[국내논문]Characterization of Fusarium oxysporum f. sp. fragariae Based on Vegetative Compatibility Group, Random Amplified Polymorphic DNA and Pathogenicity원문보기
Twenty-two isolates of Fusarium oxysporum f. sp. fragariae were obtained from diseased strawberry plants and their characteristics were investigated by vegetative compatibility group (VCG), random amplified polymorphic DNA (RAPD), and pathogenicity. Three major VCGs (A, B, and C) and one incompatibl...
Twenty-two isolates of Fusarium oxysporum f. sp. fragariae were obtained from diseased strawberry plants and their characteristics were investigated by vegetative compatibility group (VCG), random amplified polymorphic DNA (RAPD), and pathogenicity. Three major VCGs (A, B, and C) and one incompatible group were identified by nitrate reductase complementation test. The virulence pattern of the 22 isolates was studied in relation to four cultivars including Dochiodome, Red-pearl, Maehyang and Akihime. RAPD markers were used to determine genetic relationship, and created three major clusters among the 22 isolates of F. oxysporum f. sp. fragariae. Isolates belong to VCG-C were strongly pathogenic, and relatively high correlation was existed among VCG and RAPD, and virulence. In addition, VCG and RAPD pattern between pathogenic and non-pathogenic isolates were distinctly different.
Twenty-two isolates of Fusarium oxysporum f. sp. fragariae were obtained from diseased strawberry plants and their characteristics were investigated by vegetative compatibility group (VCG), random amplified polymorphic DNA (RAPD), and pathogenicity. Three major VCGs (A, B, and C) and one incompatible group were identified by nitrate reductase complementation test. The virulence pattern of the 22 isolates was studied in relation to four cultivars including Dochiodome, Red-pearl, Maehyang and Akihime. RAPD markers were used to determine genetic relationship, and created three major clusters among the 22 isolates of F. oxysporum f. sp. fragariae. Isolates belong to VCG-C were strongly pathogenic, and relatively high correlation was existed among VCG and RAPD, and virulence. In addition, VCG and RAPD pattern between pathogenic and non-pathogenic isolates were distinctly different.
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제안 방법
The program included an initial denaturation at 94℃ for 4min, 40 cycles with denaturation at 94℃ for 1 min, annealing 56℃ for 1 min, an extension at 72℃ for 2 min, and a final extension at 72℃ for 7 min. Negative controls (no template DNA) were used for each set of experiment to test for the presence of nonspecific reaction. All experiments were repeated at least three times.
The most intense bands were considered for the analysis. PCR amplifications were performed in a total volume of 20 pl containing 20 ng genomic DNA, 50 mM KC1, 1.5 mM MgCL 10 mM Tris/ HC1 (pH 8.3), 200 μM dATP, dCTP, dGTP and dTTP, primer 100 ng, and 1U of Taq DNA polymerase (Promega Corp. Madison, WI). Each reaction was overlaid: with 1 drop of mineral oil.
The primers URP-5 and URP-12 were selected among 12 primers tested on the basis of reproducible bands obtained. Preliminary amplification was conducted to determine the optimal concentration of the component in the PCR reaction mixture. The most intense bands were considered for the analysis.
대상 데이터
Individual isolates were purified through the subculture of a single conidium repeatedly. All isolates used in this study were deposited in Korean Agricultural Culture Collection, National Institute of Agricultural Biotechnology, Suwon.
Isolates were screened for pathogeni city on four strawberry cultivars in Nonsan Strawberry Experiment Station, ARES, Korea. The cultivar Dochio-dome, Redpearl, Maehyang and Akihime were used and five plants were inoculated per isolate.
Therefore, these isolates were considered to pathogenic to strawberry plants. The 20 isolates were collected from diseased strawberry in different strawberry cultivating area in Korea. There was a significant association between the pathogenicity of the isolate collected (Table 2).
sp. fragariae used in this study were collected from different major strawberry cultivating areas in Korea (Table 1). Individual isolates were purified through the subculture of a single conidium repeatedly.
데이터처리
A dendrogram was constructed a representing phenetic relationship between the isolates from the matrix of dissimilarities by the unweighted pair-group method algorithm (UPGMA). All calculations were conducted using the computer program NTSYS-pc.
이론/모형
Pairwise comparison was made between all isolates and the values used to generated a similarity matrix (Sneath and Sokal, 1973). A dendrogram was constructed a representing phenetic relationship between the isolates from the matrix of dissimilarities by the unweighted pair-group method algorithm (UPGMA). All calculations were conducted using the computer program NTSYS-pc.
