Acta Sci. Pol. Hortorum Cultus, 16(4) 2017, 113–126
www.acta.media.pl
ISSN 1644-0692
DOI: 10.24326/asphc.2017.4.12
ORIGINAL PAPER
Accepted: 5.04.2017
FUNGI INHABITING FRUIT TREE SHOOTS WITH SPECIAL REFERENCE
TO THE Diaporthe (Phomopsis) GENUS
Ewa Dorota Król1, Barbara Anna Abramczyk2, Ewa Dorota Zalewska1,
Beata Zimowska1
1
University of Life Sciences in Lublin, Poland
2
Institute of Soil Science and Plant Cultivation – State Research Institute, Puławy, Poland
ABSTRACT
The purpose of the study was to determine which fungal species colonize the shoots of apple, pear, cherry,
plum, apricot and peach in south-eastern Poland and whether Diaporthe (Phomopsis) isolates there are
among them. The study was conducted in 2010–2012 and the plant material was collected from five orchards. In three of these chemical controls were carried out, while there was no protection applied in the
other two. The results showed that species composition of the fungi was very similar regardless of host
plant and year of study. Alternaria spp., Fusarium spp. and Phoma spp. were isolated most frequently from
the studied shoots. Fungi cultures known for pathogenicity towards fruit trees such as Botrytis cinerea,
Colletotrichum gloeosporioides, Leucostoma spp., Monilinia spp. and Neofabraea spp. were also isolated.
Moreover, there were isolates of Diaporthe (Phomopsis) among the fungi present in the shoots. Cultures of
these fungi were obtained both from shoots with disease symptoms and from visually healthy ones but
mainly from shoots originating from unprotected orchards, what indicates their greater threat to fruit trees
grown without chemical protection. This is the first documented report of the occurrence of Diaporthe
(Phomopsis) on fruit trees in Polish conditions.
Key words: orchard plants, biodiversity of fungi, occurrence
INTRODUCTION
The shoots of fruit trees are inhabited by numerous
fungi species including bark and wood pathogens.
Among them, both worldwide and in Poland, the most
frequently mentioned are: Neonectria galligena (Bres.)
Rosman et Samuels, Leucostoma spp., Neofabraea
spp., Botryosphaeria spp. causing cankers and Chondrostereum purpurem causing “silver leaf”. [Regner et
al. 1990, Spotts et al. 1990, Grabowski 1993, Grabowski and Leś 1996, Adams et al. 2002, Grabowski
2002, Latorre et al. 2002, Valiuškaite and Raudonis
2008, Gramaje et al. 2012, Romanazzi et al. 2012].
Recently, diseases caused by Diaporthe (Phomopsis) have become an increasing problem in the or-
chard regions of the world. These fungi are common
in different climatic zones and colonize many plant
species. Diaporthe (Phomopsis) species are associated with bark necrosis, shoot blight and cancer, decay,
wilting, fruit rot and mummification [Farr et al. 1999,
Kanematsu et al. 2000, Farr et al. 2002, Karaoglanidis and Bardas 2006, Živkovič et al. 2007, Król
and Kowalik 2010, 2011, Udayanga et al. 2011,
Gomes et al. 2013]. Among the species most dangerous for fruit plants are Phomopsis viticola Sacc.,
P. perniciosa Grove, P. ambigua (Sacc.) Trav.,
P. amygdali (Del.) Tuset et Portilla comb. nov.,
P. mali Roberts, P. juglandina (Sacc.) Höhn,
ewa.krol@up.lublin.pl
© Copyright by Wydawnictwo Uniwersytetu Przyrodniczego w Lublinie
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
P. ampelina (Berk. et Curt.) Grove., P. oblonga
(Desm.) Traverso and P. vaccinii Shear. [Machowicz-Stefaniak 1993, Farr et al. 1999, Mostert et al.
2001, Król 2002, Arsenijević and Gavrilović 2005,
Karaoglanidis and Bardas 2006, Michailides and
Thomidis 2006, Król and Kowalik 2010, 2011,
Udayanga et al. 2011, Mirabolfathy et al. 2013].
Though only some of the Phomopsis species produce their sexual stage Diaporthe, because of changes in the taxonomy of these fungi, the name
Diaporthe has been conserved over Phomopsis and
Diaporthe eres has been recognized as the main
pathogen of woody plants from different botanical
families including the family Rosaceae, Juglandaceae, Vitaceae and Ericaceae [Rossman et al.
2014, Udayanga et al. 2014].
