuber-helvella Helvella +
/helotiales Helotiales +
/elaphomyces Elaphomyces +
Sordariomycetes Nectriaceae +
Chaetosphaeria +
Cercophora +
Sordariomycete +
Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Hypocreaceae +
Leotiomycetes Leptodontidium +
Leohumicola +
Hyaloscyphaceae +
Dothideomycetes Botryosphaericeae +
Leptosphaeria +
Zygomycota Zygomycetes Mortierellaceae +
Ectomycorrhizal Fungi in African Tropical Rainforests 9
Fig. 2. Relative abundance of the EcM-forming lineages revealed from EcMs and sporocarps
collected from each forest type.
10 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Tedersoo et al. 2010a, 2011, Jairus et al. 2011). In regard to this observation
and because the members of this lineage produce inconspicuous sporocarps,
one could argue that they might have been missed in the mixed forest where
relatively few samples were collected. However, some EcM-forming lineages
such as /pisolithus-scleroderma and /tricholoma were more abundant
in the mixed forest (accounting for 11.98% and 5.53%, respectively) than
in the monodominant forest where they accounted for < 1% each (Fig.
2). Thus, although possibly underestimating the number and abundance
of EcM-forming lineages in the mixed forest, our results show that the
monodominant forest tends to harbor more EcM-forming lineages.
In terms of species richness, the lineages of /russula-lactarius (86 spp.),
/tomentella-thelephora (30 spp.), /sebacina (19 spp.), and /boletus (13 spp.)
dominated in the monodominant forest, while the /russula-lactarius (42
spp.), /tomentella-thelephora (21 spp.), /boletus (20 spp.), and /amanita (15
spp.) dominated in the mixed forest. Other species-rich lineages included
the /pisolithus-scleroderma (10 spp.) and /tricholoma (10 spp.) in the
mixed forest and the /clavulina (11 spp.) and /amanita (10 spp.) in the
monodominant forest (Fig. 3).
The EcM-forming lineages included 189 and 126 putative EcM species
for the monodominant forest and mixed forest, respectively (Table 2).
The high EcM species richness observed from these African forests is a
Fig. 3. Number of putative EcM fungal species identified from EcMs and sporocarps collected
from each forest type.
12 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 2. Number (No.) of EcM samples observed and estimated EcM species richness in each
forest type. *The numbers appearing in bold were obtained after rarefaction to 217.
common trend in Neotropical (Morris et al. 2009, Smith et al. 2011, 2013,
Henkel et al. 2012) and Paleotropical forests (Peay et al. 2010, Tedersoo et
al. 2010b, 2011). The calculation of sample-based rarefaction curves and
minimal species richness estimates showed that the sampling efforts were
insufficient in both types of forests. Indeed, the rarefied accumulation
curves of species and minimal species richness estimates (Chao2, Jack2 and
ICE) did not reach a clear asymptote with increasing sample size (Fig. 4).
The Chao2, Jack2 and ICE richness estimators predicted 249.08, 259.25 and
282.2 putative EcM species, respectively for the mixed forest and 575.42,
440.97 and 675.07 putative EcM species, respectively for the monodominant
forest (Table 2).
Surprisingly, when rarefying the samples from the monodominant
forest to n = 217 to match the sample size of the mix from the mixed forest,
we found no significant difference in terms of richness of putative EcM
fungal species between the two forests types (Table 2, Fig. 4). Indeed, the
95% CI (confidence intervals) for Chao2 are overlapping and the indices of
species diversity, Fisher’s alpha and Shannon are not significantly different
between the two types of forests (Table 2). This lack of difference in richness
of EcM fungal species between the mixed forest and monodominant forest
may be in part related to the prevalence of multi-host EcM fungi (Onguene
and Kuyper 2002, Diédhiou et al. 2010) which could mask the influence of
plant host species on EcM fungal community structure. Interestingly, this
rejects the generalization of the idea that host diversity is the sole driver
of diversity in EcM community (Dickie 2007, Ishida et al. 2007, Tedersoo
Ectomycorrhizal Fungi in African Tropical Rainforests 13
Fig. 4. Rarefied accumulation curves of (a) EcM fungal species and (b) minimal species richness
estimates for the monodominant forest (MoF, black) and mixed forest (MiF, grey). Dotted lines
in (a) indicate 95% CI.
14 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
et al. 2008). This idea may be supported by the finding that African EcM
fungal communities are not strongly structured by soil horizon and host
at the plant species and family levels (Tedersoo et al. 2011).
5. Conclusion
This study has provided valuable information on the diversity and
community structure of EcM fungi from mixed and monodominant forests
in tropical Africa. The monodominant forest tends to harbor more EcM-
forming lineages than the mixed forest, while there was no significant
difference in terms of richness of EcM fungal species between these two
forest types. The dominant EcM-forming lineages are similar in the two
forest types. On the other hand, a large number of fungi of unknown trophic
status were recovered from healthy EcMs; some of them may represent new,
overlooked tropical EcM-forming lineages. From this finding, it should be
evident that further research is necessary to (i) determine the EcM lifestyle
of the latter fungal taxa, (ii) rigorously assess and compare the phylogenetic
community composition between the two forest types, and (ii) highlight
the role of EcM fungi in the recruitment and establishment of seedlings of
EcM tree species in monodominant and mixed forests.
Acknowledegments
We thank the anonymous referees for their valuable comments on this study,
and Krista L. McGuire and Caitlyn Gillikin for improving the language.
References
Alexander, I.J. 1989. Mycorrhizas in tropical forests. In: J. Proctor (ed.). Mineral Nutrients in
Tropical Forest and Savanna Ecosystems. Blackwell Scientific Publications, Oxford, pp.
169–188.
Alexander, I.J. 2006. Ectomycorrhizas-out of Africa? New Phytol. 172: 589–591.
Alexander, I.J. and S.S. Lee. 2005. Mycorrhizas and ecosystem processes in tropical rain
forest: implications for diversity. In: D. Burslem, M. Pinard and S. Hartley (eds.). Biotic
Interactions in the Tropics. Cambridge University Press, Cambridge, pp. 165–203.
Altschul, S.F., W. Gish, W. Miller, E.W. Myers and D.J. Lipman. 1990. Basic local alignment
search tool. J. Mol. Biol. 215: 403–410.
Avis, P.G., D.J. McLaughlin, B.C. Dentinger and P.B. Reich. 2003. Long-term increase in nitrogen
supply alters above- and below-ground ectomycorrhizal communities and increases the
dominance of Russula spp. in a temperate oak savanna. New Phytol. 160: 239–253.
Bâ, A.M., J. Garbaye and J. Dexheimer. 1991. Influence of fungal propagules during the
early stade of the time sequence of ectomycorrhizal colonization on Afzelia africana Sm.
Seedlings. Can. J. Bot. 66: 2442–2447.
Bâ, A.M. and D. Thoen. 1990. First syntheses of ectomycorrhizas between Afzelia africana Sm.
(Caesalpinioideae) and native fungi from West Africa. New Phytol. 103: 441–448.
Ectomycorrhizal Fungi in African Tropical Rainforests 15
Bâ, A.M., R. Duponnois, B. Moyersoen and A.G. Diedhiou. 2012. Ectomycorrhizal symbiosis
of tropical African trees. Mycorrhiza 22: 1–29.
Bonito, G.M, A.P. Gryganskyi, J.M. Trappe and R. Vilgalys. 2010. A global meta-analysis
of Tuber ITS rDNA sequences: species diversity, host associations and long-distance
dispersal. Mol. Ecol. 19: 4994–5008.
Brundrett, M.C. 2009. Mycorrhizal associations and other means of nutrition of vascular plants:
understanding global diversity of host plants by resolving conflicting information and
developing reliable means of diagnosis. Plant Soil 320: 37–77.
Bruns, T.D., M.I. Bidartondo and D.L. Taylor. 2002. Host specificity in ectomycorrhizal
communities: what do the exceptions tell us? Integr. Comp. Biol. 42: 352–359.
Burleigh, J.G. and S. Matthews. 2004. Phylogenetic signal in nucleotide data from seed plants:
implications for resolving the seed plant tree of life. Am. J. Bot. 91: 1599–1613.
Buyck, B., D. Thoen and R. Walting. 1996. Ectomycorrhizal fungi of the Guinea–Congo region.
Proc. R. Soc. Edinb. 104: 313–333.
Colwell, R.K. 2006. Estimates: Statistical Estimation of Species Richness and Shared Species
from Samples, Version 8 [WWW document]. URL http://purl.oclc.org/estimates.
Comandini, O., A.C. Rinaldi and T.W. Kuyper. 2012. Measuring and estimating ectomycorrhizal
fungal diversity: a continuous challenge. In: M.C. Pagano (ed.). Mycorrhiza Occurrence
in Natural and Restored Environments. Nova Science Publishers, Hauppauge, NY, pp.
165–200.
de Roman, M., V. Claveria and A.M. de Miguel. 2005. A revision of the descriptions of
ectomycorrhizas published since 1961. Mycol. Res. 109: 1063–1104.
Dickie, I.A. 2007. Host preference, niches and fungal diversity. New Phytol. 174: 230–233.
Diédhiou, A.G., A.M. Bâ, S. Sylla, B. Dreyfus, M. Neyra and I. Ndoye. 2004. The early-stage
ectomycorrhizal Thelephoroid fungal sp. is competitive and effective on Afzelia africana
Sm. in nursery conditions in Senegal. Mycorrhiza 14: 313–322.
Diédhiou, A.G., O. Guèye, M. Diabaté, Y. Prin, R. Duponnois, B. Dreyfus and A.M. Bâ. 2005.
Contrasting responses to ectomycorrhizal inoculation in seedlings of six tropical African
tree species. Mycorrhiza 16: 11–17.
Diédhiou, A.G., J.-L. Dupouey, M. Buée, E. Dambrine, L. Laüt and J Garbaye. 2009. Response
of ectomycorrhizal communities to past Roman occupation in an oak forest. Soil Biol.
Bioch. 41: 2206–2213.
Diédhiou, A.G., M.-A. Selosse, A. Galiana, M. Diabaté, B. Dreyfus, A.M. Bâ, S.M. de Faria
and G. Béna. 2010. Multi-host ectomycorrhizal fungi are predominant in a Guinean
tropical rain forest and shared between canopy and tree seedlings. Environ. Microbiol.
2: 2219–2232.
Hart, T.B. 1995. Seed, seedling and sub-canopy survival in monodominant and mixed forests
of the Ituru forest, Africa. J. Trop. Ecol. 11: 443–459.
Hart, T.B., J.A. Hart and P.G. Murphy. 1989. Monodominant and species-rich forests of the
humid tropics: causes for their occurrence. Am. Nat. 133: 613–633.
Henkel, T.W., M.C. Aime, M.M.L. Chin, S.L. Miller, R. Vilgalys and M.E. Smith. 2012.
Ectomycorrhizal fungal sporocarp diversity and discovery of new taxa in Dicymbe
monodominant forests of the Guiana Shield. Biodivers. Conserv. 21: 2195–2220.
Högberg, P. and G.D. Piearce. 1986. Mycorrhizas in Zambian trees in relation to host taxonomy,
vegetation type and successional patterns. J. Ecol. 74: 775–785.
Hughes, K.W., R.H. Peterson and E.B. Lickey. 2009. Using heterozygosity to estimate a
percentage DNA sequence similarity for environmental species’ delimitation across
basidiomycete fungi. New Phytol. 182: 795–798.
Ishida, T.A., K. Nara and T. Hogetsu. 2007. Host effects on ectomycorrhizal fungal communities:
insight from eight host species in mixed conifer-broadleaf forests. New Phytol. 174:
430–440.
Jairus, T., R. Mpumba, S. Chinoya and L. Tedersoo. 2011. Invasion potential and host shifts
of Australian and African ectomycorrhizal fungi in mixed eucalypt plantations. New
Phytol. 192: 179–187.
16 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Rinaldi, A.C., O. Comadini and T.W. Kuyper. 2008. Ectomycorrhizal fungal diversity: separating
the wheat from the chaff. Fungal Divers 33: 1–45.
Rivière, T., A.G. Diédhiou, M. Diabaté, G. Senthilarasu, K. Natarajan, A. Verbeken, B. Buyck, B.
Dreyfus, G. Bena and A.M. Bâ. 2007. Genetic diversity of ectomycorrhizal basidiomycetes
from African and Indian tropical forests. Mycorrhiza 17: 415–428.
Sanon, K.B., A.M. Bâ and J. Dexheimer. 1997. Mycorrhizal status of some fungi fruiting beneath
indigenous trees in Burkina Faso. For. Ecol. Manag. 98: 61–69.
Selosse, M.-A., R. Bauer and B. Moyersoen. 2002. Basal hymenomycetes belonging to the
Sebacinaceae are ectomycorrhizal on temperate deciduous trees. New Phytol. 155:
183–195.
Selosse, M.-A., F. Richard, X. He and S.W. Simard. 2006. Mycorrhizal networks: des liaisons
dangereuses? Trends Ecol. Evol. 21: 621–628.
Selosse, M.-A., S. Setaro, F. Glatard, F. Richard, C. Urcelay and M. Weiß. 2007. Sebacinales are
common mycorrhizal associates of Ericaceae. New Phytol. 174: 864–878.
Selosse, M.-A., M.-P. Dubois and N. Alvarez. 2009. Do Sebacinales commonly associate with
plant roots as endophytes? Mycol. Res. 113: 1062–1069.
Smith, M.E., G.W. Douhan and D.M. Rizzo. 2007. Intra-specific and intrasporocarp ITS variation
of ectomycorrhizal fungi as assessed by rDNA sequencing of sporocarps and pooled
ectomycorrhizal roots from a Quercus woodland. Mycorrhiza 18: 15–22.
Smith, M.E., T.W. Henkel, M.C. Aime, A.K. Fremier and R. Vilgalys. 2011. Ectomycorrhizal
fungal diversity and community structure on three co-occurring leguminous canopy tree
species in a Neotropical rainforest. New Phytol. 192: 699–712.
Smith, M.E., T.W. Henkel, J.K. Uehling, A.K. Fremier, H.D. Clarke and R. Vilgalys. 2013.
The Ectomycorrhizal Fungal Community in a Neotropical Forest Dominated by the
Endemic Dipterocarp Pakaraimaea dipterocarpacea. PLoS ONE 8: e55160. doi:10.1371/
journal.pone.0055160.
Smith, S. and J. Read. 2008. Mycorrhizal Symbiosis. Third edition: 800pp.
Tedersoo, L. and K. Nara. 2010. General latitudinal gradient of biodiversity is reversed in
ectomycorrhizal fungi. New Phytol. 185: 351–354.
Tedersoo, L., T. Suvi, K. Beaver and U. Kõljalg. 2007. Ectomycorrhizal fungi of the
Seychelles: diversity patterns and hosts shifts from the native Vateriopsis seychellarum
(Dipterocarpaceae) and Intsia bijuga (Caesalpinioideae) to the introduced Eucalyptus
robusta (Myrtaceae), but not Pinus caribea (Pinaceae). New Phytol. 175: 321–333.
Tedersoo, L., J. Teele, B.M. Horton, K. Abarenkov, T. Suvi, I. Saar and U. Kõljalg. 2008. Strong
host preference of ectomycorrhizal fungi in a Tasmanian wet sclerophyll forest as revealed
by DNA barcoding and taxon-specific primers. New Phytol. 180: 479–490.
Tedersoo, L., R. Nilsson, K. Abarenkov and T. Jairus. 2010a. 454 Pyrosequencing and Sanger
sequencing of tropical mycorrhizal fungi provide similar results but reveal substantial
methodological biases. New Phytol. 188: 291–301.
Tedersoo, L., T.W. May and M.E. Smith. 2010b. Ectomycorrhizal lifestyle in fungi: global
diversity, distribution, and evolution of phylogenetic lineages. Mycorrhiza 20:
217–263.
Tedersoo, L., M. Bahram, T. Jairus, E. Bechem, S. Chinoya, R. Mpumba, M. Leal, E.
Randrianjohany, S. Razafimandimbison, A. Sadam, T. Naadel and U. Kõljalg. 2011. Spatial
structure and the effects of host and soil environments on communities of ectomycorrhizal
fungi in wooded savannas and rain forests of Continental Africa and Madagascar. Mol.
Ecol. 20: 3071–3080.
Tedersoo, L., M. Bahram, M. Toots, A.G. Diédhiou, T.W. Henkel, R. Kjoller, M.H. Morris,
K. Nara, E. Nouhra, K.G. Peay, S. Põlme, M. Ryberg, M.E. Smith and U. Kõljalg. 2012.
18 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
1
Département de Biologie Végétale, Université Cheikh Anta Diop (UCAD), BP 5005 Dakar,
Sénégal.
2
Laboratoire Commun de Microbiologie IRD/UCAD/ISRA, Centre de Recherche de Bel Air,
BP. 1386, Dakar, Sénégal.
3
Centre d’Ecologie Fonctionnelle et Evolutive (CNRS, UMR 5175), 1919 Route de Mende, 34
293 Montpellier cedex 5, France.
4
Muséum national d’Histoire naturelle, Département Systématique et Evolution (UMR 7205
OSEB), 45 rue Buffon, 75005 Paris, France.
5
Institut de Recherche Agronomique pour le Développement, BP. 2123, Yaoundé,
Cameroun.
6
Laboratoire des Symbioses Tropicales et Méditerranéennes (LSTM), UMR113, Campus de
Baillarguet, A10/J, 34398 Montpellier, Cedex 5, France.
*Corresponding author: fungalmolecol@gmail.com
CHAPTER
2
Ectomycorrhizas of Three
Species of Nyctaginaceae in
the Tropical Mountain Rain
Forest of South Ecuador
Ingeborg Haug,1,* Ingrid Kottke1 and Juan Pablo Suárez2
1. Introduction
Arbuscular mycorrhizas are the most ancient mycorrhizal association
(Taylor 1995) and are found in more than 80% of the plant species (Smith
and Read 2008). In a limited number of plant groups, the arbuscular
association was replaced by an ectomycorrhizal association (Smith and
Read 2008). Certain families such as Pinaceae and Fagaceae exclusively have
ectomycorrhizal associations, but there are also several isolated lineages
of ectomycorrhization in other families (Brundrett 2002). For instance, in
the Nyctaginaceae, only a few ectomycorrhiza forming species are known;
these are found in Neea Ruiz & Pav. (Harley and Smith 1983), Guapira Aubl.
(syn Torrubia, Harley and Smith 1983) and Pisonia L. (Ashford and Allaway
1982). All three genera belong to the tribe Pisonieae (Bittrich and Kühn
1
Institute of Evolution and Ecology, Plant Evolutionary Ecolology, Eberhard-Karls-University
Tübingen, Auf der Morgenstelle 1, D-72076 Tübingen, Germany.
2
Departamento de Ciencias Naturales, Universidad Técnica Particular de Loja, Loja,
Ecuador.
*Corresponding author: ingeborg.haug@uni-tuebingen.de
20 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
The root system consists of long and short roots. Ectomycorrhizas are
formed on short roots (diameter 0.2 to 0.6 mm). Five different mycorrhizal
morphotypes were distinguished (Fig. 1a–e). All morphotypes revealed the
typical features of ectomycorrhizas with a hyphal mantle and Hartig net
located between rhizodermal cells (Haug et al. 2005).
Fig. 1. Micrographs of mycorrhizas of Neea species 1 (a-e), Neea aff. floribunda, Pisonia sp. (g-h),
a. Russula puiggarii − Neea species 1, b. Russula-Lactarius − Neea species 1, c. Tomentella-Thelephora
species 1 − Neea species 1, d. Tomentella-Thelephora species 2 − Neea species 1, e. Ascomycete −
Neea species 1, f. Tomentella-Thelephora species 3 − Neea aff. floribunda, g.,h. Tomentella-Thelephora
species 3 − Pisonia sp., g. overview: long roots partially with hyphal mantle (arrows), partially
with root hairs and no hyphal mantle (*), h. one root shown at higher magnification: distal
portion with root hairs (*), proximal portion with hyphal mantle (arrows). Scale bars: a.,b.,f.,h.
1 mm; c.-e. 0,5 mm; g. 5 mm. Data from Haug et al. 2005.
Color image of this figure appears in the color plate section at the end of the book.
22 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
The root system of Neea aff. floribunda consists of brown, straight long
roots which bear bright, unramified fine roots. The fine roots (diameter
0.3 to 0.6 mm) are partially covered with a hyphal mantle (Fig. 1f). Only
one morphotype with a brown and smooth hyphal mantle was found. The
outer mantle layers are loosely plectenchymatous (Fig. 2a) while the middle
Fig. 2. Longitudinal sections of mycorrhizas of Neea aff. floribunda. (a) Outer hyphal mantle:
loose plectenchyma, (b) middle and inner hyphal mantle: dense plectenchyma, (c) tangential
section of the epidermal layer: epidermal outgrowths (*) and Hartig net, d,e. median section
through mycorrhiza: hyphal mantle, epidermal cells with outgrowths (*) and Hartig net, f.
hyphal mantle, Hartig net and intracellular hyphae (with clamps, →) in epidermal and cortical
cells (scale bars: a. - f. 15 µm). Data from Haug et al. 2005.
Ectomycorrhizas of Nyctaginaceae 23
The fine root systems of Pisonia consist of only long roots. There are sections
of the long roots (diameter 0.3 to 0.6 mm) covered with a hyphal mantle
and sections without hyphal mantle but with root hairs (Figs. 1g–h). Root
tips covered with a hyphal mantle are rare (Fig. 1h) and many roots have
no hyphal mantle at all. Only one morphotype is distinguishable. The
hyphal mantle is dark brown with silvery patches on the surface (Figs. 1g,
h). Emanating hyphae are colourless and bear clamps. The hyphal mantle
is plectenchymatous throughout, where the outer layers are loosely (Fig.
3a) and the middle layers are densely plectenchymatous (Fig. 3b). The
epidermal layer develops a Hartig net (Figs. 3c–e). Root segments without
hyphal mantle have root hairs. Loose hyphae and intracellular colonization
in the root hairs and in the epidermal cells are visible (Figs. 3f–g). Since
ultrastructural details are the same as in the hyphae of the Hartig net and
hyphal mantle (Figs. 3h-i), we conclude that the intracellular infection is due
to the ectomycorrhizal fungus. All mycorrhizas from four individuals of the
species of Pisonia had identical ITS sequences (AY667420-423). Phylogenetic
analyses of the rDNA ITS region revealed a membership within the genera
Thelephora/Tomentella (Haug et al. 2005).
Morphological and molecular results from ectomycorrhizas of the three
Nyctaginacean species are summarized in Table 1.
Fig. 3. Longitudinal sections of mycorrhizas of Pisonia sp. (a) Outer hyphal mantle: loose
plectenchyma, (b) middle hyphal mantle: dense plectenchyma, (c,d). tangential sections of the
Hartig net layer adjacent to the outer walls of the epidermal cells, (e) median section through
mycorrhiza: hyphal mantle and prominent Hartig net, (f) tangential section of the epidermal
layer: intracellular hyphae, (g) median section: intracellular hyphae in epidermal cells and
root hairs; (h,i) transmission electron micrographs of hyphal mantle hyphae, intracellular
hyphae in cortical cells and root hairs (scale bars: a. - g. 15 µm, h. - i. 2,5 µm). Data from Haug
et al. 2005.
(Ashford and Allaway 1985). The differences in the root systems correspond
to differing degrees of mycorrhization. While mycorrhization of rootlets
was close to 100% in species forming short root systems (Neea species 1,
Neea obovata, Guapira sancarlosiana; Moyersoen 1993 and results of this
study), species with long root systems showed an incomplete development
of the hyphal mantle and no suppression of root hair formation (Pisonia
grandis, Neea robusta, Neea species 2, Pisonia sp., Ashford and Allaway 1985,
Moyersoen 1993, and results presented here). The combination of long root
systems that are only partly transformed into ectomycorrhizas, with root
Ectomycorrhizas of Nyctaginaceae 25
Read 2008, Bahram et al. 2012). This may be because of the low amount of
ectomycorrhizal trees in these tropical forests (3 out of 115 investigated tree
species; Kottke and Haug 2004) and because of the scattered distribution
of individual trees. The sampling effort is still low, but species of Neea,
Pisonia, and Guapira have fungal partners that mainly belong to three
lineages (Tomentella-Thelephora, Russula-Lactarius, Clavulina) and several
of the ectomycorrhizal fungi preferred one host genus (Haug et al. 2005,
Tedersoo et al. 2010), a feature uncommon in boreal forests.
The high frequency of Thelephoraceae as fungal partners is
conspicuous. Many species of Thelephoraceae are saprotrophic (Brundrett
et al. 1996) and it is possible that ectomycorrhizal species may also exhibit
saprotrophic activity in order to survive periods without a host. Some of
the thelephoraceous sequences of the different Nyctaginacean hosts were
scattered throughout the phylogram of Thelephoraceae but others were
clustered as sister species (Suvi et al. 2010).
3. Conclusion
Although the common ancestor of Nyctaginaceae may have been
involved with one thelephoroid taxon, coevolution of Nyctaginaceae and
ectomycorrhizal Thelephoraceae in different habitats may have led to the
evolution of several different species. It will be a future challenge to study
the mycorrhizal status of additional species of Nyctaginaceae in South
America and identify their mycobionts to clarify the question of potential
co-evolution.
Acknowledgements
T h i s re s e a rc h w a s g e n e ro u s l y s u p p o r t e d b y t h e D e u t s c h e
Forschungsgemeinschaft (DFG project FOR 402, 816). We thank the
Fundacíon Científica San Francisco, Ecuador, the NCI for providing
research facilities, Jürgen Homeier and Jeremy Hayward for identification
of Nyctaginaceae, and New Phytologist for the permission to reprint the
figures (copyright 2005). The skilful technical assistance of Jutta Bloschies
is much appreciated. We would like to thank Tanja Schuster for editing
the manuscript. We also thank Krista L. McGuire and Caitlyn Gillikin for
improving the English.
Ectomycorrhizas of Nyctaginaceae 27
References
Alexander, I.J. and P. Högberg. 1986. Ectomycorrhizas of tropical angiospermous trees. New
Phytol. 102: 541–549.
Ashford, A.E. and W.G. Allaway. 1982. A sheathing mycorrhiza on Pisonia grandis R. Br.
(Nyctaginaceae) with development of transfer cells rather than a Hartig net. New
Phytologist 90: 511–519.
Ashford, A.E. and W.G. Allaway. 1985. Transfer cells and Hartig net in the root epidermis
of the sheathing mycorrhiza of Pisonia grandis R. Br. from seychelles. New Phytol. 100:
595–612.
Bahram, M., S. Polme, U. Koljalg, S. Zarre and L. Tedersoo. 2012. Regional and local patterns of
ectomycorrhizal fungal diversity and community structure along an altitudinal gradient
in the Hyrcanian forests of northern Iran. New Phytol. 193: 465–473.
Béreau, M., M. Gazel and J. Garbaye. 1997. Les symbioses mycorrhiziennes des arbres de la
forêt tropicale humide de Guyane française. Can. J. Bot. 75: 711–716.
Bittrich, V. and U. Kühn. 1993. Nyctaginaceae. In: K. Kubitzki, J.G. Rohwer and V. Bittrich
(eds.). The families and Genera of Vascular Plants, vol. 2. Springer Verlag, Berlin, pp.
473–486.
Brundrett, M.C. 2002. Coevolution of roots and mycorrhizas of land plants. New Phytol. 154:
275–304.
Brundrett, M., N. Bougher, B. Dell, T. Grove and N. Malajczuk. 1996. Working with mycorrhizas
in forestry and agriculture. ACIAR Monograph 32, 374 pp.
Chambers, S.M., J.M. Sharples and J.W.G. Cairney. 1998. Towards a molecular identification
of the Pisonia mycobiont. Mycorrhiza 7: 319–321.
Chambers, S.M., C.J. Hitchcock and J.W.G. Cairney. 2005. Ectomycorrhizal mycobionts of
Pisonia grandis on coral cays in the Capricorn-Bunker group, Great Barrier Reef, Australia.
Mycol. Research 109: 1105–1111.
Daly, D.C. and A.S. Roberts. 2004. Nyctaginaceae. In: N. Smith, S.A. Mori, A. Henderson, D.W.
Stevenson and S.V. Heald (eds.). Flowering plants of the Neotropics. Princeton University
Press, Princeton, pp. 269–271.
Harley, J.L. and S.E. Smith. 1983. Mycorrhizal Symbiosis. Academic Press, London.
Haug, I., M. Weiß, J. Homeier, F. Oberwinkler and I. Kottke. 2005. Russulaceae and
Thelephoraceae form ectomycorrhizas with members of Nyctaginaceae (Caryophyllales)
in the tropical mountain rain forest of southern Ecuador. New Phytol. 165: 923–936.
Janos, D. 1980a. Vesicular-arbuscular mycorrhizal infection in an amazonian rainforest. Acta
Amazonica 10: 527–533.
Janos, D. 1980b. Mycorrhiza influence tropical succession. Biotropica 12(Suppl.): 56–64.
Khan, A.G. 1974. The occurrence of mycorrhizas in Halophytes, Hydrophytes and Xerophytes
and of Endogone spores in adjacent soils. Journal of General Microbiology 81: 7–14.
Koske, R.E., J.N. Gemma and T. Flynn. 1992. Mycorrhizae in Haiwaiian angiosperms: A survey
with implications for the origin of the native flora. American J. Bot. 79: 853–862.
Kottke, I. and I. Haug. 2004. The significance of mycorrhizal diversity of trees in the tropical
mountain forest of southern Ecuador. Lyonia 7: 49–56.
Moyersoen, B. 1993. Ectomicorrizas y micorrizas vesicula-arbusculares en Caatinga Amazónica
del Sur de Venezuela. Scientia Guianae 3, Caracas.
Muthukumar, T. and K. Udaiyan. 2000. Arbuscular mycorrhizas of plants growing in the
Western Ghats region, Southern India. Mycorrhiza 9: 297–313.
Rachel, E.K., S.R. Reddy and S.M. Reddy. 1989. VA mycorrhizal colonization of different
angiospermic plant species in the semi-arid soils of Andhra Pradesh. Acta Bot. India
17: 225–228.
Singer, R. and I. Araujo. 1979. Litter decomposition and ectomycorrhiza in amazonian forests.
Acta Amazonica 9: 25–41.
28 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Smith, S.E. and D.J. Read. 2008. Mycorrhizal Symbiosis. 3rd ed., Academic Press, San Diego,
London.
St. John, T.V. 1980a. Una lista de espécies de plantas tropicais brasileiras naturalmente infectadas
com micorriza vesicular-arbuscular. Acta Amazonica 10: 229–234.
St. John, T.V. 1980b. A survey of mycorrhizal infection in an amazonian rain forest. Acta
Amazonica 10: 527–533.
Suvi, T., L. Tedersoo, K. Abarenkov, K. Beaver, J. Gerlach and U. Koljalg. 2010. Mycorrhizal
symbionts of Pisonia grandis and P. sechellarum in Seychelles: identification of mycorrhizal
fungi and description of new Tomentella species. Mycologia 102: 522–533.
Taylor, D.W. 1995. Cretaceous to tertiary geology and angiosperm paleogeographic history
of the Andes. In: S.P. Churchill, H. Blslev, E. Forero and J.L. Luteyn (eds.). Biodiversity
and Conservation of Neotropical Montane Forests. New York Botanical Garden, Bronx,
New York, pp. 3–9.
Tedersoo, L., A. Sadam, M. Zambrano, R. Valencia and M. Baharam. 2010. Low diversity
and high host preference of ectomycorrhizal fungi in Western Amazonia, a neotropical
biodiversity hotspot. The ISME Journal 4: 465–471.
Tester, M., S.E. Smith and F.A. Smith. 1987. The phenomenon of “nonmycorrhizal” plants.
Can. J. Bot. 65: 419–431.
Wang, B. and Y.-L. Qiu. 2006. Phylogenetic distribution and evolution of mycorrhizas in land
plants. Mycorrhiza 16: 299–363.
CHAPTER
3
Diversity and Abundance of
Ectomycorrhizal Associations
in Rain Forests of Cameroon
under Different Disturbance
Regimes
Onguene Awana Nérée,1,* Judith Marthiale Tsamo,2
Christelle Michaella Ebenye,3 Amadou Mustapha Bâ3
and Thomas Kuyper4
1. Introduction
In the rainforests of tropical Africa, two kinds of mycorrhizal associations
occur: arbuscular mycorrhiza (AM) and ectomycorrhiza (Alexander 1989,
Onguene and Kuyper 2001, Bâ et al. 2011). Whereas most timber trees form
AM, the ectomycorrhizal (ECM) habit occurs in a very limited number of
plant families, viz. Asterpeiaceae, Caesalpiniaceae (a family that is partly
arbuscular mycorrhizal), Dipterocarpaceae, Papilionoideae, Phyllanthaceae
1
Institute of Agricultural Research for Development, Soil, Water and Atmosphere department,
Yaoundé, Cameroun.
2
Department of Plant biology and physiology, University of Yaoundé I, Cameroon.
3
Laboratoire Commun de Microbiologie (LCM) IRD/UCAD/ISRA, Centre de Recherche de
Bel Air, BP. 1386, CP. 18524 Dakar, Senegal.
4
Department of soil quality, University of Wageningen, the Netherlands.
*Corresponding author: nereeoa678@yahoo.fr
30 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
following land use changes. The objectives of this study were to report on
the inventory of ectomycorrhizal associations in humid forests of Cameroon
including their habitats (soil and litter characteristics), host tree species,
ectomycorrhizas and sporocarps, and assessing the effect of logging and
agriculture practices on ECM inoculum potential.
flow while immersed in tap water. Seven to ten selected root tips for each
morphotype representative were removed from the root sample and
morphologically and anatomically observed. The key morphological and
anatomical features examined under a dissecting microscope at 40x included
root tip branching, shape of branches, mantle colour and surface texture,
presence of rhizomorphs, emanating hyphae. Tips were also observed
under a photonic microscope between 25 and 40x to confirm the presence
of a mantle and a Hartig net, various layers of hyphal arrangement, and
to determine the presence of specialized cells, rhizomorphs and cystidia,
using both cross—and longitudinal sections (Agerer 1995).
that of Afzelia 11.5 g (6.2–17.4 g). Seedlings of both species possess coarsely
branched roots with few root hairs (Onguene and Kuyper 2001).
Seeds were germinated for a week in steam-sterilized sand without
pregermination treatment. One 1-week old seedling of each tree was
placed in a small hole at the center of the soil core. Cores were placed on
benches and plants grown for four months under natural light conditions
in a greenhouse in Kribi in a randomized complete block design, watered
every three days to maintain soils at field capacity, without nutrient
amendments.
To determine whether ECM and EM fungal propagules share the same
niche, a local variety of cowpea, Vigna unguiculata (Fabaceae), was grown in
soil cores from ECM clumps. In addition, four intact soil cores were collected
around the stem base of ECM tree species, at 5 m and 10 m distance away
from the stem base of A. bipindensis and Brachystegia cynometroides at the
Ebom site, T. bifoliolata and Paraberlinia bifoliolata at the Bityili site. Cowpea
plants were raised for a month in the greenhouse under the same conditions
as previously described. At harvest, ECM fractional colonization was
assessed in water without staining while portions of Afzelia root samples
stained with acid fuchsine were assessed by the gridline intersect method
(Onguene and Kuyper 2001).
Table 3. Basal stem diameter in different forest clump types in 1000 m² area (Average
observations of three years during the dry season).
Table 4. contd....
Ectomycorrhizal Associations in Rain Forests of Cameroon 37
Table 4. contd.
38
CM 417 Brown maroon monopennate Cotton like to Felty Net 104 Absent Absent
Uapaca acuminata à maroon woolly prosenchyma prosenchyma
CM 418 Brown monopyramidal Cotton like Felty Synenchyma 403–502.3 Absent Present
Touabouate prosenchyma
brevipaniculata
Ectomycorrhizal Associations in Rain Forests of Cameroon 39
Table 5. List of putative ectomycorrhizal mushroom species by family, genus and species in
humid forests of South Cameroon.
Sclerodermataceae
Scleroderma sinnamariense Mont
S. roseacarneum sp.
Coltriciaceae
Coltricia spathulata (Hooker)
Murill
C. pyrophila (Wakef) Ryvarden
Table 5. contd....
40 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 5. contd.
100
A
)
80
Percent ECM root tips (%)
%
(
sp
it
to
or 60 a
M
C
E
tn 40
ec a
re
P
20
b
b
0 b b b
FP FI FA FG ES LS EF
Disturbance stages
100
B
80
Percent ECM root tips (%)
a
60
ab
40
ab
20
bc
c
d d
0
FP CO FI FA ES LS EF
Disturbance stages
fallow) and decreased in soils from late-successional stages. Soil cores taken
under mature Afzelia trees resulted in even higher fractional ECM root
colonization. In soils from forestry practices and ECM clumps, no ECM
colonization was observed (Fig. 1). Fractional AM colonization was always
lower than 5%; no colonization by indigenous AM fungi was observed in
soil cores from forestry practices and from ECM clumps.
Most soil cores did not produce abundant arbuscular mycorrhizal
fungal colonization on roots of Vigna unguiculata. AM colonization was
detected in 56% (54 out of 96) soil cores from forest clumps. The sparse AM
colonization varied with sites: very low to low in clumps in Ebimimbang
and Ebom, and completely absent in Nyangong. No AM colonization was
observed in soil cores around the stem base of Afzelia, Brachystegia, and
Paraberlinia, but AM colonization varied from 2 to 22% in the vicinity of
Tetraberlinia trees.
In humid forests of south Cameroon, five types of ectomycorrhizal
forest clumps exist but no longer regenerate, as observed by the dominance
of only the adult size class of individuals: Gilbertiodendron monodominant,
Microberlinia monodominant in the Korup National Park (Newbery et
al. 1988), Uapaca monodominant, Uapaca oligo-dominant, mixed oligo-
dominant ceasalps, mixed ceasalp and Uapaca trees. If six ECM timber
species out of 24 occur isolated in the midst of arbuscular mycorrhizal plant
species, most ECM host species formed clumps, either canopy dominant
or oligo-dominant. They varied in size from small to medium in the
Bipindi-Lolodorf-AkomII zone, large in the Korup National Park and very
large in the forests of south-east Cameroon, like the 10 km long stretch of
monodominant Gilbertiondendron clumps along the road from Dja River to
Ngoïla (Onguene, Amadou and Ebenye, pers.obs.). Botanical inventories
in Gabon, Congo, Central African Republic, and the Democratic Republic
of Congo also depicted such clumps of ceasalp and Uapaca tree species (De
Saint Aubin 1963, Ndong et al. 2011). On the other hand, ECM associations
forming clumps are limited elsewhere in the Neotropics. Only two genera,
Dycimbe and Aldinia have consistently been shown to form such associations
(Singer et al. 1983, Smith et al. 2011). ECM fungi have also been described
in humid forest relics of West Africa (Diédhiou et al. 2010) and in the open
forest of the Zambesian region (Buyck 1994). Such a variety of habitat
conditions cannot be explained by environmental context and floristics
alone (Smith et al. 2011).
In tropical Africa, most timber trees form arbuscular mycorrhiza (AM) but
the ectomycorrhizal habit occurs in a very limited number of plant families.
44 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
large contribution of ECM trees to basal area (Onguene and Kuyper 2001)
and to inventories of ECM fungi (Onguene and Kuyper 2012), where Bityili
was highest and Ebom lowest.
10. Conclusion
Cameroonian ectomycorrhizal fungi abundantly fruited mostly in five
types of clumps including Gilbertiodendron monodominant, Uapaca
monodominant, monodominant Microbelinia in Korup National Park,
oligo-dominant ceasalp clumps locally called “Ekop” and mixed clumps
of “Ekop” with Uapaca spp., independent of elevation, rainfall, topography
and soil texture. Though poor in plant diversity and no longer or barely
regenerating, these habitats recruited abundant and various ECM fungal
species on which native ECM tree species depend for survival, and some
others such as chanterelles which serve as a ready source of protein for local
people during harsh periods and could be a cash flow. Therefore, forest
clumps should be conserved as biodiversity sanctuaries. The presence
48 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Acknowledgements
We thank the anonymous referees for their valuable comments on this
study, and Krista L. McGuire and Caitlyn Gillikin for improving the
English. Financial support from NWO (Priority programme—Biodiversity
in disturbed ecosystems) and logistic support by the Tropenbos Cameroon
Programme and the Government of Cameroon through financial support
to the Institute of Agricultural Research for Development (IRAD) are
gratefully acknowledged. Field assistance was provided by Jean-Baptiste
Mva, Edouard Nsomoto, Serge Aba’a Aba’s, Roger Eyene, and Thomas
Mba. The technical assistance by Veronique O. Anaba during the project is
very much appreciated. Thanks to the reviewer.
References
Agerer, R. 1995. Anatomical characteristics of identified ectomycorrhizas: an attempt towards a
natural classification. In: A.K. Varma and B. Hock (eds.). Mycorrhiza: Structure, Function,
Molecular Biology and Biotechnology. Springer Verlag, Heidelberg, pp. 685–734.
Alexander, I.J. 1989. Mycorrhizas in tropical forests. In: J. Proctor (ed.). Mineral Nutrients in
Tropical Forest and Savanna Ecosystems. Blackwell Scientific Publications, Oxford, pp.
169–188.
Bâ, A.M., R. Duponnois, M. Diabaté and B. Dreyfus. 2011. Les champignons ectomycorhiziens
des arbres forestiers en Afrique de l’Ouest. Méthodes d’étude, diversité, écologie,
utilisation en foresterie et comestibilité. Collections Actiques IRD, IRD Editions 268pp.
Boerner, R.E.J., B.G. Demars and P.N. Leicht. 1996. Spatial patterns of mycorrhizal infectiveness
of soils along a successional chronosequence. Mycorrhiza 6: 79–90.
Buyck, B., D. Thoen and R. Watling. 1996. Ectomycorrhizal fungi of the Guineo-Congo Region.
Proc. R. Soc. Edinb. Sect. B 104: 313–333.
