ORIGINAL ARTICLE
RAPD-PCR of Trichoderma isolates and in vitro antagonism against Fusarium wilt pathogens of Psidium guajava L.
More details
Hide details
1
Department of Biotechnology, FASC, MITS University, Lakshmangarh-332311, Sikar, Rajasthan, India
2
Molecular Plant Pathology Lab., CISH, Lucknow-227 017, India
3
Mahatma Jyotiba Phule Rohilkhand University, Bareilly, UP, India
Journal of Plant Protection Research 2010;50(3):256-262
KEYWORDS
TOPICS
ABSTRACT
The paper concerns randomly amplified polymorphic DNA RAPD-PCR analysis of seven Trichoderma species isolates and their in vitro antagonism against wilt pathogens of Psidium guajava L. viz. Fusarium oxysporum f. sp.
psidii (F. o. f. sp. psidii) and Fusarium solani . Out of 10 RAPD oligonucleotides (OPA 1–OPA 10) tested, seven markers OPA 1, 3, 5, 7, 8, 9 and 10 efficiently differentiated the isolates of Trichoderma and showed reproducible banding patterns. A total of 248 bands were obtained from these markers along with a 61.84% per cent similarity among the seven isolates of
Trichoderma. From the seven isolates of Trichoderma, isolates T. harzianum,
T. virens and T. viride were evaluated for in vitro efficacy against F. o. f. sp. psidii and F. solani. Per cent inhibition was maximum by direct use of Trichoderma spp. in dual cultures against both pathogens, F. o. f. sp. psidii
and F. solani. Two species, T. virens and T. viride were superior in inhibiting the growth of both Fusarium spp. Fusarium isolates showed intra species variability.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
REFERENCES (33)
1.
Abd-Elsalam K.A., Schnieder F., Guo J.R. 2003. A modified DNA extraction minipreparation protocol for Fusarium isolates. J. Rapid Methods Aut. Microbiol. 11: 75–79.
2.
Arisan-Atac I., Heidenreich E., Kubicek C.P. 1995. Randomly amplified polymorphic DNA fingerprinting identifies subgroups of Trichoderma viride and other Trichoderma sp. capable of chestnut blight biocontrol. FEMS Microbiol. Lett. 126: 249–256.
3.
Bardakci F. 2001. Random amplified polymorphic DNA (RAPD) markers. Turkey J. Biol. 25: 185–196.
4.
Booth C. 1971. The Genus Fusarium. Commonwealth Mycological Institute, Kew, Surrey, England, 276 pp.
5.
Chet I., Inbar J. 1994. Biological control of fungal pathogens. Appl. Biochem. Biotechnol. 48: 37–43.
6.
Dennis C., Webster J. 1971. Antagonistic properties of species, groups of Trichoderma-II production of volatile antibiotics. Trans. Br. Mycol. Soc. 57: 41–48.
7.
Dubey S.C., Suresh M. 2006. Randomly amplified polymorphic DNA markers for Trichoderma species and antagonism against Fusarium oxysporum f. sp. ciceris causing chickpea wilt. J. Phytopathol. 154: 663–669.
8.
Eziashi E.J., Uma N.U., Adekunle A.A., Airede C.E. 2006. Effect of metabolites produced by Trichoderma species against Ceratocystis paradoxa in culture medium. African J. Biotechnol. 5: 703–706.
9.
Fujimori F., Okuda T. 1993. Aplication of the random amplified polymorphic DNA using the polymerase chain reaction for efficient elimination of duplicate strains in microbial screening. J. Antibiot. 47: 173–182.
10.
Gomez I., Chet I., Herreraestrela A. 1997. Genetic diversity and vegetative compatibility among Trichoderma harzianum isolates. Mol. Gen. Genet. 256: 127–135.
11.
Gopal K., Sreenivasulua Y., Gopia V., Prasadbabua G., Kumarb T.B., Madhusudhana P., Ahemeda S.K., Palanivel S.G. 2008. Genetic variability and relationships among seventeen Trichoderma isolates to control dry root rot disease using RAPD markers. Z. Naturforsch. Sect. C. 63: 740–746.
12.
Grondona I., Hermosa R., Tejada M., Gomis M.D., Mateos P.F., Bridge P.D., Monte E., García-acha I. 1997. Physiological and biochemical characterisation of Trichoderma harzianum, a biological control agent against soil-borne fungal plant pathogens. Appl. Environ. Microbiol. 63: 3189–3198.