The purpose of this study was to determine which
fungi species inhabit the shoots of fruit trees in southeastern Poland and to examine whether Diaporthe
isolates are also present among the other species.
Świętokrzyskie province (Winiary and Suliszów near
Sandomierz).
In three orchards (Lublin, Winiary and Suliszów)
chemical controls were performed in accordance with
the recommendations of plant protection. In the other
two domestic orchards (Bełżyce and Wojciechów),
no chemical protection was applied. The studies included shoots of different cultivars of the studied
plants showing disease symptoms in the form of necrosis or canker. Moreover, for comparative purposes, healthy shoots with no visible disease symptoms
were sampled).
The plant material was collected twice during the
growing season, i.e. the spring and autumn in 2010–
2012. Twenty diseased shoots of each species of fruit
trees were randomly chosen from the protected orchards while ten shoots were taken from the unprotected ones. In addition, five visually healthy shoots
were also collected from each orchard and fruit tree
(tab. 1).
MATERIALS AND METHODS
Mycological analysis
The fungi were isolated using the artificial culture
method [Król 2006]. First, shoot fragments were cut
into pieces the size of a match, and the external tissue
(cortex) was separated from the internal tissue (wood).
Next, the plant material was washed under running tap
water, surface-disinfected for 30 s in a 10% sodium
hypochlorite solution and washed three times for 3 min
in sterile distilled water. Each shoot was treated as
a separate object from which 20 small pieces, 10 of
The research materials were shoots of apple (Malus domestica Borkh.), pear (Pyrus communis L.),
cherry (Prunus cerasus Mill.) and plum (Prunus
domestica L) originating from three orchards located
in the Lublin province (Lublin, Bełżyce,
Wojciechów) and shoots of apple, pear, cherry, plum,
apricot (Prunus armeniaca L.) and peach (Prunus
persica (L.) Batsch) from two orchards in the
Table 1. Number of shoots collected for testing, depending on the host plant and the orchard location (one-time trial)
Host plant
Protected orchards
Unprotected orchards
Lublin
Suliszów
Winiary
Bełżyce
Wojciechów
Apple
Pear
Cherry
Plum
Apricot
Peach
20d + 5vh
20d + 5vh
20d + 5vh
20d + 5vh
20d + 5vh
–
20d + 5vh
–
20d + 5vh
20d + 5vh
–
20d + 5vh
–
20d + 5vh
20d + 5vh
20d + 5vh
10d + 5vh
10d + 5vh
10d + 5vh
10d + 5vh
10d + 5vh
10d + 5vh
10d + 5vh
10d + 5vh
Total
80d + 20vh
80d + 20vh
80d + 20vh
40d + 20vh
40d + 20vh
d – diseased shoots; vh – visually healthy shoots
114
www.hortorumcultus.actapol.net
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
Table 2. Fungi isolated from diseased shoots of fruit plants in the years 2010–2012 (protected orchards)
Number of
shoots
Percentage
(%)
Number of
isolates
Percentage
(%)
Acremonium charticola (Lindau) W. Gams
Alternaria alternata (Fr.) Keissl.
Aspergillus niger Tiegh.
Aureobasidium pullulans (de Bary et Löwenthal) G. Arnaud
Botrytis cinerea Pers.
Chaetomium globosum Kunze
Cladosporium carpophilum Thüm.
Cladosporium cladosporioides (Fres.) de Vries
Cladosporium herbarum (Pers.) Link
Cladosporium sphaerospermum Penz.
Clonostachys rosea (Link) Schroers, Samuels, Seifert et W.
Gams
Colletotrichum gloeosporioides (Penz.) Sacc.
Diaporthe spp. (Phomopsis spp.)
Epicoccum nigrum Link
Fusarium oxysporum Schltd.
Fusarium poae (Peck.) Wollenw.
Fusarium sporotrichioides Sherb.
Gibberella baccata (Wallr.) Sacc.
Gibberella fujikuroi (Sawada) Wollenw.
Haematonectria haematococca (Berk. et Broom) Samuels et
Rossman
Leucostoma personii (Nitschke) Höhn.
Monilinia sp.
Neofabraea alba (E.J. Guthric) Verkley
Neofabraea perennans Kienholz
Paraconiothyrium fuckelii (Sacc.) Verkley et Gruyter
Penicillium expansum Link
Peyronellaea pomorum (Thüm) Aveskamp, Gruyter et Verkley
Phoma glomerata (Corda) Wollenw. et Hochapfel
Phoma herbarum Westend.