Boa, E. 2006. Champignons comestibles sauvages—Vue d’ensemble sur leurs utilisations et
leur importance pour les populations. FAO, Rome.
Buyck, B. 1994. Ubwoba: les champignons comestibles de l’Ouest du Burundi. Adm. Gén.
Coop. Dév. Publi. Agric. Bruxelles 34: 123.
Connell, J.H. and M. Lowman. 1989. Low-diversity tropical rainforests: some possible
mechanisms for their existence. Am. Nat. 134: 88–119.
De Kessel, A., J.T.C. Codjia and N.S. Yorou. 2002. Guide des champignons comestibles du
Bénin. Cotonou, République du Bénin, Jardin Botanique National de Belgique et Centre
International d’Ecodéveloppement Intégré (CECODI). Impr. Coco-Mutimedia 275pp.
De Saint Aubin, G. 1963. La forêt du Gabon—Centre Techn. For. Trop., Nogent-sur-Marne.
Diédhiou, A.G., M.-A. Selosse, A. Galiana, M. Diabaté, B. Dreyfus, A.M. Bâ, S. de Faria
and G. Béna. 2010. Multi-host ectomycorrhizal fungi are predominant in a Guinean
tropical rainforest and shared between canopy and seedlings. Environ. Microbiol. 12:
2219–2232.
Ectomycorrhizal Associations in Rain Forests of Cameroon 49
Eneke, E.T.B and I.J. Alexander. 2012. Mycorrhiza status of Gnetum spp. in Cameroon:
evaluating diversity with a view to ameliorating domestication efforts. Mycorrhiza 22:
99–108.
Fassi, B. and M. Moser. 1991. Mycorrhizae in the natural forests of tropical Africa and the
Neotropics. In: A. Fontana (ed.). Funghi, Piante e Suolo, Centro di studio Micologia del
Consiglio nazionalle delle Ricerche, Torino, Italy, pp. 183–202.
Grubb, P.J. 1977. The maintenance of species richness in plant communities: the importance
of the regeneration niche. Biol. Reviews 52: 107–145.
Henkel, T.W., J. Terborgh and R.J. Vilgalys. 2002. Ectomycorrhizal fungi and their leguminous
hosts in the Pakaraima Mountains of Guyana. Mycol. Res. 106: 515–531.
Janos, D.P. 1996. Mycorrhizas, succession and rehabilitation of deforested lands in the humid
tropics. In: J.C. Frankland, N. Magan and G.M. Gadd (eds.). Fungi and Environmental
Change. Cambridge University Press, Cambridge, pp. 129–161.
Jonsson, L., A. Dahlberg, M.-C. Nilsson, O. Karén and O. Zackrisson. 1999. Continuity
of ectomycorrhizal fungi in self-regenerating boreal Pinus sylvestris forests studied
by comparing mycobiont diversity on seedlings and mature trees. New Phytol. 142:
151–162.
Letouzey, R. 1968. Etude phytogéographique du Cameroon. Paris, Editions P. le Chevalier.
Letouzey, R. and R. Mouranche. 1952. Ekop du Cameroun. Centre Technique Forestier Tropical.
Nogent-sur-Marne.
Malaisse F. 1967. Se nourrir en forêt claire africaine—Approche écologique et nutritionnelle.
Les Presses Agronomiques de Gembloux, Gembloux, Belgique.
Moyersoen, B., A.H. Fitter and I.J. Alexander. 1998. Spatial distribution of ectomycorrhizas
and arbuscular mycorrhizas in Korup National Park, Cameroon, in relation to edaphic
parameters. New Phytol. 139: 311–320.
Ndong, E.H., J. Degreef and A. De Kesel. 2011. Champignons comestibles des forêts denses
d’Afrique centrale. Taxonomie et identification. Vol. 10. ABC Taxa. ISSN 1784–1291:
262pp.
Newbery, D.M., I.J. Alexander, D.W. Thomas and J.S. Gartlan. 1988. Ectomycorrhizal rain
forest legumes and soil phosphorus in Korup National Park, Cameroon. New Phytol.
109: 433–450.
Newbery, D.M., I.J. Alexander and J.A. Rother. 1997. Phosphorus dynamics in lowland African
rainforest: the influence of ectomycorrhizal trees. Ecol. Monographs 67: 367–409.
Onguene, N.A. 2000. Diversity and dynamics of mycorrhizal associations in tropical rain
forests with different disturbance regimes in south Cameroon. Tropenbos Cameroon
Ser. 3: 1–167.
Onguene, N.A. and T.W. Kuyper. 2012. Habitat and diversity of ectomycorrhizal mushrooms
in humid forests of South Cameroon. Cameroon J. Exp. Biol. 8: 26–34.
Onguene, N.A. and T.W. Kuyper. 2001. Mycorrhizal associations in the rain forest of South
Cameroon. For. Ecol. Manag. 140: 277–287.
Onguene, N.A. and T.W. Kuyper. 2002. Importance of the ectomycorrhizal network for the
seedling survival and ectomycorrhizal formation in rain forests of south Cameroon.
Mycorrhiza 12: 13–17.
Onguene, N.A. and T.W. Kuyper. 2005. Growth response of three timber species to soils with
different arbuscular mycorrhizal inoculum potentials in South Cameroon. Indigenous
inoculum and effect of addition of grass inoculum. For. Ecol. Manag. 210: 283–290.
Rammeloo, J. and R. Walleyn. 1993. The edible fungi of Africa south of Biafra: a literaturesurvey.
Scripta Bot. Belg. 5: 1–62.
Richards, P.W. 1996. The tropical rain forest—an ecological study, 2nd edition. Cambridge,
Cambridge University Press.
Rivière, T., A.G. Diédhiou, M. Diabaté, G. Senthilarasu, K. Natarajan, A. Verbeken, B. Buyck, B.
Dreyfus, G. Béna and A.M. Bâ. 2007. Genetic diversity of ectomycorrhizal basidiomycetes
from African and Indian tropical forests. Mycorrhiza 17: 415–428.
50 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Sanon, K.B., A.M. Bâ and J. Dexheimer. 1997. Mycorrhizal status of some fungi beneath
indigenous trees in Burkina Faso. For. Ecol. Manag. 98: 61–69.
Singer, R. and I. Araujo. 1986. Litter decomposition and ectomycorrhizal Basidiomycetes in
an igapo forest. Plant Syst. Evol. 153: 107–117.
Singer, R., I. Araujo and M.H. Ivory. 1983. The ectotrophically mycorrhizal fungi of the
neotropical lowlands, especially Central Amazonia. Nova Hedw. 77: 1–352.
SAS Inc. 2004. SAS—X user’s guide, 4th edition. Gorinchem.
Smith, M.E., T.W. Henkel, M.C. Aime, A.K. Fremier and R. Vilgalys. 2011. Ectomycorrhizal
fungal diversity and community structure on three co-occurring leguminous canopy tree
species in a Neotropical rainforest. New Phytol. 192: 699–712.
Smits, W. 1994. Dipterocarpaceae: mycorrhizae and regeneration. Tropenbos Series 9: 1–243.
Taylor, D.L. and T.D. Bruns. 1999. Community structure of ectomycorrhizal fungi in a Pinus
muricata forest: minimal overlap between the mature forest and resistant propagule
communities. Mol. Ecol. 8: 1837–1850.
Tedersoo, L., A. Sadam, M. Zambrano, R. Valencia and M. Bahram. 2009. Low diversity and
high host preference of ectomycorrhizal fungi in Western Amazonia, a neotropical
biodiversity hotspot. Int. Soc. Micro. Ecol. 1–7.
Van Dijk, H., N.A. Onguene and T.W. Kuyper. 2003. Knowledge and utilization of edible
mushrooms by local populations of the rain forest of South Cameroon. Ambio. 32:
19–23.
Walleyn, R. and J. Rammeloo. 1994. The poisonous and useful fungi of Africa south of the
Sahara. Scripta Bot. Belg. 10: 1–56.
CHAPTER
4
Mycorrhizal Fungi Diversity
and their Importance on
the Establishment of Native
Species Seedlings within
Madagascarian Degraded
Sclerophyllous Forest
Rondro Harinisainana Baohanta,1,* Herizo
Andrianantoandro Randriambanona,1 Marc Ducousso,2
Christophe Nirina Rakotoarimanga,1 Yves Prin,2
Heriniaina Ramanankierana1 and Robin Duponnois1
1. Introduction
The impact of human activities on tropical ecosystems has increased
dramatically in recent decades leading to a global reduction of primary
forests (Laurance 1999, Morris 2010). For tree species, fragmentation
of forests into patches has led to degradation of both their habitat and
1
Laboratoire de Microbiologie de l’Environnement, Centre National de Recherches sur
l’Environnement BP 1739 Fiadanana Antananarivo 101, Madagascar.
2
Laboratoire des Symbioses Tropicales et Méditerranéennes (LSTM), UMR 113 CIRAD/INRA/
IRD/SupAgro/UM2, Campus International de Baillarguet, TA A-82/J, Montpellier, France.
*Corresponding author: ninish.rondro@yahoo.fr
52 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
A and B) and deciduous forest (site C). The main chemical characteristics
of the upper soil layer (2–20 cm) of these sites are shown in Table 1.
These investigations showed that sclerophyllous woodlands dominated
by U. bojeri contain a wide range of sporocarps belonging to at least four
different fungal families: Russulaceae, Cantharellaceae, Boletaceae, and
Amanitaceae. Overall, 94 sporocarps of putative epigeous ECM fungi
were collected in three survey sites. They were identified as belonging
to the ECM genera Afroboletus, Amanita, Boletus, Cantharellus, Leccinum,
Gyroporus, Rubinoboletus, Russula, Scleroderma, Suillus, Tricholoma, and
Xerocomus (Table 2). Russula was the most frequent ECM genus recorded
(32.9% of the above-ground sporocarp diversity) followed by the genera
Amanita (17.1%) and Cantharellus (Fig. 1a). At species level, 21 species were
recorded for Russula followed by 14 Amanita species,10 Cantharellus species,
and 10 Boletus species (Fig. 1a, b). The fungal richness of the above-ground
sporocarps decreased from site A (40 species) to site C and B with 29 and
27 species, respectively.
The results of Ramanankierana et al. (2007) showed a large diversity of
sporophores recorded under ectomycorrhizal tree and shrub species in the
sclerophyllous forests of Madagascarian highlands. Different fungal families
(Russulaceae, Cantharellaceae, Boletaceae, and Amanitaceae) observed at
this forest have been recorded in other tropical forests under Afzelia africana,
Monotes kerstingii, Uapaca guineensis, and U. somon in Africa (Thoen and Bâ
1989, Sanon et al. 1997) and under dipterocarps in Asia (Lee 1998). It has
been already demonstrated that Russulaceae are often dominant in the
tropical rainforests of Africa, Asia, and Madagascar (Buyck et al. 1996, Lee
et al. 1997, Walting and Lee 1998, Rivière et al. 2006).
In the northern part of this sclerophyllous forest (Site A), three
ectotrophic plant species (U. bojeri, Leptolaena pauciflora Baker and Leptolaena
bojeriana (Baill.) Cavaco. were identified by Baohanta et al. (2012). The
highest fungal diversity of the above-ground sporocarpswas recorded at
Table 1. Main chemical characteristics of the upper soil layer (2–20 cm). Data from
Ramanankierana et al. (2007).
Table 2. contd....
Table 2 .contd.
56
and dark stem, red-reticulated becoming yellow at the base like individuals
rhizomorph, flesh and pores turning blue by wound
Leccinum sp1 Small grey boletus (1.8 to 3 cm diameter by 3 to 4cm high), yellow Patch of 3 to 4 x x x
pores, red hairy scales on the stem, base of the stem yellow like the individuals
rhizomophs
Boletus sp2 Big brownish-brown wet cap (7 to 8 cm diameter to 12 to 15 cm Patch of 5 to 6 x
high), white and smooth flesh individuals
Xerocomus sp1 Brown scaly cap (8.5 cm diameter) showing white flesh between Solitary, scarce x
scales, white stem (1.4 cm diameter by 5 to 6 cm high) with some
red zone
Leccinum sp2 Yellow grey scaled boletus (4.5 to 6 cm diameter by 6 to 7 cm high), Solitary, scarce x
stem yellow at the base and red in its upper part, yellow blueishing
pores
Boletus sp3 Brown cap (7.5 cm diameter) with red-pink pigments, yellow and Solitary, scarce x
red pores, greenishing and blueishing tubes, yellowish stem with
some red pigments
Leccinum sp3 Red purplish-blue wet cap (7 cm diameter), yellow burnishing stem Solitary, scarce x
(0.8 cm diameter by 6 cm high), concolored yellow flesh and pores,
blueishing after air exposure
Boletus sp4 Big smooth and shiny red boletus (8 to 12 cm diameter by 7 to 8 cm Patch of 2 to 3 x x x
high), yellowish stem with some pink pigments, concolored flesh individuals
(1.6 cm thick)
Xerocomus sp2 Pale to dark brown scaly dry cap (5 cm diameter), white dirty stem Solitary, scarce x
(0.8 cm diameter by 4 cm high) with a white-yellowish flesh, yellow
greenish and pink pores
Boletus sp5 Yellowish brown cap (8 cm diameter) with flat partially pink scales, Solitary, scarce x
yellow pores and stem (1.2 cm diameter by 6 cm high), white flesh
(1.6 cm thick)
Boletus sp6 Dark brown scaly cap showing yellow flesh, pale concolored pores Solitary, scarce x
and stem
Boletus sp7 Brown boletus with dry and silky cap (4.5 cm diameter), concolored Solitary, scarce x
dark stem (2.2 cm diameter by 5.2 cm high), white flesh (1.6 cm
thick) rapidly turning to red, then black after air exposure
Boletus sp8 Pale brown boletus with silky cap (5 cm diameter), white stem Solitary, scarce x
(1.5 cm diameter by 5.2 cm high) and flesh (1.3 cm thick) turning
purplish-blue after air exposure
Table 2 .contd....
Mycorrhizal Fungi Diversity and Native Plant Regeneration in Madagascar 57
Table 2 .contd.
58
Russula sp11 Small purple to purple-reddish umbilicate when young to flat when Solitary to patch of x
ageing cap (2 to 7 cm diameter), sticky surface, regular margin, 3 individuals
adnate white to yellowish closely spaced gills, white flesh
Russula sp13 Brown-reddish convex and smooth glutinous cap (6 to 15 cm Solitary x
diameter), decurrent gills, white flesh turning greyish by air
exposure
Russula sp14 Dark yellow to brown convex to flat sticky cap (4 to 10 cm Patch of 3 to 5 x
diameter), adnate closely spaced gills individuals
Russula sp15 Yellow to orange-yellow flat slightly umbilicated with an Patch of 2 to 5 x
involucrated yellow margin cap (2 to 8 cm diameter) with a smooth individuals
surface with small strias
Russula sp16 Pink to reddish (darker at the centre) fragile convex glutinous cap, Patch of 2 to 4 x
(2 to 6 cm diameter) with a smooth or dusty surface, white flesh individuals
Russula sp17 Slightly umbilicated convex and glutinous cap (4 to 10 cm Solitary to patch of x
diameter), dark yellow tending to brown, yellow to pale orange 4 individuals
closely spaced gills
Strobilomycetacea
Afroboletus sp1 Brown-purple scaly cap (3 to12 cm diameter), fibrous stem, pale Patch of 3 to 5 x
yellow flesh turning purplish by air exposure individuals
Afroboletus sp2 Flat-convex dusty cap (3 to 10 cm diameter) with dark-brown to Patch of 2 to 3 x
black scales, fibrous stem inflated at the base, greyish-yellow flesh individuals
Sclerodermataceae
Scleroderma sp1 Whitish to yellowish small pyriformic fruit bodies, size below 3 cm Solitary to patch of x
in diameter, dark grey gleba 5 individuals
Scleroderma sp2 Whitish to yellowish 3 to 7cm diameter fruit bodies with grey spots Solitary, rarely x
at the top, dark grey gleba patchy
Tricholoma sp2 Yellow cap (3 to 9 cm diameter), dry surface, involucrated margin, Solitary to patch of x
thick widely spaced gills, yellow flesh keeping yellow even after 4 individuals
exposure to air
Tricholoma sp3 Yellow-greyish cap (3 to 12 cm diameter), dry surface, white Solitary to patch of x
yellowish stalk, white flesh 4 individuals
Tricholoma sp4 Dark-grey cap (3 to 15 cm diameter), smooth dry surface, thick gills, Solitary x
white flesh
Mycorrhizal Fungi Diversity and Native Plant Regeneration in Madagascar 61
62 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
us
let
us
o
or
ob
op
n
bi
yr
Ru
G
b
25
20
Species per genus
15
10
5
0 us
us
ph
et
ol
om
ob
G
in
ub
R
Genus
Fig. 1. Structure of the ectomycorrhizal community (above-ground species richness) expressed
as genus relative frequency of the most abundant genera (a) and total species per genus (b).
this site (40 species). With other tropical ectomycorrhizal tree species, Lee
et al. (1997) recorded only 28 fungal species under Shorea leprosula, while
Sanon et al. (1997) have identified 14 fungal species under U. guineensis
and 11 species under U. somon in Burkina Faso. However, numerous
studies in temperate areas indicate little correlation between above-ground
(sporocarps) and below-ground (morphotypes of ectomycorrhizas) fungal
diversity (Buscot et al. 2000, Horton and Burns 2001). Further investigations
using molecular approaches are needed to entirely describe the fungal
community associated with plants.
Mycorrhizal Fungi Diversity and Native Plant Regeneration in Madagascar 63
100
Mycorrhizal colonization (%)
90
80
70
60
50
40
30
20
10
0
0 1 2 3 4 5 6
Time (months)
Fig. 2. Mycorrhizal colonization and sequence of ectomycorrhizal morphotypes on U. bojeri
seedlings during 5 months growing time on soil collected from the native stand of this tree,
: AM colonization, : Total ectomycorrhizal colonization, ▲: morphotype 1,•: morphotype
2, x : morphotype 3. Data from Ramanankierana et al. (2007).
64 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 4. Mycorrhizal status of early established shrub species in the degraded areas of the
Arivonimamo and Ambatofinandrahana forests. Data from Baohanta (2011).
Fig. 3. Vegetation structure (A and C), global microbial activity and phosphatase activities of soil
(B and D) at degraded forest edge (from an individual adult tree of U. bojeri) of sclerophyllous
forest in high plateau of Madagascar.
and on U. bojeri soil (0 m) (Fig. 3B, D). Correlation between these two soil
parameters and the distribution of shrub species was observed in the
Arivonimamo forest. It has been shown that the abundance of L. pauciflora
and L. bojeriana was associated with a high value of global microbial and
phosphatase activities of soil (Fig. 3).
Some authors (Gómez-Aparicio et al. 2005, Montané et al. 2010) have
already reported that the propagation of some shrub species enhance litter
quality and the amount of organic matter in soil. Such situations could lead
to a definite increase in soil fertility. However, more experiments must be
undertaken in order to confirm that modified soil conditions by pioneer
shrub species facilitate both the establishment of tree species as well as
the development of late successional species in Madagascarian highland
sclerophyllous forest. On degraded soil, mycorrhizal fungi constitute one of
the well-known biological components that can improve host performance
by enhancing nutrient and water uptake from the soil and protecting host
roots from pathogens and toxic compound (Smith and Read 2008).
Fig. 4. Results of the PCA on the data table of soil, plant, and microbial activity parameters.
(A) Correlation circle of all the parameters. The 12 variables are: pH=pH, P=total phosphorus
(mg Kg–1), N=total nitrogen (%), OM=total organic matter (%), FDA= global microbial activity;
AcP=acid phosphatase, AlkP=alkaline phosphatase, SB=shoot biomass (g), RB= root biomass
(g), ER=ectomycorrhizal rate (%), PN=leaf nitrogen (%), PP=leaf phosphorus (mg Kg–1), H=
Shannon diversity index of ectomycorrhizal fungi. (B) Map of sample scores on the first two
principal components. Samples are coded as follows. The first three characters correspond to
the soil origin; Eca=soil collected under E. camaldulensis, Ppa= soil collected under P. patula,
Ubo=soil collected under U. bojeri, BaS=bare soil. The treatment applied to the U. bojeri seedlings
is coded as follows. U=Uapaca plant alone, UL=Uapaca plant + L. bojeriana, Ulc=Uapaca plant
+ L. bojeriana cut after 4 months cultivation. For example, a sample coded “EcaULc” is a U.
bojeri seedling grown in soil collected under E. camaldulensis in which a plant of L. bojeriana
was grown and cut after 4 months of cultivation.
72 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
in fungal rDNA amplification. A total of 2025 ECM root tips were collected
from 15 soil cores and classified into 165 ECM morphotypes without
mixing root samples from different soil cores. DNA extraction and RFLP
analysis were carried out on 495 ectomycorrhizas and revealed 137 RFLP
types (Table 6). Considering that each RFLP type corresponds to a single
ECM fungal species, the highest diversity of ectomycorrhizas expressed by
Simpson’s diversity index and by Shannon-Wiener information index was
found on mixed vegetation. These diversity indexes were similar between
S. oblongifolia and mixed host plant vegetation. However, diversity indexes
were lower for U. bojeri, suggesting that in homogeneous vegetation this tree
species harbors more rare species than it does with the multiple assemblage
community.
Overall, 52.3% of identified RFLP types were found in both U. bojeri
and S. oblongifolia root systems. The remaining RFLP types were found
exclusively on either U. bojeri (19%) or S. oblongifolia (28.5%) (Fig. 5). Previous
studies have reported that ECM fungi are not uniformly distributed in forest
ecosystems but rather with variability in terms of presence, abundance,
and community composition (Baylis 1980, Perry et al. 1989, Dickie et al.
2004, Dickie and Reich 2005). Environmental factors can influence patterns
of fungal distribution such as soil characteristics and soil microbiota
(Smith and Read 2008), but it is largely admitted that mycorrhized plant
communities are the main factor influencing the availability of ECM fungal
propagules in soil (Dickie and Reich 2005). These biological processes
provide large implications for the establishment of ECM plant cover and
for fungal community ecology.
Table 6. Sampling design, host plant colonization by ectomycorrhizal fungal and fungal
diversity.
Origins
U. bojeri S. oblongifolia Mixture
vegetation
Number of soil cores 5 5 5
Number of ectomycorrhizal root tips 665 580 780
Number of RFLP types (without soil core 38 46 53
comparison)
Percentage of shared identified taxa* 60 58,33 76,92
between the two host plant species (%)
Simpson’s diversity index 6.75 10.58 11.22
Shannon-Wiener information index 0.87 1.05 1.07
*Taxa are identified as RFLP types based on similarity of fragmented size in GenBank.
Mycorrhizal Fungi Diversity and Native Plant Regeneration in Madagascar 73
In the present study, two ECM plant species (U. bojeri and S. oblongifolia)
can support patches of ECM fungi. The same study also illustrated that
the growth of U. bojeri seedlings was significantly higher in soils sampled
from areas with established adult trees of U. bojeri than in other soils
(Table 7). Well-developed seedlings on these soils were accompanied by
higher ECM colonization rate and by significant taxa richness of ECM
communities. The stimulation effect of external fungal mycelium radiating
from mature trees on young seedling ECM formation was evidenced in
a Guinean tropical forest (Diédhiou et al. 2010), in Mediterranean holm
oak (Richard et al. 2005), and in temperate coniferous forests (Jonsson
et al. 1999, Kranabetter and Friesen 2002). However, in the experimental
site, the ECM shrub S. oblongifolia displayed an important effect on ECM
fungal infection of young seedlings. In this part of the Madagascarian
sclerophyllous forest, S. oblongifolia was naturally established especially
at the forest edge or in degraded surface areas free of U. bojeri adult trees.
This investigation demonstrates that more than half of collected ECM taxa
were shared between S. oblongifolia and U. bojeri grown separately or in
mixed formation. It suggests that S. oblongifolia, as a pioneering species,
may persist on disturbed sites and facilitate the survival of ECM fungi
that could potentially infect U. bojeri seedling roots. This nurse effect was
particularly efficient by enhancing the ECM fungal diversity associated with
young U. bojeri seedlings and the rate of ECM infection. These results are
similar to reports on facilitation of ectomycorrhizal infection of Pseudotsuga
74 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
8. Conclusion
This chapter shows that the development of U. bojeri, an endemic tree of
the Malagasysclerophyllous forest, strongly requires the occurrence of
compatible ECM fungi. These ECM symbionts could be from an adult
tree of this plant where various putative ECM fungi were observed, or
from pioneer shrub species that persist on disturbed sites and facilitate
the survival of ECM fungi propagules that could potentially infect roots
of U. bojeri seedling. In this way, two species of endemic shrub species
(L. bojeriana and S. oblongifolia) could be of great interest. These data
suggest that these nurse plant species should be considered to enhance
the performance of reforestation efforts in Madagascar by providing ECM
inoculum potential.
Mycorrhizal Fungi Diversity and Native Plant Regeneration in Madagascar 75
Acknowledgments
We thank the anonymous referees for their valuable comments on this study,
and Krista L. McGuire and Caitlyn Gillikin for improving the language.
References
Agarwal, D.K., J.A. Silander, A.E. Gelfand, R.E. Dewar and J.G. Mickelson. 2005. Tropical
deforestation in Madagascar: analyses using hierarchical, spatially explicit, Bayesian
regression models. Ecol. Modelling 185 : 105–131.
Baohanta, R.H. 2011. Facilitation de la régénération d’Uapaca bojeri L. par la gestion des
communautés de champignons mycorhiziens associés aux espèces pionnières de la zone
dégradée de la forêt sclérophylle d’Arivonimamo. Thèse de doctorat en Science de la
vie. Université d’Antananarivo, p. 114.
Baohanta Rondro, H., J. Thioulouse, H. Ramanankierana, Y. Prin, R. Rasolomampianina,
E. Baudoin, N. Rakotoarimanga, A. Galiana, H. Randriambanona, M. Lebrun and R.
Duponnois. 2012. Restoring native forest ecosystems after exotic trees plantation in
Madagascar: combination of the local ectotrophic species Leptolena bojeriana and Uapaca
bojeri mitigates the negative influence of the exotic species Eucalyptus camaldulensis and
Pinus patula. Biol. Invasion. DOI 10.1007/s10530-012-0238-5.
Baylis, G.T.S. 1980. Mycorrhizas and the spread of beech. New Zeal J. Ecol. 3: 151–153.
Borchers, S. and D. Perry. 1987. Early successional hardwoods as refugia for ectomycorrhizal
fungi in clearcut douglas-fir forests of southwestern Oregon. In: D.M. Sylvia, L.L. Hung
and J.H. Graham (eds.). Mycorrhizae in the Next Decade: Practical Applications and
Research Priorities. University of Florida, Gainesville, Florida, p. 84.
Brooker, R.W., F.T. Maestre, R.M. Callaway, C.L. Lortie, L.A. Cavieres, G. Kunstler, P. Liancourt,
K. Tielbörger, J.M.J. Travis, F. Anthelme, C. Armas, L. Coll, E. Corcket, S. Delzon, E.
Estelle Forey, Z. Kikvidze, J. Olofsson, F. Pugnaire, C.L. Quiroz, P. Saccone, K. Schiffers,
M. Seifan, B. Touzard and R. Michalet. 2008. Facilitation in plant communities: the past,
the present and the future. J. Ecol. 96: 18–34.
Buscot, F., J.C. Munch, J.Y. Charcosset, M. Gardes, U. Nehls and R. Hampp. 2000. Recent
advances in exploring physiology and biodiversity of ectomycorrhizas highlight the
functioning of these symbioses in ecosystems. FEMS Microbiol. Rev. 24: 601–614.
Buyck, B., D. Thoen and R. Walting. 1996. Ectomycorrhizal fungi of the Guinea-Congo region.
Proc. R. Soc. Edinb. 104: 313–333.
Callaway, R.M. 1995. Positive interactions among plants. Bot. Rev. 61: 306–349.
Callaway, R.M. 1997. Positive interactions in plant communities and the individualistic-
continuum concept. Oecologia1 12: 143–149.
Castro, J., R. Zamora and J.A. Hódar. 2006. Restoring Quercus pyrenaica forests using pioneer
shrubs as nurse plants. Applied Vegetation Sc. 9(1): 137–142.
Cazares, E. and J.M. Trappe. 1993. Vesicular endotrophytes in roots of the Pinaceae. Mycorrhiza
2: 153–156.
Chilvers, G.A., F.F. Lapeyrie and D.P. Horan. 1987. Ectomycorrhizal vs endomycorrhizal Fungi
within the same root system. New Phytologist 107: 441–448.
Cooke, J.C. and M.W. Lefor. 1998. The mycorrhizal status of selected plant speciesfrom
Connecticut wetlands and transition zones. Restor. Ecol. 6(2): 214–222.
Dhillion, S.S. 1994. Ectomycorrhizae, arbuscular mycorrhizae, and Rhizoctonia sp. of alpine
and boreal Salix spp. in Norway. Arct. Alp. Res. 26: 304–307.
Dickie, I.A., R.C. Guza, S.E. Krazewski and P.B. Reich. 2004. Shared ectomycorrhizal fungi
between a herbaceous perennial (Helianthemum bicknellii) and oak (Quercus) seedlings.
New Phytol. 164: 375–382.
76 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Dickie, I.A., R.T. Koide and K.C. Steiner. 2002. Influence of established trees on mycorrhizas,
nutrition, and growth of Quercus rubra seedlings. Ecol. Monographs 72: 505–521.
Dickie, I.A. and P.B. Reich. 2005. Ectomycorrhizal fungal communities at forest edges. J. Ecol.
93: 244–255.
Diédhiou, A.G., M.A. Selosse, A. Galiana, M. Diabaté, B. Dreyfus, A.M. Bâ and G. Béna. 2010.
Multi-host ectomycorrhizal fungi are predominant in a Guinean tropical rainforest and
shared between canopy trees and seedlings. Environ. Microbiol. 12: 2219–2232.
Dos Santos, V.L., R.M. Muchovej, A.C. Borges, J.C.L. Neves and M.C.M. Kasuya. 2001. Vesicular-
arbuscular-/ecto-mycorrhiza succession in seedlings of Eucalyptus spp. Brazilian J.
Microbiol. 32: 81–86.
Duchesne, L.C., R.L. Peterson and B.E. Ellis. 1988. Pine root exudates stimulate the synthesis
of antifungal compounds by the ectomycorrhizal fungus Paxillus involutus. New Phytol.
108: 471–476.
Ducousso, M., C. Bourgeois, B. Buyck, G. Eyssartier, M. Vincelette, R. Rabevohitra, G. Béna, L.
Randrihasipara, B. Dreyfus and Y. Prin. 2004. The last common ancestor of Sarcolaenaceae
and Asian Dipterocarp trees was ectomycorrhizal before the India—Madagascar
separation, about 88 million years ago. Mol. Ecol. 13(1): 231–236.
Ducousso, M., H. Ramanankierana, R. Duponnois, R. Rabévohitra, L. Randrihasipara, M.
Vincelette, B. Dreyfus and Y. Prin. 2008. Mycorrhizal status of native trees and shrubs from
eastern Madagascar littoral forests with special emphasis on one new ectomycorrhizal
endemic family, the Asteropeiaceae. New Phytol. 178: 233–238.
Duponnois, R., S. Diédhiou, J.L. Chotte and M.O. Sy. 2003. Relative importance of the
endomycorrhizal and/or ectomycorrhizal associations in Allocasuarina and Casuarina
genera. Can. J. Microbiol. 49(4): 281–287.
DuPuy, D. and J. Moat. 1996. A refined classification of the primary vegetation of Madagascar
based on the underlying geology: using GIS to map it distribution and to assess its
conservation status. Proceedings of the International Symposium on the “Biogeography
de Madagascar”, Lourenço WR., 205–218. Paris.
Egerton-Warburton, L. and M.F. Allen. 2001. Endo- and ectomycorrhizas in Quercus agrifolia
Nee. (Fagaceae): patterns of root colonization and effect on seedling growth. Mycorrhiza
11: 283–290.
Founoune, H., R. Duponnois and A.M. Bâ. 2002. Influence of dual arbuscular endomycorrhizal/
ectomycorrhizal symbiosis on the growth of Acacia holosericea in glasshouse conditions
(A. Cunn. Ex. G. Don). Ann. For. Sci. 59: 93–98.
Gómez-Aparicio, L., J.M. Gómez, R. Zamora and J.L. Boettinger. 2005. Canopy vs soil effects
of shrubs facilitating tree seedlings in Mediterranean montane ecosystems. J. Vegetation
Sc. 16(2): 191–198.
Haselwandter, K. 1997. Soil micro-organisms, mycorrhiza, and restoration ecology. In: K.M.
Urbanska, N.R. Webb and P.J. Edwards (eds.). Restoration Ecology and Sustainable
Development. University Press, Cambridge, pp. 65–80.
Högberg, P. 1982. Mycorrhizal associations in some woodland and forest trees and shrubs in
Tanzania. New Phytol. 92: 407–415.
Holl, K.M. 2002. Effects of shrubs on tree seedling establishment in an abandoned tropical
pasture. J. Ecol. 90: 179–187.
Horton, T.R. and T.D. Burns. 2001. The molecular revolution in ectomycorrhizal ecology:
peeking into the black-box. Mol. Ecol. 10: 1855–1871.
Jonsson, L., A. Dahlberg, M.C. Nilsson, O. Zackrisson and O. Kren. 1999. Ectomycorhizal
fungal communities in late-successional Swedish boreal forests, and the composition
following wildfire. Mol. Ecol. 8: 205–215.
Kranabetter, J.M. and J. Friesen. 2002. Ectomycorrhizal community structure on western
hemlock seedlings transplanted from forests into openings. Can. J. Bot. 80: 861–868.
Kull, C.A., J. Ratsirarson and G. Randriamboavonjy. 2002. Les forêts de tapia des Hautes Terres
malgaches. J. Cult. Geog. 19(2): 22–58.
Mycorrhizal Fungi Diversity and Native Plant Regeneration in Madagascar 77
Ruiz-Jaen, M.C. and T.M. Aide. 2005. Restoration success: how is it being measured? Restor.
Ecol. 13: 569–577.
Sanon, K., A.M. Bâ and J. Dexheimer. 1997. Mycorrhizal status of some fungi fruiting beneath
indigenous trees in Burkina Faso. For. Ecol. Manag. 98: 61–69.
Schatz, G.E., P.P. LowryII and A.E. Wolf. 2001. Endemic families of Madagascar.VII. A synoptic
revision of Leptolaena thouars sensu strict (Sarcolaenaceae).Adansonia série 3. 23(2):
171–189.
Sicardi, M., F. Garcia-Prechac and L. Frioni. 2004. Soil microbial indicators sensitive to land use
conversion from pastures to commercial Eucalyptus grandis (Hill ex maide) plantations
in Uruguay. Appl. Soil Ecol. 27: 125–133.
Simard, S.W. and D.M. Durall. 2004. Mycorrhizal networks: a review of their extent, function,
and importance. Can. J. Bot. 82: 1140–1165.
Smith, O.H., G.W. Peterson and B.A. Needelman. 2000. Environmental indicators of
agroecosystems. Adv. Agron. 69: 75–97.
Smith, S.E. and D.J. Read. 2008. Mycorrhizal symbiosis. 3rd edition. Academic Press Ltd.,
Cambridge, UK, p. 787.
Smith, M.R., I. Charvat and R.L. Jacobson. 1998. Arbuscular mycorrhizae promote establishment
of prairie species in a tallgrass prairie restoration. Can. J. Bot. 76: 1947–1954.
Thoen, D. and A.M. Bâ. 1989. Ectomycorrhizae and putative ectomycorrhizal fungi of Afzelia
Africana and Uapaca guineensis in Southern Senegal. New Phytol. 113: 549–559.
Walting, R. and S.S. Lee. 1998. Ectomycorrhizal fungi associated with members of the
Dipterocarpaceae in Peninsular Malaysia-II. J. Trop. For. Sci. 10: 421–430.
Weaver, M. and M. Kellman. 1981. The effect of fragmentation on tree woodlot biota in Southern
Ontario. J. Biogeography 8: 199–210.
Williams, A., H.J. Ridway and D.A. Norton. 2012. Different arbuscular mycorrhizae and
competition with an exotic grass affect the growth of Podocarpus cunninghamii Colenso
cuttings. New For. DOI: 10.10007/s11056-012-9309-9.
CHAPTER
5
Morpho-anatomical
Characterization of Three
Sebacinales Ectomycorrhizal
Species from a Pakaraimaea
dipterocarpacea ssp. nitida
(Dipterocarpaceae) Forest in
Southern Venezuela
Bernard Moyersoen
1. Introduction
Sebacinales are a genetically diverse group of basidiomycetes with a broad
range of lifestyles including a diversity of mycorrhizal and endophytic
associations (Weiß et al. 2011). These fungi are widespread and the same
species can colonize a wide range of host plants and be involved in different
symbiotic associations (Selosse et al. 2002a, Weiß et al. 2011). Anatomically,
Sebacinales are characterized by longitudinally septate basidia, imperforate
parenthesomes, and a lack of both clamp connections and hymenial cystidia
(Weiß et al. 2004). A very high diversity within the Sebacinales has been
described from host plant and soil DNA samples and the observed global
diversity was estimated to be 365 ITS taxonomic units (Setaro et al. 2012).
Molecular-phylogenetically, Sebacinales are divided into two groups
called A and B (Weiß et al. 2004). Up to date, most ectomycorrhizal (EcM)
associations have been documented from group A (Weiß et al. 2004, 2011)
and only one clade in group B included EcM strains (Hynson et al. 2013).
Only a few EcM fruitbody species belonging to Sebacina, Tremellodendron
and Tremelloscypha have been described to date (Weiß et al. 2004). These
morphospecies include the cosmopolitan cryptic species S. incrustans
(Pers.) Tul. & C. Tul. and S. epigea (Berk. & Broome) Bourdot & Gazin and
the polymorphic Tremelloscypha gelatinosa (Murrill) Oberw. & K. Wells (Weiß
et al. 2004, Wells and Oberwinkler 1982). In addition to the fruitbodies,
EcM anatomotypes can be used to hypothesize phylogenetic relationships
(Agerer 2006). Amongst Sebacinales, only seven EcM anatomotypes
have been described to date and all of these EcMs were from temperate
areas (Selosse et al. 2002b, Urban et al. 2003, Azul et al. 2006, Twieg and
Durall 2009, Wei and Agerer 2011). A precise taxonomy should rely on a
combination of characters (Dayrat 2005). Morphological and anatomical
descriptions, in combination with other data, are a prerequisite for accurate
EcM biodiversity surveys. A greater knowledge of Sebacinales EcM morpho-
anatomy could be particularly useful for the taxonomy and biodiversity
surveys of this fungal group with cryptic or unknown fruitbody species.
This study is part of a phylogeographic and taxonomic analysis
of Pakaraimaea dipterocarpacea Maguire & Ashton ssp. nitida Maguire &
Steyerm. EcM fungi in Southern Venezuela (Moyersoen 2006, 2012a,b,c,d). P.
dipterocarpacea, an endemic tree species from Guayana region (Maguire et al.
1977, Maguire and Ashton 1980, Moyersoen 2006), is one of the few known
locally dominant EcM tree species in the Neotropical lowland forests. This tree
is phylogenetically related to the most important EcM tropical tree family in
SE Asia: the Dipterocarpaceae. The phylogeny of P. dipterocarpacea EcM fungi
might reflect the disjunct distribution of this endemic tree species. A diverse
EcM fungal community was observed in a 400 m2 P. dipterocarpacea forest plot
(Moyersoen 2012a). In total, 40 EcM fungal species were found and up to 12
different EcM morphotypes could colonize a single 125cm3 soil core. Floristic
comparisons and a molecular-phylogenetic analysis of two newly reported
Inocybe species (Moyersoen 2012a,d) indicated that host jump and fungal
dispersion (Pirozynski 1983) were probably important radiation mechanisms
for P. dipterocarpacea fungi (Moyersoen 2012a). Possible EcM endemic fungi
in the forest indicated that P. dipterocarpacea fungal communities might also
include old relictual species (Moyersoen 2012a). Similar trends regarding the
great EcM fungal diversity, the dominance of host-generalist fungi and the
presence of possible unique Agaricales were observed in a P. dipterocarpacea
ssp. dipterocarpacea forest in Guyana (Smith et al. 2013).
Tropical Sebacinales Ectomycorrhizas 81
Anatomical characters of mantle in plan views (Figs. 1b–h): Outer mantle layers
(Figs. 1b–e), a transition between a dense plectenchyma of thick-walled
ramified hyphae and a pseudoparenchyma (type E/C/H, Agerer 2006),
with a matrix, covered by a loose network of ramified, thick-walled hyphae
from which emanating hyphae and rhizomorphs originate; hyphae from
the network mostly cylindrical, with frequent short protrusions and
triangular inflation at ramifications, sometimes aculeate; ramifications
Y-shaped or at 90°, sometimes polytomies with three branches; cells 2.2–3.5
µm diam.; cell walls light orange, smooth, irregularly thick (0.5–1.7 µm);
septa simple, frequent, of the same thickness or thinner than the walls,
sometimes incomplete; anastomoses lacking; hyphae deeper in the surface
layer variably shaped; walls difficult to distinguish from the matrix at 1000X
with transmission light microscope; walls and matrix yellowish. Middle
mantle layers (Figs. 1f–g) a dense plectenchyma of ramified, cylindrical
hyphae; cells 1.7–2.7 µm diam.; ramifications Y-shaped; cell walls smooth,
yellowish, thin; septa simple, frequent, of the same thickness or thicker
than the hyphal walls, sometimes incomplete; anastomoses scarce, short,
open. Inner mantle layers (Fig. 1h) a dense plectenchyma of short, ramified
hyphae, in a matrix; cells mostly cylindrical or of variable shape; walls thin,
yellowish; septa simple, frequent, of the same thickness or thicker than the
hyphal walls, sometimes incomplete; anastomoses scarce, short, open. Very
tip with the same arrangement as in other parts of mantle.
Anatomical characters of emanating elements (Figs. 2a–d): Rhizomorphs (Figs.
