Research in Plant Disease (식물병연구)

The Korean Society of Plant Pathology (한국식물병리학회)
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Detection of Colletotrichum acutatum and C. gloeosporioides by Real Time PCR

Real Time PCR을 이용한 Colletotrichum acutatum과 C. gloeosporioides의 검출

  • Kim, Seung-Han (Bonghwa Alpine Medicinal Plant Research Station) ;
  • Kwon, Oh-Hun (Yeongyang Pepper Research Station of Kyeongbuk Agricultural Research and Extension Service)
  • 김승한 (경상복도농업기술원 봉화고냉지약초시험장) ;
  • 권오훈 (영양고추시험장)
  • Published : 2008.12.01
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Abstract

Real time PCR was used to discriminate Colletotrichum acutatum and C. gloeosporioides for analysis of population density. Two primers, caInt2 and cgint, used for conventional PCR to discriminate two species were modified with fluorescent dye to make probe for real time PCR. Fluorescence signals were successfully detected by fCaInt2 and vCgint probe coupled with primer pair Unicon and Unicor1 resulting in discrimination of C. acutatum and C. gloeosporioides by comparison of delta Rn value.

C. gloeosporioides와 C. acutatum의 개체군 밀도분석을 위해 기존 ITS부위를 이용한 PCR방법에 사용한 caInt2와 cgint 프라이머에 형광을 표지하여 C. acutatum에 특이적인 fcaInt2와 C. gloeosporioides에 특이적인 vcgint의 두 probe를 제작하였다. 이 두개의 프라이머와 Unicof1, Unicor1 primer를 이용 real time PCR을 수행하였을 때 C. acutatum은 fcaInt2 probe에, C. gloeosporioides는 vcgint에 특이적인 형광증폭곡선을 나타냄에 따라 delta Rn 값을 비교함으로 두 종의 구분이 가능하였다.

Keywords

References

  1. Adaskaveg, J. E. and Hartin, R. J. 1997. Characterization of Colletotrichum acutatum isolates causing anthracnose of almond and peach in California. Phytopathology 87: 979-987 https://doi.org/10.1094/PHYTO.1997.87.9.979
  2. Afanador-Kafuri, L., Minz, D. , Maymon, M. and Freeman, S. 2003. Characterization of Colletotrichum isolates from tamarillo, passiflora, and mango in Colombia and identification of a unique species from the genus. Phytopathology 93: 579-587 https://doi.org/10.1094/PHYTO.2003.93.5.579
  3. Chung, W.-H., Ishii, H., Nishimura, K., Fukaya, M., Yano, K. and Kajitani, Y. 2006. Fungicide sensitivity and phylogenetic relationship of anthracnose fungi isolated from various fruit crops in Japan. Plant Dis. 90: 506-512 https://doi.org/10.1094/PD-90-0506
  4. Fraaije, B. A., Cools, H. J., Fountaine, J., Lovell, D. J., Motteram, J., West, J. S. and Lucas, J. A. 2005. Role of ascospores in further spread of QoI-resistant cytochrome b alleles (G143A) in field populations of Mycosphaerella graminicola. Phytopathology 95: 933-941 https://doi.org/10.1094/PHYTO-95-0933
  5. Freeman, S., Pham, M. and Rodriguez, R. J. 1993. Molecular genotyping of Colletotrichum species based on arbitrarily primed PCR, A+Trich DNA, and nuclear DNA analyses. Exp. Mycol. 17: 309-322 https://doi.org/10.1006/emyc.1993.1029
  6. Freeman, S. and Rodriguez, R. J. 1995. Differentiation of Colletotrichum species responsible for anthracnose of strawberry by arbitrarily primed PCR. Mycol. Res. 99: 501- 504 https://doi.org/10.1016/S0953-7562(09)80653-9
  7. Freeman, S., Shalev, Z. and Katan, J. 2002. Survival in soil of Colletotrichum acutatum and C. gloeosporioides pathogenic on strawberry. Plant Dis. 86: 965-970 https://doi.org/10.1094/PDIS.2002.86.9.965
  8. Haugland, R. A., Brinkman, N. and Vesper, S. J. 2002. Evaluation of rapid DNA extraction methods for the quantitative detection of fungi using real-time PCR analysis. J. Microbiol. Methods 50: 319-323 https://doi.org/10.1016/S0167-7012(02)00037-4
  9. Hollick, P. S., Taylor, R. J., Mccomb, J. A., Dixon, K. W. and Krauss, S. L. 2004. Optimisation of DNA extraction for AFLP analysis of mycorrhizal fungi of terrestrial orchids Caladeniinae and Drakaeinae. Plant Mol. Biol. Reptr. 22: 307a-307h https://doi.org/10.1007/BF02773143
  10. Mccartney, H. A., Foster, S. J., Fraaije, B. A. and Ward, E. 2003. Molecular diagnostics for fungal plant pathogens. Pest Manag. Sci. 59: 129-142 https://doi.org/10.1002/ps.575
  11. Peres, N. A. R., Souza, N. L., Peever, T. L. and Timmer, L. W. 2004. Benomyl sensitivity of isolates of Colletotrichum acutatum and C. gloeosporioides from citrus. Plant Dis. 88: 125-130 https://doi.org/10.1094/PDIS.2004.88.2.125
  12. Ward, E. and Akrofi, A. Y. 1994. Identification of fungi in the Gaeumannomyces-Phialophora complex by RFLPs of PCRamplified ribosomal DNAs. Mycol. Res. 98: 219-224 https://doi.org/10.1016/S0953-7562(09)80189-5