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Development of a Rapid Molecular Detection Marker for Colletotrichum species with AFLP 원문보기

Mycobiology, v.32 no.3, 2004년, pp.123 - 127  

Eom, Seung-Hee (Plant Molecular Biology & Microbial Biotechnology Laboratory, Division of Applied Plant Sciences, College of Agriculture and Life Sciences, Kangwon National University) ,  Kim, Kwon-Jong (Plant Molecular Biology & Microbial Biotechnology Laboratory, Division of Applied Plant Sciences, College of Agriculture and Life Sciences, Kangwon National University) ,  Jung, Hee-Sun (Plant Molecular Biology & Microbial Biotechnology Laboratory, Division of Applied Plant Sciences, College of Agriculture and Life Sciences, Kangwon National University) ,  Lee, Sang-Pyo (Plant Molecular Biology & Microbial Biotechnology Laboratory, Division of Applied Plant Sciences, College of Agriculture and Life Sciences, Kangwon National University) ,  Lee, Youn-Su (Plant Molecular Biology & Microbial Biotechnology Laboratory, Division of Applied Plant Sciences, College of Agriculture and Life Sciences, Kangwon National University)

Abstract ▼ AI-Helper 아이콘AI-Helper

Sweet persimmons have been increasingly cultivated in the southern part of Korea. However, anthracnose disease caused by Colletotrichum species is one of the major hindrances in cultivation and productions. In this study, we used polymerase chain reaction(PCR) to detect Colletotrichum species with t...

주제어

#AFLP   #Anthracnose   #Colletotrichum species   #Sweet persimmons  

AI 본문요약
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제안 방법

  • The specific AFLP DNA fragment was labelled using DIG-11-dUTP (digoxigenein-3-O-methylcarbomyl-(-amino-caproyl~5-(~3-aminoallyl)-uridine-5'-triphosphate) according to the manufacturers, instructions (Specific PCR Reaction, Boehringer-Mannheim, Gemiany). A total of 100 jjd of the PCR labeling reaction mixture containing 10 ng of template DNA, 05 /jM of each primer (PUC/ M13 forward and PUC/M13 reverse), 1 xPCR buffer, 5 units of Taq polymerase (Quantum, USA), 0.1 mM of dGTP, dCTP and dATP, 0.09 mM dTTP and 1 mM dUTP was prepared and reacted in a program of 90 s # 94℃, 90 s at 50"C, 2 min at 72"C for 24 cycles and 4 min at 72℃. PCR products amplified by designed primer set (Co.
  • In this study, sequencing and analysis of AFLP fragment of Colletotrichum species allowed us to design specific PCR primer. These primer sets, Co.

대상 데이터

  • , 2001; Table 1). Several isolates of Colletotrichum species provided from Korean Agricultural Culture Collection (KACC) and Korean Collection for type Cultures (KCTC) were included in the experiment (Table 2). The Colletotrichum species isolates were cultivated in PDA (Potato Dextrose Agar) for 7 days at 25℃.

이론/모형

  • They were transferod onto Hybond N membranes (Quantum, USA) using a # transfer system. Identification of homologous sequences of DNA, usin형 a labelled probe was possible with the DIG-detec-tion chemiluminescent method (Boehringer-Mannheim).
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참고문헌 (22)

  1. Agostini, J. P., Timmer, L. Wand Mitchell, D. J. 1992. Morphological and Pathological characteristics of strains of Colletotrichum gloeosporioides from citrus. Phytopathology 82: 1377-1382 

    상세보기 crossref

  2. Agrios, G. N. 1998. Plant Pathology. 4th ed. Pp. 331-333. Academic Press, New York 

  3. Gunnell, P. S. and Gubler, W D. 1992. Taxonomy and morphology of Colletotrichum species pathogenic to strawberries. Mycologia. 84: 157-165 

    crossref

  4. Guthrie, P. A. I, Magill, C. W, Frederiksed, R. A. and Odvody, G. N. 1992. Randomly amplified polymorphic DNA markers: a system for identifying and differentiating isolates of Colletotrichum graminicola. Phytopathology 82: 832-835 

    상세보기 crossref

  5. Innis, M. A. and Gelfand, D. H. 1990. Optimization of PCRs; in PCR protocols. A guide to methods and applications, pp. 3-12, Academic press, Inc., Harcourt Brace Joranovich, New York. M. A., Innis, D. H, Gelfand, I. J., Sninsky, and T. J., Whitc,(eds.) 