13.
Gupta R.P., Yadava B.C., Singh R.V., Rai O. 2003. Evaluation of Bioagents Against Fusarium wilt of Pigeonpea. p. 1–63. In: Proc. BS-57. National Symposium of Pulses for Crop-Diversification and Natural Resource Management, Kanpur, India, 148 pp.
14.
Hutchinson S.A., Cowan M.E. 1972. Identification of biological effects of volatile metabolites from culture filtrate of Trichoderma harzianum. Trans. Br. Mycol. Soc. 59: 71–77.
15.
Jaccard P. 1908. Nouvelles recherches sur la distribution florale. Bull. Soc. Vaudoise Sci. Nat. 44: 223–270.
16.
Kidwai M.K., Vikas, Srivastava S., Singh H.B. 2006. Compatibility of Trichoderma harzianum to selected fungicide. J. Eco- Friend. Agric. 1: 156–161.
17.
Kumar P., Misra A.K., Pandey B.K. 2007. In vitro evaluation of Trichoderma spp. against vegetative mango malformation pathogen Fusarium moniliformae var. subglutinans. J. Eco- Friend. Agric. 2: 187–189.
18.
Maththeis J.P., Roberts R.G. 1992. Identification of geosmin as a volatile metabolite of Penicillium expansum. Appl. Environ. Microbiol. 58: 3170–3172.
19.
Misra A.K., Pandey B.K. 1999. Pathogenicity and evaluation of fungicides against guava wilt pathogens. J. Mycol. Pl. Pathol. 29: 274–275.
20.
Misra A.K. 2006. Wilt of guava-a disease of national importance. Indian Phytopathol. 59: 269–280.
21.
Misra A.K., Gupta V.K. 2009. Trichoderma: biology, biotechnology and biodiversity. J. Eco-Friend. Agric. 4: 99–117.
22.
Muthumeenakshi S., Brown A.E., Mills P.R. 1998. Genetic comparision of aggressive weed mould strains of Trichoderma harzianum from mushroom compost in North America and the British Isles. Mycol. Res. 4: 385–390.
23.
Prasad N., Mehta P.R., Lal S.B. 1952. Fusarium wilt of guava (Psidium guajava L.) in Uttar Pradesh, India. Nature 169, p. 753.
24.
Raper K.B., Thom C. 1949. A Manual of Penicillia. Williams and Wilkins, Baltimore, 691 pp.
25.
Rifai M.A. 1969. Revision of the genus Trichoderma. Mycol. Pap. 116: 1–56.
26.
Rohlf F.J. 1998 NTSYS-pc. Numerical Taxonomy and Multivariate Analysis System, Version 2.0. – Applied Biostatistics, New York, 37 pp.
27.
Schlick A., Kuhls K., Meyer W., Lieckfeldt E., Börner T., Messner K. 1994. Fingerprinting reveals gamma-ray induced mutations in fungal DNA: implications for identification of patent strains of Trichoderma harzianum. Curr. Genet. 26: 74–78.
28.
Simon C., Sivasithaparam M. 1988. Interaction among Gaeumannomyces graminum var. tritci, Trichoderma koningii and soil bacteria. Can. J. Microbiol. 34: 871–876.
29.
Singh A., Singh H.B. 2004. Control of color rot in mint (Mentha sp.) caused by Sclerotium rolfsii using biological mean. Curr. Sci. 87: 362–366.
30.
Sneath P.H.A., Sokal R.R. 1973. Numerical Taxonomy. Freeman, San Francisco, 513 pp.
31.
Tapwal A., Sharma Y.P., Lakhanpal T.N. 2004. Effect of volatile compound released by Gliocladium virens and Trichoderma spp. on growth of Alternaria mellea. J. Mycol. Pl. Pathol. 34: 308–310.
32.
Watts S., Dahiya J., Chaudhary K. 1988. Isolation and characterization of new antifungal metabolite of Trichoderma reseii. Plant Soil 107: 81–84.
33.
Zimand G., Valinsky L., Elad Y., Chet I., Manulis S. 1994. Use of the RAPD procedure for the identification of Trichoderma strains. Mycol. Res. 98: 531–534.