Seimatosporium spp.
Trichoderma koningii Oudem.
Trichoderma viride Pers.
Trichothecium roseum (Pers.) Link
Truncatella truncata (Lév.) Steyaert
Saccharomyces spp.
White no sporulating mycelium
Black no sporulating mycelium
32
788
9
21
129
43
48
40
8
3
70
2.22
54.72
0.62
1.49
8.96
2.97
3.33
2.78
0.56
0.21
4.86
128
3758
39
36
368
126
72
120
24
16
143
1.32
38.79
0.40
0.37
3.80
1.30
0.74
1.24
0.25
0.17
1.48
126
134
78
99
6
125
9
283
58
8.75
9.31
9.42
6.88
0.42
8.68
0.62
19.65
4.03
268
295
193
174
17
308
30
887
149
2.77
3.05
1.99
1.80
0.18
3.18
0.31
9.16
1.54
41
19
32
19
16
111
76
10
370
1
71
1
24
2
75
2
9
2.85
1.32
2.22
1.32
1.11
7.71
5.28
0.69
25.69
0.07
4.93
0.07
1.67
0.14
5.21
0.14
0.62
76
42
68
39
33
246
257
26
1178
5
220
3
64
6
246
5
22
0.78
0.43
0.70
0.40
0.34
2.54
2.65
0.26
12.16
0.05
2.27
0.03
0.66
0.06
2,54
0.05
0.23
Total
1440
100
9687
100
Fungus
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115
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
Table 3. Fungi isolated from diseased shoots of fruit plants in the years 2010–2012 (unprotected orchards)
Fungus
Number of
shoots
Percentage
(%)
Number of
isolares
Percentage
(%)
Acremonium charticola (Lindau) W. Gams
Alternaria alternata (Fr.) Keissl.
Aspergillus niger Tiegh.
Botrytis cinerea Pers.
Chaetomium globosum Kunze
Cladosporium cladosporioides (Fres.) de Vries
Cladosporium herbarum (Pers.) Link
Cladosporium sphaerospermum Penz.
Clonostachys rosea (Link) Schroers, Samuels, Seifert et W. Gams
Colletotrichum gloeosporioides (Penz.) Sacc.
Cylindrocarpon obtusisporum (Cook et Harkn.) Wollenw.
Diaporthe spp. (Phomopsis spp.)
Elsinoë ampelina Shear
Epicoccum nigrum Link
Fusarium oxysporum Schltd.
Fusarium poae (Peck.) Wollenw.
Fusarium sporotrichioides Sherb.
Gibberella avenacea (R.J Cook)
Gibberella baccata (Wallr.) Sacc.
Gibberella fujikuroi (Sawada) Wollenw.
Gibberella pulicaris (Kunze) Sacc.
Haematonectria haematococca (Berk. et Broom) Samuels et Rossman
Leucostoma personii (Nitschke) Höhn.
Neofabraea alba (E.J.Guthric) Verkley
Neofabraea perennans Kienholz
Paraconiothyrium fuckelii (Sacc.) Verkley et Gruyter
Penicillium expansum Link
Pestalotiopsis spp.
Peyronella pomorum (Thüm.) Aveskamp, Gruyter et Verkley
Phoma glomerata (Corda) Wollenw. Et Hochapfel
Phoma herbarum Westend.
Sordaria spp.
Stachybotrys chartarum (Ehrenb.) S. Hughes
Trichoderma harzianum Rifai
Trichoderma koningii Oudem.
Trichoderma viride Pers.
Trichothecium roseum (Pers.) Link
Truncatella truncata (Lév.) Steyaert
Saccharomyces spp.
