UK Journal of Pharmaceutical and Biosciences Published its Most Recent Issue; Vol 3 (2)

1. UK Journal of Pharmaceutical and Biosciences Vol. 3(2), 42-45, 2015 RESEARCH ARTICLE
Mycoflora Associated with Different Varieties of Cotton (Gssypium arboreum, L.)
Anil Kumar Kushwaha
Department of Botany, D.B.S. College, Kanpur - 208001, Uttar Pradesh, India
Article Information
Received 3 March 2015
Received in revised form 23 April 2015
Accepted 28 April 2015
Abstract
Different varieties of Gossypium arboreum,L. viz; Lohit, RG-8, LD-327, and Sanjay seeds were
evaluated for field and storage fungi from large number of samples from different state of India.
Thirty seven fungal species were isolated from cotton seeds by different methods.
Keywords:
Gossypium arboreum,
Mycoflora,
PDA,
Blotter method
*
Corresponding Author:
E-mail: anilrania@gmail.com
Mob.: 09956203404
1 Introduction
Cotton occupies a prominent place in the national economy of our
country and prosperity of millions of country's rural and urban
population, who depend upon a beautiful harvest of this vegetative
fibre and oil yielding cash crop. Several fungi found on market seeds
used in foods are known to cause considerable damage either
directly to those that carry them or to the crops that are raked from
such contaminated seed stocks.
2 Materials and Methods
Different varieties of Gossypium arboreum, L. viz; Lohit, RG-8, LD-
327, and Sanjay were collected from different state of India (Uttar
Pradesh,Rajasthan, Punjab and Gujrat) from 2010 to 2011
Isolation of seed mycoflora
The seed mycoflora was isolated by using standard moist blotter
method (SBM) and Agar plate methods (APM) as recommended1-4
.
a) Standard blotter method (SBM)
Pair of white blotter papers of 8.5cm diameter was jointly soaked in
sterile distilled water and were placed in pre-sterilized petriplates of
10cm iameter. Ten seeds of test samples per petriplates were placed
at equal distance on the moist blotters. One hundred seeds were
tested for each treatment. The plates were incubated at 25±2 °C
under diurnal conditions for 7 days.
b) Agar plate method (APM)
In this method, pre-sterilized corning glass petriplates of 10cm
diameter were medium. On cooling the medium, ten seeds per
petriplates of the test sample were placed at equal distance
aseptically. Incubation conditions and other details were same as
described for the blotter method. In order to isolate only internal
mycoflora, seeds were pretreated with 0.1% solution of mercuric
chloride for two minutes and subsequently thoroughly washed thrice
with sterile distilled water and placed on agar plates. Seeds without
any such pre-treatment were employed for the total seed mycoflora
(control).
C) Identification of seed-borne fungi
The fungi occurring on each and every seed in the plates were
identified preliminary on the basis of sporulation characters like
sexual or asexual spores with the help of stereoscopic binocular
microscope. The identification and further confirmation of seedborne
fungi was made by preparing slides of the fungal growth and
observing them under compound microscope. The identification was
made with the help of manuals. Pure cultures of these fungi were
prepared and maintained on potato dextrose agar (PDA) slants
poured with 15ml of autoclaved PDA .Percentage occurrence of
each genus was calculated by using the formula.
UK Journal of Pharmaceutical and Biosciences
Available at www.ukjpb.com
ISSN: 2347-9442

2. Kushwaha Mycoflora Associated with Different Varieties of Cotton
UK J Pharm & Biosci, 2015: 3(2); 43
3 Results
The results obtained on prevalence, isolations and identification of
seed mycoflora of different varieties of Gossypium arboreum,.
conducted on storage fungi parasitizing from 2010 to 2011. These
seeds were brought into the laboratory and analyzed.
3.1 Prevalance of storage fungi from seeds of different cotton
varieties
The study on prevalence of different group of fungi on seeds of Lohit
(Table 1), RG-8 (Table 2), LD-327 (Table 3) and Sanjay (Table 4)
have been carried out through blotter test and agar plating methods.
The percentage infection by fungi from each sample was calculated
and depicted in the following tables.
