RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) ANALYSIS TO DETERMINE THE GENETIC VARIABILITY AMONG VIRULENT AND LESS VIRULENT ISOLATES OF FUSARIUM MONILIFORME, FUSARIUM OXYSPORUM AND FUSARIUM SOLANI ISOLATED FROM INFECTED COTTON SEEDLINGS
Root-rot of cotton (Gossypium spp.) is one of the most important diseases in Upper Egypt. Isolation has been done from diseased cotton roots and seeds which were collected from 11 counties in Assiut province, Egypt. Identification procedures of the isolated fungi confirmed that the isolated fungi were for Fusarium solani, F. moniliforme and F. oxysporum and Rhizoctonia solani. Thirty six isolates of Fusarium spp. and 10 isolates of Rhizoctonia solani were tested for their pathogenicity on both Giza 80 and Giza 83 cotton seedlings to verify their virulence on seedlings. The pathogenicity test results have grouped the Fusarium spp. isolates into three groups; highly virulent that caused 91-100% mortalities; moderately virulent that caused 81-90% mortalities and low virulent that caused lower than 81.0% mortalities. Data also shows that, in general Giza 80 cotton cultivar was more susceptible for infection with Fusarium spp. when compared with Giza 83 cotton cultivar. In case of Rhizoctonia solani, data revealed that the infection percentage was significantly affected by isolates while cotton cultivars had no significant influence on infection. Four 10-mer primers (1:6-d, 2:6-d, 4:6-d and 5:6-d) were used in RAPD-PCR to determine the genetic variability between six isolates, one virulent and one less virulent, of F. moniliforme, F. oxysporum and F. solani. Our results showed that the primer 2:6-d clearly separated F. moniliforme, F. oxysporum and F. solani and proved to be quite powerful in distinguishing the three different species and isolates of Fusarium spp.
Abd-El-Moneem, H. M. K. 1996. Effect of micronutrients on incidence of sesame charcoal root rot wilt disease complex. Assiut J. of Agric. Sci. 27 (3): 181-195.
Ainsworth, G. C. 1971. Dictionary of the fungi. Commonwealth Mycological Institute, Kew, Surrey, England. 412 pp.
Altomare, C. and O. Petrini and A. Logrieco and A. Bottalico. 1997. Taxonomic relationships among the toxigenic species Fusarium acuminatum, Fusarium sporotrichioides and Fusarium tricinctum by isozyme analysis and RAPD assay. Canadian J. of Botany 75 (10):1674-1684.
Assigbetse, K. B. and D. Fernandez and M. P. Dubois and J. P. Geiger. 1994. Differentiation of Fusarium oxysporum f. sp. vasinfectum races on cotton by random amplified polymorphic DNA (RAPD) analysis. Phytopath. 84 (6): 622-626.
Booth, C. 1977. Fusarium laboratory guide to the identification of the major species. Commonwealth Mycological Institute, Kew, Surrey, England.1-58pp.
Castrillo, L. A. and W. M. Brooks. 1998. Differentiation of Beauveria bassiana isolates from the darkling beetle Alphitobius diaperinus, using isozyme and RAPD analyses. J. of Invertebrate Pathol. 72: 190-196.
Crowhurst, R. N. and B. T. Hawthorne and E. H. A. Rikkerink and M. D. Templeton. 1991. Differentiation of Fusarium solani f. sp. cucurbitae races 1 and 2 by random amplication of polymorphic DNA. Curr. Genet. 20:391-396.
Edwards, K. J. 1998. Random amplified polymorphic DNAs (RAPDs). In: Karp A., Issac P. G., Ingram D. S. (eds): Molecular Tools for Screening Biodiversity. Chapman and Hall, Cambridge 1:171-175.
Goodwin, P. A. and S. L. Annis. 1991. Rapid identification of genetic variation and pathotype of Leptosphaeria maculans by random amplified polymorphic DNA assay. Appl. Environ. Microbiol. 57:2482-2486.