Black no sporulating mycelium
3
118
10
14
13
12
6
2
12
7
1
145
1
49
6
5
12
1
3
65
1
87
3
5
2
4
37
1
20
1
104
1
1
1
34
1
1
2
36
1
0.63
24.58
2.08
2.92
2.71
2.5
1.25
0.42
2.5
1.46
0.21
30.21
0.21
10.21
1.25
1.04
2.50
0.21
0.63
13.54
0.21
18.13
0.63
1.04
0.42
0.83
7.71
0.21
4.17
0.21
21.70
0.21
0.21
0.21
7.08
0.21
0.21
0.42
7.50
0.21
10
300
33
27
49
23
11
3
35
27
1
550
1
113
29
18
55
3
9
228
1
296
3
15
11
11
78
3
71
2
292
3
3
1
88
5
2
4
145
5
0.39
11.70
1.29
1.05
1.91
0.90
0.43
0.12
1.37
1.05
0.04
21.45
0.04
4.41
1.13
0.70
2.14
0.12
0.35
8.89
0.04
11.54
0.12
0.58
0.43
0.43
3.04
0.12
2.77
0.08
11.38
0.12
0.12
0.04
3.43
0.2
0.08
0.15
5.65
0.20
Total
480
100
2564
100
116
www.hortorumcultus.actapol.net
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
Table 4. Fungi isolated from visually healthy shoots of fruit plants in the years 2010–2012 (protected and unprotected orchards)
Number of
shoots
Percentage
(%)
Number of
isolates
Percentage
(%)
Acremonium charticola (Lindau) W. Gams
Alternaria alternata (Fr.) Keissl.
Alternaria tenuissima (Kunze) Wiltshire
Aspergillus niger Tiegh.
Aureobasidium pullulans (de Bary et Löwenthal) G. Arnaud
Botrytis cinerea Pers.
Chaetomium globosum Kunze
Cladosporium cladosporioides (Fres.) de Vries
Cladosporium sphaerospermum Penz.
Clonostachys rosea (Link) Schroers, Samuels, Seifert et W. Gams
Colletotrichum gloeosporioides (Penz.) Sacc.
Diaporthe spp. (Phomopsis spp.)
Epicoccum nigrum Link
Fusarium sporotrichioides Sherb
Gibberella fujikuroi (Sawada) Wollenw.
Haematonectria haematococca (Berk. et Broom) Samuels et
Rossman
Leucostoma personii (Nitschke) Höhn.
Neofabraea alba (E.J.Guthric) Verkley
Penicillium expansum Link
Peyronella pomorum (Thüm.) Aveskamp, Gruyter et Verkley
Phoma herbarum Westend.
Sordaria spp.
Trichoderma koningii Oudem.
Saccharomyces spp.
13
205
1
3
3
28
3
6
5
10
8
34
9
8
5
8
2.17
34.17
0.17
0.50
0.50
4.67
0.50
1.00
0.83
1.67
1.33
5.67
1.50
1.33
0.83
1.33
79
911
5
8
13
74
7
17
31
51
33
95
17
23
20
39
4.23
48.79
0.27
0.43
0.70
3.96
0.38
0.91
1.66
2.73
1.77
5.09
0.91
1.23
1.07
2.09
9
5
6
12
28
4
13
33
1.50
0.83
1.00
2.00
4.67
0.67
2.17
5.50
11
16
21
50
131
10
42
163
0.59
0.86
1.12
2.68
7.02
0.53
2.25
8.73
Total
600
100
1867
100
Fungus
the outer and inner tissues, were placed in Petri dishes with Czapek-Dox agar (BD Difco). Plates were
incubated at 24°C in the dark for 12 days. Growing
fungal colonies were transferred to 2% potatodextrose agar (PDA, BD Difco) slants for further
identification. In the case of Diaporthe isolates,
which sporulate with difficulty, they were cultured in
a poor PDA medium containing carnation leaf to
stimulate sporulation [Castillo-Pando 1997]. Single
spore colonies were prepared for morphological identification using the serial dilution method.
www.hortorumcultus.actapol.net
The system by Kirk et. al. [2008] was adopted as
a basis for fungi classification and the names of fungi
were given according to the current taxonomic status
of the species in the Index Fungorum database.
RESULTS
Mycological analysis of fruit trees resulted in
14.118 fungal isolates belonging to more than
30 species within three years of research. Among
them, 9.687 isolates originated from shoots showing
117
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
various disease symptoms and harvested from chemically protected orchard, 2.564 from unprotected
shoots and 1.867 cultures from apparently healthy
ones (tabs 2, 3 and 4). The species composition of the
fungi colonizing shoots of examined host plants was
similar in the years of the study. Alternaria alternata
and Fusarium spp. were isolated most frequently,
regardless of the orchard and the host plant.
Table 5. Fungi most frequently colonizing shoots of fruit plants in protected orchards in the years 2010–2012
Percentage of fungi
Fungus
2010
2011
2012
et
it
et
it
et
it
Alternaria alternata
50.82
37.04
44.43
29.10
49.75
18.42
Botrytis cinerea
2.05
0.59
3.64
7.15
3.44
3.90
Cladosporium spp.