  6. Mesquita, A. G. G., Paula, T J., Moreira, Jr., M. A. and Barros, E. G. 1998. Identification of races of Colletotrichum lindemuthianum with the aid of PCR-based molecular markers. Plant Disease. 82: 1084-1087 

    상세보기 crossref

  7. Mills, P. R., Sreenivasaprasad, S. and Brown, A. E. 1992. Detection and differentiation of Colletotrichum gloeosporioides isolates using PCR. FEMS Microbiol. Lett. 98: 137-144 

    상세보기 crossref

  8. Old, R. W and Primrose, S. B. 1994. Polymerase chain reaction; in Principles of Gene Manipulation. Pp. 178-190. Blackwell Science, Cambridge, MA. USA. 473Pp 

  9. Paran, I. Land Michelmore, R. W 1993. Development of reliable PCR-bascd markers linked to downy mildew resistance genes in lettuce. Theoretical and Applied Genetics. 85: 985-993 

  10. Raeder, U. and Broda, P. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol. 1: 17-20 

    상세보기 crossref

  11. Reineke, A. and Karlovsky, P. 2000. Simplified AFLP protocol: replacement of primer labeling by the incorporation of alphalabeled nucleotides during PCR. Biotechniques. 28: 622-623 

  12. Rusell, J. R., Fuller, J. D., Macaulay, M., B. Hatz, G., Jahoor, A., Powell, W and Waugh, R. 1997. Direct comparison of levels of genetic variation among barely accessions detected by RFLPs, AFLPs, SSRs and RAPDs. Theor. Appl. Genet. 95: 714-722 

    상세보기 crossref

  13. Sambook, J., Fristch, E. F. and Maniatis, T. A. 1989. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 

  14. Schilling, A. G., Mller, E. M. and Geiger, H. H. 1996. Polymerase chain reaction based assays for species-specific detection of Fusarium culmorum, F. graminareum and F. avenaoceum. Phytopathology 86: 515-522 

    상세보기 crossref

  15. Sreenivasaprasad, S., Brown, A. E. and Mills, P. R. 1992. DNA sequence variation and interrelationships among Colltotrichum species causing strawberry anathracnose. Physiol. Mol. Plant Pathol. 41: 265-281 

    상세보기 crossref

  16. Sreenivasaprasad, S. Sharada, K., Brown, A. E. and Mills, P. R. 2000. PCRbased detection of Colletotrichum acutattum on stawberry. Plant Pathol. (In press) 

  17. Kim, K. H., Cheong, B. G., Chang, T. H., Yim, T. H., Lee, Y. S. and Song, W D. 2001. Develpoment of method of diagnosis, forecast and integrate control on the major disease of Sweet Persimmom (Diospyros kaki L.). Ministry of agricultural and Forestry. Pp. 17-40 

  18. Vaillancourt, L. J. and Hanau, R.. M. 1992. Genetic and morphological comparisons of Glomorella (Colletotrichum) isolates from maize and from sorghum. Exp. Mycol. 16: 2219-2293 

  19. Vakalounakis, D. J. and Fragkiakakis, G. A. 1999. Genetic diversity of Fusarium oxysporum isolates from cucumbers: Differentiation by pathogenicity, vegetative compatibility, and RAPD fingerprinting. Phytopathology 89: 161-168 

    상세보기 crossref

  20. Vilarinhos, A. D., Paula, Jr., T. T, Barros, E. G. and Moreia, M. A. 1995. Characterizations of races of Colletotrichum lindermuthianum by the random amplified polymorphic DNA technique. Braz: Phytopathol. 20: 194-198 

  21. Vos, P., Hoger, R., Bleeker, M., Reijans, M., Van de Lee, T, Homes, M., Fijters, A., Pot, J., Peleman, J., Kuiper, M. and Zabeau, M. 1995. AFLP: A new technique for DNA fingerprinting. Nuel. Acids Res. 23: 4407-4414 

    상세보기 crossref

  22. William, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A. and Tingey, S. V. 1990. DNA polymorphic amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18: 6531-6535. 

    상세보기 crossref

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