2a–b) abundant, 5–25 µm diam., uniform-compact (uniform hyphae, densely
packed and glued together, type B in Agerer 2006), often with ramified
hyphae at distal end; hyphae 2.6–3.6 µm diam.; hyphal ramifications at
rhizomorph distal end, Y-shaped, at 90°, or oriented backwards; walls
yellowish, smooth, 0.7–1.5 µm thick, thin at hyphal distal end; septa simple,
frequent, of the same thickness or thinner than the hyphal walls, sometimes
incomplete; rhizomorph apex simple or with ramified, tortuous hyphae
with protrusions, and with a protuberance at the tip; anastomoses open,
short or with a bridge, only observed at rhizomorph distal end. Emanating
hyphae (Figs. 2c–d) abundant, with or without short protrusions at proximal
end, 2.8–3.2 µm thick; septa simple, frequent, evenly distributed, generally
thinner than the hyphal walls, sometimes incomplete; hyphal ends simple;
walls yellowish, 1–1.5 µm thick, sometimes thinner at distal end, surface
smooth; ramifications lacking; anastomoses lacking; shorter emanating
hyphae similar to bent awl-shaped cystidia (type A, Agerer 2006), 52–135
µm long, diam. between 1.7–2.5 and 2.6–3.7 µm, and wall thickness between
1–1.5 and 0.5–1 µm at proximal and distal end, respectively.
Anatomical characters, longitudinal section (Fig. 3): Mantle plectenchymatous,
15–25 µm wide, with two layers; outer layer 7–9 µm thick, with thick-walled
Tropical Sebacinales Ectomycorrhizas 85
Fig. 2. Sebacinales BM03M3 + Pakaraimaea dipterocarpacea ssp. nitida. a–b. Rhizomorphs, thick-
walled hyphae of uniform diameter running in parallel (a), apex simple (a) or with ramified,
tortuous hyphae with protrusions and protuberance at the tip (b), septa frequent, of the same
thickness or thinner than the walls (a, b), anastomosis open, short or with a hyphal bridge (a).
c–d. Emanating hyphae, thick-walled, with or without short protrusion at proximal end (c),
frequent simple septa, generally thinner than hyphal walls, shorter emanating hyphae similar
to bent awl-shaped cystidia (d). (Bars = 10 µm).
Fig. 10. Sebacinales PD10 + Pakaraimaea dipterocarpacea ssp. nitida. a. Plan view of middle
mantle layer, dense plectenchyma of thinner-walled, ramified hyphae, with a matrix. b. Plan
view of inner mantle layer, dense plectenchyma of ramified hyphae, septa frequent, sometimes
incomplete. c–d. Emanating hyphae, thick-walled, distal end simple, sometimes thinner-
walled and with a protuberance at the tip (c), septa infrequent, thinner than the hyphal walls,
short protrusions occurring throughout (d), ramifications Y-shaped or at 90° with triangular
inflation (d). (Bar = 10 µm).
Fig. 11. Sebacinales 6MM2 + Pakaraimaea dipterocarpacea. a–b. Plan views of outer layer,
ramified, thick-walled hyphae (a) from which many emanating hyphae originate (b), inflations
intercalary or triangular at ramifications (a), short protrusions (b), ramifications Y-shaped or
at 90° (a), polytomies with three branches (b), septa simple, frequent, of the same thickness or
thinner than the hyphal walls (a–b). c. Plan view of middle mantle layer; dense plectenchyma,
walls generally thinner than in outer layer. d. Plan view of inner mantle layer, a transition
between a plectenchyma and a pseudoparenchyma; thin walls, palmetti type lobes, anastomosis
short, open. e–g. Rhizomorph, thick-walled hyphae of uniform diameter running in parallel,
apex simple, cell walls thinner at distal end, septa straight or curved, clustered at distal end,
ramification Y-shaped (f), anastomosis open, short (g). h–i. Emanating hyphae, thick-walled,
thinner-walled at distal end, with or without short protrusion at proximal end (i), simple septa
frequent, of the same thickness or thinner than the hyphal walls, straight or curved, ramification
Y-shaped, shorter emanating hyphae similar to awl-shaped cystidia (i). (Bar = 10 µm).
hyphal net with variably shaped hyphae as in PD10 was also reported on
Sebacina incrustans + Picea abies (Urban et al. 2003), sample Y62M1 (Urban
et al. 2003), Quercirhiza dendrohyphidiomorpha (Azul et al. 2006) and Pinirhiza
multifurcata (Wei and Agerer 2011). No structures similar to PD10 staghorn-
like hyphae (thick-walled, often swollen hyphae with numerous finger-
like outgrowths and ramifications including polytomies with numerous
branches) have been previously described on Sebacinales EcM. Schematic
representations of cystidia with finger-like outgrowths (type E and I) were
reported by Agerer (1991). The term hyphae is preferred for PD10 outermost
mantle hyphal cells because these structures were bearing a variable
number of outgrowths, with variable length and sometimes septa. PD10
variably shaped hyphae are called staghorn-like hyphae for the similarity
with staghorn hyphae on Pseudotsuga menziesii (Mirb.) Franco + Byssoporia
(Poria) terrestris var. lilacinorosea M.J. Larsen & Zak EcM anatomotype (Zak
and Larsen 1978, Agerer and Rambold 2004–2011). Differences between B.
terrestris and PD10 staghorn-like hyphae included the thin walls, and the
lack of inflations and conspicuous polytomies in the first anatomotype.
6MM2 anatomotype shared features of Sebacina incrustans + Picea abies
(sample Y129M5) (Urban et al. 2003, Agerer and Rambold 2004–2011) and
Sebacinaceae sp. + Betula papyrifera Marshall (Twieg and Durall 2009). A
matrix was lacking and the thick-walled plectenchymatous outer mantle
layer was covered by numerous thick-walled emanating elements in these
three anatomotypes. Rhizomorphs were lacking in Sebacina incrustans + Picea
abies (L.) H. Karst. (sample Y129M5) (Urban et al. 2003) and Sebacinaceae sp.
+ Betula papyrifera (Twieg and Durall 2009). Emanating elements in Sebacina
incrustans + Picea abies (Urban et al. 2003, Agerer and Rambold 2004–2011)
and Sebacinaceae sp. + Betula papyrifera (Twieg and Durall 2009) were
interpreted as awl-shaped cystidia and emanating hyphae. 6MM2 shorter
emanating hyphae were similar to awl-shaped cystidia (Agerer 2006). The
thinner wall at cell distal end and the frequent septa suggest they are a
transitional growing stage of emanating hyphae. Despite similarities with
Sebacina incrustans anatomotype, the presence of rhizomorphs and the
absence of cystidia indicate that 6MM2 belongs to a different species.
5. Conclusions
This study confirmed that morpho-anatomical features are important
information, together with ultrastructure and DNA sequences to confirm
the EcM status of Sebacinales species. This first study of tropical Sebacinales
EcM anatomotypes corroborated the earlier conclusion (Agerer 2006) that
Sebacinales anatomotypes share general morpho-anatomical features.
96 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Ackowledgements
The Instituto Venezolano de Investigaciones Científicas and University
of Tübingen provided support including lab facilities. I am particularly
grateful to R Bauer for his help in ultrastructural analyses, and M. Weiß
and I. Kottke for facilitating molecular and light microscope work at an
early stage of this project. Thanks to R. Agerer, anonymous reviewers for
helpful comments on the manuscript and Krista L. McGuire and Caitlyn
Gillikin for improving the language.
Tropical Sebacinales Ectomycorrhizas 97
References
Agerer, R. 1991. Characterization of ectomycorrhiza. Methods Microbiol. 23: 25–73.
Agerer, R. 2001. Exploration types of ectomycorrhizae. A proposal to classify ectomycorrhizal
mycelial systems according to their patterns of differentiation and putative ecological
importance. Mycorrhiza 11: 107–114.
Agerer, R. 2006. Fungal relationships and structural identity of their ectomycorrhizae. Mycol.
Progress 5: 67–107.
Agerer, R. and G. Rambold. 2004–2011. DEEMY—an information system for characterization
and determination of ectomycorrizae. www.deemy.de Mϋnchen, Germany.
Altschul, S.F., LM. Thomas, A.S. Alejandro, Z. Jinghui, M. Webb and J.L. David. 1997. Gapped
BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic
Acids Research 25: 3389–3402.
Azul, A.M., R. Agerer and H. Freitas. 2006. “Quercirhiza dendrohyphidiomorpha” + Quercus
suber L. Descr. Ectomyc. 9/10: 87–91.
Dayrat, B. 2005. Towards integrative taxonomy. Biol. J. Linn. Soc. 85: 407–415.
Henkel, T.W., P. Roberts and M.C. Aime. 2004. Sebacinoid species from the Pakaraima
mountains of Guyana. Mycotaxon 89(2): 433–439.
Hynson, N.A., M. Weiß, K. Preiss, G. Gebauer and K.K. Treseder. 2013. Fungal host specificity
is not a bottleneck for the germination of Pyroleae species (Ericaceae) in a Bavarian forest.
Mol. Ecol. doi 10.1111/mec.12180.
Jairus, T., R. Mpumba, S. Chinoya and L. Tedersoo. 2011. Invasion potential and host shifts
of Australian and African ectomycorrhizal fungi in mixed eucalypt plantations. New
Phytol. 192: 179–187.
Kennedy, A.H., D.L. Taylor and L.E. Watson. 2011. Mycorrhizal specificity in the full
mycoheterotrophic Hexalectris Raf. (Orchidaceae: Epidendroideae). Mol. Ecol. 20:
1303–1316.
Kõljalg, U., K.-H. Larsson, K. Abarenkov, R.H. Nilsson, I.J. Alexander, U. Eberhardt, S. Erland,
K. Høiland, R. Kjøller, E. Larsson, T. Pennanen, R. Sen, A.F.S. Taylor, L. Tedersoo, T.
Vrålstad and B.M. Ursing. 2005. UNITE: a database providing web-based methods for
the molecular identification of ectomycorrhizal fungi. New Phytol. 166(3): 1063–1068.
Maguire, B., P.S. Ashton, C. de Zeeuw, D.E. Giannasi and K.J. Niklas. 1977. Pakaraimoideae,
Dipterocarpaceae of the Western hemisphere. Taxon 26(4): 341–385.
Maguire, B. and P.S. Ashton. 1980. Pakaraimaea dipterocarpacaea II. Taxon 29(2/3): 225–231.
Maguire, B. and J.A. Steyermark. 1981. Pakaraimaea dipterocarpacea. III. Memoires of the New
York Botanical Garden 32: 306–309.
Morris, M.H., M.A. Pérez-Pérez, M.E. Smith and C.S. Bledsoe. 2008. Multiple species of
ectomycorrhizal fungi are frequently detected on individual oak root tips in a tropical
cloud forest. Mycorrhiza 18: 375–383.
Morris, M.H., M.A. Pérez-Pérez, M.E. Smith and C.S. Bledsoe. 2009. Influence of host species
on ectomycorhizal communities associated with two co-occurring oaks (Quercus spp.)
in a tropical cloud forest. FEMS doi:10.1111/j.1574-6941.2009.00704.x.
Moyersoen, B. 2006. Pakaraimea dipterocarpacea is ectomycorrhizal, indicating an ancient
Gondwanaland origin for the ectomycorrhizal habit in Dipterocarpaceae. New Phytol.
172: 753–762.
Moyersoen, B. 2012a. Dispersion, an important radiation mechanism for ectomycorrhizal
fungi in neotropical lowland forests? In: P. Sudarshana, M. Nageswara–Rao and J.R.
Soneji (eds.). Tropical Forests, pp. 93–116.
Moyersoen, B. 2012b. Clavulina sp. + Pakaraimaea dipterocarpacea Maguire & Ashton ssp. nitida
Maguire & Steyerm. Descr. Ectomyc. 13: 16–23.
Moyersoen, B. 2012c. Coltriciella sp. + Pakaraimaea dipterocarpacea Maguire & Ashton ssp. nitida
Maguire & Steyerm. Descr. Ectomyc. 13: 24–30.
98 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Moyersoen, B. 2012d. Inocybe sp. + Pakaraimaea dipterocarpacea Maguire & Ashton ssp. nitida
Maguire & Steyerm. Descr. Ectomyc. 13: 42–47.
Nilsson, R.H., E. Kristiansson, M. Ryberg, N. Hallenberg and K.-H. Larsson. 2008. Intraspecific
ITS variability in the kingdom fungi as expressed in the international sequence databases
and its implications for the molecular species identification. Evol. Bioinf. 4: 193–201.
Peay, K.G., P.G. Kennedy, S.J. Davies, S. Tan and T.D. Bruns. 2009. Potential link between
plant and fungal distributions in a dipterocarp rainforest: community and phylogenetic
structure of tropical ectomycorrhizal fungi across a plant and soil ecotone. New Phytol.
185: 529–542.
Pirozynski, K.A. 1983. Pacific mycogeography: an appraisal. Aust. J. Bot., Suppl. Ser. 10:
137–159.
Rivière, T., A.G. Diedhiou, M. Diabate, G. Senthilarasu, K. Natarajan, A. Verbeken, B. Buyck, B.
Dreyfus, G. Bena and A.M. Bâ. 2007. Genetic diversity of ectomycorrhizal Basidiomycetes
from African and Indian tropical rain forests. Mycorrhiza 17: 415–428.
Roy, M., S. Watthana, A. Stier, F. Richard, S. Vessabutr and M.-A. Selosse. 2009. Two
mycoheterotrophic orchids from Thailand tropical dipterocapacean forests associate
with a broad diversity of ectomycorrhizal fungi. BMC Biology 7: 51 doi:10.1186/1741-
7007-7-51.
Selosse, M.-A., M. Weiß, J.-L. Jany and A. Tillier. 2002a. Communities and populations of
Sebacinoid basidiomycetes associated with the achlorophyllous orchid Neottia nidus-avis
(L.) L.C.M. Rich. and neighbouring tree ectomycorrhizae. Mol. Ecol. 11: 1831–1844.
Selosse, M.-A., R. Bauer and B. Moyersoen. 2002b. Basal hymenomycetes belonging to the
Sebacinaceae are ectomycorrhizal on temperate deciduous trees. New Phytol. 155:
183–195.
Setaro, S.D., S. Garnica, P.I. Herrera, J.P. Suárez and M. Göker. 2012. A cluster optimization
strategy to estímate species richness of Sebacinales in the tropical Andes based on molecular
sequences from distinct DNA regions. Biodivers. Conserv. 21: 2269–2285.
Smith, M.E., T.W. Henkel, M.C. Aime, A.K. Fremier and R. Vilgalys. 2011. Ectomycorrhizal
fungal diversity and community structure on three co-occurring leguminous canopy tree
species in a Neotropical rainforest. New Phytol. doi: 10.1111/j.1469-8137.2011.03844.x.
Smith, M.E., T.W. Henkel, J.K. Uehling, A.K. Fremier, H.D. Clarke and R. Vilgalys. 2013. The
ectomycorrhizal fungal community in a Neotropical forest dominated by the endemic
dipterocarp Pakaraimaea dipterocarpacea. PLoS one 8(1): e55160. doi:10.1371/journal.
pone.0055160.
Spurr, A.R. 1969. A low-viscosity epoxy resin embedding medium for electron microscopy. J.
Ultrastruct. Res. 26: 31–43.
Tedersoo, L., R.H. Nilsson, K. Abarenkov, T. Jairus, A. Sadam, I. Saar, M. Bahram, E. Bechem,
G. Chuyong and U. Kõljalg. 2010a. 454 Pyrosequencing and Sanger sequencing of tropical
mycorrhizal fungi provide similar results but reveal substantial methodological biases.
New Phytol. doi:10.1111/j.1469-8137.2010.03373.x.
Tedersoo, L., A. Sadam, M. Zambrano, R. Valencia and M. Bahram. 2010b. Low diversity
and high host preference of ectomycorrhizal fungi in Western Amazonia, a neotropical
biodiversity hotspot. The ISME J. 4: 465–471.
Tedersoo, L., M. Bahram, T. Jairus, E. Bechem, S. Chinoya, R. Mpumba, M. Leal, E.
Randrianjohany, S. Razafimandimbison, A. Sadam, T. Naadel and U. Kõljalg. 2011. Spatial
structure and the effects of host and soil environments on communities of ectomycorrhizal
fungi in wooded savannas and rain forests of Continental Africa and Madagascar. Mol.
Ecol. 20: 3071–3080.
Twieg, B. and D. Durall. 2009. Sebacinaceae sp. + Betula papyrifera Marsh. In: D.M. Goodman,
D.M. Durall, J.A. Trofymow and S.M. Berch (eds.). Concise descriptions of North
American Ectomycorrhizae. Mycologue Publications, and Canada-B. C. Forest Resource
Development Agreement, Canadian Forest Service, Victoria, B. C., pp. CDE28.
Urban, A., M. Weiß and R. Bauer. 2003. Ectomycorrhizas involving sebacinoid mycobionts.
Mycol. Res. 107(1): 3–14.
Tropical Sebacinales Ectomycorrhizas 99
Wei, J. and R. Agerer. 2011. Two Sebacinoid ectomycorrhizae on Chinese pine. Mycorrhiza
21: 105–115.
Weiß, M., M.A. Selosse, K.L. Rexer, A. Urban and F. Oberwinkler. 2004. Sebacinales: a hitherto
overlooked cosm of heterobasidiomycetes with a broad mycorrhizal potential. Mycol.
Res. 108(9): 1003–1010.
Weiß, M., Z. Sýkorová, S. Garnica, K. Riess, F. Martos, C. Krause, F. Oberwinkler, R. Bauer and
D. Redecker. 2011. Sebacinales everywhere: previously overlooked ubiquitous fungal
endophyes. PLos ONE 6(2): e16793. doi: 10.1371/journal.pone.0016793.
Wells, K. and F. Oberwinkler. 1982. Tremelloscypha gelatinosa, a species of a new family
Sebacinaceae. Mycologia 74(2): 325–331.
Zak, B. and M.J. Larsen. 1978. Characterization and classification of mycorrhizae of Douglas-
fir. III. Pseudotsuga menziesii + Byssoporia (Poria) terrestris vars. lilacinorosea, parksii, and
sublutea. Can. J. Bot. 56: 1416–1424.
CHAPTER
6
Abundance, Distribution, and
Function of Pisolithus albus
and other Ectomycorrhizal
Fungi of Ultramafic Soils in
New Caledonia
Philippe Jourand,1,* Fabian Carriconde,2
Marc Ducousso,3 Clarisse Majorel,1 Laure Hannibal,1
Yves Prin3 and Michel Lebrun4
1. Introduction
Ultramafic soils, also known as “serpentine soils” in literature, are a
weathered product from ultramafic bedrock that covers less than 1% of the
earth’s surface (Coleman and Jove 1992). These soils are characterized by
high concentrations of iron oxides (up to 85% w/w), unbalanced calcium-
1
IRD, UR040 LSTM, Centre IRD, BPA5, Promenade Roger Laroque, 98848 Nouméa Cedex,
Nouvelle-Calédonie.
2
Institut Agronomique néo-Calédonien (IAC), Axe 2 ‘Diversités biologique et fonctionnelle
des écosystèmes terrestres’, Nouméa IRD research centre, BP18239, 98857 Nouméa, Nouvelle-
Calédonie.
3
CIRAD, UMR LSTM, TA A-82 ⁄ J Campus International de Baillarguet, 34398 Montpellier
Cedex 5 France.
4
Université Montpellier 2, UMR28 LSTM, TA A-82 ⁄ J Campus International deBaillarguet,
34398 Montpellier Cedex 5, France.
*Corresponding author: philippe.jourand@ird.fr
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia 101
Fig. 1. Geographical map of the New Caledonian archipelago in the South Pacific Ocean with
location of ultramafic massifs (in grey). Data from Perrier et al. (2006a).
Fig. 2. Repartition of the four distinct plant formations (sites 1 to 4) on the topographic sequence
studied by Perrier et al. (2006) at the Koniambo Massif. Most abundant and potential species
for restoration purpose within each vegetation type are shown. Plants with ECM structures
or ECM-like-structures on their root systems are indicated by an asterisk or two asterisks,
respectively. The distance in meters (m) from the valley to the plateau and the altitude are
given in abscissa and ordinate, respectively. Modified data from Perrier et al. (2006a).
104 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 1. Plant families and species in New Caledonia characterized as ECM. The biogeographical
native status is given according to Jaffré et al. (2001) (N: native, i.e., species for which their
natural distribution area extend beyond the boundaries of New Caledonia; E: endemic species;
EE: endemic genus). The types of soils on which species are encountered are also indicated (C:
calcareous; UM: ultramafic soils; VS: volcano-sedimentary). For species known to be present
on more than one type of soil, the predominant types are highlighted in bold. Modified data
from Amir and Ducousso (2010).
Nothofagus balansae E UM
Nothofagus codonandra E UM
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia 105
balansae (site 4), located on the topographic sequence at the Koniambo Massif and genotyped by sequencing of the ITS region. The host plant (putative),
the morphospecies when available, the ITS sequence length, the closest BLAST match and the related information are presented. ITS sequence data
generated by Perrier (2005) were recently analyzed.
GenBank Sequence
Plant Host plant Bases Best match GenBank
Sample reference Sample type Morphospecies accesion lenght Closest species BLAST match ‡ % Similarity
formation (putative) † matched accesion number
number (bp)
K66C Sporophore 3 Tristaniopsis guillainii Pisolithus sp FJ656011 527 Pisolithus sp 520/526 99% AF270787
K02C Sporophore 4 Nothofagus balansae Boletus sp FJ656001 609 Boletus sp 384/449 86% EU569234
K05C Sporophore 4 Nothofagus balansae nd FJ656002 551 Cortinarius subgemmeus* 498/561 89% JX000354
K06C Sporophore 4 Nothofagus balansae - FJ656003 605 Phellodon sp 544/597 91% GU222318
K09C Sporophore 4 Nothofagus balansae nd FJ656004 547 Austrogautieria macrospora* 438/504 87% GQ981492
K10C Sporophore 4 Nothofagus balansae nd FJ656005 670 Tricholoma imbricatum 628/668 94% AY573537
K10C Sporophore 4 Nothofagus balansae nd FJ656005 670 Tricholoma imbricatum 626/668 94% AY573537
K12C Sporophore 4 Nothofagus balansae nd FJ656006 572 Cortinarius austrovenetus 534/573 93% GQ890318
K14C Sporophore 4 Nothofagus balansae Inocybe sp FJ656007 506 Dermocybe largofulgens* 483/504 96% GU233324
K16C Sporophore 4 Nothofagus balansae nd FJ656008 706 Lactarius scrobiculatus 660/719 92% EU597079
K18C Sporophore 4 Nothofagus balansae Lactaroides FJ656009 598 Russula zonaria * 526/569 92% DQ421990
KC02C Sporophore 4 Nothofagus balansae nd FJ656012 436 Cortinarius lividus 390/433 90% AF539734
KC05C Sporophore 4 Nothofagus balansae nd FJ656014 580 Inocybe aeruginascens* 491/569 86% GU949591
KC08C Sporophore 4 Nothofagus balansae nd FJ656016 670 Lactarius olympianus 624/684 91% EF685079
KC11C Sporophore 4 Nothofagus balansae nd FJ656018 609 Russula sp 577/608 95% GU222292
KC12C Sporophore 4 Nothofagus balansae nd FJ656019 518 Cortinarius flammuloides 460/540 85% AF539716
KC16C Sporophore 4 Nothofagus balansae nd FJ656020 574 Cortinarius multiformis 518/589 88% AF389135
KC17C Sporophore 4 Nothofagus balansae nd FJ656021 574 Leratiomyces ceres 394/411 96% HQ604750
KC19C Sporophore 4 Nothofagus balansae nd FJ656022 583 Cortinarius singularis * 508/584 87% JQ287672
KC22C Sporophore 4 Nothofagus balansae nd FJ656023 528 Austrogautieria macrospora* 442/503 88% GQ981492
KD37C Sporophore 4 Nothofagus codonandra nd FJ656038 582 Cortinarius eutactus* 559/582 96% JX000366
KC23C Sporophore 4 Tristaniopsis guillainii nd FJ656024 414 Cortinarius elaiops* 241/262 92% JX000369
K01C Sporophore 4 Nothofagus balansae nd FJ656000 701 Tricholoma ustale 632/713 89% AF458435
K22C Sporophore 4 Nothofagus balansae nd FJ656010 582 Inocybe aeruginascens* 491/569 86% GU949591
KC03C Sporophore 4 Nothofagus balansae nd FJ656013 612 Russula sp 593/613 97% GU222292
KC06C Sporophore 4 Nothofagus balansae nd FJ656015 436 Phaeocollybia redheadii 394/411 96% JN102541
Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
KC10C Sporophore 4 Nothofagus balansae nd FJ656017 447 Cortinarius aff. austrosanguineus 427/454 94% GQ890317
KD36C Sporophore 4 Nothofagus codonandra nd FJ656037 585 Cortinarius elaiops* 554/586 95% JX000369
KD42C Sporophore 4 Nothofagus codonandra nd FJ656039 485 Tricholoma ustale 402/455 88% AF458435
KE01-2M ECM 3 Tristaniopsis guillainii - FJ656040 457 Piloderma sp 400/450 89% JQ711951
KE02M ECM 3 Tristaniopsis guillainii - FJ656041 511 Cortinarius vernicifer* 387/449 86% JX000370
KE04M ECM 3 Tristaniopsis guillainii - FJ656042 438 Piloderma sp 383/430 89% JQ711951
KD10M ECM 4 Nothofagus balansae - FJ656025 552 Oidiodendron chlamydosporicum 477/519 92% AF062789
KE06M ECM 4 Nothofagus balansae - FJ656043 584 Cortinarius amoenus 544/590 92% AF389160
KE12-1M ECM 4 Nothofagus balansae - FJ656045 620 Tricholoma ustale 557/640 87% AF458435
KD18M ECM 4 Nothofagus codonandra nd FJ656026 474 Cortinarius calyptratus* 425/476 89% EU525980
f g yp
KD29-2M ECM 4 Nothofagus codonandra - FJ656033 608 Cortinarius elaiops* 558/606 92% JX000369
KD31''-2M ECM 4 Nothofagus codonandra - FJ656034 570 Cortinarius elaiops* 518/600 86% JX000369
KD31'M ECM 4 Nothofagus codonandra - FJ656035 682 Tomentellopsis submollis 642/684 94% JQ711898
KD36-2M ECM 4 Nothofagus codonandra - FJ656036 584 Cortinarius singularis* 509/584 87% JQ287672
KD19_1S Hyphae 3 Tristaniopsis guillainii - FJ656027 698 Lycoperdon sp 682/726 94% JX029934
KD19_2S Hyphae 3 Tristaniopsis guillainii nd FJ656028 592 Cortinarius eutactus* 560/597 94% HQ533023
KD19_9S Hyphae 3 Tristaniopsis guillainii - FJ656029 410 Cortinarius sp 350/402 87% JQ287690
KD20_5S Hyphae 3 Tristaniopsis guillainii - FJ656031 585 Cortinarius sp 543/594 91% JN942302
KD20_6S Hyphae 3 Tristaniopsis guillainii nd FJ656032 615 Cortinarius sp 552/621 89% JN942302
KE12_2S Hyphae 4 Nothofagus balansae - FJ656046 608 Tricholoma ustale 554/629 88% AF458435
KE18_2S Hyphae 4 Nothofagus codonandra - FJ656047 540 Cortinarius subgemmeus* 477/560 85% JX000354
† ECM root tips were sampled by tracing the roots from the tree trunks.
‡ Voucher specimens are indicated by an asterisk.
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia
107
108 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 3. (A) Podoserpula miranda and (B) Cantharellus garnieri. Photos by courtesy from Ducousso
Marc, CIRAD.
Color image of this figure appears in the color plate section at the end of the book.
Fig. 4. Pisolithus albus from New Caledonia. A: Pisolithus albus MD07-117 from the Koniambo
massif; B: Pisolithus albus MD07-228 from the Ouen-Toro, Noumea; C: cross section of Pisolithus
albus MD07-166 from Pindjen water-fall and D: globose spores (8.77 to 9.62 µm) of Pisolithus
albus MD06-379 from Poum, erected spines (1.2 µm) are clearly visible. From Jourand et al.
(2010a).
Color image of this figure appears in the color plate section at the end of the book.
P. aurantioscabrosus, SE Asia
41
Pisolithus sp.10, Australia
95
87 Pisolithus sp.8 and P. microcarpus,
55
Australia, S America
72
NC
40
52 NC
32
100 51
P. albus
44 NC New Caledonia,
23 65 NC Australia, New-Zealand,
SE Asia, W Africa
NC
68
13
63 98
P. marmoratus, Australia
37
Pisolithus sp.3 and P. sp.4, Europe
57
46
76
P. tinctorius, N America, Europe
100
Fig. 5. Phylogenetic synthetic relationships among representative Pisolithus sp. from New
Caledonia collection sites and worldwide reference isolates. The phylogeny is based on
the analysis of the rDNA ITS1, 5.8S and ITS2 sequences. Tree shown is a 50% Majority rule
consensus of the most parsimonious trees (Tree Length = 3733) obtained with PAUP4 (see
Materials & Methods). Values indicated at tree nodes are percentage values of 1000 bootstrap
replicates under MP criterion using fast stepwise addition (only values >50% are shown).
The tree was rooted with Suillus luteus ITS sequences. Significant bootstrap frequencies are
indicated. Abbreviations: S America: South America; SE Asia: South East Asia; W Africa: West
Africa, E Africa: East Africa.
In ultramafic soils, nickel (Ni) is one the most bioavailable and phytotoxic
element: nickel content may reach up to 10 g/kg in ultramafic soils when
compared with the average 50 mg/kg in cultivated soils (Wenzel and
Jockwer 1999, Echevarria et al. 2006). This mineral element is a crucial
selecting factor for plant survival on ultramafic soils: to grow on such
high concentrations of nickel as found in serpentine environments (often
coinciding with high concentrations of other heavy metals), plants had to
develop major adaptations that include exclusion of the absorption of the
toxic metal by the roots and/or metal hyperaccumulation with internal
complexation and compartmentation (Kazakou et al. 2008). In addition,
ECM symbioses might contribute to limit the metal accessibility and uptake
by the plant (Colpaert et al. 2011).
average Ni EC50 two to three times higher than the Ni EC50 already reported
for other Pisolithus spp. mentioned above. To explain the high variability in
nickel-tolerance observations, it was first hypothesized that such variations
could be correlated to high real fluctuations of bioavailable nickel content
in ultramafic soils, which is assessed as the DTPA-Ni fraction according to
Echevarria et al. (2006). Perrier et al. (2006a) reported that the nickel-DTPA
concentrations in ultramafic soils varied in a range from 17 to 980 µmol/
kg. Assuming that the average nickel-DTPA concentration does not reflect
real fluctuations of bioavailable nickel in ultramafic soils, and considering
the range of nickel-DTPA concentrations in ultramafic soils reported by
Perrier et al. (2006a,b), it is not surprising to find isolates of P. albus with high
variations in nickel tolerance from the same ultramafic site. Similar variations
in metal-tolerant fungal populations in correlation to metal-soil content have
already been reported. For instance, in Suilloid fungi, populations displayed
zinc tolerance relative to zinc concentrations in polluted soils, suggesting
an evolutionary adaptation of fungi to the soil environment (Colpaert et
al. 2004). More recently, evidence of adaptation to nickel was provided in
isolates of Cenococcumgeophilum from ultramafic soils in Portugal and the
USA (Gonçalves et al. 2009). No clear relationship between the phenotypic
physiological response to nickel and the population genetic differentiation
observed within P. albus from soils could be established as the nickel-tolerant
isolates from ultramafic soils did not cluster in a homogeneous group. It
was thus tempting to speculate that the capacity of some P. albus to tolerate
high nickel concentrations reflects the expression of an adaptive response
to high concentrations of bioavailable nickel in soils as suggested for other
fungi in response to high heavy metal levels (Hartley et al. 1997, Colpaert
et al. 2004, Gonçalves et al. 2009). However, if New Caledonian population
of P. albus seems to be structured into one ecotype, nickel tolerance alone
might not be a sufficient feature to explain such results. Thus, the ultramafic
constraint should be considered as a whole, even if each factor (N, P, K
contents, Ca/Mg imbalance, heavy metal presence) is studied separately,
as suggested by Kazakou et al. (2008).
Fig. 7. A) Scatter plot presenting gene expression levels in Pisolithus albus Ni-tolerant
ecotypefree-living mycelium grown without or with Ni at 250 µM. The expression levels of
genes were normalized using a scale of 0 to 10,000. Each circle in the plot represents expression
of one gene. B) Functional GO terms assignment and distribution of total sequences of two
transcriptomes of Ni-tolerant P. albus with (+250 µM) and without nickel, among Gene
Ontology (GO) biological process, molecular function and cellular component. From Majorel
et al. (2012).
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia 117
80
Gene N° 1 (GPI- anchor-like) Gene N° 2 (predicted protein)
1.2x
70 4
60 1x
1.5x
mRNA accumulation
mRNA accumulation
3
Arbitrary units
Arbitrary units
50
40 2.2x
2
30
1.2x
20 1.4x 1
10 ND ND
ND ND
0 0
MD06 -337 MD09 -001 MD09 -045 MD09 -063 MD09 -078 MD06-337 MD09 -001 MD09-045 MD09 -063 MD09 -078
1x
(Arbitrary units)
Arbitrary units
1 2.5x
3
2 0,6
1x
1x
1 1.4x
ND ND 0,2 ND ND
0 0
MD06 -337 MD09- 001 MD09 -045 MD09 -063 MD09 -078 MD06 -337 MD09 -001 MD09 -045 MD09-063 MD09 -078
Gene N°6 (S - adenosylmethionine transferase) Gene N°9 (APC amino acid permease)
30
1.6x 9x
25 400
mRNA accumulation
mRNA accumulation
(Arbitrary units)
Arbitrary units
20 300
1.3x
15 60
10 40
1x
5 1.2x 20 1.3x
ND ND ND ND
0 0
MD06 -337 MD09 -001 MD09 -045 MD09 -063 MD09 -078 MD06 -337 MD09 -001 MD09-045 MD09-063 MD09-078
Fig. 8. Comparison of mRNA accumulation profiles for six selected Ni up-regulated genes in
five P. albus isolates from ultramafic soil in presence of nickel 50 µM (black columns) and in
absence of nickel (grey columns). Three nickel-tolerant isolates (MD06-337, MD09-045, and
MD07-001) and two nickel-sensitive isolates (MD09-078 and MD09-063) were compared.
Transcript accumulation was quantified by qPCR using 2-ΔΔCT method with normalization to
two reference genes, GAPDH and EF4α, and is expressed as arbitrary units. The data indicate
mean values ± S.D. values, calculated from three technical replicates with triplicate biological
samples. The fold induction by nickel is presented above the black columns in italics. ND:
mRNA non-detected (Ct values >37). From Majorel et al. (2012).
118 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
As ECM symbioses are known to play a major role in the fitness of plants
in the presence of heavy metals (Jentschke and Godbold 2000), experiments
were carried out to analyse the symbiotic interactions between P. albus
and one of its host plants in the presence of nickel. Ectomycorrhizal
Pisolithus albus isolated in nickel-rich ultramafic soils from New Caledonia
and showing in vitro adaptive nickel tolerance were inoculated to
Eucalyptus globulus Labill used as a Myrtaceae plant-host model to study
ectomycorrhizal symbiosis. Plants were then exposed to a nickel dose-
response experiment with increased nickel treatments up to 60 mg/kg soil
as maximum extractable nickel content found in ultramafic soils (Perrier
et al. 2006a). Results showed that plants inoculated with ultramafic ECM
P. albus were able to tolerate high and toxic concentrations of Ni (up to
60 mg/kg) while uninoculated controls were not (Fig. 9). At the highest
nickel concentration tested, root growths were more than 20-fold higher
and shoot growths more than 30-fold higher in ECM plants compared with
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia 119
Fig. 9. Eucalyptus globulus seedlings after 12-weeks growth. A and A’ mycorrhizal; B and B’:
non-mycorrhizal (controls). A and B: no nickel added; A’ and B’ seedlings treated with Ni.
From Jourand et al. (2010b).
Color image of this figure appears in the color plate section at the end of the book.
4. Conclusions
Overall, the observations about ECM diversity found on ultramafic soils
in New Caledonia raise very compelling questions about the evolutionary
processes involved in fungal diversification in New Caledonia and at a
regional scale. The focus on ECM Pisolithus albus isolated from soils in New
Caledonia highlighted the identification of an ultramafic nickel-tolerant
ecotype as reported in Jourand et al. (2010a), showing specific and adaptive
molecular response to this metal (Majorel et al. 2012), and having a key role
in plant host adaptation to toxic nickel concentrations as found in these soils
(Jourand et al. 2010b). Together, these results constitute an important step in
evaluating the potential of ECM symbioses for plant adaptation to ultramafic
soils containing high concentrations of heavy metals, which is a prerequisite
for their use in strategies for ecological restoration of mine sites as suggested
by Reddell et al. (1999), Perrier et al. (2006a) and, more recently, Khosla and
Reddy (2008). Further characterization of ECM fungal communities in New
Caledonia would increase knowledge about fungal diversity and identify
fungal species that might be relevant for plant inoculation purpose and
their direct implications in restoration strategies.
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia 121
Acknowledgements
Most of these studies were supported by (i) the GIP CNRT “Nickel and its
Environment” [grant number GIPCNRT98] and (ii) the ANR ECCO2005
and BIODIV2007 research programs entitled “Niko” and “Ultrabio”
respectively. The authors wish to thank Pr R. Reid (University of Adelaide,
South Australia), Dr T. Jaffré (IRD, Nouméa, New Caledonia), S. Santoni
(INRA, Montpellier, France), M.E. Soupe, J. Riss, C. Richert for their
respective contributions and/orsuggestions. They are also thankful Mr.
Pierrick Gailhbaud and Mr Antoine Leveau of Koniambo Nickel Society
(KNS), Vavouto, Koné, New Caledonia. The authors thank the anonymous
referees for their valuable comments on this study, and Krista L. McGuire
and Caitlyn Gillikin for improving the language.
References
Alexander, E., R. Coleman, T. Keeler-Wolfe and S. Harrison. 2007. Serpentine Geoecology
of Northern North America. Geology, Soils, and Vegetation. Oxford University Press,
New York.
Aggangan, N., B. Dell and N. Malajczuk. 1998. Effects of chromium and nickel on growth of
the ectomycorrhizal fungus Pisolithus and formation of ectomycorrhizas on Eucalyptus
urophylla S.T. Blake. Geoderma 84: 15–27.
Altschul, S.F., W. Gish, W. Miller, E.W. Myers and D.J. Lipman. 1990. Basic local alignment
search tool. J. Mol. Biol. 215: 403–10.
Amir, H. and M. Ducousso. 2010. Les bactéries et les champignons du sol sur roches
ultramafiques. In: L. L’huillier, T. Jaffré and A. Wulff (eds.). Mines et environnement en
Nouvelle-Calédonie: les milieux sur substrats ultramafiques et leur restauration. IAC,
Nouméa, Nouvelle-Calédonie, pp. 129–145.
Anderson, I.C., S.M. Chambers and J.W.G. Cairney. 1998. Use of molecular methods to estimate
the size and distribution of mycelial individuals of the ectomycorrhizal basidiomycete
Pisolithus tinctorius. Mycol. Res. 102: 295–300.
Bellion, M., M. Courbot, C. Jacob, D. Blaudez and M. Chalot. 2006. Extracellular and cellular
mechanisms sustaining metal tolerance in ectomycorrhizal fungi. FEMS Microbiol. Let.
254: 173–181.
Blaudez, D., C. Jacob, K. Turnau, J.V. Colpaert, U. Ahonen-Jonnarth, R. Finlay, B. Botton and
M. Chalot. 2000. Differential responses of ectomycorrhizal fungi to heavy metals in vitro.
Mycol. Res. 104: 1366–1371.
Bolchi, A., R. Ruotolo, G. Marchini, E. Vurro, L. Sanità di Toppi, A. Kohler, E. Tisserant, F.
Martin and S. Ottonello. 2011. Genome-wide inventory of metal homeostasis-related
gene product including a functional phytochelatin synthase in th hypogeous mycorrhizal
fungus Tuber melanosporum. Fungal Genet. Biol. 48: 573–584.
Bougher, N.L., B.A. Fuhrer and E. Korak. 1994. Taxonomy and biogeography of Australian
Rozites species mycorrhizal with Nothofagus and Myrtaceae. Aust. J. Bot. 7: 353–375.
Brady, K., A. Kruckberg and H. Bradshaw. 2005. Evolutionary ecology of plant adaptation to
serpentine soils. Ann. Rev. Ecol. Evol. Syst. 36: 243–266.
Branco, S. 2010. Serpentine soils promote ectomycorrhizal fungal diversity. Mol. Ecol. 19:
5566–5576.
Branco, S. and R.H. Ree. 2010. Serpentine soils do not limit mycorrhizal fungal diversity.
PLoS One 5:e11757.
122 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Brearley, F.Q. 2006. Differences in the growth and ectomycorrhizal community of Dryobalanops
lanceolata (Dipterocarpaceae) seedlings grown inultramafic and non-ultramafic soils.
Soil Biol. Biochem. 38: 3407–3410.
Brooks, R. 1987. Serpentine and its vegetation. Dioscorides Press, Portland.
Brundrett, M., N. Bougher, B. Dell, T. Grove and N. Malajczuk. 1996. Working with mycorrhizas
in forestry and agriculture. ACIAR Monogr. 32. Australian Centre for International
Agricultural Research, Canberra, Australia, p. 374.
Chiarucci, A. and A.J.M. Baker. 2007. Advances in the ecology ofserpentine soils. Plant Soil
293: 1–217.
Coleman R.G. and C. Jove. 1992. Geological origin of serpentinites. In: A.J.M. Baker, J. Proctor
and R.D. Reeves (eds.). The Vegetation of Ultramafic (Serpentine) Soils. Proceedings of
the First International Conference on Serpentine Ecology. Intercept, Hampshire, UK,
pp. 1–17.
