FUNGI ASSOCIATED WITH SORGHUM SEEDS

by

V. S. VER~IA 1) & ABRAR M. KHAN

Department o~ Botany, Aligarh Muslim University, Aligarh, India.

(15.1.1965)

A large number of plant parasitic as well as saprophytic organisms

have long been known to be transmitted through the seeds. Fungus
flora is b y far the most important which is disseminated with the
seeds of cultivated as well as natural plants. These fungi and bac-
teria m a y be present on the surface in an actively growing form
or as thick-walled resting mycelium, chlamydospores, resting
spores or as dormant mycelium within the seed itself. Some of
the reported cases of surface borne pathogens on the seed are
Sorghum Smut (Sphacelotheca sorghi), Bunt of wheat. A number
of fungi are known to be carried within the true seeds, e.g., An-
thracnose of wheat (Colletotrichum cereale), Loose smut of wheat
and barley etc. Besides this Anthracnose of cotton and beans,
Helminthosporiose of Rice, Red Rot and Smut of Sugarcane are
also seed transmitted. Such diseases are known as seed borne
diseases and are responsible for serious economic losses. These
fungi cannot be said to belong to any particular class or group.
Many of the seed borne pathogens are virulent parasites with
their existence closely linked to that of the seed plant, some are
saprophytic b u t behave as parasites under certain conditions;
others appear to exert no particular influence one w a y or the other
while a few must be regarded as beneficial. Among cultivated
crops, certain of these fungi cause destructive diseases, as for
instance seedling blight, damping off etc. and reduce the emergence
and germination of seedlings and in turn yield of crop is reduced.
Seed Certification is the most important part of Plant Quarantines,
where seeds are tested for external and internal fungi and only
those seeds are imported which are free from disease. Seed testing
assumes all the greater importance in the present context of our
country's food situation as it helps in building up improved seed
stocks, the use of which is one of the recommended methods for

i) Present address: Mycology & P l a n t Pathology Division, I n d i a n Agricultural

Research Institute, New Delhi-12. India.
 F U N G I ASSOCIATED WITH SORGHUM SEEDS 315

stepping up of food production. Seed Testing for the export of

seed samples and the issuing of the necessary health certificates
is being attended to at the Indian Agricultural Research Institute.
During the past 24 months, 1097 seed samples of different crops
were tested in the Division of Mycology and Plant Pathology for
this purpose and 61 samples were rejected owing to their infesta-
tion with pathogenic organisms. Since production of healthy crops
is the most important object of modern farming, it is necessary to
investigate seed borne pathogens.
In India, seed borne diseases of important Cereal crops cause
great losses. VASVDF~VA (Proc. Eight meeting of the Crops and
soils wing of the Board of Agric. and Animal Husbandry in India
held at Patiala, 1950) stated that the grain smut of Jowar was
estimated to cause an annual loss of 325,000 tons while the loss in
grain yield due to loose smut of wheat is about 100,000 tons. On
the basis of such estimates, it has been stated that losses due to
all seed borne diseases caused b y fungi, on our chief cereal crops
alone, would easily run into a million tons a year or in other words
they would be equivalent to more than a quarter of our estimated
annual deficit of cereal food grains in this country.
In order to avoid such fungi being carried with the seeds, attempts
have been made in some countries to prepare lists of recorded seed
borne fungi. This has received impetus due to tile efforts of organiza-
tions like the International Seed Testing Association. Critical studies
have been made of the fungal flora of seeds in Canada and special
attention has been paid to the seed-borne fungi of corn, flax,
peas and other vegetables but there is a very meagre information
about the seed borne fungi of Sorghum. BAIN (1950) has reported
a number of fungi viz., Acrothecium, AZternaria, Aspergillus, Chaeto-
mdla, Chaetomium, Cladosporium, Fusarium, Helieosporae, Hd-
minthosporium, Rhizopus, Sordaria and other unidentified fungi
from Sorghum varieties of Lousiana. In an experiment of one
Sorghum seed treatment in Georgia, HARRIS & LVTTRELL (1954)
identified moulds namely Aspergillus sp., Penicillium sp., Hdmin-
thosporium (Brachycladium) sfleci/erum. (BAIN) NICOT, Curvularia
lunata (WAKI~.) BOED. LEUKEL & MARTIN (1943)have reported
that species of Alternaria, Fusarium, Penicillium, Aspergillus and
Rhizopus are found on Sorghum seeds in abundance, besides other
Sphacelotheca & Helminthosporium.
It is evident that practically no information is available about
the seed borne fungi of Indian Sorghum varieties as well as those
being released b y the Rockefeller Foundation in India. It was,
therefore, considered desirable to identify all the fungi found on
Sorghum seeds of these different varieties. The present paper de-
scribes the different fungi recovered from some selected Indian
varieties and some of those supplied through the courtesy of the
Rockefeller Foundation in India.
316 V. S. VEI~fA ~ A. M. KHAN

