Abstract

Sexual reproduction plays an important role in the biology and epidemiology of oomycete plant pathogens such as the heterothallic species Phytophthora infestans. Recent worldwide dispersal of A2 mating type strains of P. infestans resulted in increased virulence, gene transfer, and genetic variation, creating new challenges for disease management. To develop a genetic assay for mating type identification in P. infestans, we used the Amplified Fragment Length Polymorphism (AFLP) technique. The primer combination E+AT/M+CTA detected a fragment specific to A1 mating type (Mat-A1) of P. infestans. This fragment was cloned and sequenced, and a pair of primers (INF-1, INF-2) were designed and used to differentiate P. infestans Mat-A1 from Mat-A2 strains. The Mat A1-specific fragment was detected using Southern blot analysis of PCR products amplified with primers INF-1 and INF-2 from genomic DNA of 14 P. infestans Mat-A1 strains, but not 13 P. infestans Mat-A2 strains or 8 other isolates representing several Phytophthora spp. Southern blot analysis of genomic DNAs of P. infestans isolates revealed a 1.6 kb restriction enzyme (EcoRI, BamHI, AvaI)-fragment only in Mat-A1 strains. The A1 mating type-specific primers amplified a unique band under stringent annealing temperatures of $63^{\circ}C-64^{\circ}C$, suggesting that this PCR assay could be developed into a useful method for mating type determination of P. infestans in field material.

주제어

#Phytophthora infestans   #molecular marker   #mating type A-1   #AFLP  

저자의 다른 논문

참고문헌 (39)