Colpaert, J.V., L.A.H. Muller, M. Lambaerts, K. Adriaensen and J. Vangronsveld. 2004.
Evolutionary adaptation to Zn toxicity in populations of Suilloid fungi. New Phytol.
162: 549–560.
Colpaert, J.V., J. Wevers, E. Krznaric and K. Adriaensen. 2011. How metal-tolerant ecotypes
of ectomycorrhizal fungi protect plants from heavy metal pollution. Annals of Forest
Science 68: 17–24.
Dickie, I.A., N. Bolstridge, J.A. Cooper and D.A. Peltzer. 2010. Co-invasion by Pinus and its
mycorrhizal fungi. New Phytol. 187: 475–484.
Ducousso, M., C. Contesto, M. Cossegal and Y. Prin. 2004. Cantharellus garnierii sp. nov., une
nouvelle chanterelle des maquis nickélifères de Nouvelle-Calédonie. Cryptogamie
Mycol. 25: 135–145.
Ducousso, M., S. Proust, V. Denis and G. Eyssartier. 2009. Podoserpula miranda nom prov., une
nouvelle espèce de champignon très spectaculaire découverte en Nouvelle-Calédonie.
Bois et Forêts des Tropiques 302: 73–75.
Ducousso, M., F. Carriconde, E. Frischt, F. Juillot, L. Hannibal, C. Majorel and Jourand P.
2012. Diversité des champignons ectomycorhiziens d’Acaciaspirorbis dans des habitats
très contrastés en Nouvelle-Calédonie. Journées Francophones Mycorhizes, troisième
édition, 5-7 septembre 2012, Nancy, France.
Echevarria, G., S. Massoura, T. Sterckeman, T. Becquer, C. Schwartz and J.L. Morel. 2006.
Assessment and control of the bioavailability of Ni in soils. Environ. Toxicol. Chem. 25:
643–651.
Finlay, R.D. 2004. Mycorrhizal fungi and their multifunctional roles. Mycol. 18: 91–96.
Fuchs, B.B. and E. Mylonakis. 2009. Our paths might cross: the role of the fungal cell wall
integrity pathway in stress response and cross talk with other stress response pathways.
Eukaryot. Cell 8: 1616–1625.
Garnica, S., M. Weib and F. Oberwinkler. 2003. Morphological and molecular phylogenetic
studies inSouth American Cortinarius species. Mycol. Res. 107: 1143–1156.
Gonçalves, S.C., A. Portugal, M.T. Goncalves, R. Vieira, M.A. Martins-Loucao and H.
Freitas. 2007. Genetic diversity and differential in vitro responses to Ni in Cenococcum
geophilumisolates from serpentine soils in Portugal. Mycorrhiza 17: 677–686.
Gonçalves, S.C., M.A. Martins-Louçao and H. Freitas. 2009. Evidence of adaptative tolerance
to nickel in isolates of Cenococcum geophilum from serpentine soils. Mycorrhiza 19:
221–230.
Harrington, T.J. and D.T. Mitchell. 2002. Colonisation of root systems of Carex flacca and C.
pilulifera by Cortinarius (Dermocybe) cinnamomeus. Mycol. Res. 106: 452–459.
Harrison, S. and N. Rajakaruna. 2011. Serpentine: the Evolution and Ecology of a Model System.
The University of California Press, Berkeley and Los Angeles, CA.
Hartley, J., J.W.G. Cairney and A.A. Meharg. 1997. Do ectomycorrhizal fungi exhibit adaptive
tolerance to potentially toxic metals in the environment? Plant Soil 189: 303–319.
Hawksworth, D.L. 1991. The fungal dimension of biodiversity: magnitude, significance, and
conservation. Mycol. Res. 95: 641–655.
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia 123
Hawksworth, D.L. 2001. The magnitude of fungal diversity: the 1.5 million species estimate
revisited. Mycol. Res. 105: 1422–1432.
Horak, E. and J. Mouchacca. 1998. Annoted checklist of New Caledonian Basidiomycota. I.
Holobasidiomycetes. Mycotaxon 68: 75–129.
Horak, E. and A.E. Wood. 1990. Cortinarius Fr. (Agaricales) in Australasia. 1. Subgen. Myxacium
and subgen. Paramyxacium. Sydowia 42: 88–168.
Izzo, A., J. Agbowo and T.D. Bruns. 2005. Detection of plot-level changes in ectomycorrhizal
communities across years in a old-growth mixed-conifer forest. New Phytol. 166:
619–630.
Jacob, C., M. Courbot, F. Martin, A. Brun and M. Chalot. 2004. Transcriptomic responses to
cadmium in the ectomycorrhizal fungus Paxillus involutus. FEBS Letters 576: 423–427.
Jaffre, T. 1992. Floristic and ecological diversity of the vegetation on ultramafic rocks in New
Caledonia. In: A.J.M. Baker, J. Proctor and R.D. Reeves (eds.). The Vegetation of Ultramafic
Soils. Intercept Ltd, Andover, UK, pp. 101–107.
Jaffre, T. and L. L’Huillier. 2010. La végétation des roches ultramafiques ou terrains miniers.
In: L. L’huillier, T. Jaffré and A. Wulff (eds.). Mines et environnement en Nouvelle-
Calédonie: les milieux sur substrats ultramafiques et leur restauration. IAC, Nouméa,
Nouvelle-Calédonie, pp. 45–103.
Jaffré, T., P. Morat, J.M. Veillon, F. Rigault and G. Dagostini. 2001.Composition and
characterisation of the native flora of New Caledonia. Documents Scientifiques et
Techniques—IRD: II, 121 p.
Jentschke, G. and D.L. Godbold. 2000. Metal toxicity and ectomycorrhizas. Physiol. Plant
109: 107–116.
Jourand, P., M. Ducousso, C. Loulergue-Majorel, L. Hannibal, S. Santoni, Y. Prin and M. Lebrun.
2010a. Ultramafic soils from New Caledonia structure Pisolithus albus in ecotype. FEMS
Microbiol. Ecol. 72: 238–249.
Jourand, P., M. Ducousso, R. Reid, C. Majorel, C. Richert, J. Riss and M. Lebrun. 2010b. Nickel-
tolerant ectomycorrhizal Pisolithus albus ultramafic ecotype isolated from nickel mines
in New Caledonia strongly enhance growth of the host plant Eucalyptus globulus at toxic
nickel concentrations. Tree Physiol. 30: 1311–1319.
Kazakou, E., P.G. Dimitrakopoulos, R.D. Reeves, A.J.M. Baker and A.Y. Troumbis. 2008.
Hypotheses, mechanisms, and trade-offs of tolerance and adaptation to serpentine soils:
from species to ecosystem level. Biol. Rev. 83: 495–508.
Khosla, B. and M.S. Reddy. 2008. Response of ectomycorrhizal fungi on the growth and
mineral nutrition of eucalyptus seedlings in bauxite mined soil. Amer-Eurasian J. Agri.
Environ. Sci. 3: 123–126.
L’Huillier, L., T. Jaffré and A. Wulf. 2010. Mines et environnement en Nouvelle-Calédonie
: les milieux sur substrats ultramafiques et leur restauration. IAC Ed, Noumea, New
Caledonia.
Majorel, C., L. Hannibal, M.E. Soupe, F. Carriconde, M. Ducousso, M. Lebrun and P. Jourand.
2012. Tracking nickel adaptive biomarkers in Pisolithus albus from New Caledonia using
a transcriptomic approach. Mol. Ecol. 21: 2208–2223.
Martin, F., C. Delaruelle and J.L. Hilbert. 1990. An improved ergsoterol assay to estimate fungal
biomass in ectomycorrhizas. Mycol. Res. 94: 1059–1064.
Martin, F., J. Diez, B. Dell and C. Delaruelle. 2002. Phylogeography of the ectomycorrhizal
Pisolithus species as inferred from ribosomal DNA ITS sequence. New Phytol. 153:
345–357.
Marschner, H. 1995. Mineral nutrition of higher plants 2nd edn. Academic Press, London,
UK, p. 889.
Marx, D.H. 1977. Tree host range and world distribution of the ectomycorrhizal fungus
Pisolithus tinctorius. Can. J. Microbiol. 23: 217–223.
McCoy, S.G. 1991. Edaphic controls influencing the distribution of Nothofagus aequilateralis
on ultrabasic soils at the Col the Mouirange, New Caledonia, Australian National
University.
124 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Meharg, A.A. 2003. The mechanistic basis of interactions between mycorrhizal associations
and toxic metal cations. Mycol. Res. 107: 1253–1265.
Morat, P., T. Jaffré, F. Tronchet, J. Munzinger, Y. Pillon, J.-M. Veillon and M. Chalopin. 2012.
The taxonomic database “FLORICAL” and characteristics of the indigenous flora of New
Caledonia. Adansonia 34, in press.
Moser, A.M., C.A Petersen, J.A. D’Allura and D. Southworth. 2005. Comparison of
ectomycorrhizas of Quercus garryana (Fagaceae) on serpentine and non serpentine soils
in southwestern Oregon. Am. J. Bot. 92: 224–230.
Moser, A.M., J. Frank, J.A. D’Allura and D. Southwood. 2009. Ectomycorrhizal communities
of Quercus garryana are similar on serpentine and non serpentine soils. Plant Soil 315:
185–194.
Mouchacca, J. 1998. Ascomycetes described from New Caledonia, South Pacific region.
Mycotaxon 67: 99–121.
Mouchacca, J. and E. Horak. 1998. Annotated cheklist of New Caledonia Basidiomycota. II.
Rusts and Smuts. Mycotaxon 69 : 13–30.
Moyersoen, B., Beever, R.E. and F. Martin 2003. Genetic diversity of Pisolithus in New-Zealand
indicates multiple long-distance dispersal from Australia. New Phytol. 160: 569–579.
Muller, L.A., J. Vangronsveld and J.V. Colpaert. 2007. Genetic structure of Suillus luteus
populations in heavy metal polluted and non-polluted habitats. Mol. Ecol. 16: 4728–
4737.
Myers, N., R.A. Mittermeier, C.G. Mittermeier, G.A.B. da Fonseca and J. Kent. 2000. Biodiversity
hotspots for conservation priorities. Nature 403: 853–858.
Perrier, N. 2005. Bio-géodiversité fonctionelle des sols latéritiques miniers: application à
la restauration écologique (Massif du Koniambo, Nouvelle-Calédonie). PhD thesis.
University of New Caledonia, New Caledonia.
Perrier, N., H. Amir and F. Colin. 2006a. Occurrence of mycorrhizal symbioses in the metal-rich
lateritic soils of the Koniambo Massif, New Caledonia. Mycorrhiza 16: 449–458.
Perrier, N., J.P. Ambrosi, F. Colin and R.J. Gilkes. 2006b. Biogeochemistry of a regolith: the New
Caledonian Koniambo ultramafic massif. J. Geochem. Explor. 88: 54–58.
Proctor, J. 2003. Vegetation and soil and plant chemistry on ultramafic rocks in the tropical
Far East. Perspect. Plant Ecol. 6: 105–124.
Rajkumar, M., M.N.V. Prasad, H. Freitas and N. Ae. 2009. Biotechnological applications of
serpentine soil bacteria for phytoremediation of trace metals. Crit. Rev. Biotechnol. 2:
120–130.
Ray, P., R. Tiwari, G.U. Reddy and A. Adholeya. 2005. Detecting the heavy metal tolerance
level in ectomycorrhizal fungi in vitro. World J. Microb. Biot. 21: 309–315.
Reddell, P., V. Gordon and M.S. Hopkins. 1999. Ectomycorrhizas in Eucalyptus tetrodonta
and E. miniata forest communities in tropical Northern Australia and their role in the
rehabilitation of these forests following mining. Aust. J. Bot. 47: 881–907.
Ruytinx, J., A.R. Craciun, K. Verstraelen, J. Vangronsveld, J.V. Colpaert and N. Verbruggen.
2011. Transcriptome analysis by cDNA-AFLP of Suillus luteus Cd-tolerant and Cd-sensitive
isolates. Mycorrhiza 21: 145–154.
Schoch, C.L., K.A. Seifert, S. Huhndorf, V. Robert, J.L. Spouge, C.A. Levesque, W. Chen and
F.B. Consortium. 2012. Nuclear ribosomal internal transcribed spacer (ITS) region as a
universal DNA barcode marker for Fungi. P. Natl. Acad. SCI USA 109: 6241–6246.
Smith, S. and D. Read. 2008. Mycorrhizal Symbiosis, 2nd edn. Academic Press, London.
Smith, M.E., G.W. Douhan and D.M. Rizzo. 2007. Ectomycorrhizal community structure in
a xeric Quercus woodland based on rDNA sequence analysis of sporocarps and pooled
roots. New Phytol. 174: 847–863.
Tam, P.C.F. 1995. Heavy metal tolerance by ectomycorrhizal fungi and metal amelioration by
Pisolithus tinctorius. Mycorrhiza 5: 181–187.
Tedersoo, L., U. Kõljalg, N. Hallenberg and K.-H. Larsson. 2003. Fine scale distribution of
ectomycorrhizal fungi and roots across substrate layers including coarse woody debris
in a mixed forest. New Phytol. 159: 153–165.
Ectomycorrhizal Fungi of Ultramafic Soils in New Caledonia 125
Tedersoo, L., T. Jairus, B.M. Horton, K. Abarenkov, T. Suvi, I. Saar and U. Koljalg. 2008. Strong
host preference of ectomycorrhizal fungi in a Tasmanian wet sclerophyll forest as revealed
by DNA barcoding and taxon specific primers. New Phytol. 180: 479–490.
Urban, A., M. Puschenreiter, J. Strauss and M. Gorfer. 2008. Diversity and structure of
ectomycorrhizal and co-associated fungal communities in a serpentine soil. Mycorrhiza
18: 339–354.
Wang, B. and Y.-L. Qiu. 2006. Phylogenetic distribution and evolution of mycorrhizas in land
plants. Mycorrhiza 16: 299–363.
Wenzel, W.W. and F. Jockwer. 1999. Accumulation of heavy metals in plants grown on
mineralised soils of the Austrian Alps. Environ. Pollut. 104: 145–15.
CHAPTER
7
Diversity and Function of
Ectomycorrhiza between
Scleroderma and Afzelia
Species in Burkina Faso
(West Africa)
Kadidia Bibata Sanon,1,* Amadou Mustapha Bâ2 and
Robin Duponnois3
1. Introduction
Most forest trees draw water and nutrients from the soil through fungi
associated with their root systems. This symbiotic association, called
mycorrhizas, contributes to water and mineral nutrition, and root protection.
In return, the fungus receives photosynthetic products necessary for its
growth and development (Smith and Read 2008).
1
Institut de l’Environnement et de Recherches Agricoles, Département Productions Forestières
(INERA/DPF), Laboratoire de Microbiologie Forestière, BP 7047 Ouagadougou 03, Burkina
Faso.
2
Laboratoire Commun de Microbiologie (UCAD-IRD-ISRA) BP 1386 Bel Air, Dakar,
Sénégal.
3
Laboratoire Ecologie & Environnement (Unité associée au CNRST, URAC 32). Faculté des
Sciences Semalia. Université Cadi Ayyad. Marrakech. Maroc.
*Corresponding author: sbkady@gmail.com
Diversity and Function of Scleroderma in Burkina Faso 127
Fig. 1. Climatic regions of Burkina Faso and localization of prospected areas. Data from Sanon
et al. (2009a).
Color image of this figure appears in the color plate section at the end of the book.
Diversity and Function of Scleroderma in Burkina Faso 129
with 4–5 months of rainfall between 900 and 600 mm, the Sahelian zone in
the north with about 2–3 months of rain and an annual rainfall less than
600 mm. According to Boussim (2010), two phytogeograpical zones can be
distinguished: the Soudanian zone extending from N 009°20 to N 13° making
2/3 of the land area, and the Sahelian zone (N 13° to 15°) that represent one
third of the country. The vegetation is characterized by the predominance of
mixed woody and herbaceous formations patchily distributed mostly in the
northern part (steppes, savannas, woodlands). The natural vegetation covers
60% of the country (FAO 1987). Gallery forests (1%) and woodlands (1%)
occur in the southern part. Tiger bush (1%) and steppe (4%) are common
in the North, whilst wooded savannas and shrublands (53%) can be found
in the rest the country.
Ectomycorrhizal-rich vegetations are located in the wetter southern
areas. They are mostly Caesalpinioid- and Phyllanthioid-dominated
woodlands of the Sudanian zone in the Southwestern, Southern and Eastern
parts of the country (Fig. 1). In many such woodlands, Anogeissus leiocarpus
(DC.) Guill. & Perr. dominates with other species such as Pterocarpus
erinaceus Poir., Burkea africana Hook., Afzelia africana Sm., Albizia chevalieri
Harms. and mainly Isoberlinia doka Craib. & Stapf., I. tomentosa (Harms.)
Craib. & Stapf. and Detarium microcarpum Guill. & Perr. (Thombiano et al.
2012). The presence of Isoberlinia doka as monospecific planting or associated
with Isoberlinia tomentosa also characterizes this area. Gallery forests, mainly
those of Kou and Mouhoun Reserve Forests are exclusively dominated by
EcM tree species such as Berlinia grandiflora (Vahl) Hutch. & Dalziel.
Different tropical plant families known to form EcM symbiosis have been
reported (Ducousso et al. 2008, Bâ et al. 2011, Bechem and Alexander 2012).
Compared to these data, an inventory was first performed in the field in
the Sudanian zone. It allowed identifying seven ectomycorrhizal species
sorted into three families or subfamily and five genera (Table 1).
Assessment of mycorrhizal status of these species was performed on the
roots of young plants, mature trees and natural regeneration. Arbuscular
mycorrhizal were not observed on roots of Isoberlinia spp. Some studies have
reported the presence of arbuscular mycorrhizal fungi on A. africana roots
(Thoen and Ducousso 1989), however we did not find this type of symbiosis
in our samples. For B. grandiflora, the presence or not of AM has not been
evaluated. Sites containing these EcM species were identified mainly in the
south-west and to a lesser extent in the south and east of the country (Fig.
1). These sites can be grouped as shown in Table 2.
Sites contained U. guineensis and B. grandiflora (3 sites in Southwest),
and the site of A. africana in Southwest are located in gallery forests. Apart
130 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
from the site of Isoberlinia spp. and M. kerstingii in Southwest which were
identified in 2000 and explored, all other sites were surveyed in 1994, 1995,
2000, 2005 and 2006. Sites in South and East were surveyed in 2000 and fungi
frequently encountered were Scleroderma species. Except gallery forests, the
other trees species are found in woodlands and savannas. Surveys were
conducted during the rainy season, the period of fungi fruiting, particularly
in June, July and August. All putative ECM fungi fruitbodies near these
trees were harvested. They were cleaned, photographed and described
morphologically. They were transported to the laboratory for isolation in
pure culture and/or dried in an oven at 50°C and stored in herbarium of
the Laboratory of Microbiology INERA/DPF.
The sampled specimens are sorted into 18 genera (Fig. 2) and 8 orders
(Aphyllophorales, Agaricales, Boletales, Cantharellales, Gautierales,
Hymenogastrales, Russulales and Sclerodermatales) and identified
according to the herbarium of the Microbiology Laboratory of IRD Senegal
Diversity and Function of Scleroderma in Burkina Faso 131
Fig. 2. Genera of ectomycorrhizal fungi identified by their morphology. (From Sanon et al.
1997, Sanon 1999, Sanon et al. 2012, in press).
3. Scleroderma species
Scleroderma belongs to Gasteromycete and to the Sclerodermataceae family.
This genus is widespread in tropical and temperate zone and is a symbiont
of many families of temperate and tropical plants of economic importance
such as Pinaceae, Myrtaceae, Fagaceae, Dipterocarpaceae, Gnetaceae and
subfamily Caesalpinioideae (Munyanziza and Kuyper 1995, Sims et al.
1997, Bechem and Alexander 2012). The genus Scleroderma encompasses
more than 25 accepted species (Sims et al. 1995, Sims et al. 1997, Guzman et
al. 2004). However, many species remain unidentified especially in tropical
Africa and Asia (Sanon et al. 1997, David and David 1998, Sims et al. 1999).
A revised key to the species of the genus Scleroderma was developed by
132 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Sims et al. (1995). This key is based primarily on the characteristics of the
peridium and spores. Thus, three morphotypes or sections are accepted
with the genus Scleroderma depending on the morphology of their spores:
Section Aculeatispora with spiny or warty spores; section Sclerangium with
sub-reticulate spores and the section Scleroderma with spores totally covered
with reticulum (Sims et al. 1995).
Fig. 4. Spores of Scleroderma sp2 without ornamentations (G. 100x, photonic microscope).
1999, 2002, Diédhiou et al. 2005). To attest whether the increased growth
of host plants resulted from inoculated symbionts, it was essential to
assess the persistence of these symbionts. In the absence of fructification,
study of the persistence of mycosymbionts implied that it was possible to
identify them from the mycorrhizas formed in situ. However, tracing the
identity of a fungal species from the EcM root tips was quite tricky, due
to poorly variable morphological and structural characteristics (Agerer
1987, Gardes et al. 1991, Sanon 1999, Sanon et al. 2002). To overcome these
difficulties, molecular biology techniques are increasingly used to enable
the identification of ectomycorrhizal fungi from fruiting bodies, cultured
134 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
mycelia and mycorrhizas (Gardes et al. 1991, Liang et al. 2004, Matsushita
et al. 2005, Ruiz-Diez et al. 2006, Sica et al. 2007). Also, although the
morphological characteristics allowed for the identification of a relatively
large number of species of the genus Scleroderma, molecular tools provided
more detail in the identification of ectomycorrhizal fungi (Hansen et al. 2002,
Rivière et al. 2007, Tedersoo et al. 2007). Various studies have shown that in
vitro DNA amplification (PCR) followed by the analysis of Polymorphism
Restriction Fragment Length (RFLP), or sequencing the internal transcribed
spacer (ITS) and inter-genic spacer (IGS) are among the most important
tools for analyzing inter and intra-specific fungal symbionts (Liang et al.
2004, Matsushita et al. 2005, Ruiz-Diez et al. 2006, Sica et al. 2007). The
ribosomal DNA spacers (ITS and IGS) are known to be inter-and intra-
specific variable and used as markers to distinguish multiple species or
isolates of ectomycorrhizal fungi (Karen et al. 1997, Peter et al. 2001, Horton
2002, Gomes et al. 2002).
To assess the inter- and intraspecific diversity of Scleroderma harvested
in Burkina Faso, the ITS and IGS1 regions were amplified and digested with
two restriction enzymes HinfI and MboI. DNA was extracted from isolates
and fruiting bodies using the methods described by Grube et al. (1995)
and Martin et al. (1997) or the DNeasy kit according to the manufacturer’s
instructions (Qiagen, France). The universal primers and ITS1/ITS4
and CNL12/5SA were used to amplify ITS and IGS1, respectively. The
amplification conditions, digestion and electrophoresis are described by
Sanon et al. (2009a).
Among the six morphological species identified (Table 3), the analysis of
length polymorphism restriction fragments revealed eight ribotypes noted,
A, B, C, D, E, F, G and H with 1-3 ribotypes within each morphological
species (Table 4). Two ribotypes were identified within the morphotype S.
dictyosporum (A, B), three for S. verrucosum (C, D, E), one for Scleroderma
sp1 (F) and two for Scleroderma sp2 (G, H). Some ribotypes were more
represented than others; this was the case of ribotype A of S. dictyosporum,
C of S. verrucosum and H of Scleroderma sp2. However, no relation was
established between the species origin (host plant) and ribotypes observed.
RFLP profiles of Scleroderma sp3 and Scleroderma sp4 were identical to those
of ribotype D of S. verrucosum and A of S. dictyosporum respectively (Sanon
et al. 2009a). This suggested that these two species are rather ecotypes of
S. verrucosum and S. dictyosporum, respectively. Thus, the six morphospecies
identified should be grouped into four species: S. dictyosporum, S. verrucosum,
Scleroderma sp1 and Scleroderma sp2.
Except ribotype G of Scleroderma sp2, ITS of at least two ribotypes of
each species have been sequenced. ITS sequences were obtained from 9
fruiting bodies and 16 isolates in culture. They were submitted to GenBank
database and similar sequences were identified using the algorithm Blastn.
Diversity and Function of Scleroderma in Burkina Faso 135
Table 4. Size of restriction fragments (in base pairs) of the ITS and IGS1 regions digested with
MboI and HinfI. The fruiting bodies are noted “SD, SV, SP1, SP2, SP3 or SP4” and isolates “IR”.
Fragments less than 50 bp are not shown on the table. Data from Sanon et al. (2009a).
from other species analyzed (52% bootstrap value) but was grouped with
the genus Scleroderma. The morphology of spores of this species suggested
that it might be a new species specific to Africa. However, further studies
are needed on a larger number of samples and also the sequencing of the
second ribotype of this species.
Diversity and Function of Scleroderma in Burkina Faso 137
Fig. 5. Neighbor joining phylogenetic tree of Scleroderma species based on ITS sequences.
Rhizopogon occidentalis is used as outgroup. Numerical values on the branches are the bootstrap
values of 1000 replications. Data from Sanon et al. (2009a).
The solid mycelial inoculum was used in two experiments on A. africana and
A. quanzensis in the nursery. The inoculum was produced on a solid substrate
composed of peat and vermiculite according to the method described by
Duponnois and Garbaye (1991).
Provenances of A. africana Height (cm) Roots dry Total dry Colonization MD (%) N (%) P (%) K (%)
and fungal isolates weight (g) weight (g) (%)
AaBF
IR109 41.97 b 2.83 abc 7.81 ab 69.00 a 21.25 ab 1.96 d 0.11 bc 1.41 cde
IR406 44.96 ab 2.53 bc 7.73 ab 32.00 abc 20.43 ab 2.33 ab 0.09 c 2.12 a
IR408 47.78 ab 3.29 ab 9.00 a 64.00 a 31.66 a 1.94 d 0.14 a 1.66 bc
ORSXM002 42.75 b 3.98 a 9.10 a 46.00 ab 32.24 a 1.85 d 0.10 bc 1.45 cd
Control 38.98 b 1.76 c 6.15 b 0.00 c - 2.02 cd 0.10 bc 1.02 e
AaSN
IR109 46.85 ab 2.07 bc 6.34 b 58.00 ab 0.15 b 2.03 cd 0.13 ab 1.55 c
IR406 47.80 ab 2.71 abc 6.95 ab 38.00 ab 8.92 ab 2.28 abc 0.10 bc 2.05 ab
IR408 52.40 a 2.17 bc 7.03 ab 22.00 bc 9.95 ab 2.54 abc 0.11 bc 2.11 a
ORSXM002 52.78 a 2.92 abc 8.18 ab 64.00a 22.61 ab 2.08 bcd 0.12 abc 1.80 abc
Control 52.60 a 1;73 c 6.33 b 0.00 c - 1.90 d 0.10 bc 1.13 de
Values followed by the same letter within columns are not significantly different using ANOVA (SAS, Bonferroni test, P<0.05).
Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Diversity and Function of Scleroderma in Burkina Faso 141
Based on the results of the previous study, the two promising isolates
identified were tested on A. quanzensis (Bâ et al. 2002). A. quanzensis
responded differently to inoculation depending on the isolate. Thelephora
sp. ORSXM002 efficiently colonized the root system of inoculated plants
and stimulated root biomass compared to S. dictyosporum IR408 (Table 7).
The mycorrhization rate obtained with ORSXM002 (66.40%) was similar
to that obtained with the Senegalese A. africana provenance (64%) (Bâ et al.
1999). However, for IR408, this rate was very low (19.40%). Stimulation of
root biomass by ORSXM002 was not reflected in the total biomass of plants.
Conversely, this strain had stimulated the total biomass of A. africana in
the same conditions. Therefore, mycorrhizal dependency of A. quanzensis
inoculated with these two species does not exceed 16%, which was quite
Table 7. Effect of inoculation with IR408 and ORSXM002 on growth parameters and nutrition
of A. quanzensis seedlings. MD, Mycorrhizal dependency. Data from Bâ et al. (2002).
A. quanzensis/ Height Shoot dry Root dry Colonization MD N (%) P (%) K (%)
Isolates (cm) weight (g) weight (g) (%) (%)
inoculated
IR408 69.86 b 8.84 a 2.26 a 19.40 b 3.50 a 2.09 a 0.13 a 1.63 b
ORSXM002 68.22 ab 9.14 a 3.60 b 66.40 a 16.00 a 2.14 a 0.14 a 1.56 b
Control 64.28 a 8.80 a 1.91 a 0.00 c - 2.15 a 0.12 a 0.81 a
Values followed by the same letter within columns are not significantly different using ANOVA
(SAS, Bonferroni test, P<0.05).
142 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
low compared to A. africana (Bâ et al. 1999). These data suggested that the
different origin of strains and host plant may influence the response to
inoculation with A. quanzensis compared to A. africana. As in the case of A.
africana, only the K content of the shoot was greater in plants inoculated
with both strains (Table 7).
5. Conclusion
This work allowed the identification of ectomycorrhizal fungi associated
with seven host plants in Southwest, South and East of Burkina Faso (A.
africana, B. grandiflora, I. doka, I. dalzeillii, M. kerstingii, U. guineensis, U. somon).
All harvested fungi belonged to genera already identified as ectomycorrhizal
and some were identified at the species level. An unexpected diversity of
ectomycorrhizal fungi (about 78 morphological species) could be detected
despite the geo-climatic conditions of the regions surveyed and phenological
fluctuation of the fungi from one year to another in low humidity areas.
Our inventory revealed the dominance of species of the genera Russula,
Boletelus, Leccinum, Amanita and Scleroderma. For the latter, ITS sequencing of
ribotypes showed that they belonged to at least seven phylogenetic species:
4 species for S. verrucosum morphotype, 2 for S. dictyosporum and a single
ribotype sequenced for Scleroderma sp2. The particular morphology of
spores of Scleroderma sp2 and its position on the phylogenetic tree compared
with other species of Scleroderma suggested a new species unique to Africa,
especially in West Africa.
We tested the effectiveness of four fungal isolates, S. dictyosporum,
IR109 and IR408, S. verrucosum, IR406 and Thelephora sp, ORSXM002, on
the growth and mineral nutrition on two provenances of A. africana in
the nursery. We have shown that the inoculation induced a wide range of
plant growth stimulation (0.17% to 48% of biomass increase) and improved
mineral nutrition, especially K. The use of two provenances of A. africana
has highlighted the strong influence of the genetic background of the host
plant on the beneficial effect of mycorrhization. Mycorrhizal dependency
indicated that the Senegalese origin was less dependent on mycorrhiza
for growth than that of Burkina Faso. The use of fungal strains effective to
improve A. africana plant growth in controlled mycorrhization programs
can be oriented towards S. dictyosporum IR408 and Thelephora sp. ORSXM002
for both tree provenances. These two isolates tested on A. quanzensis have
shown a limited significant effect compared to A. africana suggesting that
the origin of the host plant in relation to the fungal isolates tested may
influence the response to inoculation.
Diversity and Function of Scleroderma in Burkina Faso 143
However, these strains should also be tested under other soil conditions
because our experiment was conducted in a sandy soil unrepresentative
of A. africana forests in West Africa. Any selection of strain must therefore
be supplemented by tests under various conditions and respecting local
conditions of plantations.
Acknowledgements
We thank the anonymous referees for their valuable comments on this study,
Krista McGuire and Jean Garbaye for improving the language.
References
Agerer, R. 1987–1996. Color Atlas of ectomycorrhizae with glossary. Pp??? In: R. Agerer (ed.).
Einhorn-Verlag Eduard Dietenberger.
Alexander, I.J. 2006. Ectomycorrhizas—out of Africa? New Phytol. 172: 589–591.
Bâ, A.M. 1990. Contribution à l’étude de la symbiose ectomycorhizienne chez deux essences
forestières d’Afrique intertropicale: Afzelia africana Sm. et Uapaca guineensis Müll. Arg.
Thèse de l’Université des Sciences et Techniques du Languedoc, Montpellier, pp. 193.
Bâ, A.M. and D. Thoen. 1990. First syntheses of ectomycorrhizas between Afzelia africana Sm.
(Caesalpinioideae) and native fungi from West Africa. New Phytol. 103: 441–448.
Bâ, A.M. and T. Guissou. 1996. Rock phosphate and vesicular-arbuscular mycorrhiza effects
on growth and nutrient uptake of Acacia albida (Del.) seedlings in an alkaline sandy soil.
Agrofor. Syst. 34: 129–137.
Bâ, A.M., Y. Dalpé and T. Guissou. 1996a. Les Glomales d’Acacia holosericea Cunn. ex G.
Don. et d’Acacia mangium Willd.: diversité et abondance relative des champignons
endomycorhiziens à arbuscules dans deux types de sols de plantations au Burkina Faso.
Bois For. Trop. 250: 5–18.
Bâ, A.M., M. Bazié and T. Guissou. 1996b. Effet du phosphate naturel sur de jeunes Acacia albida
Del. en présence ou non de mycorhizes. Cahiers Scientifiques du CIRAD 12: 237–244.
Bâ, A.M., B.K. Sanon, R. Duponnois and J. Dexheimer. 1999. Growth responses of Afzelia africana
Sm. seedlings to ectomycorrhizal inoculation in nutrient-deficient soil. Mycorrhiza 9:
91–95.
Bâ, A.M., T. Guissou, R. Duponnois, C. Plenchette, O. Sacko, D. Sidibé, K. Sylla and B. Windou.
2001. Mycorhization contrôlée et fertilisation phosphatée: applications à la domestication
du jujubier (Ziziphus mauritiana Lam.). Fruits 56: 261–269.
Bâ, A.M., B.K. Sanon and R. Duponnois. 2002. Influence of ectomycorrhizal inoculation on
Afzelia quanzensis Welw. seedlings in a nutrient-deficient soil. For. Ecol. Manag. 161:
215–219.
Bâ, A.M., R. Duponnois, M. Diabaté and B. Dreyfus. 2011. Les champignons ectomycorhiziens
des arbres forestiers en Afrique de l’Ouest: méthodes d’étude, diversité, écologie,
utilisation en foresterie et comestibilité. Editions IRD pp. 264.
Bâ, A.M., R. Duponnois, B. Moyersoen and A. Diédhiou. 2012. Ectomycorrhizal symbiosis of
tropical African trees. Mycorrhiza 22: 1–29.
Bechem, E.E. and I.J. Alexander. 2012. Mycorrhizal status of Gnetum africanum in Cameroon:
Evaluating diversity with a view to ameliorating domestication efforts. Mycorrhiza 22:
99–108.
Boussim J.I. 2010. Les territoires phytogéographiques. In: A. Thiombiano and D. Kampmann
(eds.). Atlas de la Biodiversité de l’Afrique de l´Ouest, Tome II : Burkina Faso. BIOTA,
Ouagadougou/Frankfurt am Main, pp. 152–155.
144 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
David, J.F. and M.S. David. 1998. Variation in the ribosomal DNA internal transcribed spacer
of a diverse collection of ectomycorrhizal fungi. Mycol. Res. 102: 859–865.
Dianda, M., J. Bayala, T. Diop and S.J. Ouedraogo. 2009. Improving growth of shea butter tree
(Vitellaria paradoxa C.F. Gaertn.) seedlings using mineral N, P and arbuscular mycorrhizal
(AM) fungi. Biotechnol. Agron. Soc. Environ. 13: 93–102.
Diédhiou, A.G., A.M. Bâ, S.N. Sylla, B. Dreyfus, M. Neyra and I. Ndoye. 2004. The early-stage
ectomycorrhizal Telephoroid fungal sp. is competitive and effective on Afzelia africana
Sm. in nursery conditions in Senegal. Mycorrhiza 14: 313–322.
Diédhiou, A.G., O. Guèye, M. Diabaté, Y. Prin, R. Duponnois, B. Dreyfus and A.M. Bâ. 2005.
Contrasting responses to ectomycorrhizal inoculation in seedlings of six tropical African
tree species. Mycorrhiza 16: 11–17.
Ducousso, M., H. Ramanankierana, R. Duponnois, R. Rabévohitra, L. Randrihasipara, M.
Vincelette, B. Dreyfus and Y. Prin. 2008. Mycorrhizal status of native trees and shrubs from
eastern Madagascar littoral forests with special emphasis on one new ectomycorrhizal
endemic family, the Asteropeiaceae. New Phytol. 178: 233–238.
Duponnois, R. and J. Garbaye. 1991. Techniques for controlled synthesis of the Douglas-fir-
Laccaria laccata ectomycorrhizal symbiosis. Ann. Sci. For. 48: 641–650.
Duponnois, R. and A.M. Bâ. 1999. Growth stimulation of Acacia mangium Willd. by Pisolithus
sp. in some Senegalese soils. For. Ecol. Manag. 119: 209–215.
FAO. 1987. Etude sur la contribution du secteur forestier à l’économie du Burkina Faso. Rapport
de synthèse du projet préparé par le gouvernement du Burkina Faso et la FAO. Rome.
Fortin, J.A., C. Plenchette and Y. Piché. 2008. Les mycorhizes: La nouvelle révolution verte.
Edition Multimondes. Québec, Canada, pp. 131.
Garbaye, J., J.C. Delwaulle and D. Diangana. 1988. Growth response of eucalyptus in the
Congo to ectomycorrhizal inoculation. For. Ecol. Manag. 24: 151–157.
Gardes, M., T.J. White, J.A. Fortin, T.D. Bruns and J.W. Taylor. 1991. Identification of indigenous
and introduced symbiotic fungi in ectomycorrhizae by amplification of nuclear and
mitochondrial ribosomal DNA. Can. J. Bot. 69: 180–190.
Gomes, E.A., M.C.M. Kasuya, E.G. de Barros, A.C. Borges and E.F. Araujo. 2002. Polymorphism
in the internal transcribed spacer (ITS) of the ribosomal DNA of 26 isolates of
ectomycorrhizal fungi. Genetics and Mol. Biol. 25: 477–483.
Grube, M., P.T. Depriest, A. Gargas and J. Hafellner. 1995. DNA isolation from lichen ascomata.
Mycol. Res. 99: 1321–1324.
Guissou T., A.M. Bâ, J.M. Ouadba, S. Guinko and R. Duponnois. 1998. Responses of Parkia
biglobosa (Jacq.) Benth., Tamarindus indica L. and Ziziphus mauritiana Lam. to arbuscular
mycorrhizal fungi in a phosphorus-deficient sandy soil. Biol. Fert. Soils 26: 194–198.
Guissou, T., A.M. Bâ, C. Plenchette, S. Guinko and R. Duponnois. 2001. Effets des mycorhizes
à arbuscules sur la tolérance à un stress hydrique chez quatre arbres fruitiers: Balanites
aegyptiaca (L.) Del., Parkia biglobosa (Jacq.) Benth., Tamarindus indica L. et Ziziphus mauritiana
Lam. Sécheresse 12: 121–127.
Guzman, G., F. Ramirez-Guillém, O.K. Miller and D.J. Lodge. 2004. Scleroderma stellatum
versus Scleroderma bermudense: the status of Scleroderma echinatum and the first record of
Veligaster nitidum from the Virgin Islands. Mycologia 96: 1370–1379.
Hamel, C., H. Fyles and D.L. Smith. 1990. Measurement of development of endomycorrhizal
mycelium using three different vital strains. New Phytol. 115: 297–302.
Hansen, K., T. Laessoe and D. Pfister. 2002. Phylogenetic diversity in the core group of Peziza
inferred from ITS sequences and morphology. Mycol. Res. 106: 879–902.
Horton, T. 2002. Molecular approaches to ectomycorrhizal diversity studies: variation in ITS
at a local scale. Plant Soil 244: 29–39.
Jones, M.D., D.M. Durall and P.B. Tinker. 1990. Phosphorus relationships and production of
extramatrical hyphae by two types of willow ectomycorrhizas at different soil phosphorus
levels. New Phytol. 115: 259–267.
Diversity and Function of Scleroderma in Burkina Faso 145
Karen, O., N. Högberg, A. Dahlberg, L. Jonsson and J.E. Nylund. 1997. Inter- and intraspecific
variation in the ITS region of rDNA of ectomycorrhizal fungi in Fennoscandia as detected
by endonuclease analysis. New Phytol. 136: 313–325.
Kessler, J.J. and C. Geerling. 1994. Profil environnemental du Burkina Faso. Université
Agronomique, Département de l’Aménagement de la Nature. Wageningen, les Pays
Bas, pp. 63.
Kumar, S., K. Tamura and M. Nei. 2004. MEGA3: Integrated software for Molecular Evolutionary
Genetics Analysis and Sequence Alignment. Briefings in Bioinform. 5: 150–163.
Laclavère, G. 1993. Atlas du Burkina Faso. Jeune Afrique, pp. 53.
Le Tacon, F., I.F. Alvarez, D. Bouchard, B. Henrion, R.M. Jackson, S. Luff, J.I. Parlade, J. Pera,
E. Stenström, N. Villeneuve and C. Walker. 1992. Variations in field response of forest
trees to nursery ectomycorrihzal inoculation in Europe. In: D.J. Read, D.H. Lewis, A.H.
Fitter and I.J. Alexander (eds.). Mycorrhizas in Ecosystems pp. 119–134.
Lei, J., F. Lapeyrie, N. Malajczuk and J. Dexheimer. 1990. Infectivity of pine and eucalypt
isolates of Pisolithus tinctorius (Pers.) Coker & Couch on roots of Eucalyptus urophylla S. T.
Blake in vitro. II. Ultrastructural and biochemical changes at the early stage of mycorrhiza
formation. New Phytol. 116: 115–122.
Liang, Y., L.D. Guo LD and K.P. Ma. 2004. Genetic structure of a population of the
ectomycorrhizal fungus Russula vinosa in subtropical woodlands in southwest China.
Mycorrhiza 14: 235–240.
Malajczuk, N., F. Lapeyrie and J. Garbaye. 1990. Infectivity of pine and eucalypt isolates of
Pisolithus tinctorius on roots of Eucalyptus urophylla in vitro. 1. Mycorrhiza formation in
model systems. New Phytol. 114: 627–631.