성능/효과
2). From the results of pathogenicity test using 4 strawberry cultivars, VCGs and RAPD patterns between pathogenic and nonpathogenic isolates were much different.
fragariae isolates that recovered from different regions of strawberry cultivating area in Korea. On the other hand, the isolates belong to VCG-C as well as RAPD pattern II group were very strong pathogenic to be 2.65 of disease index, while the others that had weak virulence were belong to different RAPD groups, and relatively high correlation was found between VCG, RAPD, and virulence of the isolates (Table 3). Three clusters of pathogenic and nonpathogenic isolates contained five clonal individuals based on RAPD profile (Fig.
참고문헌 (42)
Annamalai, P., Ishii, H., Lalithakumari, D. and Revathi, R.1995. Polymerase chain reaction and its applications in fungal disease diagnosis. J. Plant Dis. Protect. 102:91-104
Appel, D. J. and Gordon, T. R. 1994. Local and regional variation in populations of Fusarium oxysporum from agricultural field soils. Phytopathology 84:786-791
Appel, D. J. and Gordon, T. R. 1996. Local and regional variation in populations of Fusarium oxysporum based on RFLP analysis of the intergenic spacer region of the ribosomal DNA. Mol. Plant-Microbe Interact. 9:125-138
Assigbetse, K. B., Fernandez, D., Dubois, M. P. and Geiger, J. P. 1994. Differentiation of Fusarium oxysporum f. sp. vasinfectum races on cotton by random amplified polymorphic DNA (RAPD) analysis. Phytopathology 84:622-626
Correll, J. C., Puhala, J. K. and Schneider, R. W. 1986. Identification of Fusarium oxysporum f. sp. apii on the basis of colony size, virulence, and vegetative compatibility. Phytopathology 76:396-400
Correll, J. C., Klttiich, C. J. R. and Lesile, J. F. 1987. Nitrate non-utilizing mutants of Fusarium oxysporum and their use in vegetative compatibility tests. Phytopathology 77:1640-1646
Correll, J. C. 1991. The relationship between speciales, races and vegetative compatibility groups in Fusarium oxysporum. Phytopathology 81:1061-1065
Correll, J. C., Rhoads, D. D: and Guerber, J. C. 1993. Examination of mitochondrial DNA restriction fragment length polymorphisms, DNA fingerprints, and randomly amplified polymorphic DNA of Colletotrichum orbiculare. Phytopathology 83:1199-1204
Elias, K. S., Zamir, D., Lichtman-Pleban, T. and Katan, T. 1993. Population structure of Fusarium oxysporum f. sp. lycopersici: restriction fragment length polymorphisms provide genetic evidence that vegetative compatibility group is an indicator of evolutionary origin. Mol. Plant-Microbe Interact. 6:565-572
Fernandez, D., Assigbetse, K., Dubois, M. and Geiger, J. P. 1994. Molecular characterization of races and vegetative compatibility groups in Fusarium oxysporum f. sp. vasinfectum. App. Environ. Microbiol. 60:4039-4046
Gordon, T. R. and Okamoto, D. 1992. Variation in mitochondrial DNA among vegetatively compatible isolates of Fusarium oxsporum. Exp. MycoI. 16:2445-250
Grajal-Martin, M. J., Simon, C. J. and Muehlbauer, F. J. 1993. Use of random amplified polymorphic DNA (RAPD) to characterize race 2 of Fusarium oxysporum f. sp. pisi. Phytopathology 83:612-614
Hyun, J. W., Kim, S. O. and Park, W. M. 1996a. Vegetative compatibility, isozyme polymorphisms and pathogenicity of isolates of Fusarium oxysporum f. sp. fragariae. Kor. J. Piant Pathol. 12:33-40
Hyun, J. W. and Park, W. M. 1996b. Differentiation of Fusarium oxysporum f. sp. fragariae isolates by random amplified polymorphic DNA (RAPD) analysis. Kor. J. Plant Pathol. 12:41-46
Jacobson, D. J. and Gordon, T. R.1990. Variability of mitochondrial DNA as an indicator of relationship between populations of Fusarium oxysporum f. sp. melonis. Mycol. Res. 94:734-744
Katan, T. and Katan, J. 1988. Vegetative compatibility grouping of Fusarium oxysporum f.sp. vasinfectum from tissue and rhizosphere of cotton plants. Phytopathology 78:852-855
Katan, T., Zamir, D., Sarfatti, M. and Katan, J. 1991. Vegetative compatibility groups and subgroups in Fusarium oxysporum f. sp. radicis-lycopersici. Phytopathology 81:255-256