M A T E R I A L AND M E T H O D S

Spore suspension of the Superficial fungi on different Sorghum

varieties were prepared b y shaking a few seeds of the different
varieties with a little q u a n t i t y of sterilized water obtained in Test
tubes. The suspension was poured in plates containing Potato
dextrose agar. The plates were then incubated at room temperature
(approximately 25 ° C) for a week. Colonies of the different fungi
that developed were identified and isolated in pure cultures.
In order to study the internal fungi, seeds of different varieties
were surface sterilized in rectified spirit for 45 seconds. The seeds
were then rinsed twice in sterilized water and planted in plates con-
taining Potato dextrose agar. The plates were then incubated at
room temperature (about 25 ° C) for 8--10 days. Colonies of the
different fungi t h a t developed were identified and isolated in pure
cultures.

EXPERIMENTAL RESULTS

Indian Sorghum varieties from which the superficial and Endo-

phytic fungi were recovered are given below.
Cholan Co. - - 1, Cholan Co. - - 4, Cholan Co. - - 9, Cholan Co. --- 11,
Cholan Co. - - 12, 48/8A, 48/10A, 48/10(a), 49 Local, 52/6, Malva
53/1, 53/2, 53/6, 4102, 4106(a) - - A, 4403 ]3, Milo - - 4 and Kanpur
Local.
Sorghum varieties being introduced by the Rockefeller Founda-
tion were also used ill these tests and are listed below: Broom
corn 2 black spanish, Kaoling 301, F P I 62610, Milo 332 Single
dwarf, Shallus 475, Combine Shallu, Yellow Endosperm 503, Forage
676, Miscellaneous Grain variety 849.
Misc. Grain variety 854, African variety 901, Indian variety
1013, Indian variety 1021, Indian variety 1060. These seeds had
been treated previously with a mixture of equal parts of D.D.T.
10% and copper carbonate. Complete lists of the different external
and Internal fungi recovered from all the Sorghum varieties are
given in Table I and II.

CONCLUSION

It is evident from the above experiments t h a t Sorghum seeds

carry a large number of fungi. Of these, fifteen are external fungi,
and another fifteen occur as endophytic fungi. Only three members
of Phycomycetes viz., Phycomyces, Circinella, Syncephalastrum
were recovered on two particular varieties, while none of them were
known to develop on surface sterilized seeds. Species of Penicillium
and Aspergillus occur regularly as common molds on seeds exter-
nally as well as internally. Aspergillus circinatus L. MANGIN has
been reported to cause the Milo Root Rot of Sorghum. There is
one more genus of Ascomycetes namely, Chaetomium which has
 FUNGI ASSOCIATED "WITH SORGHUM SEEDS 317

been recovered in m a n y varieties, but externally on a single variety

Milo - - 4. No representatives of Basidiomycetes were recovered as
most of them, like rusts and smuts, are obligate parasites. Majority
of the fungi recovered belong to the group Fungi Imperfecti. Nine
members of this group have been recovered as external fungi and
twelve as internal fungi. Fusarium, Curvularia, Pullularia, Tricho-
derma are recovered on a large number of varieties as external
fungi, while others, namely Cladosporium, Helminthosporium,

TABLE I.
List o/ External Fungi recovered from all sorghum varieties.

S. N o . External Fungi recovered Taxonomy of t h e f u n g i r e c o v e r e d

1. Phycomyces nitens AGARD~ Phycomycetes, Mucorales, Mucoraceae

2. Circinella simplex VAN TIXGI~EM ,,
3. Syncaphalastrum raeemosum
GCHROBTER ,, Cephalidaceae
4. Chaetimium ]imeti FUCKEL Ascomycetes, Sphaeriales, Chaetomiaceae
5. Curvutaria lunata ~OI~DIJN Imperfecti, MonJliales, Dematiaceae
6. Cladosporium /ulvum L I N K . . . .
7. Helminthosporium sativum
PAMMEL, K I N G , B A K K E ,, ,,
8. Hetminthosporium anomalum
GILMAN & ABBOTT . . . .
9. Montospora brevis MASON . . . .
10. Pullularia pullulans BBRKI~OUT ,,
11. Aspergillus candidus L I N K . . . .
12. Aspergitlus luehanensis t N u I ,,
13. A. niger VAN TIEGHEM . . . .
14. A. sydowi (B & C) THOM a n d CHIJRCH . . . .
15. Cephalosporium aspermum MARCHEL . . . .
16. Pencittium cesiae
BAINIER ~C S A R T O R Y ,, ,, Monitiaceae
17. Penieillium spinulosum THOM . . . .
18. Penieiltium Waksmani ZALESKI ,, ~J
19. Trichoderma lignorum ( T O P E ) H A E Z ,,
20. Phoma insidiosa ,, Spharopsidales Spharioidaceae
21. Fusarium conglutinans WOLLtgNWEBER . . . . Tuberculariaceae
22. Fusarium lalertium S E E S . . . .
23. Fusarium sporotriehiodes St~Bt~BAKOFF . . . .
24. Bacteria