Martin, F., G. Costa, C. Delaruelle and J. Diez. 1997. Genomic fingerprinting of ectomycorrhizal
fungi by microsatellite-primed PCR. In: A. Varma and B. Hock (eds.). Mycorrhiza Manual.
Springer Lab Manuel Berlin, Springer-Verlag, pp. 463–474.
Martin, F., J. Diez, B. Dell and C. Delaruelle. 2002. Phylogeography of the ectomycorrhizal
Pisolithus species as inferred from the nuclear ribosomal DNA ITS sequences. New
Phytol. 153: 345–358.
Matsushita, N., K. Kikuchi, Y. Sasaki, A. Guerin-Laguette, F. Lapeyrie, L.M. Vaario, M. Intini
and K. Suzuki. 2005. Genetic relationship of Tricholoma matsutake and T. nauseosum
from the Northern Hemisphere based on analyses of ribosomal DNA spacer regions.
Mycoscience 46: 90–96.
Munyanziza, E. and T.W. Kuyper. 1995. Ectomycorrhizal synthesis on seedlings of Afzelia
quanzensis Welw. using various types of inoculum. Mycorrhiza 5: 283–287.
Peter, M., U. Büchler, F. Ayer and S. Egli. 2001. Ectomycorrhizas and molecular phylogeny of
the hypogeous russuloid fungus Arcangeliella borziana. Mycol. Res. 105: 1231–1238.
Rivière, T., A.G. Diedhiou, M. Diabaté, G. Senthilarasu, K. Natarajan, M. Ducousso, A.
Verbeken, B. Buyck, B. Dreyfus, G. Bena and A.M. Bâ. 2007. Diversity of ectomycorrhizal
Basidiomycetes in West African and Indian tropical rain forests. Mycorrhiza 17:
415–428.
Ruiz-Diez, B., A.M. Rincon, M.R. de Felipe and M. Fernandez-Pascual. 2006. Molecular
characterisation and evaluation of mycorrhizal capacity of Suillus isolates from Central
Spain for the selection of fungal inoculants. Mycorrhiza 16: 465–474.
Sanon, K.B., A.M. Bâ and J. Dexheimer. 1997. Mycorrhizal status of some fungi fruiting beneath
indigenous trees in Burkina Faso. For. Ecol. Manag. 98: 61–69.
Sanon, B.K. 1999. La symbiose ectomycorhizienne chez quelques Césalpiniacées et
Euphorbiacées des forêts du Sud-Ouest du Burkina Faso. Etude morphologique et
cytologique, mycorhization contrôlée et étude de la diversité inter- et intraspécifique
de Sclérodermes ectomycorhiziens. Thèse de Doctorat de l’Université Henri Poincaré
Nancy I pp. 119.
Sanon, B.K., J. Dexheimer, A.M. Bâ, M. Dianda and J. Gerard. 2002. Structure comparée des
ectomycorhizes de Afzelia africana Sm. et Scleroderma sp. Science et Technique, série Sci.
Nat. Agro. 1: 17–28.
146 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Sanon, B.K., A.M. Bâ, C. Delaruelle, R. Duponnois and F. Martin. 2009a. Morphological and
molecular analyses in Scleroderma species associated with some Caesalpinioid legumes,
Dipterocarpaceae and Phyllanthaceae trees in southern Burkina Faso. Mycorrhiza 19:
571–584.
Sanon, B.K., M. Dianda, T. Guissou and A.M. Bâ. 2009b. Description des champignons
ectomycorhiziens du genre Scleroderma de quelques formations forestières du Sud du
Burkina Faso. Cameroon J. Experiment. Biol. 2: 69–78.
Sanon, B.K., R. Duponnois, A.M. Bâ, M. Doucousso and B. Dreyfus. 2012. Ectomycorrhizal
fungi diversity from south-west Burkina Faso using part of mitochondrial rDNA sequence
analysis. In press.
Sary, H. 1989. Essai de mycorrhization de Afzelia africana avec Pisolithus tinctorius (Pers.) Coker
et Couch. In: Trees for development in Sub-Saharan Africa, Proceedings of a regional
seminar held by the International Foundation for Science (IFS), ICRAF House, Nairobi,
Kenya.
Sica, M., L. Gaudio and S. Aceto. 2007. Genetic structure of Tuber mesentericum Vitt. Based on
polymorphism at the ribosomal DNA ITS. Mycorrhiza 17: 405–414.
Sims, K.P., R. Watling and P. Jeffries. 1995. A revised key to the genus Scleroderma. Mycotaxon:
403–420.
Sims, K.P., R. Watling, R. De La Cruz and P. Jeffries. 1997. Ectomycorrhizal fungi of the
Philippines: a preliminary survey and notes on the geographic biodiversity of the
Sclerodermatales. Biod. Conserv. 6: 45–58.
Sims, K.P., R. Sen, R. Watling and P. Jeffries. 1999. Species and population structures of Pisolithus
and Scleroderma identified by combined phenotypic and genomic marker analysis. Mycol.
Res. 103: 449–458.
Smith, S.E. and D.J. Read. 2008. Mycorrhizal symbiosis. Third edition pp. 800.
Tedersoo, L., T. Suvi, K. Beaver and U. Kôljalg. 2007. Ectomycorrhizal fungi of the
Seychelles: diversity patterns and host shifts from the native Vateriopsis seychellarum
(Dipterocarpaceae) and Intsia bijuga (Caesalpiniaceae) to the introduced Eucalyptus robusta
(Myrtaceae), but not Pinus caribea (Pinaceae). New Phytol. 175: 321–333.
Thombiano, A., M. Schmidt, S. Dressler, A. Ouédraogo, K. Hahnand and G. Ziska. 2012.
Catalogue des plantes vasculaires du Burkina Faso. Conservatoire et Jardin Botanique
de la Ville de Genève pp. 396.
Thoen, D. and M. Ducousso. 1989. Champignons et ectomycorhizes du Fouta Djalon. Bois
For. Trop. 221: 45–63.
Thomson, B.D., T.S. Grove, N. Malajczuk and G.E. St. J. Hardy. 1994. The effectiveness of
ectomycorrhizal fungi in increasing the growth of Eucalyotus globulus Labill. in relation
to root colonisation and hyphal development in soil. New Phytol. 126: 517–524.
CHAPTER
8
The Physiology of
Scleroderma sinnamariense
Mont. (Sclerodermaceae),
an Ectomycorrhizal Fungus
Associated with Gnetum spp.
(Gnetaceae)
Eneke Esoeyang Tambe Bechem
1. Introduction
Scleroderma sinnamariense Mont. is a bright yellow ectomycorrhizal fungus
with a pantropical distribution (Guzman and Ovrebo 2000) which was
originally described in French Guiana. This fungus is a gasteromycete and
produces macroscopic sporocarps called “puffballs” (Fig. 1). These yellowish
leathery, rounded structures become dry and crack at maturity, releasing
dry powdery basidiospores which are dispersed by wind. The basidiomes
are not eaten in Cameroon by humans but it is possible that they are eaten
by some animals including small mammals, birds and insects. They are,
however, eaten by humans in Asia. S. sinnamariense is commonly recorded
Department of Botany and Plant Physiology, Faculty of Science, University of Buea, P. O. Box
63 Buea, South West Region, Republic of Cameroon.
Email: tamenekeso@yahoo.co.uk
148 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 2. Molecular phylogenetic tree-diagram of single linkage analysis of RFLP patterns from
digestion of the ITS region of rDNA amplified from EM root tips from Gnetum, mycelium,
and EM fruiting bodies. Numbers at nodes indicate bootstrap indices over 50% obtained after
1,000 replicates. Data from Bechem and Alexander (2012a).
150 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 3. Yellow Gnetum EM root tip formed with S. sinnamariense, showing extensive extramatrical
mycelia. Bar is 0.3 mm.
Color image of this figure appears in the color plate section at the end of the book.
The Physiology of Scleroderma sinnamariense Mont. 151
3. Growth in Culture
Just like most mycobionts, S. sinnamariense can be isolated from EM root tips
but recovery is often less than 20% and sometimes can be as low as 5% or
less of the number of plated tips (Molina and Palmer 1982). The low success
rates could be attributed to many factors including the inability of fungi to
grow on the media used, insufficient surface sterilization, and requirement
of special nutrients. The mycobiont can also be isolated from spores and
sporocarps. Several media types have been used for the isolation of EM
fungi from root tips. These include modified Melin-Norkran (MMN) agar
(Marx 1969), Hagems agar (Modess 1941), Pachlewski agar (Pachlewski
and Pachlewska 1974), modified woody plant medium, potato dextrose
agar, etc.
S. sinnamariense was successfully isolated from Gnetum EM root tips
on MMN agar containing benomyl (1 ug/ml), chlortetracycline (30 ug/ml)
and streptomycin (10 ug/ml) (Bechem 2004). More than 350 individual EM
root tips were plated with a fungal symbiont recovery rate of 38%. This was
obtained after surface sterilization using 6% Cl2 for 4 min. In the said study,
only one fungus type was isolated from the yellow morphotype unlike other
studies such as Zak and Marx (1964), who isolated two or more fungi from
the same pine EM roots.
Scleroderma sinnamarense was a yellow culture with prominent hyphae
which had numerous clamp connections. The arrangement of hyphae in
culture was similar to that observed on the mycorrhiza tip. Growth rate
of the fungus was very slow in comparison to some isolates of Pisolithus
sp., an observation similar to that of Sims et al. (1999). Growth rarely
exceeded 35–40 mm in ten weeks, giving a growth rate of 0.5–0.57 mm d–1.
This growth rate was much slower than that observed on the same fungus
species studied by Sims et al. (1999) where growth rate was as high as 1.5
mm d–1 on some isolates. Growth was slower on Malt extract agar (MEA),
Potato dextrose agar (PDA), and on Hagem’s agar, in comparison to MMN
and Pachlewski agar (Fig. 4) (Bechem 2004).
S. sinnamariense species possessed colonies with matted mycelial growth
which sometimes had a floccose periphery, with an irregular to occasionally
feathery outline. Cultures were bright yellow in color, with the reverse
showing the same coloration throughout. Observation of this fungal isolate
was quite similar to that of Sims et al. (1999). Color was impermanent, with
cultures occasionally changing to white after prolonged storage. The reverse
side of these white cultures was whitish in the extremities but yellowish in
the centre. The fungus exuded a yellowish-green pigment and sometimes
produced faint greenish droplets into the agar. Putra et al. (2011) isolated
two known coloring constituents, methyl 4, 4’-dimethoxyvulpinate and
152 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 4. Dry weight yields (mg) of five isolates of S. sinnamariense following growth for 30 days
on liquid nutrient media, modified Melin Norkran (MMN) and Pachlewski (PACH). Vertical
bars represent standard errors of mean. Each point is a mean of three replicates.
4. Nitrogen Nutrition
Nitrogen is present in soil in diverse forms such as ammonium, nitrate,
proteins, peptides and amino acids. Studies using different fungal species
have shown that EM fungi have the ability to take up and transfer N to the
partner plant. Studies on the use of different N sources by Scleroderma spp.
are lacking but some fungi species such as Hebeloma spp. and Paxillus species
have been widely studied in this respect (Abuzinadah and Read 1986).
S. sinnamariense was studied to evaluate its ability to use different N
sources for growth (Bechem 2012a). This fungus produced measurable
biomass (Table 1) following growth on MMN liquid broth containing a
range of nitrogen sources including ammonium, nitrate, glutamic acid,
arginine, alanine, glycine, peptone, and bovine serum albumin (BSA)
(Bechem 2012a).
This Scleroderma produced greater biomass on ammonium-N than
that produced on nitrate-N. Growth in ammonium was accompanied by
a fall in pH of the medium, while an increase in pH was observed with
The Physiology of Scleroderma sinnamariense Mont. 153
Table 1. Dry weight yields (mg) after 30 days of growth of Scleroderma sinnamariense on liquid
MMN media containing different nitrogen sources.
N source S. sinnamariense
Mean ± s.e.m
Calcium nitrate 4.2 ± 0.1
Ammonium sulphate 7.2 ± 1.1
Glutamic acid 8.8 ± 1.2
Arginine 12.9 ± 1.9
Alanine 11.2 ± 0.6
Glycine 2.8 ± 0.5
Bovine Serum Albumin 3.5 ± 0.2
Peptone 16.7 ± 1.4
No Nitrogen 2.5 ± 0.1
Values are means ± standard errors of mean. Number of replicates (n) = 3. Least significant
difference at 95 % (one-way comparison) was 3.324.
growth in nitrate. This inability to use nitrate had been observed in other
ectomycorrhizal fungi species (Rangel-Castro et al. 2002, Sawyer et al.
2003).
The fungus grew best on peptone-N, producing a biomass which was
more than twice (131.9%) the yield observed on ammonium-N. Growth
on BSA was poor, when compared to ammonium. However, utilization of
this N source was better at very low pH (3.0). This low pH optimum for
proteolytic activity suggested that broad spectrum acid proteases were
involved in the catalytic process.
Glycine could not provide the N needed for growth by the fungus. Its
utilization was therefore limited. But arginine, alanine and glutamic acid
were good sources of nitrogen for the fungus. Its growth on these N sources
was higher than that observed in ammonium. It is possible to conclude from
this observation that free amino acid pools in soil as well as those released
directly by the action of fungal proteases might help to supplement the
supply of mineral N.
Ammonium has always been thought to be the N source for the growth
of microorganisms. The results of the study carried out by Bechem (2012a)
showed that organic N sources like peptone, arginine, and alanine were
better for the growth of Scleroderma sinnamariense.
In Cameroon where this fungus was collected, the pH of the soils is
mostly acidic and rarely exceeds 4.5. It is therefore possible that under such
conditions the activity of the proteinases necessary for protein uptake and
utilization would be optimal, enabling the endophyte to access organic N
from the rhizosphere and bulk soil. The phytobiont can therefore access
these N sources via the mycobiont.
154 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
5. Carbon Nutrition
There has been no published study on the growth of Scleroderma spp. on
different carbon sources, except for the study by Bechem (2012b). She
evaluated the ability of S. sinnamariense to grow on different C sources in
the presence of either ammonium-N or peptone-N. The C sources used in
the study included; glucose, sucrose, soluble starch, cellulose, cellulose
plus ‘starter’ glucose. She observed that glucose was the best carbon source
for this fungus and that growth on glucose was significantly greater when
peptone was the N source. Growth on the other C sources was relatively
poor, implying that the fungus could hardly utilize soluble starch, cellulose,
and cellulose with ‘starter’ glucose in peptone or ammonium-N. The ability
of the fungus to use sucrose only in ammonium-N might have been due to
changes in pH which resulted following ammonium uptake. Hampp and
Schaeffer (1994) had observed that sucrose hydrolysis was significant at
pH levels below 4.0. In the study by Bechem (2012b) the media were not
buffered and uptake of ammonium would have caused acidification of the
medium. The drop in pH may have favored the hydrolysis of sucrose to
its constituent glucose and fructose, which might have been used by the
fungus for growth. In this study, ‘starter’ glucose caused adaptive growth
of the fungus in cellulose in the presence of ammonium and peptone-N.
There is a possibility that the limited growth of Scleroderma on sucrose
and cellulose could restrict the fungus to the root of the host plant due to
unavailability of nutrients.
6. Phosphorus Nutrition
Some researchers had suggested that the release of organic acids from EM
fungi may solubilize recalcitrant mineral phosphates (Leyval and Berthelin,
1986). Others believed that before insoluble organic P can be broken down
by fungal phosphatases and the resulting phosphate absorbed by the roots,
they would first have to be solubilized (Lapeyrie et al. 1991).
In the experiment reported by Bechem (2011), the ability of
S. sinnamariense to use calcium tetra hydrogen di-orthophosphate CaH4(PO4)2,
hydroxyapatite Ca5(PO4)3OH, calcium phytate C6H6Ca6O24P6 and amorphous
iron phosphate FePO4 as P sources for growth was evaluated. The effect
of the internal P status of the inoculum, the medium N source and the
presence/absence of buffer on growth was also determined.
In the said study, P-sufficient S. sinnamariense showed growth on all P
sources with peptone as sole N source except on FePO4 in unbuffered media.
The source of medium P and the presence of buffer had significant effects
(P< 0.001) on colony diameter. Maximum growth of 40 mm was observed
on FePO4 in buffered conditions. Growth of fungus in buffered medium
The Physiology of Scleroderma sinnamariense Mont. 155
7. Ectomycorrhiza Formation
Scleroderma spp. have been reported generally to form white mycorrhizas
from clamped hyphae (Jeffries 1999). S. sinnamariense, however, has been
reported to form bright yellow mycorrhizas with Gnetum spp. (Bechem
and Alexander 2012a, Onguene and Kuyper 2001, Ingleby 1999). The
same fungus was reported to form dark brown EM with Shorea leprosula
Miq. but the hyphae in the rhizomorphs linking the sporocarps to the EM
were said to be bright yellow (Lee et al. 1997). This fungus seems to have a
narrow host range and has been associated only with Gnetum, Parashorea,
and Shorea. EM synthesis with rooted Gnetum cuttings under controlled
conditions was possible with the use of spores, soil, and mycorrhizal tips
as inoculum (Bechem and Alexander 2009).
house and field trials (Jeffries 1999, Castellano 1996, Bâ et al. 2002). Most
of the studies reported used species other than S. sinnamariense except for
Dodd et al. (1996) who used this fungus to inoculate seedlings of Parashorea
malanonan. Height increments of over 22% were observed on inoculated
seedlings when compared to uninoculated plants.
Bechem and Alexander (2012b) carried out an experiment in which they
evaluated the effects of mycorrhization and the addition of P (30mg/kg and
300mg/kg) on the growth of rooted Gnetum cuttings. It was noted that the
number of new EM tips decreased with an increase in P level implying that
the rate of colonization decreased with increasing P concentrations. This
finding demonstrated that the level of fertilization that can be applied to
Gnetum plants in farms and plantations is of critical importance and the
type of fertilizer must be carefully chosen so as to allow for the continuous
development and survival of the mycorrhizas. Ectomycorrhization led to an
increase in shoot, root and total plant dry weights, root: shoot ratio, and shoot
elongation as well as the total number of new leaves. Ectomycorrhization
of Gnetum in this study therefore led to improved access to added P. Non-
ectomycorrhizal plants in the said study responded to phosphorus addition
by increased root and total plant dry weights, shoot elongation and total
number of new leaves. An increase in foliar P concentration was also
demonstrated, thus suggesting that P availability may have been one of the
limiting factors under these experimental conditions. In a previous study,
Lee and Alexander (1994) had demonstrated that non-mycorrhizal Hopea
odorata responded to P addition by showing an increase in shoot dry weight
and P concentration. They also observed that ectomycorrhizal colonization
increased shoot P concentration and dry weight by about 100% as compared
to those of uncolonized plants growing on P amended soil.
Both P addition and EM colonization led to an increase in total
foliar P. Mycorrhizal colonization increased foliar N concentrations at
all P concentrations. Foliar N concentrations of mycorrhizal plants were
higher when P was added, but this was not the case for non-mycorrhizal
plants. There was a linear relationship between percent ectomycorrhizal
colonization and total foliar N at low P concentrations but this was significant
(P< 0.05) only in absence of added P. However, there was no evidence of an
association between total foliar P and percent colonization.
The relationship between ectomycorrhizal colonization and total foliar
N at low P concentration in the study by Bechem and Alexander (2012b)
gives an indication of the functional relationship between EM colonization
and nutrient (N, P) acquisition by plant species of the tropical rain forests.
Although ectomycorrhization and P addition had significant effects on
total foliar P in the said study, the analysis of data did not show any linear
relationship between percent colonization and foliar P. This observation
was inconsistent with that of Moyersoen et al. (1998) in which a clear
160 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
9. Conclusion
From the findings of this researcher and her collaborators, there is further
evidence that some tropical ectomycorrhiza fungi are able to use amino acids
and some proteins as a sole N source. The stimulatory effect of peptone
was interesting, although the implication of this observation in nature is
to be determined. There was evidence that the fungus was able to utilize
organic P and N. It expressed both extracellular and cell-bound phosphatase
activity, with activity being higher in the absence of inorganic P. There is
therefore a possibility that the fungus would be able to access some organic
P sources in nature when inorganic P is limiting, an ability which would
be beneficial to the host plant. It is conclusive that ectomycorrhizas are an
integral part of Gnetum’s physiology. Any useful and effective conservation
strategy therefore would have to incorporate the diversity and biology of
the fungal partner.
The Physiology of Scleroderma sinnamariense Mont. 161
Acknowledgements
I appreciate the assistance from colleagues at the Department of Botany and
Plant Physiology, University of Buea for their support. I am indebted to Dr
Egbe Enow Andrew for reading this manuscript. I thank the anonymous
referees for their valuable comments on this study, and Krista L. McGuire
and Caitlyn Gillikin for improving the language.
References
Abuzinadah, R.A. and D.J. Read. 1986. The role of proteins in the nitrogen nutrition of
ectomycorrhizal plants .I. Utilisation of peptides and proteins by ectomycorrhizal fungi.
New Phytol. 103: 481–493.
Antibus, R.K., R.L. Sinsabaugh and A.E. Linkins. 1992. Phosphatase activities and phosphorus
uptake from inositol phosphate by ectomycorrhizal fungi. Can. J. Bot. 70: 794–801.
Bâ, A.M., K.B. Sanon and R. Duponnois. 2002. Influence of ectomycorrhizal inoculation on
Afzelia quanzensis Welw. seedlings in a nutrient-deficient soil. For. Ecol. Manag. 161:
215–219.
Bechem, E.E. 2004. Mycorrhizal status of Gnetum spp. in Cameroon: Evaluating diversity
with a view to ameliorating domestication efforts. PhD Thesis, University of Aberdeen,
Aberdeen, Scotland.
Bechem, E.E. 2011. Growth and in vitro phosphate solubilizing ability of Scleroderma sinnamariense:
A tropical mycorrhiza fungus isolated from Gnetum africanum ectomycorrhiza root tips.
J. Yeast Fungal Res. 2: 132–142.
Bechem, E.E. 2012a. Utilisation of organic and inorganic nitrogen sources by Scleroderma
sinnamariense Mont. isolated from Gnetum africanum Welw. Afr. J. Biotechnol. 11:
9205–9213.
Bechem, E.E. 2012b. Effect of carbon and nitrogen sources on in vitro growth of Scleroderma
sinnamariense Mont., a pantropical ectomycorrhizal fungus. Int. J. Biol. Chem. Sci. 6:
1192–1201.
Bechem, E.E. and I.J. Alexander. 2009. Inoculum production and inoculation of Gnetum
africanum rooted cuttings using a range of mycorrhizal fungi. Int. J. Biol. Chem. Sci. 3:
578–586.
Bechem, E.E. and I.J. Alexander. 2012a. Mycorrhizal status of Gnetum africanum in Cameroon:
Evaluating diversity with a view to ameliorating domestication efforts. Mycorrhiza 22:
99–108.
Bechem, E.E. and I.J. Alexander. 2012b. Phosphorus nutrition of ectomycorrhizal Gnetum
africanum plantlets from Cameroon. Plant Soil 353: 379–393.
Cadiz, R.T. and H.B. Florido. 2001. Gnetum gnemon Linn. BAGO. Res. Info. Series Ecosysts.
13: 1–6.
Calleja, M., D. Mousain, B. Lecouvreur and J. d’Auzac. 1980. Influence de la carence phosphatée
sur les activitées phophatases acides de trois champignons mycorrhizens: Hebeloma edurum
Metrod., Suillus granulutus (L. ex Fr.) O. Kuntze et Pisolithus tinctorius (Pers.) Coker et
Couch. Physiol. Vég. 18: 489–504.
Cairney, J.W.G. and S.E. Smith. 1992. Influence of intracellular phosphorus concentration on
phosphate absorption by the ectomycorrhizal basidiomycete Pisolithus tinctorius. Mycol.
Res. 96(8): 673–676.
Castellano, M.A. 1996. Outplanting performance of mycorrhizal inoculated seedlings. In: K.G.
Mukerji (ed.). Concepts in Mycorrhizal Research. Kluwer, Dordrecht, pp. 223–301.
Demoulin, V. and D.M. Dring. 1971. Two new species of Scleroderma from tropical Africa.
Trans. Br. Mycol. Soc. 56: 163–165.
162 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Dodd, J.C., P. Jeffries and R. De La Cruz. 1996. The use of mycorrhiza fungi in reforestation
programmes in the Philippines. Final Report EU Contract C II*-CT91-0904, STD3
Programme, DG XII, Brussels.
Gianinazzi-Pearson, V. and S. Gianinazzi. 1989. Cellular and genetical aspects of the interactions
between host and fungal symbionts in mycorrhizae. Genome 31: 341–366.
Guzman, G. 1970. Monografia del genero Scleroderma Pers. Amend. Fr. Darwinania 16:
233–407.
Guzman, G. and C.L. Ovrebo. 2000. New observations in sclerodermataceous fungi. Mycologia
92: 174–179.
Hampp, R. and C. Schaeffer. 1994. Mycorrhiza-carbohydrate and energy metabolism. In:
A. Varma and B. Hock (eds.). Mycorrhiza-Structure, Function, Molecular biology and
Biotechnology. Springer-Verlag, Germany.
Hasselquist, N.J., L.S. Santiago and M.F. Allen. 2010. Belowground nitrogen dynamics in
relation to hurricane damage along a tropical dry forest chronosequence. Biogeochemistry
98: 89–100.
Hodge, A. 2003. N capture by Plantago lanceolata and Brassica napus from organic material: the
influence of spatial dispersion, plant competition and an arbuscular mycorrhizal fungus.
J. Exp. Bot. 54: 2331–2342.
Ingleby, K. 1999. Scleroderma sinnamariense Mont. + Gnetum africanum. In: R. Agerer, R.M.
Danielson, K. Ingleby, K. Luoma and R. Treu (eds.). Descriptions of Ectomycorrhizae
4: 127–133.
Jennings, D.H. 1997. The Physiology of Fungal Nutrition. Cambridge University Press,
Cambridge.
Jeffries, M. 1999. Scleroderma. In: J.W.G Cairney and S.M Chambers (eds.). Ectomycorrhizal
Fungi: Key Genera in Profile 7: 187–200.
Kropp, B.R. 1990. Variation in acid phosphatase activity among progeny from controlled crosses
in the ectomycorrhizal fungus Laccaria bicolor. Can. J. Bot. 68: 864–866.
Lapeyrie, F., J. Ranger and D. Vairelles. 1991. Phosphate-solubilizing activity of ectomycorrhiza
fungi in vitro. Can. J. Bot. 69: 342–346.
Lee, S.S. and I.J. Alexander. 1994. The response of seedlings of two dipterocarp species to
nutrient additions and ectomycorrhizal infection. Plant Soil 163: 299–306.
Lee, S.S., I.J. Alexander and R. Watling. 1997. Ectomycorrhizae and putative ectomycorrhizal
fungi of Shorea leprosula Miq. (Dipterocarpaceae). Mycorrhiza 7: 63–81.
Leyval, C. and J. Berthelin. 1986. Comparison between the utilization of phosphorus from
insoluble mineral phosphates by ectomycorrhizal fungi and rhizobacteria. In: V.
Gianinazzi-Pearson and S. Gianinazzi (eds.). Physiological and Genetical Aspects of
Mycorrhizae INRA: Paris, France, pp. 340–345.
Marx, D.H. 1969. The influence of ectotrophic mycorrhizal fungi on resistance of pine roots
to pathogenic infections. I. Antagonism of mycorrhizal fungi to root pathogenic fungi
and soil bacteria. Phytopathology 59: 153–163.
Martinelli, L.A., M.C. Piccolo, A.R. Townsend, P.M. Vitousek, E. Cuevas, W. Mcdowell, G.P.
Robertson, O.C. Santos and K. Treseder. 1999. Nitrogen stable isotopic composition of
leaves and soil: tropical versus temperate forests. Biogeochemistry 46: 45–65.
McElhinney, C and D.T. Mitchell. 1993. Phosphate activity of four ectomycorrhizal fungi in a
Sitka spruce-Japanese larch plantation in Ireland. Mycol. Res. 97(6): 725–732.
Modess, 0. 1941. Zur Kenntnis der Mykorrhizabildner von Kiefer and Fichte. Symb. Bot.
Upsal. 5: 1–146.
Molina, R. and J.G. Palmer. 1982. Isolation, maintenance and pure culture manipulation of
ectomycorrhizal fungi. In: N. Schenck (ed.). Methods and Principles of Mycorrhizal
Research. The American Phytopathological Society, pp. 115–129.
Moyersoen, B., I.J. Alexander and A.H. Fitter. 1998. Phosphorus nutrition of ectomycorrhiza
and arbuscular mycorrhizal tree seedlings from a lowland tropical forest in the Korup
National Park, Cameroon. J. Trop. Ecol. 14: 47–61.
The Physiology of Scleroderma sinnamariense Mont. 163
Onguene, N.A. and T.W. Kuyper. 2001. Mycorrhiza associations in the rain forest of South
Cameroon. For. Ecol. Man. 140: 277–287.
Pachlewski, R. and J. Pachlewska. 1974. Studies on symbiotic properties of mycorrhizal fungi
of pine (Pinus sylvestris) with the aid of the method of mycorrhizal synthesis in pure
culture on agar, Warsaw, Poland: Forest Research Institut, pp. 139.
Putra, D.P., I. Nurmilasari, Y. Komala, Asakawa and D. Arbain. 2011. The coloring constituents
of Scleroderma sinnamariense (Sclerodermaceae). Nat. Prod. Commun. 6: 357–360.
Rangel-Castro, J.I., E. Danell and A.F.S. Taylor. 2002. Use of different nitrogen sources by the
edible ectomycorrhizal mushroom Cantharellus cibarius. Mycorrhiza 12: 131–137.
Richter, D.L. and J.N. Bruhn. 1989. Pinus resinosa ectomycorrhizae: seven host-fungus
combinations synthesized in pure culture. Symbiosis 7: 211–228.
Sawyer, N.A., S.M. Chambers and J.W.G. Cairney. 2003. Utilisation of inorganic and organic
nitrogen sources by Amanita species native to temperate eastern Australia. Mycol. Res.
107: 413–420.
Sims, K., R. Watling and P. Jeffries. 1995. A revised key to the genus Scleroderma. Mycotaxon
56: 403–420.
Sims, K., R. Sen, R. Watling and P. Jeffries. 1999. Species and population structures of Pisolithus
and Scleroderma identified by combined phenotypic and genomic marker analysis. Mycol.
Res. 103: 449–458.
Tibbett, M., F.E. Sanders and J.W.G Cairney. 1998. The effect of temperature and inorganic
phosphorus supply on growth and acid phosphatase production in arctic and temperate
strains of ectomycorrhizal Hebeloma spp. in axenic culture. Mycol. Res. 102: 129–135.
Van Leerdam, D.M., P.A. Williams and J.W.G. Cairney. 2001. Phosphate-solubilizing abilities
of ericoid mycorrhizal endophytes of Woollsia pungens (Epacridaceae). Aust. J. Bot. 49:
75–80.
Villegas, J. and J.A. Fortin. 2002. Phosphorus solubilization and pH changes as a result of
the interactions between soil bacteria and arbuscular mycorrhizal fungi on a medium
containing NO3- as nitrogen source. Can. J. Bot. 80: 571–576.
Vitousek, P.M. and R.L. Sandford. 1986. Nutrient cycling in moist tropical forest. An. Rev.
Ecol. and Syst. 17: 137–167.
Watling, R. 1994. Taxonomic and Floristic notes on some Malaysian Larger Fungi- I. Malayan
Nat. J. 48: 67–78.
Zak, B. and D.H. Marx. 1964. Isolation of individual fungi from roots of individual slash
pines. For. Sci. 10: 214.
CHAPTER
9
Alleviation of Salt Stress by
Scleroderma bermudense in
Coccoloba uvifera Seedlings in
the French West Indies
Amadou Mustapha Bâ,1,2,3,* Raymond Avril,1
Eric Bandou,1 Seynabou Sène,2 Robin Duponnois,3 Régis
Courtecuisse,4 Samba Sylla2 and Abdala Diédhiou2
1. Introduction
More than 800 million ha of land throughout the world is affected by
salt levels. The soil types most affected by excess salt are found in 30% of
Australian soils (Flowers and Colmer 2008). Excess salt in soils can have
two origins: (i) natural causes, i.e., weathering of parent rocks, deposition
of oceanic salts carried in wind and rain (10 mg/kg of NaCl would deposit
1
Laboratoire de Biologie et Physiologie Végétales (LBPV), Faculté des Sciences Exactes et
Naturelles, Université des Antilles et de la Guyane, BP. 592, 97159, Pointe-à-Pitre, Guadeloupe,
France.
2
Laboratoire Commun de Microbiologie (LCM) IRD/UCAD/ISRA, Centre de Recherche de
Bel Air, BP. 1386, CP. 18524 Dakar, Sénégal.
3
Laboratoire des Symbioses Tropicales et Méditerranéennes (LSTM), UMR113, Campus de
Baillarguet, A10/J, 34398 Montpellier, Cedex 5, France.
4
Département de Mycologie et de Botanique, Université de Lille, BP. 83, F-59006 Lille cedex,
France.
*Corresponding author: amadou.ba@ird.fr, amadou.ba@univ-ag.fr
Alleviation of Salt Stress by Scleroderma bermudense by Coccoloba uvifera 165
10 kg/ha of salt for each 100 mm of rainfall per year); (ii) human causes,
i.e., excess amounts of fertilizers, irrigation with brackish water (20% of
irrigated lands in the world are affected), and (iii) increased sea level caused
by global climate changes. Subsequently, two main groups of salted soils
exist: saline soils (soluble salts) and alkaline sodic soils (adsorbed sodium).
Saline soils affect plant growth without changing the physical properties
of soil. Soil is classified as saline when electrical conductivity ECe is ≥
4 dS m–1 (~40 mM or 2.4‰ NaCl) and generates an osmotic pressure of
0.2 MPa. Saline soils cover 397 million ha worldwide (FAO 2005). Sodic
soils affect plant growth and soil structure by saturating the adsorption
complex by Na+, which causes soil structure to evolve to an unstable state
of dispersion. This can be overcome through the flocculant power of Ca++
from gypsum (CaSO42H2O). Soil is classified as sodic when Exchangeable
Sodium Percentage (ESP) is ≥ 15% of cation exchange capacity. Sodic soils
cover 434 million ha worldwide (FAO 2005).
Plants respond to salinity stress at two levels: (i) osmotic stress (rapid
response) due to the increase in external osmotic pressure (reduction of
shoot growth rate); this is the osmotic or water-deficit effect of salinity;
(ii) Salt-specific effect (slow response) due to the accumulation of toxic
ions (Na+, Cl–) in leaves (decrease of shoot growth rate only for sensitive
plant). This is the toxic effect of salinity (Munns 2005). The osmotic stress
not only has an immediate effect on growth, but also has a greater effect
on growth rates than the ionic stress. To overcome salt-stress problems, it
is possible to select salt-tolerance plants, to use biological processes such
as mycorrhizal interactions, or to desalinate soil by leaching excessive
salts (Parida and Das 2005). The desalinisation of soils is not economically
viable for sustainable agriculture. Although the results are somewhat
variable, salt tolerant plants develop mechanisms such as osmotic and ionic
homeostasis. Osmotic homeostasis consists of osmotic adjustment through
the active accumulation of compatible solutes (e.g., K+, sucrose, proline,
glycine betaine) in the cytosol to maintain turgor of cells (e.g., stomatal
opening) that would otherwise dehydrate. Ionic homeostasis consists of
Na+ exclusion by roots through transporters (e.g., type SOS1) to ensure that
Na+ does not accumulate to toxic concentrations in leaves, or Na+ and Cl–
compartmentation through transporters (e.g., type NHX) in the vacuoles
to avoid toxic concentrations within the cytoplasm.
The majority of terrestrial plants form symbioses with mycorrhizal fungi
(Smith and Read 2008). There is considerable evidence that mycorrhizal
fungi can improve plant growth and nutrition in soils subject to a range
of saline stress (Giri and Mukerji 2004, Parida and Das 2005, Bandou et al.
2006, Bois et al. 2006). Mycorrhiza can help moderately salt-tolerant plants
prevent Na+ and Cl– translocation to shoot and leaf tissues, and improved
salt tolerance after mycorrhizal colonization may also result from more
166 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 1. Distribution of three Coccoloba species (Coccoloba uvifera, C. swartzii and C. pubescens) and
their putative ectomycorrhizal fungi along a salinity gradient in Martinique and Guadeloupe
(Lesser Antilles).
Color image of this figure appears in the color plate section at the end of the book.
way in which salinity exerts its influence on these vital processes through
an osmotic effect or a specific ion toxicity is still not resolved (Katembe et
al. 1998).
Table 2 shows the diversity of putative ECM fungi associated with
different Coccoloba species in the Greater and Lesser Antilles. Some of them
were common with those we collected in Martinique and Guadeloupe. We
collected 19 putative ECM fungi, of which 6 were found under C. uvifera
and 13 under C. pubescens and C. swartzii along a gradient of salinity
(Fig. 1) (Bessard et al. 2008). The 11 epigeous (Russula, Lactarius, Fistulinella,
Scleroderma, Boletelus, Clavaria, Clavulina, Coltricia, Limacella, Ramaria and
Thelephora) and 2 hypogeous (Austrogautieria and Melanogaster) fungal
species were collected under C. pubescens and C. swartzii in areas without
salt, whereas 8 different fungal species were collected in C. uvifera in salted
sandy soils (0–2‰ of salinity) (Fig. 1). Diversity of ECM fungi increased
when the salinity was low. It could be structured by salinity, host preference,
and/or charcteristics of soils along the salinity gradient. Also, abundance
of sporocarps fruiting from under mature trees and seedlings in C. uvifera
coastal forest were assessed over the salinity gradient (Fig. 2). Six species
of sporocarps were identified in low salinity levels (0–2‰) and only one
of them (S. bermudense) fructified in high salinity levels (2–15‰). Thus,
sporocarps of S. bermudense were more abundant in high salt levels than
Table 2. Putative ectomycorrhizal fungi associated with different Coccoloba species in the Greater and Lesser Antilles.
170
70
60
Number of sporocarps
Salted soil (2-15‰)
50 Salted soil (0-2‰)
40
30
20
10
4
Number of species
0
1 15 29 43 57 71 85 99 113127141155169183
Number of sporocarps
Fig. 3. Species accumulation curve for sporocarps in a C. uvifera coastal forest in Guadeloupe.
Dotted lines indicate 95% CI.
9 f S. bermudense
f Melanogaster sp.
8
Scleroderma sp.
7 e
e
Radial growth (cm)
6 e
5
4 d cd
3 bc
ab
2 a a
ab ab a a
1
0
0 100 200 300 500
NaCl (mM)
Fig. 4. Radial growth of S. bermudense, Melanogaster sp. and Scleroderma sp. in the presence of
NaCl at a range of concentrations in axenic culture for 1 mon (Two-ways ANOVA, P<5%).
174 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
increasing NaCl concentration, although the decline was not significant with
incresasing NaCl up to 100 mM. Similar results were found for biomass
yield for all parameters (Fig. 5). Selection of the salt concentration ranges
used in the present study was based on field measurements of soil salinity
(Bandou 2005, Bandou et al. 2006). Our data indicated that S. bermudense
were broadly resistant to NaCl at concentrations encountered in the saline
soils of littoral forests. This fungus could also require Na+ for growth as
their radial growth and biomass yield increased at two concentrations
(100–200 mM) relative to the zero NaCl treatment. Chen et al. (2001) also
found that some isolates of Pisolithus showed clear peaks in biomass yield
at concentrations in the range of 25–100 mM NaCl. Much of the information
on fungal salt tolerance comes from studies on aquatic fungi (Clipson
and Jennings 1992). While some marine fungi require Na+ for growth i.e.
one fungal species growsat 12.8 M NaCl in the Great Salt Lake, terrestrial
fungi appear to have less of a requirement. The observed peaks could also
reflect responses to lowered external water potential generated by NaCl
in the medium. This hypothesis is supported by similar peaks observed in
experiments in which the growth of ECM fungi was measured at different
0.25
S. bermudense
Melanogaster sp.
h
0.2 Scleroderma sp.
gh
Dry mass (g)
0.15
f
f f
0.1
e
de bcd
cde cd
0.05 abc
ab
a a a
0
0 100 200 300 500
NaCl (mM)
Fig. 5. Biomass yield of S. bermudense, Melanogaster sp. and Scleroderma sp. in the presence of
NaCl at a range of concentrations in axenic culture for 1 mon (Two-ways ANOVA, P<5%).
Alleviation of Salt Stress by Scleroderma bermudense by Coccoloba uvifera 175
9000
S. bermudense
8000 e
Melanogaster sp. de
Scleroderma sp. de
7000 de de
d d
6000
Na (ppm)
5000
c
4000 bc
3000
ab
2000 a
a
1000
a a a
0
0 100 200 300 500
NaCl (mM)
Fig. 6. Na contents of S. bermudense, Melanogaster sp. and Scleroderma sp. in the presence of
NaCl at a range of concentrations in axenic culture for 1 mon (Two-ways ANOVA, P<5%).
176 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
1200
S. bermudense
800 b
a
600 a a
a a a
a a
400 a a a
200
0
0 100 200 300 500
NaCl (mM)
Fig. 7. Proline contents of S. bermudense, Melanogaster sp. and Scleroderma sp. in the presence of
NaCl at a range of concentrations in axenic culture for 1 mon (Two-ways ANOVA, P<5%).
Table 3. Effects of inoculation with Scleroderma bermudense and NaCl levels on growth variables
and ectomycorrhizal (ECM) colonization in seedlings of Coccoloba uvifera at 3 mon (p<1%)
(Bandou et al. 2006).