1.43
0.59
5.9
1.04
1.64
1.52
Clonostachys rosea
0.62
1.42
0.7
2.42
0.45
3.58
Diaporthe spp./Phomopsis spp.
0.96
0.24
3.52
3.41
1.97
6.76
Fusarium spp. + Gibberella spp. + Haematonectria haematococca
20.96
20.36
12.72
10.29
17.94
19.36
Phoma spp. + Peyronella pomorum
10.62
15.50
8.76
20.68
11.22
26.51
Trichoderma spp.
0.21
0.24
4.95
4.29
0.50
1.19
Other species of fungi
12.33
24.02
15.38
21.62
13.09
18.76
et – external tissue, it – internal tissue
Table 6. Fungi most frequently colonizing shoots of fruit plants in unprotected orchards in the years 2010–2012
Percentage of fungi
Fungus
Alternaria alternata
2010
2011
2012
et
it
et
it
et
it
17.70
4.49
12.57
7.88
11.65
11.37
0
0
1.77
2.22
1.61
0
Cladosporium spp.
0.4
0
2.16
2.96
1.79
0.58
Clonostachys rosea
1.19
6.53
0.79
2.22
0
0
Diaporthe spp./Phomopsis spp.
1.79
39.18
25.15
54.93
6.63
16.62
Fusarium spp. + Gibberella spp. + Haematonectria haematococca
44.93
13.47
17.29
6.89
32.44
24.20
Phoma spp. + Peyronella pomorum
19.68
24.49
10.02
6.89
13.26
15.45
Botrytis cinerea
Trichoderma spp.
0.99
0
4.91
2.46
6.10
5.83
Other species of fungi
13.32
11.84
25.34
13.55
26.52
25.95
et – external tissue, it – internal tissue
118
www.hortorumcultus.actapol.net
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
spring
30
autumn
25
24,7
20
14,1
15
10,4
9,8
10
8,3
7,1
6,9
5,8
5
1,7 2,1
1,4 0,8
2,1
1,2 1,1
1 0,5
0,9
other species
Trichoderma spp.
Phoma spp.
Clonostachys
rosea
Fusarium spp.,
Gibberella spp.,
Haematonectria
Diaporthe spp.
Cladosporium
spp.
A. alternata
B. cinerea
0
Fig. 1. Percentage of selected species of fungi inhabiting the shoots of fruit plants in 2010–2012 depending on term of
studies (protected orchards)
20
spring
18,5
autumn
18
16
13,9
14
12
11,4
11,1
10
8,6
8
6
7,1 7,2
5,9 5,8
4
3
2
1
0
0
A. alternata
B. cinerea
2,7
1,3
0,5 0,9
Cladosporium
spp.
1
0,1
Diaporthe spp.
Fusarium spp.,
Gibberella spp.,
Haematonectria
Clonostachys
rosea
Phoma spp.
Trichoderma
spp.
other species
Fig. 2. Percentage of selected species of fungi inhabiting the shoots of fruit plants in 2010–2012 depending on term of
studies (unprotected orchards)
www.hortorumcultus.actapol.net
119
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
Every year of the study Alternaria alternata was
isolated from diseased shoots originating from both
protected and unprotected orchards. The cultures of
this fungus were isolated from 788 (54.72%) and 118
(24.58%) of the studied shoots and 3.758 (38.79%)
and 300 (11.70%) isolates of this fungus were obtained from them, respectively (tabs 2 and 3). Alternaria alternata was isolated most frequently in 2010
from the protected orchards (tab. 5). This fungus
species mostly inhabited the external tissue of the
shoots collected in the spring season (tabs 5 and 6,
figs 1 and 2).
Among the fungi often isolated from diseased
shoots were also species of the genus Fusarium.
This genus had been previously described within
the class Deuteromycotina because the perfect stage
of Fusarium spp. was not known. Currently, according to the latest taxonomy, the genus Fusarium
is classified within the division of Ascomycota. The
species in which the perfect stage is not known
constantly belong to Fusarium, while species which
produce the teleomorph stage belong to the genus
Gibberella or Haematonectria [Kirk et al. 2008,
http://www.indexfungorum.org/names/names.asp].
The above mentioned fungi were obtained from
580 shoots (40.28%) originating from protected orchards and from 180 (37.51%) shoots collected from
unprotected ones. From these shoots, 1.565 (16.17%)
and 639 (24.91%) fungal isolates were obtained,
respectively (tabs 2 and 3). In the case of diseased
shoots from chemically protected orchards the percentage of these fungi was the highest in 2010, both
from external and internal tissue (tab. 5). Moreover,
in the years of the study, Fusarium spp. were isolated
more frequently in the autumn than in the spring (fig.