Montospora, Cephalosporium and Phoma are observed on certain

particular varieties. Fusarium, Phoma, Pullularia and Curwdaria
were recovered as internal fungi on a large number of varieties,
while others namely Hormodendron, Alternaria, Cladosporium,
Helminthosporium, Heterosporium, Blastomyces, Monilia, Phomopsis
were recovered on certain particular varieties. In this respect these
results confirm those of BAIN (1950) and HARRIS & LUTTRELL
(1954). As stated b y BAIN, Acrothecium, Chaetomella, Helicosporae,
Sordaria were not recovered, but a few new fungi were recovered
 318 v.s. VERMA & A. M, KHAN

and isolated namely Pullularia, Hetero@orium, Monilia, the patho-

genicity of which is not yet established. There are some varieties the
seeds of which carried the same fungus internally as well as exter-
nally, while others differ in this respect. Most common example is
that of variety Kanpur Local b y which Helminthosporium anomalum
has been recovered both internally as well as externally. CICCARONE
(1949) has reported the occurrence of Gloeocercospora sorghi BAIN
& EDG. on sorghum seeds in the United States. However, i t could

TABLI~ I I .
L i s t O/ Internal fungi recovered [rom all sorghum, varieties.

S. N o . Internal Fungi Recovered Taxonomy of F n n g i recovered

1. Chaetomium succineum AMES Ascomycetes, Sphaeriales, Chaetomiaceae

2. Ctadosporium lignicola CORDA Imperfecti, Mcniliales, Dematiaceae
3. Curvularia tunata (WALKER)
BO~DIJN ,, ,, J,
4. H e l m i n t h o s p o r i u m a n o m a l u m
GILMAN ~ ABBOTT j, ,, 3,
5. Heferosporium gracila JKLOTZSCH . . . . ~J
6. Hormodendron dadosporiodes
SACCARDO ,, ,,

7. P u l l u l a r i a pullulans BERKHOUT . . . .
8. Alternaria geophila DASZEWSKA . . . .
9. AspergilIus luchanensis I N t o . . . . Monitiaceae
10. Aspergillus candidus L I N K . . . . ,,

11. Aspergillus niger VAN T I E G H n M . . . .

12. Aspergiltus wentti W'EHMEN . . . . ~J

13. Aspergitlus sydowi T H o M & CH~'RCH . . . .

14. Blasto~ezyces dermatidis COST & R O L L . . . .
15. M o n i l i a grisea DASZEWSKA . . . .
16. P e n i c i l l i u m cesiae
BAtNIER ~; SATORY ,, ,,
17. Penicillium ]ellutanum BIOURGE . . . . J,

18. PeniciIlium s p i n u l o s u m Tl-lOM . . . .

19. F u s a r i u m e o n g t u t i n a n s WoLLl~NWEBI~R . . . . Tuberculariacea e
20. F u s a r i u m ehlamydosporum
"VVOLLENWEBER & REINKING ,,
21. P h o m a insidiosa ,, Sphaeropsidales Sphaerioidaceae
22. P h o m o p s i s vexans SACC. ,, ,,
32. Bacteria

not be detected on these varieties.

In the present investigation the size and diameter of spores
were not measured but much attention has been paid on the
shape and colour of spores and also on the colour of colonies for
identification. Fusarium and Penicillium could not always be
identified upto the species which are known to occur. Although
Phoma was not identified to the species, it is of interest to note
that t9. insidiosa has been reported (KocH & RUMBOLD, 1921)
causing a disease of Sorghum in Abyssinia and the fungus was
 FUNGI ASSOCIATED 'WITH SORGHUM SEEDS 319

orginally described on Sorghum seeds from the same country.