0.16
Stem
g
0.14 Leaf fg g
fg fg
0.12 Root f
0.1
e
P (%)
e
0.08 de e
de
bc cd
0.06
bc bc bc
bc bc
0.04 ab ab ab
ab ab
a
0.02
0
0 200 350 500 0 200 350 500
Controls Inoculated
NaCl (mM)
Fig. 8. Changes in phosphorus (P) concentrations in the leaves, shoots, and roots of C. uvifera
seedlings in response to NaCl levels and inoculation with the ectomycorrhizal (ECM) fungus
S. bermudense at 3 mon (Bandou et al. 2006).
Na and Cl by the ECM plants growing in the NaCl treatments. One of the
main mechanisms for enhanced salinity tolerance in ECM plant has also
been proposed to be the exclusion of Na and Cl (Giri and Mukerji 2004,
Tian et al. 2004).
ECM plants had higher concentrations of P than non-inoculated plants
particularly under salt stress conditions. This could be due to the extended
network of hyphae beyond the depletion zone around roots that acquire
178 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
more P and therefore suppress the adverse effect of salinity stress. These
results suggest that S. bermudense can alleviate the deleterious effects of NaCl
stress on plants by mechanisms which may also be related to improvement
of Puptake (Fig. 8).
If Na and Cl are sequestered into plant vacuoles, K and organic solutes
(e.g., proline) should accumulate in the cytoplasm and organelles to balance
the osmotic pressure of intravacuolar ions (Munns 2002). Na and Cl were
not accumulated under salt stress and found at low concentrations in ECM
seedlings of C. uvifera when compared to controls (Figs. 9 and 10).
It was also noteworthy that K concentrations were higher in leaves of
ECM seagrape seedlings than in controls at all salinity levels. S. bermudense
seemed to protect seedlings by increasing the K/Na ratio to maintain
higher turgor under salt stress (Figs. 11 and 12). It is well known that
high concentrations of K and low concentrations of Na can play a major
role in salt stress tolerance by regulating stomatal openings and osmotic
potential in the vacuoles (Zhu 2003, Munns 2005). Some studies indicated
that plants subjected to increased salinity accumulated less K because
Na uptake competed with the uptake of K, leading to decreased K levels
(Larcher 1995, Parida and Das 2005). Reductions in Na and Cl uptake and
2.5
i
i Stem
2 Leaf
i
Root
h
1.5 h
Na (%)
1
g
efg fg d-g fg fg
efg
0.5 a-e a-e b-f
b-f c-f
abc abc a-d a-d
ab a
0
0 200 350 500 0 200 350 500
Controls Inoculated
NaCl (mM)
Fig. 9. Changes in sodium (Na) concentrations in the leaves, shoots, and roots of C. uvifera
seedlings in response to NaCl levels and inoculation with the ectomycorrhizal (ECM) fungus
S. bermudense at 3 mon (Bandou et al. 2006).
Alleviation of Salt Stress by Scleroderma bermudense by Coccoloba uvifera 179
5 i
Stem
i
Leaf
4 i
Root
Cl (%)
h
3
h h
h gh gh
2 fg
d-f ef
c-f b-e b-e
1 a-e
abc abc b-e b-e
abc ab ab a
0
0 200 350 500 0 200 350 500
Controls Inoculated
NaCl (mM)
Fig. 10. Changes in chloride (Cl) concentrations in the leaves, shoots, and roots of C. uvifera
seedlings in response to NaCl levels and inoculation with the ectomycorrhizal (ECM) fungus
S. bermudense at 3 mon (Bandou et al. 2006).
2,5
Stem
Leaf j i ij
2
Root
h
1,5
K (%)
efg
fg g efg
1 efg d-g efg c-g efg
c-g fg
a-d a-e abc b-g b-g
a ab a-d
b-f
0,5
0
0 200 350 500 0 200 350 500
Controls Inoculated
NaCl (mM)
Fig. 11. Changes in potassium (K) concentrations in the leaves, shoots, and roots of C. uvifera
seedlings in response to NaCl levels and inoculation with the ectomycorrhizal (ECM) fungus
S. bermudense at 3 mon (Bandou et al. 2006).
180 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
24
22 Stem a
20 Leaf
18
Root
K/Na (%) 16
14
12
ab
10
c
8
6
d
d de
4 de def
de de de de de
def e
2 def def def def e e
f f f
0
0 200 350 500 0 200 350 500
Controls Inoculated
NaCl (mM)
Fig. 12. Changes in ration K/Na concentrations in the leaves, shoots, and roots of C. uvifera
seedlings in response to NaCl levels and inoculation with the ectomycorrhizal (ECM) fungus
S. bermudense at 3 mon (Bandou et al. 2006).
Table 4. Correlation coefficients (r) between K, P, Na, Cl, and proline, and total biomass
production (Bandou et al. 2006).
a a b
Inoculated Non-inoculated Inoculated plus non-inoculated
K 0,65* 0.50 0.91*
P 0.85* 0.25 0.55*
Na –0.70* –0.94* –0.74*
Cl –0.71* –0.91* –0.79*
Proline 0.95* –0.93* 0.94*
a b
* Significant (p<1%); 10 df; 22 df
Alleviation of Salt Stress by Scleroderma bermudense by Coccoloba uvifera 181
seedlings despite their higher evaporative leaf surface (Table 5). Also, it is
noteworthy that leaf proline content was higher in leaves of ECM seagrape
seedlings than in the controls at all salinity levels (Fig. 13). S. bermudense
seemed to also protect seagrape seedlings by increasing the proline content
to maintain higher turgor under salt stress.
Table 5. Effects of inoculation with Scleroderma bermudense and NaCl levels on the number of
leaves, leaf area, and minimal leaf water potential in seedlings of Coccoloba uvifera at 3 mon
(p<1%) (Bandou et al. 2006).
ECM status NaCl levels Number of Leaf area (cm2) Minimal leaf water
(mM) leaves potential (-bars)
Non-inoculated 0 3.60 d 33.7 b 9.2 e
200 2.90 c 18.7 a 13.4 bc
350 2.30 b 15.6 a 14.0 b
500 1.90 a 15.7 a 15.0 a
Inoculated 0 4.10 e 80.9 e 5.9 f
200 4.20 e 76.4 d 11.1 d
350 3.80 de 68.9 d 12.2 cd
500 3.60 d 56.05 c 15.50 a
900 d
Leaf proline content (mmol g DW-1)
800
700
600
c
500
400
300 b
200
a a a
100 a a
0
0 200 350 500 0 200 350 500
Controls Inoculated
NaCl (mM)
Fig. 13. Changes in proline concentrations in the leaves of C. uvifera seedlings in response
to NaCl levels and inoculation with the ectomycorrhizal (ECM) fungus S. bermudense at
3 mon.
182 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
5. Conclusion
Salinity had a negative impact on growth of mature trees and naturally
regenerating seedlings of C. uvifera. Salinity could contribute to the
structuration of the three Coccoloba species as well as their associated ECM
fungi. The data from the present study also suggest that S. bermudense possess
considerable resistance to salinity. Also, seagrape tolerance to salt stress was
considerably enhanced by S. bermudense. Reductions in Na and Cl uptake
and enhanced P nutrition may be significant in helping the ECM plants
to grow under saline conditions. Furthermore, the higher accumulation
of K in the leaves of ECM plants could render ECM plants more tolerant
to osmotic stress induced by exposure to salt. The overall improved water
status of seagrape seedlings related to ECM inoculation with S. bermudense
may also be a contributing factor. Further studies should be done to
elucidate the physiological mechanisms (e.g., stomatal conductance, water
use efficiency, transpiration, photosynthetic rate, osmotic adjustment) by
which S. bermudense improved the water status of seagrape seedlings. Our
results should be taken cautiously as the experiment was a greenhouse study
with 3 month-old seedlings. From an ecological point of view, the ECM
symbiosis may be an important factor for the establishment and survival
of seagrape stands under saline stress along the beaches of the Caribbean.
Moreover, transplanting ECM-inoculated seagrape to such degraded sites
not only benefits the individual plant but, more importantly, may result in
the development of ornamental plantings and coastal windbreaks along
Caribbean beaches and roadsides.
Acknowledgements
We thank the anonymous referees for their valuable comments on this study,
and Krista L. McGuire and Caitlyn Gillikin for improving the language.
References
Al-Karaki, G.N., R. Hammad and M. Rusan. 2001. Response of two tomato cultivars differing
in salt tolerance to inoculation with mycorrhizal fungi under salt stress. Mycorrhiza 11:
43–47.
Augé, R.M. 2001. Water relations, drougth and vesicular-arbuscularmycorrhizal symbiosis.
Mycorrhiza 11: 3–42.
Bandou, E. 2005. Diversité et fonctionnement des symbioses ectomycorhiziennes de Coccoloba
uvifera (L.) L. en situation de stress salin et hydrique. Master of Science thesis, UAG, p.
36.
Bandou, E., F. Lebailly, F. Muller, M. Dulormne, A. Toribio, J. Chabrol, R. Courtecuisse, C.
Plenchette, Y. Prin, R. Duponnois, M. Thiao, S. Sylla, B. Dreyfus and A.M. Bâ. 2006.
The ectomycorrhizal fungus Scleroderma bermudense alleviates salt stress in seagrape
(Coccolobauvifera L.) seedlings. Mycorrhiza 16: 559–565.
Alleviation of Salt Stress by Scleroderma bermudense by Coccoloba uvifera 183
Bessard, S., I. Boulogne, J. Chabrol, J.P. Fiard, C. Lécuru, R. Courtecuisse and A.M. Bâ. 2008.
Diversity of ectomyorrhizal fungi and ectomycorrhizas of three Coccoloba species in
Martinique. Poster presented at the Workshop « Mycorrhizas in tropical forests »,
Universidad Téchnica Particular de Loja, Ecuador.
Bois, G., A. Bertrand, Y. Piché, M. Fung and D.P. Khasa. 2006. Growth, compatible solute
and salt accumulation of five mycorrhizal fungal species grown over a range of NaCl
concentrations. Mycorrhiza 16: 99–109.
Chen, D.M., S. Ellul, K. Herdman and J.W.G. Cairnay. 2001. Influence of salinity on biomass
production by Australian Pisolithus spp. isolates. Mycorrhiza 11: 231–236.
Clipson, N.J.W. and D.H. Jennings. 1992. Dendryphiellasalina and Debaryomyceshansenii: models
for ecophysical adaptation to salinity by fungi that grow in the sea. Canadian Journal
of Botany 70: 2097–2105.
FAO. 2005. ftp://ftp.fao.org/agl/iptrid/salinity_brochure_fr.pdf.
Fournet, J. 2002. Flore illustrée des phanérogames de Guadeloupe et de Martinique. Editions
Quae, Tomes 1 and 2, p. 2538.
Flowers, T.J. and T.D. Colmer. 2008. Salinity tolerance in halophytes. New Phytol. 179:
945–963.
Giri, B. and K.G. Mukerji. 2004. Mycorrhizal inoculant alleviates salt stress in Sesbaniaaegyptiaca
and Sesbaniagrandiflora under field conditions : evidence for reduced sodium and
improved magnesium uptake. Mycorrhiza 14: 307–312.
Guzman, G., F. Ramirez-Guillén, O.K. Miller, D.J. Lodge and T.J. Baroni. 2004. Sclerodermastellatum
versus Scleroderma bermudense : the status of Scleroderma echinatum and the first record of
Veligasternitidum from the Virgin Islands. Mycologia 96: 1370–1379.
Howard, R. 1988. Flora of the lesser Antilles. (F LAnt) 4, pp. 132.
Jennings, D.H. 1995. The physiology of fungal nutrition. Cambridge University Press,
Cambridge, England.
Juniper, S. and L. Abbott. 1993. Vesicular-arbuscularmycorrhizas and soil salinity. Mycorrhiza
4: 45–57.
Katembe, W.J., I.A. Ungar and J.P. Mitchell. 1998. Effect of salinity on germination and seedling
growth of two Atriplexspecies (Chenopodiaceae). Ann. Bot. 82: 165–175.
Kernaghan, G., B. Hambling, M. Fung and D. Khasa. 2002. In vitro selection of Boreal
ectomycorrhizal fungi for use in reclamation of saline-alkaline habitats. Restor. Ecol.
10: 1–9.
Kreisel, K. 1971. Clave para la identificacion de los macromicetos de Cuba. La Habana: Ser.
A, CiencasBiologicas 16, Universidad de la Habana, p. 101.
Langenfeld-Heyser, R., J. Gao, T. Dicic, P.H. Tachd, C.F. Lu, E. Fritz, A. Gafur and A. Polle.
2007. Paxillusinvolutus mycorrhiza attenuate NaCl-stress responses in the salt-sensitive
hybrid poplar Populus x canescens. Mycorrhiza 17: 121–131.
Larcher, W. 1995. Physiological Plant Ecology: Ecophysiology and Stress Physiology of
Functional Groups. Springer, Third edition.
Miller, O.K., D.J. Lodge and T.J. Baroni. 2000. New and interesting ectomycorrhizal fungi from
Puerto Rico, Mona, and Guana Islands. Mycologia 92: 558–570.
Munns, R. 2002. Comparative physiology of salt and water stress. Plant Cell Env. 25:
239–250.
Munns, R. 2005. Genes and salt tolerance: bringing them together. New Phytol. 167:
645–663.
Parida, A.K. and A.B. Das. 2005. Salt tolerance and salinity effects on plants: a review. Ecotox.
Env. Safety 60: 324–349.
Parrota, J.A. 1994. Coccoloba uvifera (L.) L., seagrape, uva de playa. Research note SOITF-SM-
74. U.S. Department of Agriculture, Forest Service, Southern Forest Experiment Station,
New Orleans, LA pp. 5.
Pegler, D.N. 1983. Agaric flora of the Lesser Antilles. Kew Bull Add Ser 9, pp. 668.
Smith, S. And J. Read. 2008. Mycorrhizal symbiosis. Hardcover, pp. 800.
184 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Shen, B., S. Hohman, R.G. Jensen and H.J. Bohnert. 1999. Roles of sugar alcohols in osmotic
stress adaptation. Replacement of glycerol by mannitol and sorbitol in yeast. Plant
Physiol. 121: 45–52.
Tang, M., M. Sheng, H. Chen and F.F. Zhang. 2009. In vitro resistance of three ectomycorrhizal
fungi. Soil Biol. Bioch. 41: 948–953.
Tian, C.Y., G. Feng, X.L. Li and F.S. Zhang. 2004. Different effects of arbuscularmycorrhizal
fungal isolates from saline or non-saline soil on salinity tolerance of plants. Appl. Soil
Ecol. 26: 143–148.
Yano-Melo, A.M., O.J. Saggin and L.C. Maia. 2003. Tolerance of mycorrhized banana (Musa
sp. cv. Pacovan) plantlets to saline stress. Agr. Eco. Env. 95: 343–348.
Zhu, J.K. 2003. Regulation of ion homeostasis under salt stress. Curr. Opinion Plant Biol. 6:
441–445.
CHAPTER
10
The Contribution of
Ectomycorrhizal Fungal
Feedbacks to the Maintenance
of Tropical Monodominant
Rain Forests
Krista L. McGuire
1. Introduction
Tropical monodominant forests have been classically defined as areas of
forest where one tree species comprises >60% of the stems or basal area of
canopy trees (Torti et al. 2001). These forests occur throughout all tropical
regions (Fig. 1) and have been noted as interesting deviations from the
more extensive high-diversity rain forests (Leigh et al. 2004). A number of
mechanisms have been suggested to explain their existence and persistence
among the mosaics of highly diverse forest (Connell and Lowman 1989,
Hart 1990, Torti and Coley 1999), however, a single explanation for tropical
monodominance does not suffice (Torti et al. 2001, Peh et al. 2011). It
is now widely accepted that multiple mechanisms must be operating
simultaneously that lead to feedbacks, which enable a monodominant tree
species to persist and maintain site dominance (McGuire 2008).
Fig. 1. Map illustrating examples of monodominant and co-dominant forests with the tree
species or genus listed and the tree family in parentheses. This diagram is not an exhaustive
list of monodominant species, but it illustrates how monodominant forests are found in all
tropical regions.
Color image of this figure appears in the color plate section at the end of the book.
share and suggested that there must be interactions among these traits that
together promote positive feedbacks leading to monodominance.
Community feedback models clearly demonstrate that negative plant-
soil feedbacks lead to plant coexistence and positive feedbacks lead to
dominance (Bever et al. 2010, Bever et al. 2012). These models account for
individual plant responses to differing soil microbes and then scale these
interactions to community-level patterns of plant diversity. The negative
plant-soil feedbacks are predicted to originate from plant-pathogen
interactions and the positive feedbacks are derived from plant-symbiont
interactions. These theoretical models have been empirically supported by a
number of studies, although the majority of the experimental work has been
done as pot studies and herbaceous plant manipulations. Very few tropical
studies have been performed and little is known about the functioning of
plant-soil feedbacks in undisturbed systems. Tropical ECM monodominant
forests are ideal model systems for exploring these feedbacks, as they offer a
natural diversity gradient that has not been significantly altered by human
manipulation. Since the outcome of undisturbed forest dynamics in these
systems is that of single species dominance, it can be inferred that positive
plant-soil feedbacks have outweighed negative feedbacks in these systems
(Bever et al. 2010).
For the current review, I will be focusing on a subset of tropical
monodominant tree species: those that form ectomycorrhizal (ECM)
associations in lowland, aseasonal rain forests, and are not associated
with extreme edaphic conditions or periodic disturbances. The reason for
focusing on ECM monodominant forests is that the mechanisms operating
to maintain monodominance likely include ECM-mediated feedbacks that
are unique to these systems, as the ecology, evolution, and physiology of
ECM fungi are different than for fungi that form arbuscular mycorrhizal
(AM) associations, the dominant mycorrhizal type in most diverse tropical
forests. As such, there are several attributes of ECM fungi that may confer
a competitive advantage to their host trees, particularly when growing in
nutrient-poor soils (Smith and Read 2008). For example, ECM fungi can
grow much larger than AM fungi due to the septate nature of their cells.
When ECM hyphae are broken, they can begin growing again from the last
septation, enabling mycelia to grow tens and sometimes hundreds of meters
in diameter. By contrast, AM fungi have coenocytic hyphae, so following
cellular damage they must resume growth from the root-fungal interface.
Consequently, AM fungi are not thought to grow mycelia that are more than
several centimeters in diameter. This discrepancy in size places ECM fungi
at an advantage in terms of nutrient foraging area and space occupancy.
In fact, where ECM fungi occur, they can comprise up to 90% of the fungal
biomass (Wallander et al. 2001). Also, the recent evolutionary divergence
of multiple ECM fungal lineages from decomposer fungi has implications
188 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
βA CB
CA DB
Fig. 2. Incorporation of monodominant and mixed forest trees into theoretical soil-feedback
models (adapted from Bever et al. 2010, 2012). The direction and strength of the plant-soil
feedbacks should correlate with the relative abundance of the tree species. The net pairwise
effect can be quantified by evaluating ((αA + βB) – (αB + βA)).
The Contribution of Ectomycorrhizal Fungal Feedbacks 189
Fig. 3. Map showing the ECM monodominant Dicymbe corymbosa forest and mixed forest
sites in Guyana.
190 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
in the monodominant forest relative to the mixed forest was much smaller
than the advantage gained by D. corymbosa seedlings (Fig. 4). It was possible
that these mixed tree species were benefitting from lower pathogen and
seed predator densities, since the overall density of their conspecific adult
trees was lower in the monodominant forest, although these mechanisms
have not been explicitly tested. However, two seed density treatments were
implemented for all tree species to test the prevalence of seed predation in
both forest types, and there were no notable differences in seed predation
rates in either forest type for any of the density treatments. Thus, it appeared
that seedling recruitment, rather than seed survival, was more reflective
of canopy diversity patterns, and that ECM inoculum density was a major
factor driving these patterns for D. corymbosa. The reduced availability of
ECM inoculum was also reflected in a molecular study that sequenced
fungal DNA from forest floor samples in the monodominant and mixed
forests (McGuire et al. 2010). There were significantly fewer DNA sequences
aligning to ECM fungi in the mixed forest compared to the monodominant
forest, which reflects the reduced colonization levels of D. corymbosa
seedlings in the 2007 study. Whether D. corymbosa seedlings are subjected
to pathogen attack in the mixed forest because of lower ECM inoculum
has not been determined, but is a plausible hypothesis that may directly
relate to the lower ECM colonization levels. In other ECM trees, reduced
colonization can lead to increased vulnerability to root pathogen attack
(Marx 1972, Whipps 2001, 2004), and may be a mechanism preventing the
rapid spread of the ECM monodominant forest into the mixed forest.
Survivorship
Fig. 4. Survivorship of seeds and seedlings after one year in monodominant compared to
mixed forest. Seeds of Dicymbe corymbosa and four other species found in both forest types
were reciprocally transplanted. Data are reproduced from McGuire 2007a.
192 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Acknowledgements
We thank Krista L. McGuire and Caitlyn Gillikin for their valuable
comments on this study.
The Contribution of Ectomycorrhizal Fungal Feedbacks 197
References
Abuzinadah, R.A. and D.J. Read. 1986. The role of proteins in the nitrogen nutrition of
ectomycorrhizal plants .1. Utilization of peptides and proteins by ectomycorrhizal fungi.
New Phytol. 103: 481–493.
Bâ, A.M., R. Duponnois, B. Moyersoen and A.G. Diédhiou. 2012. Ectomycorrhizal symbiosis
of tropical African trees. Mycorrhiza 22: 1–29.
Barto, E.K., M. Hilker, F. Muller, B.K. Mohney, J.D. Weidenhamer and M.C. Rillig. 2011.
The Fungal Fast Lane: Common Mycorrhizal Networks Extend Bioactive Zones of
Allelochemicals in Soils. Plos One 6.
Bever, J.D., I.A. Dickie, E. Facelli, J.M. Facelli, J. Klironomos, M. Moora, M.C. Rillig, W.D.
Stock, M. Tibbett and M. Zobel. 2010. Rooting theories of plant community ecology in
microbial interactions. Trends in Ecol. Evol. 25: 468–478.
Bever, J.D., T.G. Platt and E.R. Morton. 2012. Microbial population and community dynamics
on plant roots and their feedbacks on plant communities. Ann. Rev. Microbiol. 66:
265–283.
Brearley, F.Q. 2012. Ectomycorrhizal Associations of the Dipterocarpaceae. Biotropica 44:
637–648.
Brownlee, C., J.A. Duddridge, A. Malibari and D.J. Read. 1983. The structure and function of
mycelial systems of ectomycorrhizal roots with special reference to their role in forming
inter-plant connections and providing pathways for assimilate and water transport.
Plant and Soil 71: 433–443.
Brundrett, M.C. 2009. Mycorrhizal associations and other means of nutrition of vascular plants:
understanding the global diversity of host plants by resolving conflicting information
and developing reliable means of diagnosis. Plant and Soil 320: 37–77.
Chazdon, R.L. 1988. Sunflecks and their importance to forest understorey plants. Adv. Ecol.
Res. 18: 1–63.
Colpaert, J.V. and K.K. van Tichelen. 1996. Decomposition, nitrogen and phosphorus
mineralization from beech leaf litter colonized by ectomycorrhizal or litter-decomposing
basidiomycetes. New Phytol. 134: 123–132.
Connell, J.H. and M.D. Lowman. 1989. Low-diversity tropical rain forests: some possible
mechanisms for their existence. Am. Nat. 134: 88–119.
Degagne, R.S., T.W. Henkel, S.J. Steinberg and L. Fox III. 2009. Identifying Dicymbe corymbosa
monodominant forests in Guyana using satellite imagery. Biotropica 41: 7–13.
Dickie, I.A. and B. Moyersoen. 2008. Towards a global view of ectomycorrhizal ecology. New
Phytol. 180: 263–265.
Finlay, R.D. and D.J. Read. 1986. The structure and function of the vegetative mycelium
of ectomycorrhizal plants.1. translocation of C-14 labeled carbon between plants
interconnected by a common mycelium New Phytol. 103: 143–156.
Floudas, D., M. Binder, R. Riley, K. Barry, R.A. Blanchette, B. Henrissat, A.T. Martínez, R.
Otillar, J.W. Spatafora, J.S. Yadav, A. Aerts, I. Benoit, A. Boyd, A. Carlson, A. Copeland,
P.M. Coutinho, R.P. de Vries, P. Ferreira, K. Findley, B. Foster, J. Gaskell, D. Glotzer, P.
Górecki, J. Heitman, C. Hesse, C. Hori, K. Igarashi, J.A. Jurgens, N. Kallen, P. Kersten, A.
Kohler, U. Kües, T.K.A. Kumar, A. Kuo, K. LaButti, L.F. Larrondo, E. Lindquist, A. Ling, V.
Lombard, S. Lucas, T. Lundell, R. Martin, D.J. McLaughlin, I. Morgenstern, E. Morin, C.
Murat, L.G. Nagy, M. Nolan, R.A. Ohm, A. Patyshakuliyeva, A. Rokas, F.J. Ruiz-Dueñas,
G. Sabat, A. Salamov, M. Samejima, J. Schmutz, J.C. Slot, F. St. John, J. Stenlid, H. Sun, S.
Sun, K. Syed, A. Tsang, A. Wiebenga, D. Young, A. Pisabarro, D.C. Eastwood, F. Martin,
D. Cullen, I.V. Grigoriev and D.S. Hibbett. 2012. The Paleozoic origin of enzymatic lignin
decomposition reconstructed from 31 fungal genomes. Science 336: 1715–1719.
Gadgil, R.L. and G.D. Gadgil. 1971. Mycorrhiza and litter decomposition. Nature 233: 133.
Gadgil, R.L. and P.D. Gadgil. 1975. Suppression of litter decomposition by mycorrhizal foots
of Pinus radiata. New Zealand J. For. Sci. 5: 35–41.
198 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Galante, T.E., T.R. Horton and D.P. Swaney. 2011. 95% of basidiospores fall within 1 m of the
cap: a field- and modeling-based study. Mycologia 103: 1175–1183.
Gross, N.D., S.D. Torti, D.H. Feener Jr. and P.D. Coley. 2000. Monodominance in an African
Rain Forest: Is Reduced Herbivory Important? 1. Biotropica 32: 430–439.
Hart, T.B., J.A. Hart and P.G. Murphy. 1989. Monodominant and species-rich forests of the
humid tropics: causes for their co-occurrence. American Naturalist 133: 613–633.
Hart, T.B. 1990. Monospecific dominance in tropical rain forests. Trends Ecol. Evol. 5: 6–11.
Henkel, T.W. 2003. Monodominance in the ectomycorrhizal Dicymbe corymbosa
(Caesalpiniaceae) from Guyana. J. Trop. Ecol. 19: 417–437.
Henkel, T.W., M.C. Aime, M.M.L. Chin, S.L. Miller, R. Vilgalys and M.E. Smith. 2012.
Ectomycorrhizal fungal sporocarp diversity and discovery of new taxa in Dicymbe
monodominant forests of the Guiana Shield. Biodiv. Conserv. 21: 2195–2220.
Hobbie, E.A., N.S. Weber, J.M. Trappe and G.J. van Klinken. 2002. Using radiocarbon to
determine the mycorrhizal status of fungi. New Phytol. 156: 129–136.
Ingleby, K., R.C. Munro, M. Noor, P.A. Mason and M.J. Clearwater. 1998. Ectomycorrhizal
populations and growth of Shorea parvifolia (Dipterocarpaceae) seedlings regenerating
under three different forest canopies following logging. For. Ecol. Manag. 111: 171–179.
Kiers, E.T., T.M. Palmer, A.R. Ives and J.F. Bruno. 2010. Mutualisms in a changing world: an
evolutionary perspective. Ecology 13: 1459–1474.
Koele, N., I.A. Dickie, J. Oleksyn, S.J. Richardson and P.B. Reich. 2012. No globally consistent
effect of ectomycorrhizal status on foliar traits. New Phytol. 196: 845–852.
Leigh, E.G., P. Davidar, C.W. Dick, J. Puyravaud, J. Terborgh, H. ter Steege and S.J. Wright. 2004.
Why do some tropical forests have so many species of trees? Biotropica 36: 445–473.
Lindahl, B., J. Stenlid, S. Olsson and R. Finlay. 1999. Translocation of P-32 between interacting
mycelia of a wood-decomposing fungus and ectomycorrhizal fungi in microcosm systems.
New Phytol. 144: 183–193.
Lindahl, B., J. Stenlid and R. Finlay. 2001. Effects of resource availability on mycelial
interactions and P-32 transfer between a saprotrophic and an ectomycorrhizal fungus
in soil microcosms. FEMS Microbiol. Ecol. 38: 43–52.
Marx, D.H. 1972. Ectomycorrhizae as biological deterrents to pathogenic root infections. Ann.
Rev. Phytopath. 10: 429–&.
McGuire, K.L. 2007a. Common ectomycorrhizal networks may maintain monodominance in
a tropical rain forest. Ecology 88: 567–574.
McGuire, K.L. 2007b. Recruitment dynamics and ectomycorrhizal colonization of Dicymbe
corymbosa, a monodominant tree in the Guiana Shield. J. Trop. Ecol. 23: 297–307.
McGuire, K.L. 2008. Ectomycorrhizal associations function to maintain tropical monodominance.
In: Z.A. Siddiqui, M.S. Akhtar and K. Futai (eds.). Mycorrhizae: Sustainable Agriculture
and Forestry. Springer, Netherlands, pp. 287–302.
McGuire, K.L., D.R. Zak, I.P. Edwards, C.B. Blackwood and R. Upchurch. 2010. Slowed
decomposition is biotically mediated in an ectomycorrhizal, tropical rain forest. Oecologia
164: 785–795.
Meinhardt, K.A. and C.A. Gehring. 2012. Disrupting mycorrhizal mutualisms: a potential
mechanism by which exotic tamarisk outcompetes native cottonwoods. Ecol. Appl. 22:
532–549.
Newman, E.I. 1988. Mycorrhizal Links between Plants—Their Functioning and Ecological
Significance. Adv. Ecol. Res. 18: 243–270.
Olsson, P.A., M. Chalot, E. Baath, R.D. Finlay and B. Soderstrom. 1996. Ectomycorrhizal mycelia
reduce bacterial activity in a sandy soil. FEMS Microbiol. Ecol. 21: 77–86.
Peay, K.G., M. Garbelotto and T.D. Bruns. 2009. Spore heat resistance plays an important role
in disturbance-mediated assemblage shift of ectomycorrhizal fungi colonizing Pinus
muricata seedlings. J. Ecol. 97: 537–547.
Peay, K.G., M.G. Schubert, N.H. Nguyen and T.D. Bruns. 2012. Measuring ectomycorrhizal
fungal dispersal: macroecological patterns driven by microscopic propagules. Mol. Ecol.
21: 4122–4136.
The Contribution of Ectomycorrhizal Fungal Feedbacks 199
Peh, K.S. H., B. Sonke, J. Lloyd, C.A. Quesada and S.L. Lewis. 2011. Soil Does Not Explain
Monodominance in a Central African Tropical Forest. Plos One 6: 9.
Ramachela, K. and J.M. Theron. 2010. Effect of ectomycorrhizal fungi in the protection of Uapaca
kirkiana seedlings against root pathogens in Zimbabwe. Southern For. 72: 37–45.
Read, D.J. 1991. Mycorrhizas in ecosystems. Experientia 47: 376–391.
Richards, P.W. 1952. The tropical rain forest. second edition edition. University Press,
Cambridge, UK.
Rineau, F., D. Roth, F. Shah, M. Smits, T. Johansson, B. Canback, P.B. Olsen, P. Persson, M.N.
Grell, E. Lindquist, I.V. Grigoriev, L. Lange and A. Tunlid. 2012. The ectomycorrhizal
fungus Paxillus involutus converts organic matter in plant litter using a trimmed brown-
rot mechanism involving Fenton chemistry. Environ. Microbiol. 14: 1477–1487.
Simard, S.W. and D.M. Durall. 2004. Mycorrhizal networks: a review of their extent, function,
and importance. Can. J. Bot. 82: 1140–1165.
Simard, S.W. 2009. The foundational role of mycorrhizal networks in self-organization of
interior Douglas-fir forests. For. Ecol. Manag. 258: S95–S107.
Smith, J.E. and D. Read. 2008. Mycorrhizal Symbiosis. Third Edition edition. Academic Press,
San Diego (CA).
Talbot, J.M., S.D. Allison and K.K. Treseder. 2008. Decomposers in disguise: mycorrhizal
fungi as regulators of soil C dynamics in ecosystems under global change. Func. Ecol.
22: 955–963.
Torti, S.D. and P.D. Coley. 1999. Tropical monodominance: A preliminary test of the
ectomycorrhizal hypothesis. Biotropica 31: 220–228.
Torti, S.D., P.D. Coley and T.A. Kursar. 2001. Causes and consequences of monodominance in
tropical lowland forests. Am. Nat. 157: 141–153.
Wallander, H., L.O. Nilsson, D. Hagerberg and E. Baath. 2001. Estimation of the biomass
and seasonal growth of external mycelium of ectomycorrhizal fungi in the field. New
Phytol. 151: 753–760.
Waltert, B., V. Wiemken, H.P. Rusterholz, T. Boller and B. Baur. 2002. Disturbance of forest by
trampling: Effects on mycorrhizal roots of seedlings and mature trees of Fagus sylvatica.
Plant and Soil 243: 143–154.
Whipps, J.M. 2001. Microbial interactions and biocontrol in the rhizosphere. J. Exp. Bot. 52:
487–511.
Whipps, J.M. 2004. Prospects and limitations for mycorrhizas in biocontrol of root pathogens.
Can. J. Bot. 82: 1198–1227.
Wolfe, B.E., V.L. Rodgers, K.A. Stinson and A. Pringle. 2008. The invasive plant Alliaria
petiolata (garlic mustard) inhibits ectomycorrhizal fungi in its introduced range. J. Ecol.
96: 777–783.
Wolfe, B.E., R.E. Tulloss and A. Pringle. 2012. The Irreversible Loss of a Decomposition Pathway
Marks the Single Origin of an Ectomycorrhizal Symbiosis. Plos One 7.
Wu, B.Y., H. Maruyama, M. Teramoto and T. Hogetsu. 2012. Structural and functional
interactions between extraradical mycelia of ectomycorrhizal Pisolithus isolates. New
Phytol. 194: 1070–1078.
Wu, T.H. 2011. Can ectomycorrhizal fungi circumvent the nitrogen mineralization for plant
nutrition in temperate forest ecosystems? Soil Biol. Biochem. 43: 1109–1117.
CHAPTER
11
Ectomycorrhiza in Forest
Rehabilitation in Indonesia
Hesti L. Tata
1. Introduction
The total forest area1 of Indonesia is 131.3 million ha, which is about 69.9%
of the total terrestrial territory of Indonesia. However, the forest cover
inside the state forest area is only 133,327.2 ha, which is only about 0.1%
of designated forest area (Ministry of Forestry 2012). This implies that
deforestation rates in Indonesia are very high. Based on satellite imagery,
the Ministry of Forestry reported that deforestation inside the state forest
area in Indonesia during 2009/2010 was 610,375.9 ha. Deforestation in
Indonesia is mainly caused by fire, conversion of forest and peat-land to
other land uses, mining, and forest degradation due to illegal logging and
poor forest management practices (Ministry of Forestry 2012). Deforestation
in the islands of Kalimantan and Sumatra was remarkably high compared
with other islands in Indonesia (Holmes 2002). In the period between 1985
and 2007, forest loss in Sumatra alone was reported to be more than 70%
of total forest area (Laumonier et al. 2010).
Centre for Conservation and Rehabilitation, Jalan Gunung Batu 5, Bogor 16151, Indonesia.
Email: hl.tata@gmail.com
1
Forest area is designated by the institutional framework of forestry and includes areas
normally forming part of the forest area which are temporarily cleared as a result of human
intervention, but which are expected to revert to forest.
Ectomycorrhiza in Forest Rehabilitation in Indonesia 201
2
http://www.dephut.go.id/INFORMASI/RRL/Benih/GN_RHL.html
202 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
from different soils were varied (Table 1). This observation implies that
mycorrhizal inoculum potential persists in the soils after wildfire. EcM
morphotypes from unburned forests were different compared to those from
burned forests (Tata et al. 2003a).
Table 2. Ectomycorrhizal colonization of S. selanica and S. lamellata inoculated (+) and non-
inoculated (–) with Scleroderma columnare in nurseries planted on different land use types.
6. Rehabilitation Techniques
Technically, rehabilitation of degraded dipterocarp forests can be achieved
through (i) enrichment planting with desirable dipterocarp seedlings or clonal
plant materials in order to increase timber productivity, and (ii) planting
fast-growing light demanding species as a nurse canopy and followed by
under-planting with dipterocarps (Kettle 2010). The former is established
under small opening of forest canopy covers and in the presence of gaps.
The latter is established under severely degraded forest conditions.
Many studies have shown techniques to rehabilitate degraded
dipterocarp forests caused by logging and over-exploitation, fires, and the
invasion of Imperata grassland (Adjers et al. 1996, Otsamo et al. 1996, Otsamo
2000, Mori 2001). Forest rehabilitation with enrichment planting can be
done in gaps resulted from logging activities. To rehabilitate burned forests
and Imperata grasslands, strip planting techniques can be adopted. Mixed
planting between fast growing trees (such as Acacia mangium) and native
tree species is the best option to rehabilitate Imperata grassland in South
Kalimantan (Otsamo 2000). However, no EcM inoculation was mentioned
in these publications.
Rehabilitation of degraded land in Haurbentes, West Java, with
various Dipterocarpaceae trees, showed a successful human constructed
208 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
7. Conclusions
The success of rehabilitation is determined by silvicultural and ecological
factors. Ectomycorhizas play an important role in the production of planting
stocks for forest rehabilitation. In the nursery stage, ectomycorrhizal fungal
colonization increases early growth of dipterocarp species, which benefits
planting stock production. However, after field planting, very few studies
have reported success of EcM fungal inoculations on tree growth in field
conditions. Inoculation of EcM fungi in the nursery stage requires skills and
capital in the initial nursery development. Using topsoil collected under
conspecific parent trees for planting stock productions is more applicable
for EcM fungal inoculation.
Acknowledgements
The author acknowledges Meine van Noordwijk for comments on the early
draft of manuscript, anonymous reviewers for comments to improve the
manuscript and Krista L. McGuire and Caitlyn Gillikin for improving the
language.
References
Adjers, G. and A. Otsamo. 1996. Seedling production method of dipterocarps. In: A. Schulte
and D. Schone (eds.). Dipterocarp Forest Ecosystems: Towards Sustainable Management.
World Scientific, Singapore, pp. 391–410.
Anonymous. 2004. Development of tropical reforestation techniques. Report of joint study.
Kansai Electric Power Co., Gadjah Mada University and Kanso Technos Co, Ltd p. 43.
Ashton, P.S. 1982. Dipterocarpaceae. Flora Malesiana: Vol. 9(2). MartinusNijhoffPublishers.c/o.
Kluwer Academic Publishers Groups. The Netherlands.
Ectomycorrhiza in Forest Rehabilitation in Indonesia 211
Beukema, H., F. Danielsen, G. Vincent, S. Hardiwinoto and J. van Andel. 2007. Plant and bird
diversity in rubber agroforests in the lowlands of Sumatra, Indonesia. Agrofor Syst. 70:
217–242.
Brearley, F.Q. 2012. Ectomycorrhizal associations of the Dipterocarpaceae. Biotropica 44(5):
637–648.
Ciccarese, L., A. Mattsson and D. Pettenella. 2012. Ecosystem services from forest restoration:
thinking ahead. New For. 43: 543–560.
Gouyon, A., H. de Foresta and P. Levang. 1993. Does ‘Jungle Rubber’ deserve its name? Analysis
of rubber agroforestry system in Southeast Asia. Agro for. Syst. 22: 181–20.
Hadi, S. 1987. The association of fungi in dipterocarps. In: A.J.G.H. Kostermans (ed.).
Proceedings of the Third Round Table Conference on Dipterocarps. UNESCO. Jakarta,
Indonesia, pp. 73–79.
Holmes, D.A. 2002. Where Have All the Forests Gone? Environment and Social Development,
East Asia and Pacific Region. The World Bank, Washington DC. USA.
Ingleby, K., R.C. Munro, P.A. Mason and M.J. Clearwater. 1998. Ectomycorrhizal populations
and growth of Shorea parvifolia (Dipterocarpaceae) seedlings regenerating under three
different forest canopies following logging. For. Ecol. Manag. 111: 171–179.
Joshi, L., G. Wibawa, G. Vincent, D.B. Putin, R. Akiefnawati, G. Manurung, M. van Noordwijk
and S. Williams. 2002. Jungle rubber: a traditional agroforestry system under pressure.
International Center for Research in Agroforestry (ICRAF). Bogor, Indonesia.
Ketterings, Q.M., J.M. Bigham and V. Laperche. 2000. Changes in soil mineralogy and
texture caused by slash-and-burn in Sumatra, Indonesia. Soil Science Soc. Am. J. 64:
1108–1117.
Kettle, C.J. 2010. Ecological considerations for using dipterocarps for restoration of lowland
rainforest in Southeast Asia. Biodiv. Conserv. 19: 1137–1151.
Kikuchi, J. 2006. Ectomycorrhizas of dipterocarps in logged-over forests and plantations. In: K.
Suzuki, K. Ishii, S. Sakurai and S. Sasaki (eds.). Plantation Technology in Tropical Forest
Science. Springer, Germany, pp. 207–210.
Lamb, D. and M. Tomlinson. 1994. Forest rehabilitation in the Asia-Pacific Region: Past lessons
and present uncertainties. J. Trop. For. Sci. 7(1): 157–170.
Laumonier, Y., Y. Uryu, M. Stüwe, A. Budiman, B. Setiabudi and O. Hadian. 2010. Eco-floristic
sectors and deforestation threatsin Sumatra: identifying new conservation area network
priorities for ecosystem-based land use planning. Biodiv. and Conserv. 19: 1153–1174.
Lee, S.S. 1998. Root symbiosis and nutrition. In: S. Appanah and J.M. Turnbull (eds.). A Review
of Dipterocarps: Taxonomy, Ecology and Silviculture. Center for International Forestry
Research. Bogor, Indonesia, pp. 99–114.
Marx, D.H., A.B. Hatch and J.F. Mendicino. 1977. High soil fertility decreases sucrose content
and susceptibility of loblolly pine roots to ectomycorrhizal infection by Phisolithus
tinctorius. Can. J. Bot. 55(12): 1569–1574.