1). Gibberella fujikuroi (anamorph F. verticillioides)
and F. sporotrichioides were the most often isolated.
These fungi species were isolated respectively from
283 (19.65%) and 125 (8.68%) shoots, respectively,
to give a total 9.16% and 3.18% of all fungal isolates
(tab. 2). In unprotected orchards, Fusarium spp. were
isolated most frequently in 2010 and 2012, mainly in
the period of autumn and from the external tissue of
shoots (tab. 6, fig. 2). Haematonectria haematococca
(anamorph F. solani) was isolated most frequently
120
and cultures of this fungus were isolated from 87
(18.13%) shoots, resulting in 11.54% of all fungal
isolates. Gibberella fujikuroi (anamorph F. verticilioides) was also isolated quite often. The fungus inhabited 65 (13.54%) shoots and gave a total 8.89% of
isolates (tab. 3).
Cultures of Phoma spp. (including Peyronella
pomorum) were isolated each year from all of the
studied orchards and the percentage of these fungi
constituted 15.07% and 14.23% from chemically
protected and unprotected orchards, respectively
(tabs 2 and 3, figs 1 and 2). In the first orchard Phoma spp. were isolated most frequently in 2012 and
mainly from internal tissue, while in the second ones
the highest percentage of Phoma isolates were obtained in 2010 and they originated mainly from internal tissue, too (tabs 5 and 6). Phoma herbarum was
isolated most frequently and its isolates constituted
12.16% and 11.38% of the total number of isolates in
protected and unprotected orchards, respectively
(tabs 2 and 3). Moreover, Phoma spp. were isolated
in similar numbers in the spring and autumn seasons
(figs 1 and 2).
The percentage of Botrytis cinerea cultures isolated from shoots originating from protected orchards
was higher than the percentage of isolates from unprotected ones (tabs 2 and 3). From the protected
shoots B. cinerea was isolated most frequently from
internal tissue in 2011 and the number of isolates of
the fungus predominated slightly in the autumn (tab.
5, fig. 1). In the unprotected orchards, B. cinerea was
isolated only in the last two years of research and
only in the spring season. This fungus species inhabited both external and internal tissue (tab. 6, fig. 2).
Species isolated in almost all years of the study
also included Clonostachys rosea (syn. Gliocladium
roseum, G. catenulatum), Cladosporium spp. and
Trichoderma spp. (tabs 2 and 3). Clonostachys rosea
inhabited the shoots of fruit trees with similar frequency in protected and unprotected orchards because its cultures were respectively 1.48% and 1.37%
of the total number of isolates (tabs 2 and 3).
Clonostachys rosea more frequently inhabited the
internal tissue of the examined shoots but in protected orchards the fungus was isolated especially in the
www.hortorumcultus.actapol.net
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
autumn season while in the unprotected orchards it
was found mainly during the spring season (tabs 5
and 6, figs 1 and 2).
Among fungi of the genus Cladosporium the most
isolates were represented by species C. cladosporioides, in all of the tested orchards (tabs 2 and 3). The
majority of Cladosporium cultures were obtained in
2011 both from shoots originating from protected and
unprotected orchards, especially from the external
tissue (tabs 5 and 6, figs 1 and 2).
Cultures of Trichoderma spp. were also isolated
in every year of study, which were obtained most
frequently in 2011 from chemically protected orchards and in 2012 from unprotected ones, both from
the external and internal tissues (tabs 5 and 6, figs 1
and 2).
Particularly noteworthy is the fact that Diaporthe
cultures were isolated every year of the study. These
fungi were obtained respectively from 134 (9.31%)
and 145 (30.21%) shoots originating from protected
and unprotected orchards, obtaining 3.05% and
21.45% isolates, respectively (tabs 2 and 3). Diaporthe isolates were obtained mainly in 2011 and
2012 from the protected orchards, while the isolates
were obtained in 2011 from unprotected ones. These
fungi usually inhabited the internal tissue of the
shoots and more often were isolated in the spring
than in the autumn (tabs 5 and 6, figs 1 and 2). Moreover, Diaporthe cultures were obtained especially
from shoots of apple and cherry from the protected
orchards and from shoots of pear and plum from
unprotected ones (figs 3 and 4).