Table 1: Prevalence of seed mycoflora on cotton varieties Lohit
Fungi associated No. seed infected % Seed infected
Alternaria alternata 10 5.0
A. brassicicola 15 7.5
Aspergillus candidus 18 9.0
A. flavus 20 10
A. fumigatus 19 9.5
A. nidulans 12 6.0
Botrytis cinerea 14 7.0
C. brasiliense 16 8.0
Drechslera rostrata 11 5.5
Rhizoctonia solani 15 7.5
Sclerotium rolfsii 11 5.5
Trichoderma viride 14 7.0
Curvularia lunata 10 5.0
E. purpurascens 15 7.5
Total seed examined 200
The isolation of seed mycoflora from Gssypium arboreum was done
critically and are given in the formof (Table 1, 2, 3 and 4) and that
microbial countsvary according to individual varieties, the harvest
and storage conditions prior to drying. So, the observed counts are
thus a reflection of the original fungal load, of growth, as well as of
die-off which are probably enhanced by oxidation and the presence
of active compounds. The total percent fungal count on four
Gssypium arboreum, L. varieties varies from 10 to 12.5 %
Aspergillus flavus infestation, followed to 03.00 to 09.50% for
Alternaria brassicicola, Alternaria longissima, Alternaria tenuissima,
Aspergillus terreu,s Aspergillus fumigatus, Aspergillus nidulans,
Aspergillus. niger, Aspergillus oryzae, Aspergillus candida Botrytis
cinerea, Chaetomium brasiliense, Corynespora cassiicola, Curvularia
lunata, Drechslera rostrata, Epicoccum purpurascens, Mucor
hiemalis, Nigrospora oryzae, Penicillium expansum, Penicillium
chrysogenum, Rhizopus nigricans, Rhizopus arrhizus, Rhizoctonia
bataticola, Sclerotinia sclerotiorum,Trichoderma viride and
Veriticillium albo-atrum Alternaria alternata, Aspergillus sydowii,
Curvularia lunata, Drechslera tetramera, Drechslera rostrata,
Fusarium equiseti, Fusarium equiseti, Fusarium bulbigenum, Mucor
varians Rhizoctonia solni, Stachybotrys atra Sclerotium rolfsii
infection. 15 fungal sp, were isolate from discoloured oil seeds
Alternaria tenuis, Aspergillus flavus, A. niger, A. fumigatus, Fusarium
moniliforme and Rhizopus nigricans were reported to be in
predominant5
.
Table2: Prevalence of seed mycoflora on cotton varieties RG
Fungi associated No. seed infected % Seed infected
A. tenuissima 15 7.5
Aspergillus flavus 2 10
A. oryzae 16 8.0
A. sydowii 10 5.0
C. brasiliense 10 5
Drechslera rostrata 16 8.0
D. tetramera 9 4.5
E. purpurascens 18 9.0
Fusarium bulbigenum 9 4.5
F. equiseti 11 5.5
Mucor hiemalis 6 3.0
M. varians 10 5.0
P. chrysogenum 13 6.5
Rhizoctonia solni 7 3.5
R. bataticola 20 10
Stachybotrys atra 10 5.0
Total seed examined 200
Fusarium lateritium, F. udum, F. vasintectum, F. bulbigenum, F.
moniliforme, F. oxysporum, F. roseum, F. solani and F. truncatum
were also reported to be as externally as well as internally seed

3. Kushwaha Mycoflora Associated with Different Varieties of Cotton
UK J Pharm & Biosci, 2015: 3(2); 44
borne reported6,7
. Blight caused by Alternaria brassicicola, A.
macrospora and A. tenuissima were earlier reported8
.
Table 3: Prevalence of seed mycoflora on cotton varieties LD-
327
Fungi associated No. seed infected % Seed infected
Alternaria longissima 16 8.0
Aspergillus flavus 25 12.5
A. terreus 18 9.0
Chaetomium
brasiliense
16 8.0
Corynespora
cassiicola
14 7.0
Curvularia lunata 14 7.0
Fusarium bulbigenum 10 5.0
F. equiseti 10 5.0
Mucor hiemalis 13 6.5
P. expansum 14 7.0
Rhizopus arrhizus 12 6.0
R. nigricans 13 6.5
E. purpurascens 12 6.0
Rhizoctonia bataticola 13 6.5
Total seed examined 200
In table 5 (1) Range from 0.87 to 2.00 % (Alternaria alternata,
Alternaria brassicicola, Alternaria longissima, Alternaria tenuissima,
Aspergillus nidulans, Aspergillus nigerAspergillus, oryzae
Aspergillus sydowii, Botrytis cinerea, D. tetramera, F. oxysporum, M.
varians, Nigrospora oryzae, P. oxalicum, Penicillium chrysogenum,
R. nigricans, Rhizopus arrhizus, Sclerotinia sclerotiorum,
Sclerotium rolfsii , Trichoderma viride, and Veriticillium albo-atrum (2)
Range from 2.12 to 3.00% (Aspergillus candidus, Aspergillus
terreus, Corynespora cassiicola, Curvularia lunata, Fusarium
equiseti, Fusarium bulbigenum, Penicillium expansum, Rhizoctonia
solani and Stachybotrys atra (3) Range from 3.27 to 6.75%
(Aspergillus fumigatus, Chaetomium brasiliense, Drechslera
rostrata, Epicoccum purpurascens, Mucor hiemalis and Rhizoctonia
bataticola (4) Range from 6.76 to 11.25 % Aspergillus flavus.
Aspergillus flavus, A. niger, Fusarium moniliforme and Penicillium
spp, for maize seeds6
. Curvularia lunata9
, Colletotrichum
graminicola10
for jowar have been recorded.