Masano, H. Alrasjid and Z. Hamzah. 1987. Planting trials of dipterocarp species outside their
natural distribution range in the Haurbentes experimental forest, West Java. In: A.J.G.H.
Kostermans (ed.). Proceedings of the Third Round Table Conference on Dipterocarps.
UNESCO. Jakarta, Indonesia, pp. 19–37.
Matsune, R. Soda, T. Tange, S. Sasaki and Suparno. 2006. Planting techniques and growth of
dipterocarps in abandoned secondary forest in East Kalimantan. In: K. Suzuki, K. Ishi, S.
Sakurai and S. Sasaki (eds.). Plantation Technology in Tropical Forest Science. Springer,
Tokyo, pp. 221–229.
Michon, G.M. and J.M. Bompard. 1987. The dammar gardens (Shorea javanica) in Sumatra.
In: A.J.G.H. Kostermans (ed.). Proceedings of the Third Round Table Conference on
Dipterocarps. UNESCO. Jakarta, Indonesia, pp. 3–17.
Mindawati, N., Hendromono, M. Hiratsuka, T. Toma, Y. Morikawa and A.N. Gintings. 2004a.
Tree trunk volume of Shorea species case study in Darmaga and Haurbentes Research
Forest in West Java, Indonesia. J. Forest. Res. 1(1): 17–24.
212 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Mindawati, N., I. Heriansyah, M. Hiratsuka, T. Toma, A.N. Gintings and Y. Morikawa. 2004b.
Tree growth of dipterocarp plantation forest in Java, Indonesia. Info Hutan 1(2): 53–83.
Ministry of Forestry. 2012. Forestry Statistics of Indonesia 2011. Ministry of Forestry.
Jakarta.
Mori, T. 2001. Rehabilitation of degraded forests in lowland Kutai, East Kalimantan, Indonesia .
In: S. Kobayashi, J.W. Turnbull, T. Toma, T. Mori and N.M.N.A. Majid (eds.). Rehabilitation
of Degraded Tropical Forest Ecosystems: Workshop Proceedings, 2–4 November 1999,
Bogor, Indonesia. CIFOR, Bogor, Indonesia, Bogor, Indonesia, pp. 17–36.
Nawir, A.A., Murniati and L. Rumboko. 2007. Forest Rehabilitation in Indonesia: Where to
After More Than Three Decades? Center for International Forestry Research (CIFOR),
Bogor, Indonesia, p. 269.
Nuhamara, S.T. 1987. Mycorrhiza in agroforetsry: a case study. Biotropia 1(1): 53–57.
Ohta, S. and E. Syarif. 1996. Soils under lowland dipterocarp forests—characteristics and
classification. In: A. Schulte and D. Schone (eds.). Dipterocarp Forest Ecosystems: Towards
Sustainble Management. World Scientific, Singapore, pp. 29–51.
Omon, R.M. 2002. Dipterocarpaceae: Shorea leprosula Miq. cuttings, mycorrhizae and nutrients.
Ph.D. Thesis. Wageningen University, the Netherlands. Tropenbos Kalimantan Series 7.
Balikpapan, Indonesia.
Okimori, Y, J. Kikuchi and S. Hardiwinoto. 2006. Restoring the logged-over dipterocarps in
tropical rainforests of Central Sumatra. In: K. Suzuki, K. Ishi, S. Sakurai and S. Sasaki
(eds.). Plantation Technology in Tropical Forest Science. Springer, Tokyo, pp. 231–238.
Otsamo, A. 2000. Early development of three planted indigenous tree species and natural
understoreyvegetation in artificial gaps in an Acacia mangium stand on an Imperata
cylindrica grassland site in South Kalimantan, Indonesia. New For. 19: 51–68.
Otsamo, R., L. Kurniati and A. Otsamo. 1996. Dipterocarp specieson Imperata cylindrica
dominated grasslands: a case study from South Kalimantan, Indonesia. In: I. Suhardi
(ed.). Proceeding of Seminar on Ecology and Reforestation of Dipterocarp Forest in
Gadjah Mada University, Yogyakarta, Indonesia, 24-25 January 1996. Faculty of Forestry,
Gadjah Mada University and Kansai Environmental Engineering Centre, Indonesia, pp.
147–157.
Penot, E. 2004. From shifting agriculture to sustainable rubber complex agroforestry systems
(jungle rubber) in Indonesia: a history of innovations production and adoption process.
In: D. Babin (ed.). Beyond Tropical Deforestation. UNESCO/CIRAD, Montpellier, pp.
221–250.
Priadjati, A. 2002. Dipterocarpaceae: Forest fire and forest recovery. Ph.D. Thesis Wageningen
University. Tropenbos-Kalimantan Series 8. Tropenbos International, Wageningen, the
Netherlands.
Read, D.J. 1991. Mycorrhizas in ecosystems. Experientia 47: 376–391.
Riniarti, M. 2010. Dinamika kolonisasi tiga fungi ektomikoriza Scleroderma spp. dan
hubungannya dengan pertumbuhan tanaman inang. Dissertation. Institut Pertanian
Bogor, Bogor, Indonesia.
Roshetko, J.M., D.J. Snelder, R.D. Lasco and M. Van Noordwijk. 2008. Future challenge: A
paradigm shift in the forestry sector. In: D.J. Snelder and R.D. Lasco (eds.). Smallholder
Tree Growing for Rural Development and Environmental Services: Lessons from Asia.
Advances in Agroforestry Volume 5. Springer, Berlin, pp. 451–483.
Santoso, E. 1988. Pengaruh mikoriza terhadap diameter batang dan bobot kering anakan
Dipterocarpaceae. Bul Pen Hutan 504: 11–21.
Santoso, E. and M. Turjaman. 2003. Tipe-tipe struktur ektomikoriza pada Shorea selanica,
S. stenoptera, S. pinanga, dan S. palembanica (Dipetocarpaceae) di Hutan Penelitian
Haurbentes Jawa Barat. Bul Pen Hutan. 636: 33–37.
Shiva, M.P. and I. Jantan. 1998. Non-timber forest products from dipterocarps. In: S. Appanah
and J.M. Turnbull (eds.). A Review of Dipterocarps: Taxonomy, Ecology and Silviculture.
Center for International Forestry Research. Bogor, Indonesia, pp. 187–196.
Ectomycorrhiza in Forest Rehabilitation in Indonesia 213
Smits, W.T.M. 1992. Mycorrhizal studies in dipterocarp forests in Indonesia. In: D.J. Read,
D.H. Lewis, A.H. Fitter and I.J. Alexander (eds.). Mycorrhizas in Ecosystems. CAB
International. UK, pp. 283–392.
Smits, W. 1994. Dipterocarpaceae: Mycorrhizae and regeneration. Tropenbos Series 9. The
Tropenbos Foundation, Wageningen.
Soekotjo. 2005. Sistem Silvikultur Intensif. Gadjah Mada University, Yogyakarta.
Sofiyuddin, M., Janudianto and A. Perdana. 2012. Potensi pengembangan dan pemasaran
jelutung di Tanjung Jabung Barat. Brief 23. World Agroforestry Centre—ICRAF, SEA
Regional Office. Bogor, Indonesia.
Subiakto, A., C. Sakai, S. Purnomo and Taufiqurahman. 2005. Cutting propagation as an
alternative technique for mass production of dipterocarp planting stocks in Indonesia.
Presentation paper at 8th Round-table Conference on Dipterocarps: Dipterocarp
enhancing capacities in sustainable development and poverty alleviation. 15-17 November
2005. Ho Chi Minh City, Vietnam. [online] URL: http://www.apafri.org/8thdip/
Session%202/S2_Atok.doc.
Subiakto, A., Hendromono and Sunaryo. 2001. Ex situ conservation of dipterocarp species
in West Java and Banten. In: Thielges, B.A., S.D. Sastrapradja, A. Rimbawanto (eds.). In
situ and ex situ Conservation of Commercial Tropical Trees. Faculty of Forestry, Gadjah
Mada University and International Tropical Timber Organization. Yogyakarta, Indonesia,
pp. 183–191.
Sutisna, M. 2001. Taungya experiment for rehabilitation of burnt-over forest in East Kalimantan,
Indonesia. In: S. Kobayashi, J.W. Turnbull, T. Toma, T. Mori and N.M.N.A Majid (eds.).
Rehabilitation of Degraded Tropical Forest Ecosystems. Workshop Proceedings, 2-4
November 1999, Bogor, Indonesia. Center for International Forestry Research, Bogor,
Indonesia, pp. 115–122.
Tata, M.H.L. 2008. Mycorrhizae on dipterocarp in rubber agroforests (RAF) in Sumatra.
Dissertation. Utrecht University, the Netherlands. Wohrmann Print Service, Zupthen,
the Netherlands.
Tata, M.H.L., S. Hadi, C. Kusmana and Achmad. 2003a. Effect of forest fire on the survival
of ectomycorrhizal fungi on dipterocarps. In: H. Aminah, S. Ani, H.C. Sim and B.
Krishnapillay (eds.). Proceedings of the Seventh Round-Table Conference on Dipterocarps.
7–10 October 2002. Asia Pacific Association of Forestry Research Institutions (APAFRI).
Kuala Lumpur, Malaysia, pp. 173–178.
Tata, M.H.L., S. Hadi, C. Kusmana and Achmad. 2003b. Putative ectomycorrhizal fungi at
Sungai Wain Protection Forest, East Kalimantan. Proceedings the National Workshops
on Conservation and Sustainable Management of Belowground Biodiversity. May 30-31,
2003. Bogor, Indonesia.
Tata, H.L., M. Van Noordwijk and M. Werger. 2008. Trees and regeneration in rubber
agroforests and other forest-derived vegetation in Jambi (Sumatra, Indonesia). J. For.
Res. 5(1): 1–20.
Tata, H.L., M. Van Noordwijk, M.J.A. Werger and R.C. Summerbell. 2010. Limited response
to nursery-stage ectomycorrhiza inoculation of Shorea seedlings planted in rubber
agorofrests in Jambi, Indonesia. New For. 39(1): 51–74.
Tata, H.L. and M. Van Noordwijk. 2011. Farmer participation on dipterocarp tree planting
in small holder rubber plantation. In: E.B. Hardiyanto, S. Solberg and M. Osaki (eds.).
Proceedings of International Conference on New Perspectives of Tropical Forest
Rehabilitation for better Forest Functions and Management.Faculty of Forestry, Gadjah
Mada University, Yogyakarta, Indonesia, pp. 38–42.
Torquebiau, E.F. 1984. Man-made dipterocarp forest in Sumatra. Agrofor. Syst. 2: 103–127.
Turjaman, M., Y. Tamai, H. Segah, S.H. Limin, J.Y. Cha, M. Osaki and K. Tawaraya. 2005.
Inoculation with the ectomycorrhizal fungi Pisolithus arhizus and Scleroderma sp. improves
early growth of Shorea pinanga nursery seedlings. New For. 30: 67–73.
214 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Turjaman, M., Y. Tamai, H. Segah, S.H. Limin, M. Osaki and K. Tawaraya. 2006. Increase in
early growth and nutrient uptake of two Shorea seminis seedlings inoculated with two
ectomychorrizal fungi. J. Trop. For. Sci. 18: 243–249.
Turjaman, M., E. Santoso, A. Susanto, S. Gaman, S.H. Limin, Y. Tamai, M. Osaki and K.
Tawaraya. 2011. Ectomycorrhizal fungi promote growth of Shorea balangeran in degraded
peat swamp forests. Wetland Ecol. Manag. 19: 331–339.
Van Noordwijk, M., D. Murdiyarso, K. Hairiah, U.R. Wasrin, A. Rachman and T.P. Tomich.
1998. Forest soils under alternatives to slash-and-burn agriculture in Sumatra, Indonesia.
In: A. Schulte and D. Ruhiyat (eds.). Soils of Tropical Forest Ecosystems: Characteristics,
Ecology and Management. Springer-Verlag, Berlin, pp. 175–185.
Whitmore, T.C. 1984. Tropical rain forests of the Far East. 2nd edition. Clarendon Press,
Oxford, pp. 352.
Yasman, I. 1995. Dipterocarpaceae: Tree-mycorrhizae-seedling connections. Ph.D. thesis,
Wageningen Agriculture University, the Netherlands.
CHAPTER
12
The Controlled
Ectomycorrhization Practices
in Tropical Areas: Fungal
Inoculum Biotechnology,
Field Results and Research
Perspectives
Robin Duponnois,1,* Hervé Sanguin,1 Amadou Mustapha
Bâ,1,2 Antoine Galiana,1 Marc Ducousso,1 Ezékiel
Baudoin,1 Michel Lebrun1 and Yves Prin1
1. Introduction
Overexploitation of forest resources resulting from excessive industrial
exploitation, clearing for industrial purposes, and collection of firewood has
led to a dramatic deforestation during recent decades in the Mediterranean
and tropical areas (Piéri 1991). One of the detrimental effects of deforestation
is soil degradation and desertification processes resulting in alterations of
1
UMR 113 CIRAD/INRA/IRD/SUP-AGRO/UM2, Laboratoire des Symbioses Tropicales
et Méditerranéennes (LSTM). TA A-82/J, Campus International de Baillarguet. Montpellier
Cedex 5, France.
2
Laboratoire Commun de Microbiologie (LCM) IRD/UCAD/ISRA, Centre de Recherche de
Bel Air, BP. 1386, CP. 18524 Dakar, Sénégal.
*Corresponding author: Robin.Duponnois@ird.fr
216 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Ref. (1) Fungal strain Plant species Inoculum Effects on plant growth parameters (%) and ectomycorrhizal
formulation colonization
Height Shoot Root Ecto. colonization (%)
biomass biomass
A Pisolithus arhizus Shorea pinanga Ectomycorrhizal spore + 46.1 (2) + 66.7 nd(3) 87
pellets
A Scleroderma sp. S. pinanga Ectomycorrhizal spore + 41.3 + 60.5 nd 86
pellets
B P. tinctorius Acacia mangium Ectomycorrhizal spore nd + 29.1 + 40.7 52
tablets
C Rhizopogon roseolus Pinus halepensis Spore suspension –30.1 nd nd 48
C Suillus collinitus Pinus halepensis Spore suspension + 27.8 nd nd 75
D Scleroderma albidum Eucalyptus globulus Spore suspension + 32.1 + 19.7 + 42.5 nd
D S. areolatum E. globulus Spore suspension + 17.5 + 9.1 +33.5 nd
D S. cepa E. globulus Spore suspension + 30.8 + 8.8 + 2.0 nd
D S. albidum E. urophylla Spore suspension – 3.6 + 3.1 –0.7 nd
D S. areolatum E. urophylla Spore suspension + 7.2 + 4.9 + 4.0 nd
D S. cepa E. urophylla Spore suspension + 6.3 + 13.0 + 1.4 nd
E P. tinctorius P. halepensis Spore suspension + 37.1 + 44.9 + 41.7 55.4
E R. roseolus P. halepensis Spore suspension + 38.3 + 44.6 + 28.6 39.5
E Suillus collinitus P. halepensis Spore suspension + 35.1 + 28.2 + 30.6 28.9
A: Turjaman et al. 2005. B: Aggangan et al. 2010. C: Rincon et al. 2007. D: Chen et al. 2006. E: Torres and Honrubia 1994
(1)
Ref.: Reference. (2) (mean value of ectomycorrhizal plants—mean value of the non ectomycorrhizal plants) x 100)/(mean value of the ectomycorrhizal
plants). (3)nd: not determined.
Controlled Ectomycorrhization Practices 219
220 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 2. Composition of nutrient media commonly used for the isolation and culture of
ectomycorrhizal fungi. Data from Brundrett et al. 1996.
Vitamins (Og.l–1)
Thiamine HCl 0.1 0.1
Agar (g.l–1) 20 20 20
pH
Adjusted pH to 5.8 5.4 5.0
(1)
MMN: Modified Menin Norkrans medium (Marx 1969). (2) Pachlewski medium (Pachlewski
and Pachlewski 1974). (3) Ferry and Das (1968). (4) Ammonium tartrate
Waring blender for about 10s, and suspended in sterile distilled water.
This type of fungal inoculum is quantified by measuring the fungal dry
weight per ml or by counting living propagules (determination of colony
forming units by spreading 1 ml of suspension on a 6-cm Petri dish with
nutrient agar). The hyphal fragment suspension is then mixed (1:1, v:v) with
distilled water containing 20 g.l–1 sodium alginate and 50 g.l–1 autoclaved
dry powdered sphagnum peat. The final solution is pumped through a
pipe with 2-mm holes. The drops fall into a 100 g.l–1 CaCl2 solution and
form beads of reticulated calcium alginate gel (Mauperin et al. 1987). The
beads are kept in CaCl2 for 24h at room temperature to ensure complete
reticulation. Then they are washed with tap water to eliminate NaCl and
CaCl2 and stored in air-tight containers at 4°C to prevent drying. This type
of inoculum can be kept up to 9 months in these conditions. The beads
are prepared with 1–2 g mycelium (dry weight) per liter of final solution
(Mortier et al. 1988).
Table 3. Growth of some Australian Acacia species inoculated with different ectomycorrhizal
fungal strains (peat-vermiculite formulation) after 4 months culturing in glasshouse
conditions.
Fig. 1. Field performance of A. holosericea trees (expressed in tons of wood biomass per ha with
1200 planted trees per ha) inoculated or not with Pisolithus albus strain IR100 in field trials
conducted in Senegal after 18 months of plantation. Data from Duponnois et al. 2007.
Glomus clarum Azotobacter diazotrophicus, Klebsiella Ipomoea batatas Paula et al. 1992
sp.
Glomus deserticola Klebsiella pneumoniae, Alcaligenes Unicola paniculata Will and Sylvia 1990
denetrificans
Glomus fasciculatum Azotobacter chroococcum Lycopersicum esculentum Bagyaraj and Menge 1978
Glomus fasciculatum, Glomusmosseae Rhizobium meliloti Medicago sativa Azcón-Aguilar et al. 2003
Glomus fasciculatum, Glomusmosseae, Bacillus coagulans Morus alba, Carica papaya Mamatha et al. 2002
Glomus caledonium
Glomus fistulosum Pseudomonas putida Zea mays, Solunum tuberosum Vosatka and Gryndler (1999)
Glomus intraradices Bacillus subtilis, Enterobacter sp. Allium cepa Toro et al. 1997
Glomus intraradices Pseudomonas monteilii Acacia holosericea Duponnois and Plenchette 2003
Glomus intraradices Rhizobium Anthyllis cytisoides Requena et al. 1997
Glomus intraradices Agrobacterium rhizogenes, Hordeum vulgare, Triticum aestivum Fester et al. 1999
Pseudomonas fluorescens, Rhizobium
leguminosarum
Glomus intraradices Streptomyces coelicolor Sorghum Abdel-Fattah and Mohamedin
2000
Glomus mosseae Paenibacillus sp. Sorghum bicolor Budi et al. 1999
Glomus mosseae Pseudomonas sp. Lycopersicum esculentum Barea et al. 1998
Glomus mosseae Bradhyrhizobium japonicum Glycine max Xie et al. 1995
Glomus mosseae Pseudomonas fluorescens Lycopersicum esculentum Gamalero et al. 2004
Glomus mosseae Brevibacillus sp. Trifolium pratense Vivas et al. 2003
Glomus mosseae, Glomus intraradices Paenibacillus brasilensis Trifolium Artursson 2005
Complex of AMF (1) Pseudomonas putida Trifolium Meyer and Linderman (1986)
Complex of indigenous AMF Pseudomonas sp. Triticum aestivum Babana and Antoun 2005
Complex of AMF Bacillus mycoides Herbaceous plant species von Alten et al. 1993
(1)
AMF: Arbuscular Mycorrhizal Fungi.
Controlled Ectomycorrhization Practices 227
228 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Ectomycorrhizal isolates
The MHB P. monteilii strain HR13 has also significantly increased the
ectomycorrhizal colonization for all the Australian Acacia species tested
in this experiment (A. auriculiformis, A. eriopoda, A. holosericea, A. mangium
and A. platycarpa) (Fig. 3).
MHB have also been tested for their influence on ectomycorrhizal
formations between Douglas and L. bicolor strain S238 in nursery conditions.
One bacterial isolate (Pseudomonas fluorescens isolate BBc6) increased
mycorrhiza formation from around 60% (per cent of short roots mycorrhized
with L. bicolor S238N without bacterial inoculation) to 87%. This experiment
was performed with a peat-vermiculite fungal inoculum. Similar positive
effects were recorded when this bacterial strain was mixed together with the
fungus in alginate beads (Duponnois 1992). After 4 months of plantation,
BBc6 stimulated ectomycorrhizal infection from 42% to 75%. Frey-Klett et
al. (1999) showed that five months after inoculation and sowing, bacterial
inoculation significantly improved the percentage of ectomycorrhizal short
roots on plants inoculated with two doses of fungal inoculum of L. bicolor
(50 and 100 mg m–2 dry weight mycelium entrapped in alginate beads
mixed into the soil at a constant dose of 1 litre m–2) in a nursery (Fig. 4). The
Controlled Ectomycorrhization Practices 229
Acacia species
Fig. 3. Effect of Pseudomonas monteilii isolate HR13 on ectomycorrhiza formation with Pisolithus
albus isolate IR100 of five Australian Acacia species after 4 months of culturing in glasshouse
conditions. For the legend, see Fig. 2.
Fig. 4. Dose effect of P. fluorescens isolate BBcR8 on the percentage of Douglas-fir short roots
mycorrhizal with L. bicolor S238, 23 weeks after fungal and bacterial inoculations. : Fungal
inoculation dose: 50 mg dry weight mycelium m–2. : Fungal inoculation dose: 100 mg dry
weight mycelium m–2. For each fungal inoculation dose, columns indexed by the same letters
are not significantly different according to a Scheffe test. Data from Frey-Klett et al. (1999).
230 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 7. Effect of the Pseudomonas monteilii strain (isolate KR9) and/or Scleroderma disctyosporum
IR 412 on mycorrhiza formation, rhizobial development, and growth of Acacia holosericea after
4 months of culturing under glasshouse conditions.
Treatments
Control P. monteilii S. dictyosporum KR9 +
KR9 IR412 IR412
Shoot biomass (mg dry weight) 532 a (1) 553 a 1236 b 1786 c
Root biomass (mg dry weight) 184 a 198 a 536 b 868 c
Number of nodules per plant 4.2 a 4.6 a 8.3 b 12.4 c
Total nodule weight per plant (mg) 6.8 a 7.1 a 15.9 b 25.3 c
Ectomycorrhizal colonization (%) 0 0 28.3 a 48.5 c
(1)
Data in the same line followed by the same letter are not significantly different according
to the Student’s “t” test (p < 0.05).
232 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
FIR x TAR NS S S
S: significant (p < 0.05), NS: not significant (p < 0.05). (1) Standard error of the mean. (2) Data in
the same column and for each factor followed by the same letter are not significantly different
according to the Newman-Keuls test (p < 0.05). (A) Values are means of 20 replicates for fungal
inoculum and termite mound amendment rates. Fungal inoculum rate factor is for all termite
mound amendment rate treatments combined; the termite amendment factor is for all fungal
inoculum rate treatments combined.
growth) and (ii) the addition via the termite mound of a bacterial group
(i.e., fluorescent pseudomonads) that could act as Mycorrhization Helper
Bacteria (MHB) (Duponnois and Plenchette 2003). The data recorded from
this study were similar to those obtained from the cultural practice using
the conventional process of controlled mycorrhization (Table 10).
One of the main problems encountered with the conventionally
controlled mycorrhization of forest planting stocks is the large quantity of
fungal inoculum required for the production of high quality mycorrhizal
plants in nursery conditions. In tropical and Mediterranean areas, the
mycorrhizal inoculum dose added per plant to the cultural substrate is
234 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 9. Effect of ectomycorrhizal inoculation and termite mound powder amendment rates
on A. holosericea growth and on mycorrhiza formation after 2 months of culturing in pots of
1 liter.
the same column and for each factor followed by the same letter are not significantly different
according to the Newman-Keuls test (p < 0.05). (A) Values are means of 20 replicates for fungal
inoculum and termite mound amendment rates. Fungal inoculum rate factor is for all termite
mound amendment rate treatments combined; the termite amendment factor is for all fungal
inoculum rate treatments combined.
AM inoculation
Uninoculated control 648.1 a (1) 312.2 a 0
Glomus intraradices 1834.1 b 546.3 b 59.3
Ectomycorrhizal inoculation
Uninoculated control 550 a 290 a 0
Pisolithus albus IR100 1120.1 b 560.2 b 35.6
(1)
For each type of fungal inoculation, data in the same column followed by the same letter
are not significantly different according to the Newman Keuls test (p< 0.05).
6. Conclusion
Numerous studies have reported the benefits that result from using fungal
inoculants to enhance the development of tree seedlings in glasshouse and
nursery conditions. In the same way, numerous technical procedures have
been described to produce cost-effective mycorrhizal inocula. The results
presented in this chapter suggest that inoculation with ectomycorrhizal
fungi can improve the early growth of the most planted tree species in
tropical and Mediterranean forests and that this technique will accelerate the
rehabilitation of degraded forests. Unfortunately, mycorrhizal inoculation
remains underexploited in nursery practices, although the management of
tree mycorrhizal status could have very important implications for tropical
reforestation programs in developing countries. Controlled mycorrhization
could be highly improved if the fungal inoculation is associated with
Mycorrhization Helper Bacteria or termite mound powder of M. subhyalinus.
These innovative procedures decrease the necessary amount of fungal
inoculum while keeping the same mycorrhizal establishment that is usually
measured with the conventional process of controlled mycorrhization.
Hence the use of these cultural practices could be facilitated and encouraged
in forest nurseries. Future research should prioritize the generalizability of
these controlled experiments to long-term applications in tree plantations
and reforestation projects.
236 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Acknowledgements
We thank the anonymous referees for their valuable comments on this
study, and Krista L. McGuire and Caitlyn Gillikin for improving the
language.
References
Abdel-Fattah, G.M. and A.H. Mohamedin. 2000. Interactions between a vesicular-arbuscular
mycorrhizal fungus (Glomus intraradices) and Streptomyces coelicolor and their effects on
sorghum plants grown in soil amended with chitin of brawn scales. Biol. Fert. Soils 32:
401–409.
Aggangan, N.S., H.K. Moon and S.H. Han. 2010. Growth response of Acacia mangium Willd.
seedlings to arbuscular mycorrhizal fungi and four isolates of the ectomycorrhizal fungal
Pisolithus tinctorius (Pers.) Coker and Couch. New For. 39: 215–230.
Allen, E.B., M.F. Allen, D.J. Helm, J.M. Trappe, R. Molina and E. Rincon. 1995. Patterns and
regulation of mycorrhizal plant and fungal diversity. Plant and Soil 170: 47–62.
Artursson, V. 2005. Bacterial–fungal interactions highlighted using microbiomics: potential
application for plant growth enhancement. PhD Thesis, University of Uppsala, Uppsala,
Sweden.
Azcón, R., R. Rubio and J.M. Barea. 1991. Selective interactions between different species of
mycorrhizal fungi and Rhizobium meliloti strains, and their effects on growth, N2-fixation
(15N) and nutrition of Medicago sativa L. New Phytol. 117: 399–404.
Azcon-Aguilar, C., J. Palenzuela, A. Roldan, S. Bautista, R. Vallejo and J.M. Barea. 2003. Analysis
of the mycorrhizal potential in the rhizosphere of representative plant species from
desertification-threatened Mediterranean shrublands. App. Soil Ecol. 14: 165–175.
Babana, A.H. and H. Antoun. 2005. Biological system for improving the availability of Tilemsi
phosphate rock for wheat (Triticum aestivum L.) cultivated in Mali. Nutr. Cycling Agroeco.
72: 147–157.
Bagyaraj, D.J. and J.A. Menge. 1978. Interaction between a VA mycorrhiza and Azotobacter and
their effects on the rhizosphere microflora and plant growth. New Phytol. 80: 567–573.
Barea, J.M., G. Andrade, V. Bianciotto, D. Dowling, S. Lohrke, P. Bonfante, F. O’Gara and C.
Azcón-Aguilar. 1998. Impact on arbuscular mycorrhiza formation of Pseudomonas strains
used as inoculants for biocontrol of soil-borne fungal plant pathogens. Appl. Environ.
Microbiol. 64: 2304–2307.
Bending, G.D., E.J. Poole, J.M. Whipps and D.J. Read. 2002. Characterisation of bacteria from
Pinus sylvestris-Suillus luteus mycorrhizas and their effects on root-fungus interactions
and plant growth. FEMS Microbiol. Ecol. 39: 219–227.
Brundrett, M., N. Bougher, B. Dell, T. Grove and N. Malajczuk. 1996. Working with mycorrhizas
in Forestry and Agriculture. ACIAR Monograph pp. 373.
Brundrett, M.C. 2002. Coevolution of roots and mycorrhizas of land plants. New Phytol. 154:
275–304.
Budi, S.W., D. van Tuinen, G. Martinotti and S. Gianinazzi. 1999. Isolation from the Sorghum
bicolor mycorrhizosphere of a bacterium compatible with arbuscular mycorrhiza
development and antagonistic towards soilborne fungal pathogens. Appl. Environ.
Microbiol. 65: 5148–5150.
Castellano, M.A. and R. Molina. 1989. Mycorrhizae. In: The Biological Component: Nursery
Pest and Mycorrhizae Manual, Vol. 5. T.D. Landis (ed.). Agric. Handbook 674. USDA
Forest Service, Washington, DC, pp. 101–167.
Castellano, M.A. 1994. Current status of outplanting studies using ectomycorrhiza-inoculated
forest trees. In: F. Pfleger and B. Linderman (eds.). A Reappraisal of Mycorrhizae in Plant
Health. The American Phytopathological Society, St Paul, pp. 261–281.
Controlled Ectomycorrhization Practices 237
Duponnois, R., C. Plenchette, Y. Prin, M. Ducousso, M. Kisa, A.M. Bâ and A. Galiana. 2007.
Use of mycorrhizal inoculation to improve reafforestation process with Australian Acacia
in Sahelian ecozones. Ecol. Engineering 29: 105–112.
Ferry, B.W. and M. Das. 1968. Carbon nutrition of some mycorrhizal Boletus species. Trans.
Brit. Mycol. Soc. 51: 795–798.
Fester, T., W. Maier and D. Strack. 1999. Accumulation of secondary compounds in barley
and wheat roots in response to inoculation with an arbuscular mycorrhizal fungus and
co-inoculation with rhizosphere bacteria. Mycorrhiza 8: 241–246.
Founoune, H., R. Duponnois, J.M. Meyer, J. Thioulouse, D. Masse, J.L. Chotte and M. Neyra.
2002a. Interactions between ectomycorrhizal symbiosis and fluorescent pseudomonads
on Acacia holosericea: isolation of mycorrhiza helper bacteria (MHB) from a soudano-
Sahelian soil. FEMS Microbiol. Ecol. 41: 37–46.
Founoune, H., R. Duponnois, A.M. Bâ, S. Sall, I. Branget, J. Lorquin, M. Neyra and J.L. Chotte.
2002b. Mycorrhiza helper bacteria stimulated ectomycorrhizal symbiosis of Acacia
holosericea with Pisolithus alba. New Phytol. 153: 81–89.
Frey-Klett, P., J.-L. Churin, J.-C. Pierrat and J. Garbaye. 1999. Dose effect in the dual inoculation
of an ectomycorrhizal fungus and a mycorrhiza helper bacterium in two forest nurseries.
Soil Biol. Biochem. 31: 1555–1562.
Frey-Klett, P., J. Garbaye and M. Tarkka. 2007. The mycorrhiza helper bacteria revisited. New
Phytol. 176: 22–36.
Gamalero, E., A. Trotta, N. Massa, A. Copetta, M.G. Martinotti and G. Berta. 2004. Impact of
two fluorescent pseudomonads and an arbuscular mycorrhizal fungus on tomato plant
growth, root architecture and P acquisition. Mycorrhiza 14: 185–192.
Garbaye, J. and J.-L. Churin. 1997. Growth stimulation of young oak plantations inoculated
with the ectomycorrhizal fungus Paxillusinvolutus with special reference to summer
drought. For. Ecol. Manag. 98: 221–228.
Garbaye, J. and G.D. Bowen. 1989. Stimulation of ectomycorrhizal infection of Pinus radiata
by some microorganisms associated with the mantle of ectomycorrhizas. New Phytol.
112: 383–388.
Garcia, C., A. Roldan and T. Hernandez. 1997. Changes in microbial activity after abandonment
of cultivation in a semi-arid Mediterranean environment. J. Environ. Qual. 26: 285–
291.
Holt, J.A. and M. Lepage. 2000. Termites and soil properties. In: T. Abe, D.E. Bignell and M.
Higashi (eds.). Termites: Evolution, Sociality, Symbioses, Ecology. Kluwer Academic
Publishers, Dordrecht, pp. 389–407.
Jones, M.D., D.M. Durall and J.W.G. Cairney. 2003. Ectomycorrhizal fungal communities in
young forest stands regenerating after clearcut logging. New Phytol. 157: 399–422.
Kropp, B.R. and C.G. Langlois. 1990. Ectomycorrhizae in reforestation. Can. J. For. Res. 20:
438–451.
Le Tacon, F., G. Jung, P. Michelot and J. Mugnier. 1983. Efficacité en pépinière forestière d’un
inoculum de champignon ectomycorhizien produit en fermenteur et inclus dans une
matrice de polymères. Ann. For. Sci. 40: 165–176.
Le Tacon, F., G. Jung, J. Mugnier, P. Michelot and C. Mauperin. 1985. Efficiency in a forest
nursery of an ectomycorhizal fungus inoculums produced in a fermentor and entrapped
in polymeric gels. Can. J. Bot. 63: 1664–1668.
Lesueur, D. and R. Duponnois. 2005. Relations between rhizobial nodulation and root
colonization of Acacia crassicarpa provenances by an arbuscular mycorrhizal fungus,
Glomusintraradices Schenk and Smith or an ectomycorrhizal fungus, Pisolithus tinctorius
Coker & Couch. Ann. For. Sci. 62: 467–474.
Mamatha, G., D.J. Bagyaraj and S. Jaganath. 2002. Inoculation of field-established mulberry
and papaya with arbuscular mycorrhizal fungi and a mycorrhiza helper bacterium.
Mycorrhiza 12: 313–316.
Martin F., C. Delaruelle and J.-L. Hilbert. 1990. An improved ergosterol assay to estimate the
fungal biomass in ectomycorrhizas. Mycol. Res. 94: 1069–1074.
Controlled Ectomycorrhization Practices 239
Marx, D.H. 1969. The influence of ectotrophic mycorrhizal fungi on the resistance of pine roots
to pathogenic infections. I. Antagonism of mycorrhizal fungi to root pathogenic fungi
and soil bacteria. Phytopathology 59: 153–163.
Marx, D.H. and W.C. Bryan. 1975. Growth and ectomycorrhizal development of Loblolly
pine seedlings in fumigated soil infested with the fungal symbiont Pisolithus tinctorius.
For. Sci. 21: 242–254.
Marx, D.H., K. Jarl, J.L. Ruehle and W. Bell. 1984. Development of Pisolithus tinctorius
ectomycorrhizae on pine seedlings using basidio-encapsulated seed. For. Sci. 30:
897–907.
Marx, D.H. 1991. The practical significance of ectomycorrhizae in forest establishment.
Ecophysiology of Ectomycorrhizae of Forest Trees, Marcus Wallenberg Foundation
Symposia Proceedings 7: 54–90.
Mauperin, C., F. Mortier, J. Garbaye, F. Le Tacon and G. Carr. 1987. Viability of an
ectomycorrhizal inoculum produced in a liquid medium and entrapped in a calcium
alginate gel. Can. J. Bot. 65: 2326–2329.
Meyer, J.R. and R.G. Linderman. 1986. Response of subterranean clover to dual-inoculation
with vesicular-arbuscular mycorrhizal fungi and a plant growth-promoting bacterium,
Pseudomonas putida. Soil Biol. Biochem. 18: 185–190.
Molina, R. and J.G. Palmer. 1982. Isolation, maintenance and pure culture manipulation of
ectomycorrhizal fungi. In: N.C. Schenck (ed.). Methods and Principles of Mycorrhizal
Research. The American Phytopathological Society, St. Paul, pp. 115–129.
Mortier, F., F. Le Tacon and J. Garbaye. 1988. Effect of inoculum type and inoculation dose
on ectomycorrhizal development, root necrosis and growth of Douglas fir seedlings
inoculated with Laccaria laccata in a nursery. Ann. For. Sci. 45: 301–310.
Pachlewski, R. and J. Pachlewski. 1974. Studies on Symbiotic Properties of mycorrhizal Fungi
of Pine (Pinus silvestris L.) with the aid of the method of mycorrhizal synthesis in pure
culture on agar. Forest Research Institute. Warsaw. Poland.
Paula, M.A., S. Urquiaga and J.O. Siqueira. 1992. Synergistic effects of vesicular-arbuscular
mycorrhizal fungi and diazotrophicus bacteria on nutrition and growth of sweet potato
(Ipomoea batatas). Biol. Fert. Soils 14: 61–66.
Piéri, C. 1991. Les bases agronomiques de l’amélioration et du maintien de la fertilité des
terres de savanes au sud Sahara. In: Savanes d’Afrique, terre fertile? Actes des Rencontres
Internationales, Montpellier, France, pp. 43–74.
Poole, E.J., G.D. Bending, J.M. Whipps and D.J. Read. 2001. Bacteria associated with Pinus
sylvestris–Lactarius rufus ectomycorrhizas and their effects on mycorrhiza formation in
vitro. New Phytol. 151: 743–751.
Rao, N.S.S., K.V.B.R. Tilak and C.S. Singh. 1985. Effect of combined inoculation of Azospirillum
brasilense and vesicular-arbuscular mycorrhiza on pearl millet (Pennisetum americanum).
Plant and Soil 84: 283–286.
Read, D.J., J.G. Duckett, R. Francis, R. Ligrone and A. Russell. 2000. Symbiotic fungal
associations in “lower” land plants. Philos. Trans. R. Soc. Lond. Ser. B-Biol. Sci. 355:
815–830.
Requena, N., I. Jimenez, M. Toro and J.M. Barea. 1997. Interactions between plant-growth-
promoting rhizobacteria (PGPR), arbuscular mycorrhizal fungi and Rhizobium spp. in
the rhizosphere of Anthyllis cytisoides, a model legume for revegetation in mediterranean
semi-arid ecosystems. New Phytol. 136: 667–677.
Requena, N., E. Perez-Solis, C. Azcon-Aguilar, P. Jeffries and J.M. Barea. 2001. Management
of indigenous plant–microbe symbioses aids restoration of desertified ecosystems. Appl.
Environ. Microbiol. 67: 495–498.
Rincon, A., M.R. de Felipe and M. Fernandez-Pascual. 2007. Inoculation of Pinus halepensis
Mill. with selected ectomycorrhizal fungi improves seedling establishment 2 years after
planting in a degraded gypsum soil. Mycorrhiza 18: 23–32.
240 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Roldan, A., I. Querejeta, J. Albadalejo and V. Castillo. 1996. Growth response of Pinus halepensis
to inoculation with Pisolithus arhizus in a terraced rangeland with urban refuse. Plant
and Soil 179: 35–43.
Rózycki, H., M. Kampert, E. Strzelczyk, C.Y. Li and D.A. Perry. 1994. Effect of different soil
bacteria on mycorrhizae formation in pine (Pinus sylvestris L.). Folia Forest. Polon. series
A (Forestry) 36: 91–102.
Schreiner, R.P., K.L. Mihara, K.L. McDaniel and G.J. Bethlenfalvay. 2003. Mycorrhizal fungi
influence plant and soil functions and interactions. Plant and Soil 188: 199–209.
Schrey, S.D., M. Schellhammer, M. Ecke, R. Hampp and M.T. Tarkka. 2005. Mycorrhiza
helper bacterium Streptomyces AcH 505 induces differential gene expression in the
ectomycorrhizal fungus Amanita muscaria. New Phytol. 168: 205–216.
Smith, S. and D. Read. 2008. Mycorrhizal Symbiosis, 2nd edn. Academic Press, London.
Terwilliger, J. and J. Pastor. 1999. Small mammals, ectomycorrhizae, and conifer succession
in beaver meadows. Oikos 85: 83–94.
Toro, M., R. Azcón and J.M. Barea. 1997. Improvement of arbuscular mycorrhiza development
by inoculation of soil with phosphate-solubilizing rhizobacteria to improve rock phosphate
bioavailability (32P) and nutrient cycling. Appl. Environ. Microbiol. 63: 4408–4412.
Torres, P. and M. Honrubia. 1994. Inoculation of containerized Pinus halepensis (Miller) seedlings
with basidiospores of Pisolithus arhizus (Pers) Rauschert, Rhizopogon roseolus (Corda) and
Suillus collinitus (Fr) O Kuntze. Ann. For. Sci. 51: 521–528.
Turjaman, M., Y. Tamai, H. Segah, S.H. Limin, J.Y. Cha, M. Osaki and K. Tawaraya. 2005.
Inoculation with the ectomycorrhizal fungi Pisolithus arhizus and Scleroderma sp. improves
early growth of Shorea pinanga nursery seedlings. New For. 30: 67–73.
Valentine, L.L., T.L. Fieldler, A.A. Hart, C.A. Petersen, H.K. Berninghausen and D. Southworth.
2004. Diversity of ectomycorrhizas associated with Quercus garryana in southern Oregon.
Can. J. Bot. 82: 123–135.
Van der Heijden, M.G.A., J.N. Klironomos, M. Ursic, P. Moutoglis, R. Streitwolf-Engel, T.
Boller, A. Wiemken and I.R. Sanders. 1998. Mycorrhizal fungal diversity determines plant
biodiversity ecosystem variability and productivity. Nature 396: 69–72.
Vignon, C., C. Plassard, D. Moussain and L. Salsac. 1986. Assay of fungal chitin and estimation
of mycorrhizal infection. Physiol. Végét. 24: 201–207.