Kim, C. H., Seo, H. D., Cho, W. D. and Kim, S. B. 1982. Studies on varietal resistance and chemical control to the wilt of strawberry caused by Fusarium oxysporum. Kor. J. Plant Protect. 21:61-67
Kistler, H. C., Alabouvette, C., Baayen, R. P., Bentley, S., Brayford, D., Coddington, A., Correll, J., Daboussi, M. J., Elias, K., Fernandez, D., Gordon, T. R. Katan, T., Kim, H. G., Lesile, J. F., Martyn, R. D., Migheli, Q., Moore, N. Y, O'Donnell, K., Ploetz, R. C., Rutherford, M. A., Summerell, B., Waalwijk, C. and Woo, S. 1998. Systematic numbering of vegetative compatibility groups in the plant pathogenic fungi Fusarium oxysporum. Phytopathology 88:30-32
Koenig, R. I., Ploetz, R. C. and Kistler, H. C. 1997. Fusarium oxysporum f. sp. cubense consists of a small number of divergent and globally distributed clonal lineages. Phytopathology 87:915-923
Lee, S. B. and Taylor, J. W. 1990. Isolation of DNA from fungal mycelia and single spores. In: PCR protocols: a guide to methods and applications, ed. by M. A. Innis, D. H. Gelfand, J. J. Sninsky, and T. J. White, pp 282-287. Academic press, SanDiego
Leslie, J. F.1993. Fungal vegetative compatibility. Ann. Rev. Phytopathol. 31:127-150
Manicom, B. Q. and Baayen, R. P. 1993. Restriction fragment length polymorphism in Fusarium oxysporum f.sp. dianthi and other fusaria from Dianthus species. Plant Pathol. 42:851-857
Manicom, B. Q., Bar-Joseph, M., Kotze, J. M. and Becker, M. M. 1990. A restriction fragment length polymorphism probe relating vegetative compatibility groups and pathogenicity in Fusarium oxysporum f. sp. dianthi. Phytopathology 80:336-339
Manulis, S., Kogan, N., Reuven, M. and Benyephet, Y. 1994. Use of the RAPD technique for identification of Fusarium oxysporum f. sp. dianthi from carnation. Phytopathology 84:98-101
Mes, J. J., van Doom J., Roebroeck, E. J. A., van Egmond, E., van Aartrijk, J. and Boonekamp, P. M. 1994. Restriction fragment length polymorphisms, races and vegetative compatibility groups within a worldwide collection of Fusarium oxysporum f. sp. gladioli. Plant Pathol. 43:362-370
Mes, J. J., Weststeijin, E. A., Herlaar, F., Lambalk, J. J. M., Wijbrandi, J., Haring, M. A. and Comelissem, B. J. 1998. Biological and molecular characterization of Fusarium oxysporum f. sp. lycopersici divides race 1 isolates into separate virulence groups. Phytopathology 89:156-160
Nagarajan, G., Nam, M. H., Song, J. Y., Y,oo, S. J. and Kim, H. G. 2004. Genetic variation in Fusarium oxysporum f. sp. fragariae populations based on RAPD and rDNA RFLP analyses. Plant Pathol. J. 20:264-270
Nam, M. H., Jung, S. K., Kim, N. G., Yoo, S. J .and Kim, H. G. 2005. Resistance analysis of cultivars and occurrence survey of Fusarium wilt on strawberry. Plant Pathol. J. 11:35-38
Nelson, P. E., Toussoun, T .A. and Marasas, W. F. O. 1981. Fusarium: Biology and Taxonomy. Pennsylvania State University Press: University Park
Nelson, A. J., Elias, K. S., Arevalo, E., Darlington, L. C. and Baily, B. A. 1997. Genetic characterization of RAPD analysis of isolates of Fusarium oxysporum f. sp. erythroxyli associated with an emerging epidemic in Peru. Phytopathology 87:1220-1225
Sneath, P. H. A. and Sokal, R. R. 1973. Numerical taxonomy, W. H., Freeman & Co., San Francisco
Tantaoui, A., Quinten, M., Geiger, J. P. and Fernandez, D. 1996. Characterization of a single clonal lineage of Fusarium oxysporum f. sp. albedinis causing Bayoud disease of date palm in Morocco. Phytopathology 86:787-792
Wilhelm, S. 1998. Fusarium wilt (Fusarium Yellows). In: Compendium of strawberry, disease, ed. by J. L. Maas, pp 52-53, APS Press, St. Paul
Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A. and Tingey, S. V. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18:6531-6535
Winks, B. L. and Williams, Y. N. 1965. A wilt of strawberry caused by a new form of Fusarium oxysporum. Queen land J. Agri. Ani. Sci. 22:475-479
Woo, S. L., Zonia, A., Del Sorbo, G., Lorito, M., Nanni, B., Scala, F. and Noviello, C. 1996. Characterization of Fusarium oxysporum f. sp. phaseoli by pathogenic races, VCGs, RFLPs and RAPD. Phytopathology 86:966-973
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