shoots
90
isolates
79
80
73
70
60
50
45
37
40
39
34
31
30
25
25
21
20
11
10
9
0
apple
pear cherry plum apricot peach
Fig. 3. The number of shoots and isolated from them cultures of Diaporthe
(Phomopsis) in 2010–2012 depending on the plant host (protected orchards)
www.hortorumcultus.actapol.net
121
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
shoots
200
isolates
178
180
156
160
133
140
120
100
83
80
60
39
43
39
40
24
20
0
apple
pear
cherry
plum
Fig. 4. The number of shoots and isolated from them cultures of Diaporthe
(Phomopsis) in 2010–2012 depending on the plant host (unprotected orchards)
Other species of fungi were rare but among them
were species known for their pathogenic abilities
towards fruit trees, such as Colletotrichum gloeosporioides, Leucostoma personii, Monilinia spp. and
Neofabraea spp. (tabs 2 and 3).
Among fungi isolated from apparently healthy
shoots were also Diaporthe isolates, which were
obtained from 34 (5.67%) shoots, giving 5.09% isolates (tab. 4). In addition, Alternaria alternata was
isolated most frequently because it inhabited 34.17%
of the studied shoots and its isolates constituted
48.79%. Numerous cultures of yeast as well as Acremonim charticola, Botrytis cinerea and Phoma
herbarum were also obtained, whose isolates were
4.23%, 3.96% and 7.02%, respectively (tab. 4).
DISSCUSION
The results of the mycological analysis of apple,
pear, cherry, plum, apricot and peach shoots confirm
previous reports of the colonization of fruit tree
122
shoots by numerous species of fungi, among which
there are saprotrophic species, potential plant pathogens and antagonistic ones [Grabowski 1993,
1996, Grabowski and Leś 1996, Grabowski 2002].
The small variation in the species composition of
fungi found on different host plants shows their ability to colonize different species of fruit trees growing
in the orchard.
In the present study, obtaining the greater percentage of most fungi from shoots showing different
disease symptoms than from apparently healthy ones
indicates the preferences of saprotrophic species as
well as some of the pathogens to colonize decaying
wood, which is treated as a source of nutrients.
According to Grabowski [2002], undamaged shoots
are less attractive substrates due to the presence of
tannins and phenols, which are unfavourable for fungi development.
The evident dominance of Alternaria alternata on
shoots originating from protected orchards is probably due to the common occurrence [Tylkowska et al.
www.hortorumcultus.actapol.net
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
2003, Stuart et al. 2009] and resistance of this fungus
on some of the preparations used in plant control
against a variety of pathogens, as previously reported
by other researchers [Timmer and Zitko 1997,
Osowski 2004, Pres and Timmer 2006, Stuart et al.
2009].
These authors pointed out a complete lack of benzimidazole efficiency (thiophanate-methyl) in protection against A. alternata [Timmer and Zitko 1997,
Stuart et al. 2009] explaining this phenomenon as the
ability of the fungus to metabolize the ingredients of
the listed fungicides. Furthermore, Timmer and Zitko
[1997] proved the resistance of A. alternata to strobilurin. In turn, Pres and Timmer [2006] have shown
little effect of mancozeb on reducing the development of A. alternata, although Osowski [2004] reported that the efficacy of fungicides containing
mancozeb was better in controlling of potato antracnose when it was used in a mixture with another active ingredient.
It seems, that A. alternata had favourable conditions for development on the fruit tree shoots in protected orchards despite the intensive chemical control, especially since at the same time phyllosphere
decreased the number of the other species of fungi in
the shoots, that could compete with it. This hypothesis seems to be confirmed by the fact that at the same
time a smaller number of A. alternata cultures were
isolated from the shoots originating from unprotected
orchards, where a greater diversity of fungi species
was obtained.
Obtaining numerous isolates from the widely understood genus Fusarium, both from the shoots originating from the protected and unprotected orchards,
indicates their importance in the shoots’ environment,
especially because among them there are numerous
pathogens of many plant species [Kawchuk et al.
2002, Frużyńska-Jóźwiak and Andrzejczak 2010,
Kiecana and Mielniczuk 2010, Kiecana et al. 2012,
Danielewicz et al. 2013, Walkowiak and KrzyśkoŁupicka 2014].