Bacteria were found to occur on many Sorghum varieties both on
the surface as well as internally, but they could not be identified.
It has been observed that they destroy the seed and check the
growth of other pathogens. However there are some seed borne-
fungi which are considered to be beneficial for the emergence and
growth of the seedling. For instance species of A spergillus, Penicil-
lium, Chaetomium and certain bacteria have recently been found
to produce antibiotics and protect the seedling from root rot causing
fungi and other soil-borne pathogens. Some of the seed borne fungi,
chiefly species of Fusarium, A@ergillus, Rhizopus, Penicillium,
Hdminthosporium have been reported to invade and destroy the
endosperm or starchy tissue of the seed (LEUKEL~: MARTIN, 1955).
Some race of Fusarium have been reported b y LEUKEL & MARTIN
(1955) to be more harmful than others. They have reported that
one species of Penicillium, namely P. oxalicum not only attacks
the endosperm and arrests germination but also kills the seedlings.
It is evident from the experimental results that the fungi recov-
ered form Indian varieties were more in number than those re-
covered from the varieties recently released b y the Rockefeller
Foundation. This can be explained as the varieties released b y
the Rockefeller Foundation might be improved or it might have been
due to the treatment with fungicide (D.D.T. 10% and copper
carbonate). The variety 4403 B of the Old Collection was known
to give a poor germination in separate seed germination tests.
It might be due to a large number of seed borne fungi. An Indian
variety 1060 released b y the Rockefeller Foundation, variety 503
yellow endosperm have been found to carry a large number of seed
borne fungi. There is a variety Black Spanish I.S.2 which is quite
free from both superficial and internal seed borne fungi. There
are certain other varieties which are free from endophytic fungi
but possess external fungi only. The examples of such varieties
are Kaoling 301, African variety 901 and Shallus 475.

Summary
A study of the external and internal fungi associated with dif-
ferent varieties of Sorghum seeds has been made. The varieties
tested included eighteen local varieties and twelve new Sorghum
varieties obtained from the Rockefeller Foundation. The external
fungi were studied b y preparing suspensions of superficial fungi
and growing on potato dextrose agar. The internal fungi were
studied b y planting surface sterilized seeds on P.D.A. and pure
cultures of atI these fungi were prepared.
The external fungi found to be associated with the different
varieties included different species of Phycomyces, Circinella, Syn-
cephalastrum, Chaetomium, Curvularia, Cladosporium, Helmintho-
sporium, Monto@ora, Pullularia, A slbergillus, Penicillium, Cepha-
320 v.s. WRMA & A. M. ~HAN

losporium, Trichoderma, Photon, Fusarium.

The internal fungi recovered from these varieties included species
of Chaetomium, Cladosporium, Curvularia, Helminthosporium,
Heterosporium, Hormodendron, Pullularia, Alternaria, Aspergillus,
Blastomyces, Monilia, Penicillium, Fusarium, Photon, Phomopsis.
Varieties 4403B, 1060 and 503 were found to carry a large number
of fungi. Variety Black spanish was found to be entirely free from
any internal or external fungus. Varieties which were free from
endophytic fungi but possess external fungi only were Kaoling 301,
African variety 901 and Shallus 475. Three fungi viz., Pullularia,
Heterosporium, Monilia have been recovered from the Sorghum
seeds for the first time.

Acknowledgement

The writers are greatly i n d e n t e d to Dr. K. A. CHOWDHURY,

Head of the Department of Botany for offering them necessary
facilities and encouragement during the com-se of this investigation.
Thanks are also due to Dr. Kenneth O. RACHIE, Pit. D., of the
Indian Agricultural Research Program, Rockefeller Foundation
for kindly supplying the seeds of Sorghum varieties used in these
investigation.

References

BAIN, D. C. (1950) Fungi recovered from seeds of Sorghum vulgate. Phytopathoiogy

11: 521--522.
BARNETT, H. L. (1955) Illustrated genera of Imperfect fungi. Burgess Publishing
Co., Minnesota.
CHILTON, S. J. P. (1940) The occurrence of Hetminthosporium turcicum in the seed
a n d glumes of Sudan grass. Phytopathology 3 0 : 533.
ClCCARONE, A. (1949) Zonate Leaf Spot of Sorghum in Venezuela. Phytopathology
3 9 : 760--761.
DICKSON, J. G. (1950) Diseases of Field crops, Me. Graw Hill Book Co., London.
DoYER, L. (1938) Manual for determination of Seed borne diseases. Edited b y
the I n t e r n a t i o n a l Seed Testing Association.
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MUSKETT, A. E. (1950) Seed borne fungi and their Significance. Trans. Brit. mycol.
Soc. 33: 1.
LEUKEL, R. W. & J. H. MARTIN (1955) Sorghum diseases a n d their control. U.S.
Dept. Agric. Bull., 1959.
SKOLKO, A. J. & J. W. GRovEs (1953) Notes on Seed Borne fungi Chaetemium.
Canad. J. Res. 31: 779.