Vivas, A., A. Marulanda, J.M. Ruiz-Lozano, J.M. Barea and R. Azcón. 2003. Influence of a
Bacillus sp. on physiological activities of two arbuscular mycorrhizal fungi and on plant
responses to PEG-induced drought stress. Mycorrhiza 13: 249–256.
von Alten, H., A. Lindemann and F. Schönbeck. 1993. Stimulation of vesicular-arbuscular
mycorrhiza by fungicides or rhizosphere bacteria. Mycorrhiza 2: 167–173.
Vósatka, M. and M. Gryndler. 1999. Treatment with culture fractions from Pseudomonas putida
modifies the development of Glomus fistulosum mycorrhiza and the response of potato
and maize plants to inoculation. Appl. Soil Ecol. 11: 245–251.
Will, M.E. and D.M. Sylvia. 1990. Interaction of rhizosphere bacteria, fertilizer, and vesicular-
arbuscular mycorrrhizal fungi sea oats. Appl. Environ. Microbiol. 56: 2073–2079.
Xie, Z.P., C. Staehelin, H. Vierheilig, A. Wiemken, S. Jabbouri, W.J. Broughton, R. Vogeli-Lange
and T. Boller. 1995. Rhizobial nodulation factors stimulate mycorrhizal colonization of
nodulating and nonnodulating soybeans. Plant Physiol. 108: 1519–1525.
CHAPTER
13
Biodiversity and Sustainable
Use of Wild Edible Fungi in the
Sudanian Centre of Endemism:
A Plea for Valorisation
Nourou Soulemane Yorou,1,2* N’Golo Abdoulaye Koné,3
Marie-Laure Guissou,4 Atsu Kudzo Guelly,5 Dao Lamèga
Maba,2,5 Marius R.M. Ekué6 and André De Kesel7
1. Introduction
Wild Edible Fungi (WEF) play an important role in the livelihood of local
inhabitants (Boa 2004). In tropical Africa, picking wild edible mushrooms is a
lucrative activity and involves hundreds of rural women (Boa 2004, Degreef
et al. 1997, Buyck 1994a). Over 300 edible mushrooms are recorded in sub-
Saharan Africa (Rammeloo and Walleyn 1993, Walleyn and Rammeloo 1994).
Still, mycophagy greatly varies from one country to another, even within
ethnic groups of the same area (Yorou and De Kesel 2002, Guissou et al.
2008). Annual consumptions of over 30 kg per habitant have been recorded
in Central and East Africa (Degreef et al. 1997). In rural tropical Africa, WEF
are important in terms of species richness and consumed quantities, but
also as a potential source of proteins, vitamins, and minerals (Degreef et
al. 1997, Adewusi et al. 1993, Ogundana and Fagade 1981, 1982, De Kesel
and Malaisse 2010).
others (Bâ et al. 2012, Ducousso et al. 2002). Unfortunately, many such
EcM trees are disappearing at an alarming rate (Neuenschwander et al.
2011), either through large scale deforestation or selective logging of timber
species. The process of deforestation is due mostly to shifting cultivation,
timber exploitation, land conversion to pastures, and increasing urban
demands of wood fuel (charcoal). Shifting cultivation usually results in
the slashing, burning and conversion of large forest areas into croplands,
whilst timber exploitation consists primarily of selective logging the
economically important aforementioned EcM trees. In any case, the removal
and destruction of partner tree species not only erodes the diversity of the
area, but also severely hampers the natural production of locally exploited
WEF. In Benin, about 28 WEF are threatened with extinction (Yorou and De
Kesel 2011), chiefly because of forest fragmentation, habitat disturbance, and
timber exploitation for charcoal. In a similar way, numerous fungi-rich WA
ecosystems are undergoing an unrestrained process of fragmentation (Hahn-
Hadjali et al. 2010, Wegmann et al. 2010), along with the disappearance
of economically important timber species and their associated WEF. This
process deeply affects the lives of the people living in surrounding villages
and cities. Due to such uncontrolled deforestation, the forest ecosystems
of WA are not only losing their potential as providers of Non Timber
Forest Products, but also their ability to improve the livelihood of rural
communities. As WEF are diverse and of local economic importance, the
installation of an economic sector based on WEF can serve as a model to
promote awareness among rural people, forest officers/administration,
and decision makers. The main goal of establishing an economic sector
of WEF is to face poverty and to conserve the hosts and the ecosystems
of fungal species all alike. In the present contribution, we are attempting
(1) to explore the diversity of WEF and fungal hotspots in West Africa
and (2) to highlight the natural productions, phenology, nutritional, and
socio-economic exploitation of WEF in West Africa. Data are assembled
from intensive mycological investigations in some West African countries
(Benin, Togo, Burkina Faso…) over the last 10 years, but also from extensive
mycological exploration in various ecosystems from other countries like
Ghana, Mali, Ivory Coast, and Guinea (started in 2009). Using a case study
from Benin, we will analyse different steps towards the establishment of
an economic sector based on WEF in West Africa.
forests of the coastal provinces of countries like Benin, Togo, Ghana, and
Ivory Coast exhibit a rather low diversity of edible, mostly saprotrophic
fungi. Most of the native edible taxa are members of Pleurotus, Marasmiellus,
Lentinus, Cookeina, and Armillaria. Plantations of exotic tree species, man-
made ecosystems, fallows, and to some extent disturbed habitats generally
harbour few edible fungi of the genera Chlorophyllum, Macrolepiota,
Volvariealla, and Agaricus. Towards the North, i.e., entering the ecological
zone where woodlands and gallery forests of the Guineo-Sudanian type start
to dominate (White 1986), a high diversity of edible EcM fungi is observed
(Bâ et al. 2011, De Kesel et al. 2002, Ducousso et al. 2002, Yorou et al. 2002a,b).
For landlocked countries such as Burkina Faso, Niger, and Mali, WEF are
most diverse in the southern Sudanian woodlands (Guissou 2005, Sanon
et al. 1997). The Sudanian Centre of Endemism (SCE) is home to hundreds
of WEF including notably representatives of Russula, Lactarius, Lactifluus,
Amanita, and Boletus. Numerous termites’ fungi (Termitomyces spp.) are
present in West Africa. These taxa are considered an important resource
and they play a seasonal but substantial role in the local diet, medicine,
and cash incomes for native villagers. The SCE is a narrow vegetation band
stretching from East (Soudan) to West (Senegal) between 9° and 11° North
latitudes (White 1986). Vegetation is composed of a mosaic of woodlands,
savannahs and gallery forests, mostly dominated by tree species belonging
to the Caesalpiniaceae, Phyllantaceae and Dipterocarps. In this area, some
timber species such as Afzelia spp., Isoberlinia spp., Uapaca spp., Berlinia
grandiflora and Anthonotha spp. form EcM symbiosis with numerous WEF.
Ectomycorrhizal fungi are predominant not only in terms of species richness,
but also in terms of fresh biomass, a large proportion of which is consumed
by local inhabitants (Table 1). Though woodlands form vast areas in the SCE,
they are actually fragmented and regressing annually (FAO 2010). More
conserved and semi-natural woodland ecosystems are located in numerous
forest reserves. Due to their conservation status as natural or semi-natural
ecosystems, Caesalpinioid and Phyllantioid-dominated woodlands and
gallery forests of the SCE emerge as fungi-rich ecosystems in West Africa.
The presence in their flora of important EcM tree species such as Berlinia spp.
and Uapaca spp. makes them remarkable WEF-rich ecosystems, generally
acting as refuge ecosystems for numerous unique and rare species (Yorou
and De Kesel 2011). Their generally fragmented size (Natta et al. 2002,
Sparovek et al. 2002) coupled with the unique and rare mycoflora they host
make the gallery forests important fungal hotspots with top conservation
priority. Because of the presence in its flora of numerous EcM trees such as
Anthonotha spp., Gilbertiodendron spp. Paramacrolobium coeruleum (to quote
only a few), Guinea harbours the highest diversity of potential WEF that
have been scantily documented ethnomycologically.
Wild Edible Fungi of West Africa 245
Table 1. Contrasting relative dominance of fungal flora between two dense forests (FD =
dense forest and FR = riparian forest) lacking EcM trees and 4 woodland ecosystems (SA =
shrub savannah, SB = wooded savannah, FC = Woodland of Uapaca spp. and FI = woodland
of Isoberlinia spp.) dominated by EcM trees in Sudanian area.
FD FR SA SB FC FI
Total Biomass 8.6 ± 5.2 26.7 ± 4.7 241.7 ± 13.88 223.7 ± 293.9 ± 285.7 ± 16
(kg/ha/year) 15.4 16,1
% Ectomycorrhizal 6 6.8 88.3 98.9 95.0 90.0
fungi from the total
biomass
% Saprotrophic 92.0 93.2 11.7 1.1 4.0 10.0
species from the total
biomass
% Wild edible 2.3 4.1 85.4 87.7 73.1 78.9
Fungi from the total
biomass
Source: Yorou NS (2000), unpublished data.
Table 2. contd....
Wild Edible Fungi of West Africa 247
Table 2. contd.
Table 2. contd....
248 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Table 2. contd.
Source (De Kesel et al. 2002, Guissou et al. 2008, Yorou et al. 2002b). Symbiotic species include
ectomycorrhizal and termite fungi.
Fig. 1. Frequency of some WEF genera in West Africa (data from Table 2).
Wild Edible Fungi of West Africa 249
Fig. 2. Some edible Lactarius species from West Africa. a. Lactarius flammans (Kou Forest Reserve,
Burkina Faso), b. L. gymnocarpus (Baro Forest Reserve, Upper Guinea), c. L. foetens (Mouhoun
Forest Reserve, Burkina Faso), d. L. medusae (Fazao National Park, Togo), e. L. cf. pelliculatus
(Forest Guinea), f. L. tenellus (from Benin).
Color image of this figure appears in the color plate section at the end of the book.
Fig. 3. Some edible Russula species from West Africa. a. Russula congoana (Moussaya Forest
Reserve, Upper Guinea), b. Russula sp1. (Ziama National Park, Guinea), c. Russula oleifera
(Bui national Park, Ghana).
252 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 4. Some edible Cantharellus species from West Africa. a. Cantharellus addaiensis (Wari-Maro
Forest Reserve, Benin), b. Cantharellus congolensis (Aledjo Forest Reserve, Togo), Afrocantharellus
platyphyllus (Kota gallery Forest, Benin), d. Cantharellus cf. conspicuus (Bassila gallery forest,
Benin).
Color image of this figure appears in the color plate section at the end of the book.
Fig. 5. A few edible Termitomyces from West Africa. a. Termitomyces sp. (Fazao National Park),
b. Termitomyces microcarpus, c. Termitomyces schimperi (Adjengré region, Togo), d. Termitomyces
letestui (Pèrèré region, Benin).
Koné (2013) and taking into account all available data on fungus growing
termites and their symbiotic fungi, a conservative diversity of at least 20
species can be estimated in West Africa. In addition, the published sequence
data (COI and ITS rDNA sequences) for both fungus-growing termites and
their symbiontes allocate the centre of termite agriculture to western Africa
(Nobré et al. 2011, Koné et al. 2011).
Termitomyces range among the top 3 most appreciated WEF in tropical
Africa (Rammeloo and Walleyn 1993). They remain a delicacy in West
Africa where many species are highly appreciated because of their taste
(De Kesel et al. 2002, Heim 1977). In West Africa, Termitomyces sporophores
are commonly used as food, but highly appreciated species are commonly
traded at local and regional markets (Fig. 6), contributing to secure cash
incomes for local people. In Ivory Coast for example, fruit bodies of
Termitomyces letestui are intensively collected and sold, with approximate
annual harvests amounting 7000 kg per year. Seasonal income (i.e., February
to April each year) of participants in the marketing chain varies from
US$58.38 to US$341.62, thus playing a crucial role for the livelihood of rural
communities (Koné et al. 2010b).
Fig. 6. Trade of Termitomyces letestui in Ivory Coast. a. from harvesters to dwellers, b and c.
Trade by women along roadside.
256 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 7. Proportion of EcM versus saprotrophic WEF biomass within six different vegetation
types. Caesalpinioid-dominated woodlands (SA, SB, FC, FI) are dominated (up to 99%) by
EcM edible fungi in comparison to Caesalpinioid-poor dense forests (FD and FR), where
saprotrophic fungi are predominant. Source Yorou: 2000 (unpublished).
Wild Edible Fungi of West Africa 257
and Lactifluus are not only the most common in term of species richness,
but the most dominant in woodlands and gallery forests as well (Yorou et
al. 2002a). In Sudanian ecozones, a single hectare of woodlands dominated
by Isoberlinia spp. and/or Uapaca spp. may produce over 6000 sporophores,
and up to 121 kg fresh biomass of Lactifluus gymnocarpoides annually, making
it the most dominant WEF in Sudanian woodlands.
The Sudanian ecozones are characterised by a rainy season of up to 5–6
months (May to October) which contrasts strongly with a long severe dry
season. While the dry season is generally the harvest period of many crops,
the rainy season is particularly a difficult time for numerous farmers because
of the lack of harvested crops. The beginning of the rainy/wet season (May
to early July) represents a difficult period for local inhabitants. It is known
as a shortage period because stocks are empty or sold out and new crops
still unavailable. It is during this period that WEF play an important role
as suppliers of food (De Kesel and Malaisse 2010, Yorou and De Kesel 2002)
and essential proteins (Adewusi et al. 1993, Guissou et al. 2005). Yorou et
al. (2002a) demonstrated that more than 70% of total annual productions
of WEF are obtained at the beginning of the raining season (May to early
July), which coincides with the aforementioned shortage period, thus
contributing substantially to improve the food security of vast numbers
of WA people. Natural productions of WEF significantly vary throughout
the fruiting period (F(10,107) =157,57; p<0,01) and fit with an J-inverse curve
(Fig. 8). The first production peak is obtained with the first intense rains
(usually in May and June). Ecologically, the first rains generally break
Fig. 8. Temporal variability (F(10,107) =157,57; p<0,01) over 6 months (11 sampled weeks I to XI,
2-week interval from May till October) of total natural productions of ectomycorrhizal WEF
in Sudanian woodlands (SA, SB, FC and FI). Natural production fits a J-inverse curve with
two peaks of productions. Source: Yorou 2000 (unpublished).
258 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 9. Use of wild mushrooms as food by local people in Ivory Coast. a. Processing dried
mushrooms before cooking, b and c. Cleaning Termitomyces letestui for cooking.
The purpose of this activity is to identify the habitats of the identified priority
species, phenology, and natural availability. This is of great importance since
knowing target species’ preferred habitat, distribution, and phenology will
greatly reduce harvest effort of villagers. Except for Psathyrella tuberculata,
all above mentioned species include termite fungi (T. letestui, T. schimperi
and T. clypeatus) and EcM ones (Lactifluus flammans, L. gymnocarpoides
Lactifluus medusae and Cantharellus aff. rufopunctatus). Ecologically, Bassila
region harbours Caesalpinioid- and Phyllantioid-dominated woodlands
for which quantitative data on natural production and species distribution
are available (Yorou et al. 2002a). Annual productions of Termitomyces
are difficult to assess since their fructification is sporadic and relies upon
many environmental factors. Our numerous attempts to assess natural
productions of Termitomyces within permanent plots were unsuccessful.
However, Koné et al. (2010a,b) reported an annual estimation of over 7000
kg for T. letestui in southern Ivory Coast. Cantharellus aff. rufopunctatus is a
species that is confined to gallery forests dominated by Berlinia grandiflora.
Annual productions of this species amounts 178 kg/ha, on the basis of direct
weekly biomass measure of 8.9 kg during 5 fruiting months. In summary,
woodlands and gallery forests dominated by Isoberlinia spp. and Uapaca
Wild Edible Fungi of West Africa 261
spp. were identified during the study as important target habitats to deploy
sustainable harvesting in the context of a WEF-based local economical
activity ( in Central Benin).
Market surveys are aiming at identifying species that are jointly appreciated
by the harvesters and the end users as well. Market surveys should
allow making an economic examination of the sector, taking into account
harvesting effort, but also costs analysis related to mushroom drying,
labelling, and transportation from the forest villages to the location of
potential users. Such factors, coupled with the variability of environmental
factors, remain the most challenging aspects in the establishment and
continuity of a regional economic sector based on WEF. To be efficient and
visible, this economic activity not only needs to identify good target species,
but also needs to be available throughout the year. It is clear that both fresh
and dried specimens should be available and that they should meet the
users’ expectation. This goes from personal criteria and requirements about
transformed fungi (dried, in oil) to the way the edible fungi are packed,
labelled and presented. In addition, costs for collecting in the forest, drying
and transportation from rural to urban markets should be as low as possible
to allow a rapid flow at reasonable price.
In this particular case (Bassila region), though we were able to identify
priority species along with their production and distribution, a successful
mushroom-based sector could not be initiated because of (1) the lack of
potential buyers within and outside the villages, (2) lack of assistantship to
promote awareness and to coach villagers from harvesting to the selling of
WEF and finally (3) consumers’ preferences of fresh mushrooms over dried
ones. Because mushrooms are available in their immediate environment,
no villager from the study area can imagine buying mushrooms except the
very rare and highly prized species such as Termitomyces. Many villagers
harvest the little quantities of WEF needed for their own personal uses, with
very limited lucrative ambition. As WEF are collected only for personal use,
and that they are mostly appreciated in fresh conditions, there is no need
for collecting and drying more than daily required. We hardly observed
villagers drying surplus specimens for a long term use (neither for personal
consumption nor for sales). However, drying mushrooms seems not to
be a major worry in tropical rural areas. Wild edible mushrooms can be
either dried under the sun or smoked together with bush meat (Fig. 10). In
the neighbouring country Burkina Faso, the highly appreciated Phlebopus
sudanicus is harvested and commonly dried under the sun by Bobo people
for long term exploitation (Guissou et al. 2005).
262 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 10. Drying wild edible mushrooms. a and b. Drying of Psathyrella tuberculata and Volvariella
volvacea under sun, c. Smoking of Termitomyces letestui together with bush meat.
nature to justify a long lasting value chain. Unless grown under controlled
conditions, it will remain a moderately predictable but still highly seasonal
edible product from the forest.
One important challenge in the study area, and probably in many West
African cities, is to revive culinary habits of citizens so as to introduce WEF
in their daily dishes. Though WEF are valuable by villagers throughout the
study areas, they hardly gained attention of citizens of the neighbouring
cities. Generally, there is a decline in the use of WEF from one generation
to another and when moving from forest villages to the cities (Yorou and
De Kesel 2002, Degreef et al. 1997). This is probably a general fact in the
whole of West Africa (Guissou et al. 2008), and it may negatively influence
the promotion of value chain of WEF. Reversing, changing, or widening
culinary habits of citizens is probably the most tedious task in the actual West
Africa context, as firstly, many imported foodstuffs are actually regarded as
the best, and secondly, wild fungi are usually considered to be poisonous.
If Termitomyces can attract the interest of many villagers and citizens, much
effort is needed to increase the attention of the people in cities towards EcM
edible mushrooms. This can be successfully performed through large scale
awareness-raising programme about the culinary and nutritional benefits
of WEF, and importance of WEF to improve food security among villagers
and citizens.
factors
Fig. 11. Simplified diagram showing different steps and most needed investigations to be
considered in erecting an economic sector based on WEF.
6. Concluding Remarks
Wild edible fungi are mostly EcM fungi and they are a seasonal but abundant
and important source of food. In parts of West Africa, they represent a
modest but stable income for rural people. The socio-economic potential
of these wild edible fungi is high because our studies have indicated that
the naturally produced quantities largely surpass the demand and local
consumption. Wild edible fungi are under-utilized in West Africa. This
potential is however currently being hampered or even annihilated by
wide scale and devastating human activities. Let there be no doubt that the
massive production of charcoal has become the biggest threat to all EcM
forests in the whole of tropical Africa. Although forest products, including
edible fungi, are often highly valued by local inhabitants, much effort is still
needed to promote awareness among people that harvesting and marketing
of edible fungi can only become an income generating activity if EcM forests
are maintained and preserved. Multi-disciplinary investigations including
taxonomists, ecologists, food scientists, food technologists, and economists
should be implemented in order to secure a better exploitation of EcM
Wild Edible Fungi of West Africa 265
forests and their generally associated edible fungi. There is very little doubt
that sustainable use and preservation of West African forests, rather than
cutting them down for charcoal,will yield a long-term improvement in the
livelihood of millions of rural people.
Key words: Wild Edible Fungi, diversity, natural productions, phenology,
local uses, economic sector, West Africa
7. Aknowledgement
We are grateful to local communities who provided valuable ethnomycological
information. Yorou NS is grateful to the German Research Foundation for
financial supports (DFG project Ag7/19-1 and Yo 174/2-1). Koné NA is
undebted to the International Foundation for Science (Grant D/4982-1). The
authors acknowledge Krista L. McGuire and Caitlyn Gillikin for improving
the language.
8. References
Aanen, D.K. and P. Eggleton. 2005. Fungus-growing termites originated in African rain forests.
Curr. Biol. 15: 851–855.
Adewusi, S.R.A., F.V. Alofe, O. Ademeyi, O.A. Alofabi and O.L. Oke. 1993. Studies on some
edible wild mushrooms from Nigeria. I. Nutritional, teratogenic and toxic considerations.
Plant Foods Human Nutri. 43: 115–121.
Bâ, A.M., R. Duponnois, M. Diabaté and B. Dreyfus. 2011. Les champignons ectomycorrhiziens
des arbres forestiers en Afrique de l´Ouest. Méthodes d’étude, diversité, écologie,
utilisation en foresterie et comestibilité. Editions IRDpp. 264.
Bâ, A.M., R. Duponnois, B. Moyersoen and A.G. Diédhiou. 2012. Ectomycorrhizal symbiosis
of tropical African trees. Mycorrhiza 22: 1–29.
Bas, C. 1969. Morphology and subdivision of Amanita and a monograph of its section Lepidella.
Persoonia 5: 285–579.
Boa, E. 2004. Wild Edible Fungi. A Global Overview of Their Use and Importance to people.
Non-Wood Forest Products. FAO, Rome 17: 1–147.
Buyck, B. 1994a. Ubwoba, champignons comestibles de l’Ouest de Burundi. Administration
Générale Coopération pour le Développent, Agriculture volume 34: 123.
Buyck, B. 1994b. Russula I (Russulaceae). Flore Illustrée des Champignons de l’Afrique Centrale
15: 335–408.
Buyck, B. 1994c. Russula II (Russulaceae). Flore Illustrée des Champignons d’Afrique Centrale
16: 411–542.
Buyck, B. 1997. Russula III (Russulaceae). Flore Illustrée des Champignons de l’Afrique Centrale
volume 17: 545–597.
Buyck, B. and B. Nzigidahera. 1995. Ethnomycological notes from Western Burundi. Belgian
J. Bot. 128: 131–138.
Buyck, B., G. Eyssartier and A. Kivaisi. 2000. Addition to the inventory of the genus Cantharellus
(Basidiomycota, Cantharellaceae) in Tanzania. Nova Hedwigia 73: 3–4.
Buyck, B., F. Kauff, C. Cruaud and V. Hofstetter. 2013. Molecular evidence for new Cantharellus
(Cantharellalles, Basidiomycota) from tropical African miombo woodlands and a key to
all tropical African Cantherelles. Fungal Diversity 58: 281–298.
266 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
De Kesel, A. and F. Malaisse. 2010. Edible wild food: Fungi. In: F. Malaisse (ed.). How to
Live and Survive in Zambesian Open Forest (Miombo Ecoregion). Gembloux, Presses
agronomiques: 422pp. + CD rom, pp. 41–56.
De Kesel, A., N.S. Yorou and B. Buyck. 2011. Cantharellus solidus, a new species from Benin
(West-Africa) with a smooth hymenium. Cryptog. Mycol. 32: 277–283.
De Kesel, A., J.T.C. Codjia and N.S. Yorou. 2002. Guide des champignons comestibles du
Bénin. Cotonou, République du Bénin, Jardin Botanique National de Belgique et Centre
International d’Ecodéveloppement Intégré (CECODI). Impr. Coco-Multimedia, pp.
275.
Degreef, J., F. Malaisse, J. Rammeloo and E. Baudart. 1997. Edible mushrooms of Zambezian
woodland area: A nutritional and ecological approach. Biotechnology Agronomy Society
and Environment 1: 221–231.
Diédhiou, A.G., M.A. Selossé, A. Galiana, M. Diabaté, B. Dreyfus, A. M. Bâ, S. Miana and G.
Béna. 2010. Multi-host ectomycorrhizal fungi are predominant in a Guinean tropical
rainforest and shared between canopy trees and seedlings. Environ. Microbiol. 12:
2219–2232.
Ducousso, M., A.M. Bâ and D. Thoen. 2002. Les champignons ectomycorrhiziens des
forêts naturelles et des plantations d’Afrique de l’Ouest: une source de champignons
comestibles. Bois For. Trop. 275: 14.
Eyi-Ndong, H. 2009. Etude des champignons de la forêt dense humide consommés par les
populations du nord du Gabon. PhD thesis, Université Libre de Bruxelles, pp. 271.
Eyi-Ndong, H. and J. Degreef. 2009. Inventaire des champignons consommés par les Pygmés
du nord du Gabon. XVII Congrès AETFAT, 28 Fév-2 Mars 2007, Yaoundé Cameroun.
Eyi-Ndong, H. and J. Degreef. 2010. Diversité des espèces de Cantharellus, Lentinus et
Termitomyces consommés par les Pygmés du Nord du Gabon. In: J. van der Burgt, J. van
der Maesen and J.M. Onana (eds.). Systématique et Conservation des plantes Africaines.
Kew Royal Botanic Garden, pp. 133–141.
Eyi-Ndong, H., J. Degreef and A. De Kesel. 2011. Champignons comestibles des forêts denses
d´Afrique Centrale. Taxonomie et identification. ABC taxa, Bruxelles 10: 255.
Eyssartier, G. 2003. The genus Cantharellus. PhD thesis, Muséum National d´Histoire Naturelle,
Paris.
Eyssartier, G., B. Buyck and A. Verbeken. 2002. Cantharellus conspicuus sp. nov. Crypto. Mycol.
23: 95–102.
Eyssartier, G. and B. Buyck. 1998. Contribution à la systématique du genre Cantharellus en
Afrique tropicale: Etude de quelques espèces rouges. Belgian J. Bot. 131: 139–149.
FAO. 2010. FAOSTAT, FAO Statistical Databases. http,//faostat.fao.org/, dernier accès le 21
janvier 2010.
Guissou, K.L.M. 2005. Les Macromycètes du Burkina Faso. Thèse de Doctorat, Université de
Ouagadougou, Burkina Faso.
Guissou, K.M.L., P. Sankara and S. Guinko. 2005. Phlebopus sudanicus ou la viande des Bobos,
un champignon comestible dans le Département de Satiri au Burkina Faso. Crypto.
Mycol. 3: 195–204.
Guissou, K.M.L., A.M. Lykke, P. Sankara and S. Guinko. 2008. Declining wild mushrooms
recognition and usage in Burkina Faso. Economic Bot. 62: 530–539.
Hahn-Hadjali, K., R. Wittig, M. Schmidt, G. Zizka, A. Thiombiano and B. Sinsin. 2010. La
végétation de l´Afrique de l’Ouest. In: B. Sinsin and D. Kampmann (eds.). Biodiversity
Atlas of West Africa, volume I: Benin. Cotonou/Frankfurth/Main pp. 78–85.
Hama, O., E. Maes, K.M.L. Guissou, D. Ibrahim, M. Baragé, L.A. Parra Sánchez, O. Raspé and
A. De Kesel. 2010. Agaricus subsaharianus, une nouvelle espèce comestible et consommée
au Niger, au Burkina Faso et en Tanzanie. Crypto. Mycol. 31: 221–234.
Härkönen, M. 1992. Wild mushrooms, a delicacy in Tanzania. Univ. Helsingensis 2: 29–31.
Härkönen, M., T. Saarimäki and L. Mwasumbi. 1994. Edible and poisonous mushroom of
Tanzania. The African J. Mycol. Biotech. 2: 99–130.
Wild Edible Fungi of West Africa 267
Härkönen, M., T. Saarimäki and L. Mwasumbi. 1995. Edible mushrooms of Tanzania. Karstenia
35 (supplement): 1–92.
Härkönen, M., T. Niemelä and L. Mawasumbi. 2003. Edible, harmful and other fungi. The
Finnish-Tanzanian Friendship Society. Norrlinia 10: 200.
Heim, R. 1977. Termites et champignons. Les champignons termitophiles d’Afrique Noire et
d’Asie méridionale. Paris.
Koné, N.A. 2013. Symbiose termite-champignon: Origine, co-diversification et fructification
saisonnière du symbiote fongique (Termitomyces spp.). Thèse Unique de Doctorat de
l’Université Nangui Abogoua, Côte d´Ivoire : 47p.
Koné, N.A., K. Dosso, S. Konaté, Y.J. Kouadio and K.E. Linsenmair. 2011. Environmental and
biological determinants of Termitomyces species seasonal fructification in central and
southern Côte d’Ivoire. Insectes Soc. 58: 371–382.
Koné, N.A., D. Koné and P. Nicot. 2010a. State of knowledge of fungal diversity in Côte
d’Ivoire. In: S. Konaté and D. Kampmann (eds.). Biodiversity Atlas of West Africa Tome
III, pp. 172–177.
Koné, N.A., S. Konaté and E.K. Linsenmair. 2010b. Socio-economic importance of Termitomyces
in Côte d’Ivoire. In: S. Konaté and D. Kampmann (eds.). Biodiversity Atlas of West Africa
Tome III, pp. 177–178.
Malaisse, F. 1997. Se nourrir en forêts claires africaines. Approches écologiques et nutritionnelles.
CTA. Wagenigen, pp. 384.
Malaisse, F., A. De Kesel, G. N’Gasse and G. Lognay. 2008. Diversité des champignons
consommés par les Bofi de la Lobaye (République centrafricaine). Geo-Eco-Trop 32:
1–8.
Martin, G.J. 1995. Ethnobotany: A methods manual. London. Champman & Hall. Chapter
anthropology.
Morris, B. 1986. Notes on the genus Termitomyces Heim in Malawi. Soc. Malawi J 39: 40–49.
Morris, B. 1984. Macrofungi of Malawi. Some ethnobotanical notes. Bull Br. Mycol Soc 18:
48–57
Morris, B. 1994. Bowa: Ethnobotanical notes on the macrofungi of Malawi. In: J.H. Senya
and A.C. Chikuni. Proceedings of the XIIIth Plenary meeting. AETFAT, Malawi, pp.
635–793.
Mossebo, D.C. and A. Amougou. 2002. Contribution à l’étude du genre Termitomyces
(Basidiomycètes) au Cameroun: écologie et systématique. Bull. Soc. Mycol. Fr. 118:
195–249.
Mossebo, D.C., A. Amougou and R.E. Atangana. 2002. Contribution à l’étude du genre
Termitomyces (Basidiomycètes) au Cameroun : écologie et systématique. Bull. Soc. Mycol.
Fr. 118: 195–249.
Natta, A.K., B. Sinsin and L.J.G. van der Maesen. 2002. Riparian forests, a unique but
endangered ecosystem in Benin. Notulae Florae Beninensis 4. Botanische Jahrbücher
124: 55–69.
Neuenschwander, P., B. Sinsin and G. Georgen. 2011. Protection de la nature en Afrique de
l´Ouest: Une Liste rouge pour le Benin. Nature Conservation in West Africa: Red List for
Benin. International Institute of Tropical Agriculture, Ibadan, Nigeria, pp. 365.
Nobré, T., N.A. Koné, S. Konaté, K.E. Linsenmair and D.K. Aanen. 2011. On the origin and
co-diversification of fungus-growing termites and their fungal symbionts. Mol. Ecol.
20: 2619–2627.
Ogundana, S.K. and O.E. Fagade. 1981. Nutritional value of some Nigerian edible mushrooms.
Mushroom Sc. 11: 123–131.
Ogundana, S.K. and O.E. Fagade. 1982. Nutritional value of some Nigerian edible mushrooms.
Food Chem. 8: 263–268.
Parent, G. and D. Thoen. 1977. Food value of edible mushrooms from Upper-Shaba Region.
Economic Bot. 31: 436–445.
Pegler, D.N. and G.D. Piearce. 1980. The edible mushrooms of Zambia. Kew Bull. 35:
475–491.
268 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Yorou, N.S. 2010. Larger fungi. In: B. Sinsin and D. Kampmann (eds.). Biodiversity Atlas of
West Africa, volume I: Benin. Cotonou/Frankfurth/Main pp. 324–331.
Yorou, N.S. and A. De Kesel. 2002. Connaissances ethnomycologiques des peuples Nagot du
centre du Bénin (Afrique de l’Ouest). Proceeding of XVI the AETFAT congress, Brussels
2000. Syst. Geogr. Plants 71: 627–637.
Yorou, N.S. and A. De Kesel. 2011. Champignons supérieurs. Larger fungi. In: P.
Neuenschwander, B. Sinsin and G. Goergen (eds.). Protection de la Nature en Afrique de
l’Ouest: Une Liste Rouge pour le Bénin. Nature Conservation in West Africa: Red list for
Benin. International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria, pp. 47–61.
Yorou, N.S., A. De Kesel, B. Sinsin and J.T.C. Codjia. 2002a. Diversité et productivité des
champignons comestibles de la forêt classée de Wari-Maro (Bénin, Afrique de l’Ouest).
Proceedings of XVIth AETFAT Congress, Brussels 2000. Syst. Geogr. Plants 71: 613–
625.
Yorou, S.N., A. De Kesel, J.T.C. Codjia and B. Sinsin. 2002b. Biodiversité des champignons
comestibles du Bénin. Proceedings of the Symposium-Workshop on Biodiversity in Benin.
Abomey-Calavi (Benin) October 30th to November 18th 2002, pp. 231–240.
1
Department of Natural Resources Management, Faculty of Agronomy, University of Parakou,
BP 123, Parakou, Benin.
2
Department Biology I, Ludwig-Maximilians-Universität München, Menzinger Str. 67, 80638,
Munich, Germany.
3
URF des Sciences de la Nature et de l’Environnement, Station d’Écologie de Lamto, Université
Nangui Abrogoua (Côte d’Ivoire), BP 28 N’Douci.
4
École Normale Supérieure, Université de Koudougou, BP 376, Koudougou, Burkina Faso.
5
Département de Botanique et Écologie Végétale, Faculté des Sciences, Université de Lomé,
081 BP 1515 Lomé, Togo.
6
Laboratoire d´Écologie Appliquée, Faculté des Sciences Agronomiques, Université d’Abomey-
Calavi, 01 BP 526, Cotonou, Bénin.
7
Department of Cryptogamy (Bryophyta & Thallophyta), National Botanic Garden of Belgium,
Domein van Bouchout, B-1860, Belgium.
*Corresponding author: n.s.yorou@bio.lmu.de
Color Plate Section
276 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Chapter 1
Fig. 1. Distribution of EcM trees in tropical Africa: (1) rainforests in the Guinea-Congo region; (2)
open forests in the Sudanian and Zambezian regions; (3) savanna woodlands in the Sudanian
and Zambezian regions (Bâ et al. 2012).
Color Plate Section 277
Chapter 2
Fig. 1. Micrographs of mycorrhizas of Neea species 1 (a-e), Neea aff. floribunda (g-h), Pisonia sp.
(g-h), a. Russula puiggarii − Neea species 1, b. Russula-Lactarius − Neea species 1, c. Tomentella-
Thelephora species 1 − Neea species 1, d. Tomentella-Thelephora species 2 − Neea species 1, e.
Ascomycete − Neea species 1, f. Tomentella-Thelephora species 3 − Neea aff. floribunda, g.,h. .
Tomentella-Thelephora species 3 − Pisonia sp., g. overview: long roots partially with hyphal
mantle (arrows), partially with root hairs and no hyphal mantle (*), h. one root shown at higher
magnification: distal portion with root hairs (*), proximal portion with hyphal mantle (arrows).
Scale bars: a.,b.,f.,h. 1 mm; c.-e. 0,5 mm; g. 5 mm. Data from Haug et al. 2005.
278 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Chapter 6
Fig. 3. (A) Podoserpula miranda and (B) Cantharellus garnieri. Photos by courtesy from Ducousso
Marc, CIRAD.
Color Plate Section 279
Fig. 4. Pisolithus albus from New Caledonia. A: Pisolithus albus MD07-117 from the Koniambo
massif; B: Pisolithus albus MD07-228 from the Ouen-Toro, Noumea; C: cross section of Pisolithus
albus MD07-166 from Pindjen water-fall and D: globose spores (8.77 to 9.62 µm) of Pisolithus
albus MD06-379 from Poum, erected spines (1.2 µm) are clearly visible. From Jourand et al.
(2010a).
280 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 9. Eucalyptus globulus seedlings after 12-weeks growth. A and A’ mycorrhizal; B and B’:
non-mycorrhizal (controls). A and B: no nickel added; A’ and B‘ seedlings treated with Ni.
From Jourand et al. (2010b).
Color Plate Section 281
Chapter 7
Fig. 1. Climatic regions of Burkina Faso and localization of prospected areas. Data from Sanon
et al. (2009a).
282 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Chapter 8
Fig. 3. Yellow Gnetum EM root tip formed with S. sinnamariense, showing extensive extramatrical
mycelia. Bar is 0.3 mm.
Color Plate Section 283
Chapter 9
Fig. 1. Distribution of three Coccoloba species (Coccoloba uvifera, C. swartzii and C. pubescens) and
their putative ectomycorrhizal fungi along a salinity gradient in Martinique and Guadeloupe
(Lesser Antilles).
284 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Chapter 10
Fig. 1. Map illustrating examples of monodominant and co-dominant forests with the tree
species or genus listed and the tree family in parentheses. This diagram is not an exhaustive
list of monodominant species, but it illustrates how monodominant forests are found in all
tropical regions.
Color Plate Section 285
Chapter 13
Fig. 2. Some edible Lactarius species from West Africa. a. Lactarius flammans (Kou Forest Reserve,
Burkina Faso), b. L. gymnocarpus (Baro Forest Reserve, Upper Guinea), c. L. foetens (Mouhoun
Forest Reserve, Burkina Faso), d. L. medusae (Fazao National Park, Togo), e. L. cf. pelliculatus
(Forest Guinea), f. L. tenellus (from Benin).
286 Ectomycorrhizal Symbioses in Tropical and Neotropical Forests
Fig. 4. Some edible Cantharellus species from West Africa. a. Cantharellus floridulus (Wari-Maro
Forest Reserve, Benin), b. Cantharellus congolensis (Aledjo Forest Reserve, Togo), Afrocantharellus
platyphyllus (Kota gallery Forest, Benin), d. Cantharellus cf. conspicuus (Bassila gallery forest,
Benin).
Ectomycorrhizal (ECM) symbiosis is a mutualistic plant-
Ectomycorrhizal
Ectomycorrhizal
Ectomycorrhizal
Ectomycorrhizal (ECM) symbiosis a mutualistic
and Neotropical
maintenance and evolution of biodiversity and ecosystems
Neotropical
and
ecology of forest trees,trees,
affecting growth, waterwater
and nutrient
ecology of forest affecting growth, and nutrient
absorption and protection against pathogens. It is a research
absorption and protection against pathogens. It is aItresearch
Symbioses
absorption and protection against pathogens. is a research
imperative in tropical and neotropical forest ecosystems
imperative in tropical
imperative and neotropical
in tropical and neotropical forestforest
because they concern an ecologically and economically
ecosystems
ecosystems Neotropical
Neotropical
Neotropical Forests
Forests
Forests
Forests
because they they
concern an ecologically and and economically
because concern an ecologically economically
Forests
important tree species (e.g. Ceasalpinioid subfamily in Africa,
important tree species (e.g. Ceasalpinioid subfamily in Africa,
important tree species (e.g. Ceasalpinioid subfamily in Africa,
in Tropical
Dipterocarpaceae in Asia). The book is an overview of the
Dipterocarpaceae in Asia). The book is an isoverview of theof the
Dipterocarpaceae in Asia). The book an overview
knowledge of ECM symbioses in tropical and neotropical
knowledge of ECM symbioses in tropical and neotropical
knowledge of ECM symbioses in tropical and neotropical
ecosystem forests. The contents address diversity and function
ecosystem forests. The contents address diversity and function
ecosystem forests. The contents address diversity and function
of ectomycorrhiza associated with forest plants, impacts of
of ectomycorrhiza associated with with
forestforest
plants, impacts of of
of ectomycorrhiza associated plants, impacts
ectomycorrhiza on plant diversity and composition,
ectomycorrhiza on plant diversity and and composition,
ectomycorrhiza on plant diversity composition,
regeneration and dynamics of ecosystems, and biomass
regeneration and anddynamics of ecosystems, and andbiomass
regeneration dynamics of ecosystems, biomass
Editors Krista
Editors
production in forestry, adaptation of ectomycorrhiza to
EditorsAmadou
production in forestry, adaptation of ectomycorrhiza to to
production in forestry, adaptation of ectomycorrhiza
nutrient deficient, salt and ultramafic soils.
Abdala G.
nutrient deficient, salt andsaltultramafic soils. soils.
Krista
nutrient deficient, and ultramafic Abdala G. Diédhiou
Amadou
Krista M.
Abdala
Amadou M. Bâ
Editors
Editors
L. McGuire Editors
L. Diédhiou
L. McGuire
AmadouM. M.Bâ
Bâ
G. Diédhiou
Amadou
M.
McGuire
Amadou M. Bâ
Bâ
Bâ
KristaL.L.McGuire
Krista McGuire
Krista L. McGuire
AbdalaG.G.Diédhiou
Abdala Diédhiou
K20685
Abdala G. Diédhiou
6000 Broken Sound Parkway, NW
Suite 300, Boca Raton, FL 33487
711 Third Avenue
New York, NY 10017 9 781 466 59 468 5
an informa business 2 Park Square, Milton Park 9 781 466 59 468 5
Abingdon, Oxon OX14 4RN, UK 9 781 466 59 468 5 A SCIENCE PUBLISHERS BOOK
w w w. c rc p r e s s . c o m