The occurrence of Fusarium spp. on the shoots of
fruit trees is probably due to their common presence
in nature, their fast growth, which makes it easier to
substrate colonize, competition with the other species
www.hortorumcultus.actapol.net
of fungi as well as frequently appearing resistance of
these fungi to fungicides, which was observed by
Frużyńska-Jóźwiak and Andrzejczak [2010], Danielewicz et al. [2013] and Walkowiak and KrzyśkoŁupicka [2014], among others. They found that the
proper selection of plant protection products affects
the effectiveness of reducing the occurrence of
Fusarium spp., because some of them are very effective while others, such as thiophanate-methyl and
azoxystrobin, only slightly inhibit the growth of these
fungi. On the other hand, Kawchuk et al. [2002] suggest that the resistance of isolates results from their
origin from intensively protected plants.
The isolation of fungi species known for pathogenic abilities to fruit trees, i.e., Botrytis cinerea,
Coniothyrium fuckeli, Colletotrichum gloeosporioides, Neofabraea spp., Fusarium spp. and Cytospora leucostoma and numerous saprotrophic species as:
Epicoccum nigrum, Chaetomium globosum, Phoma
herbarum, Trichoderma koningii and Clonostachys
rosea both from shoots originating from protected
and unprotected orchards, shows a large role of
shoots in the transmission of pathogenic factors. Very
important is the fact that some of these fungi cause
not only bark and wood diseases but also threaten the
fruit during growth and storage. Alternaria spp.,
Fusarium avenaceum and Botrytis cinerea cause,
among others, leaf blotch and fruit spot, mouldy core,
core browning and core rot, Neofabraea spp. bitter
rot of apples and Glomerella cingulata (C. gloeosporioides) bitter rot of cherry [Gao et al. 2013,
Wenneker et. al. 2016].
Particularly noteworthy is the isolation of fungi of
the genus Diaporthe, the importance of which systematically increases [Machowicz-Stefaniak 1993,
Uddin et al. 1997, 1998, Farr et al. 1999, 2002,
Kačergius et al. 2004, Karaoglanidis and Bardas
2006, Król and Kowalik 2010, Mirabolfathy 2013].
The isolation of these fungi from shoots of apple,
pear, cherry, plum, apricot and peach trees indicates
the presence of these fungi in orchards in the provinces of Lublin and Świętokrzyskie, which is the first
documented announcement of their occurrence in the
conditions of our country. Thus, there is an increased
number of fungi from the genus Diaporthe on fruit
123
Król, E.D., Abramczyk, B.A., Zalewska, E.D., Zimowska, B. (2017). Fungi inhabiting fruit tree shoots with special reference to the
Diaporthe (Phomopsis) genus. Acta Sci. Pol. Hortorum Cultus, 16(4), 113–126. DOI: 10.24326/asphc.2017.4.12
plants in Polish conditions, because previously their
existence had been documented only on grapevine
[Machowicz- Stefaniak 1993, Król 2002, 2005, 2006,
2007, Król and Kowalik 2010] and blueberry
[Szmagara and Machowicz-Stefaniak 2005, Szmagara 2009].
However, non-specific symptoms caused by these
fungi creates problems with their detection in fruit
crops, because often Diaporthe isolates were obtained from shoots showing various blotch symptoms, necrosis or shoot damping, which were similar
to the symptoms caused by the other bark and wood
pathogens. Fungus conidiomata formation on diseased shoots could be useful in its identification, but
their occurrence in natural conditions was not observed in the years of the study. In addition, colonization by Diaporthe of also visually healthy shoots, as
demonstrated during the research, indicates the ability of these fungi for asymptomatic growth in the
shoots of fruit trees, which creates the danger of unknowingly carrying them with the nursery material
on long distances, as reported earlier by Mostert et al.
[2000], Halleen et al. [2003] and Król [2006].
CONCLUSIONS
1. The shoots of fruit trees are inhabited by many
species of fungi among which are both saprotrophic
ones as well as species known for pathogenic abilities
to bark and wood and to fruit during the vegetation
period and storage.
2. The isolation of Diaporthe cultures from the
shoots of apple, pear, cherry, plum, peach and apricot
indicates the presence of these fungi in the investigated orchards of the Lubelskie and Świętokrzyskie
provinces, which is the first information about the
occurrence of Diaporthe on fruit trees in Poland.
3. The more frequent isolation of Diaporthe from
shoots originating from unprotected orchards than
protected ones indicates their greater threat to fruit
trees grown without chemical protection.
4. Non-specific disease symptoms caused by Diaporthe hinder the identification of these fungi on the
shoots of fruit trees.
124
ACKNOWLEDGMENTS
The project was financed by the Polish Ministry
of Science and Higher Education within the framework of the KBN research project: N N310 774940.
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