Table 4: Prevalence of seed mycoflora on cotton varieties
Sanjay
Fungi associated No. seed infected % Seed infected
Alternaria alternata 10 1.25
Alternaria brassicicola 15 1.87
Alternaria longissima 16 2.00
Alternaria tenuissima 15 1.87
Aspergillus candidus 18 2.25
Aspergillus flavus 90 11.25
Aspergillus fumigatus 33 4.12
Aspergillus nidulans 12 1.5
Aspergillus niger 16 2.00
Aspergillus oryzae 11 1.37
Aspergillus sydowii 10 1.25
Aspergillus terreus 18 2.25
Botrytis cinerea 14 1.75
Chaetomium brasiliense 37 4.62
Corynespora cassiicola 23 2.87
Curvularia lunata 24 3.00
Drechslera. tetramera 9 1.12
Drechslera rostrata 27 3.27
Epicoccum purpurascens 54 6.75
Fusarium equiseti 21 2.62
Fusarium oxysporum 14 1.75
Fusarium bulbigenum 19 2.37
Mucor varians 10 1.25
Mucor hiemalis 34 4.34
Nigrospora oryzae 12 1.5
Penicillium expansum 17 2.12
Penicillium oxalicum 7 .87
Penicillium chrysogenum 13 1.62
Rhizoctonia nigricans 13 1.62
Rhizoctonia solani 22 2.75
Rhizoctonia bataticola 43 5.37
Rhizopus arrhizus 12 1.5
Sclerotinia sclerotiorum 13 1.62
Sclerotium rolfsii 11 1.37
Stachybotrys atra 18 2.25
Trichoderma viride 14 1.75
Veriticillium albo-atrum 14 1.75

4. Kushwaha Mycoflora Associated with Different Varieties of Cotton
UK J Pharm & Biosci, 2015: 3(2); 45
Table 5: Total storae fungi isolated from 4 different samples collected
around UP, Rajasthan, Punjab and Gujrat
Fungi associated No. seed infected % Seed infected
Alternaria tenuissima 15 7.5
Aspergillus flavus 20 10
A. niger 16 8.0
A. fumigatus 14 7.0
A. oryzae 11 5.5
C. cassiicola 9 4.5
Mucor hiemalis 15 7.5
Nigrospora oryzae 12 6.0
Penicillium oxalicum 7 3.5
P. expansum 17 8.5
Rhizoctonia bataticola 10 5.0
Veriticillium albo-atrum 14 7.0
Sclerotinia sclerotiorum 13 6.5
E.purpurascens 9 4.5
Rhizoctonia bataticola 10 5.0
Stachybotrys atra 8 4.0
Total seed examined 200
4. Conclusion
In the various fungi isolated from the different seed samples of
Gossypium arboreum, Aspergillus flavus was predominant and
recorded in 6.76 to 11.25 % percentage with well developed growth.
To conclude the finding suggest that there is a need for proper
storage of cotton seed to minimize the fungal growth. There also is a
clear need to increase public awareness on aspects related to seed
health and to develop suitable management practices for improving
the quality of the seeds.
8 References
1. ISTA. International Rules for Seed Testing. Seed Sci.
Tech. 1996; 24: 335-337.
2. De Tempe J. Testing cereals seeds for Fusarium infection
in Netherlands. Proc. Int. Seed Test. Ass. 1970; 35: 193-
206.
3. Neergaard Paul. Detection of seed born pathogen by
culture tests. Seed science and technology. 1973; 1: 217-
254.
4. Agarwal, V.K. Technique for the detection of seed born
fungi. Seed Res. 1976; 4: 24-31
5. Papavizas, G.C. and Lewis J.A. Effect of Cotton seed
treatments with systemic fungicides on seedling disease.
Pl. Dis. Repor. 1977; 61(7): 538-542.
6. [Kamal, Verma A. K. Seed borne mycoflora of arhar (T-
21), effect of culture filtrate of some isolates on seed
germination and fungicidal treatment. Indian J. Mycol. Pl.
Pathol. 1979; 9: 41-45
7. Chavan A.M., and Sunita P. Dani. Fungi occurring on
discloured seeds of pulses and oil seeds. India Bot. Reptr.
1993; 12(1+3) 87-90.
8. Aulakh K.S, Grawal R.K. and Goel R.C. Detection of seeds
borne fungi of maize and their role in causing seed rot and
seedling infection. Indian Phytopath. 1976; 29:241-245
9. Bhale, M.S. and M.N. Khare. Seed borne fungi of sorghum
in Madhyapradesh and their significance. Indian
Phytopath. 1982;35(4):676-679.
10. Basuchodhary. K.C. and Mathur S.B. Infection of Sorghum
seeds by Colletotrichum graminicola L. Survey, Location in
seed and transmission of the pathogen. Seed Sci. and
Techno. 1979; 7(1):87-93.