Hadrys, H. and M. Balick and B. Schierwater. 1992. Application of random amplified polymorphic DNA (RAPD) in molecular ecology. Mol. Ecol. 1:55-63.
Hamelin, R. C. and G. B. Ouellette and L. Bernier. 1993. Diagnosis and phylogeny of Gremmeniella spp. using molecular markers. Page 39 in Abstracts, 6th International Congress on Plant Pathology. Montreal, Quebec. National Research Council of Canada, Ottawa.
Hashem, M. and A. M. Hamada. 2002. Induction of resistance to root rot disease of wheat grown under field conditions. Rostlinna Vyroba. 48 (7): 312-31.
Johnson, L. F. and D. D. Baird and A. Y. Chambers and N. B. Shamiyeh. 1978. Fungi associated with post-emergence seedling disease of cotton in three soils. Phytopathol. 68: 917-920.
Mattila, Y. T. and S. Paavanen and A. Hannukkala and P. Parikka and R. Tahvonen and R. Karjalainen. 1996. Isozyme and RAPD-PCR analyses of Fusarium avenaceum strains from Finland. Plant Pathol. 45 (1): 126-134.
Murray, M. and W. Thompson. 1980. The isolation of high weight plant DNA. Nucleic Acid Res. 8: 4321-4325.
Ovesna, J. and K. Polakova and L. Leisova. 2002. DNA analyses and their application in plant breeding. Czech. J. Genet. Plant Breed. 38: 29-40.
Perez-Artes, E. and D. Maria and G. Pedrajas and J. Bejarano-Alcazar and R. Jimenez-Diaz. 2000. Differentiation of cotton-defoliating and nondefoliating pathotypes of Verticillium dahliae by RAPD and specific PCR analyses. European Journal of Plant Pathol. 106 (6): 507-517.
Ramsay, J. R. and D. S. Multani and B. R. Lyon. 1996. RAPD-PCR identification of Verticillium dahliae isolates with differential pathogenicity to cotton. Aust. J. Agric. Res. 47: 681–693.
Sneath, P. H. A. and R. R. Sokal. 1973. Numerical taxonomy: The principles and practice of numerical classification. Freeman, San Francisco. 573 pp.
Soleymani, M. J. and G. A. Hedjaroude and J. Zad, 1993. Studies on pathogenicity of some seedborne Fusarium species on cotton seedling. Iranian Journal of Plant Pathol. 29: 19-20.
Su, G. and S. O. Suh and R. W. Schneider and J. S. Russin, 2001. Host specialization in the charcoal rot fungus, Macrophomina phaseolina. Phytopathol. 91: 120-126.
Tojo, M. E. and E. Nakazono and S. Tsushima and T. Morikawa and N. Matsumoto, 1998. Characterization of two morphological groups of isolates of Pythium ultimum var. ultimum in a vegetable field. Mycosci. 39: 135-144.
Wahid, A. and M. S. Javed and A. H. Tariq and M. Ldrees, 1994. Fungi associated with seed of different cotton varieties grown in the Punjab and their control. Pak. J. Phytopathol. 6 (2): 97-102.
Watkins, G. M. 1981. Compendium of cotton diseases. The American Phytopathological Society. St. Paul, Minnesota, 87 pp.
Williams, J. G. K. and A. R. Kubelik and K. J. Livak and J. A Rafalski and S. V. Tingey. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18: 6531-6535.
Zheng, Q. and R. Ploetz. 2002. Genetic diversity in the mango malformation pathogen and development of a PCR assay. Plant Pathol. 51 (3): 208-216.
- There are currently no refbacks.
Copyright (c) 2016 Ahmed Abdelmagid, Abdel-Moneim Amein, Mohamed Hassan, Hamdy E. Hares
This work is licensed under a Creative Commons Attribution 4.0 International License.