1. Deahl, K. L., S. P. Demuth, and A. Rivera-Pena. 1995. Identification of mating types and metalaxyl resistance in North American populations of Phytophthora infestans. Am. Potato J. 72: 35-49 
2. Fry, W. E. and S. B. Goodwin. 1997. Re-emergence of potato and tomato late blight in the United States and Canada. Plant Dis. 81: 1349-1357 
3. Innis, M. A. and D. H. Gelfand. 1990. Optimization of PCRs; In Innis, M. A., D. H. Gelfand, and J. Sninsky (eds.), PCR Protocols. A Guide to Methods and Applications 
4. Kim, Y. T., B. K. Park, E. I. Hwang, N. H. Yim, S. H. Lee, and S. U. Kim. 2004. Detection of recombinant marker DNA in genetically modified glyphosate-tolerant soybean and use in environmental risk assessment. J. Microbiol. Biotechnol. 14: 390-394 
5. Lee, Y. S., S. H. Eom, K. J. Kim, H. S. Jung, and S. P. Lee. 2003. Identification of DNA markers linked to metalaxyl insensitivity loci in Phytophthora infestans. Mycobiology 31: 229-234  원문보기 상세보기
6. Old, R. W. and S. B. Primrose. 1994. Polymerase chain reaction, pp. 178-190. In S. B. Primrose, R. M. Twyman, and R. W. Old (eds.), Principles of Gene Manipulation. Blackwell Science. Cambridge, MA, U.S.A 
7. Ristaino, J. B. 2002. Tracking historic migrations of the Irish potato famine pathogen, Phytophthora infestans. Microbes Infect. 4: 1369-1377 
8. Lee, Y S. and K. S. Kim. 2000. Rapid and accurate speciesspecific detection of Phytophthora infestans through analysis of ITS regions in its rDNA. J. Microbiol. Biotechnol. 10: 651-655 
9. Van der Lee, T., I. D. Witte, A. Drecth, C. Alfonso, and F. Govers. 1997. AFLP linkage map of the oomycete Phytophthora infestans. Fungal Genet. Biol. 21: 278-291 
10. Gallegly, M. E. 1968. Genetics of pathogenicity of Phytophthora infestans. Annu. Rev. Phytopathol. 6: 375-396 
11. Rusell, J. R., J. D. Fuller, M. Macaulay, B. G. Hatz, A. Jahoor, W. Powell, and R. Waugh. 1997. Direct comparison of levels of genetic variation among barley accessions detected by RFLPs, AFLPs, SSRs and RAPDs. Theor. Appl. Genet. 95: 714- 722 
12. Judelson, H. S., L. J. Spielman, and R. C. Shattock. 1995. Genetic mapping and non-Mendelian segregation of matingtype loci in the oomycete, Phytophthora infestans. Genetics 141: 503-512 
13. Muller, H. J. 1918. A case of balanced lethal factors. Genetics 3: 422-499 
14. Shaw, D. S. 1988. The Phytophthora species, pp. 27-51. In Sidhu G. S. (ed.), Advances in Plant Pathology, vol. 6. Academic Press, New York, U.S.A 
15. Chang, T. T. and W. H. Ko. 1990. Resistance to fungicides and antibiotics in Phytophthora parasitica: Genetic nature and use in hybrid determination. Phytopathology 80: 1414-1421 
16. Kamoun, S. 2003. Molecular genetics of pathogenic oomycetes. Eukaryotic Cell 2: 191-199 
17. Shaw, D. S. 1991. Genetics, pp. 131-170. In Ingram, D. S. and D. H. Williams (eds.), Phytophthora infestans, vol. 7. Academic Press, New York, U.S.A 
18. Judelson, H. S. 1996. Recent advances in the genetics of oomycete plant pathogens. Mol. Plant-Microbe Interact. 9: 443-449 
19. Kim, M. K., H. S. Kim, B. O. Kim, S. Y Yoo, J. H. Seong, D. K. Kim, S. E. Lee, S. J. Choe, J. C. Park, B. M. Min, M. J. Jeong, D. K. Kim, Y. K. Shin, and J. K. Kook. 2004. Multiplex PCR using conserved and species-specific 16S rDNA primers for simultaneous detection of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans. J. Microbiol. Biotechnol. 14: 110-115 
20. Tooley, P. W. 1988. Use of uncontrolled freezing for liquid nitrogen storage of Phytophthora species. Plant Dis. 72: 680-682 
21. Lee, Y. S. and K. J. Kim. 2002. Genetic DNA marker for A2 mating type in Phytophthora infestans. J. Microbiol. 40: 254-259 
22. Reineke, A. and P. Karlovsky. 2000. Simplified AFLP protocol: Replacement of primer labeling by the incorporation of alpha-labeled nucleotides during PCR. Biotechniques 28: 622-623 
23. Griffith, G. W., R. Snell, and D. S. Shaw. 1995. Late blight (Phytophthora infestans) on tomato in the tropics. Mycologist 9: 87-89 
24. Smart, C. D. and W. E. Fry. 2001. Invasions by the late blight pathogen: Renewed sex and enhanced fitness. Biol. Invas. 3: 235-243 
25. Paabo, S. 1990. Amplifying ancient DNA; PCR Protocols, pp. 159-166. In Innis, M. A., D. H. Gelfand, J. J. Sninsky, and T. J. White (eds.), A Guide to Methods and Applications. Academic Press, Inc., Harcourt Brace Jovanovich, NY, U.S.A. 
26. Birch, P. R. J. and S. C. Whisson. 2001. Phytophthora infestans enters the genomics era. Mol. Plant Pathol. 2: 257-263 
27. Vos, P., R. Hoger, M. Bleeker, M. Reijans, T. Van de Lee, M. Hornes, A. Fijters, J. Pot, J. Peleman, M. Kuiper, and M. Zabeau. 1995. AFLP: A new technique for DNA fingerprinting. Nucl. Acid Res. 23: 4407-4414 
28. Gisi, U. and Y. Cohen. 1996. Resistance to phenylamide fungicides: A case study with Phytophthora infestans involving mating type and race structure. Annu. Rev. Phytopathol. 34: 549-572 
29. Ko, W. H. 1988. Hormonal heterothallism and homothallism in Phytophthora. Annu. Rev. Phytopathol. 26: 57-73 
30. Goodwin, S. B., J. L. Spielman, J. M. Matuszak, S. N. Bergeron, and W. E. Fry. 1992. Clonal diversity and genetic differentiation of Phytophthora infestans population in Northern and Central Mexico. Phytopathology 82: 955-961 
31. Lui, S. Y, K. L. Ling, and B. Ho. 2003. rDNA gene in an unlikely marker for metronidazole resistance in the Asian Helicobacter pylori isolates. J. Microbiol. Biotechnol. 13: 751-758 
32. Tooley, P. W., W. E. Fry, and M. J. Villarreal Gonzalez. 1985. Isoenzyme characterization of sexual and asexual Phytophthora infestans population. J. Hered. 76: 431-435 
33. Bishop, H. 1940. A study of sexuality in Sapromyces reinschii. Mycologia 32: 505-529 
34. Brasier, C. M. 1992. Evolutionary biology of Phytophthora. Part I: Genetic system, sexuality and the generation of variation. Annu. Rev. Phytopathol. 30: 153-177 
35. Lee, Y. S. and S. P. Lee. 2002. Development of rapid molecular detection marker for Colletotrichum spp. in leaf and fruit tissues of sweet persimmom. J. Microbiol. Biotechnol. 12: 989-992 
36. Fisher, D. J. and M. D. Hayes. 1984. Studies of mechanisms of metalaxyl fungitoxicity and resistance to metalaxyl. Crop Prot. 3: 117-185 
37. Judelson, H. S. 1996. Physical and genetic variability at the mating type locus of the oomycete, Phytophthora infestans. Genetics 144: 1005-1013 
38. Choi, J. Y., Y. H. Je, J. G. Kim, Y. C. Choi, W. T. Kim, and K. Y. Kim. 2004. Simultaneous detection and differentation of Vairimorpha spp. and Nosema spp. by multiplex polymerase chain reaction. J. Microbiol. Biotechnol. 14: 737-744 
39. McPherson, M. J., R. J. Oliver, and S. J. Gurr. 1992. The polymerase chain reaction, pp. 123-145. In Gurr, S. J., M. J. McPherson, and D. J. Bowles (eds.), Molecular Plant Pathology. A Practical Approach, vol. 1. IRL Press, NY, U.S.A. 

이 논문을 인용한 문헌 (4)

1. 2006. "" Journal of microbiology and biotechnology, 16(3): 487~490  원문보기 상세보기
2. 2006. "" Journal of microbiology and biotechnology, 16(5): 724~730  원문보기 상세보기
3. 2007. "" Journal of microbiology and biotechnology, 17(6): 1041~1044  원문보기 상세보기
4. Lee, Kang-Hyeong ; Song, Hong-Gyu 2008. "Production of Antifungal Materials by Bacillus sp. Which Inhibit Growth of Phytophthora infestans and Fusarium oxysporum" Korean journal of microbiology = 미생물학회지, 44(3): 258~263  원문보기 상세보기