Journal of Korean Society of Forest Science (한국산림과학회지)

Korean Society of Forest Science (한국산림과학회)
권호 표지

Genetic Diversity of Mulberry Dwarf Phytoplasma(MD) by SSCP Technique

SSCP기법에 의한 뽕나무오갈병 파이토플라스의 유전적 다형성 분석

  • Han, Sangsub (Department of Forest Enviroment Sciences, Chonbuk National University)
  • 한상섭 (전북대학교 산림환경과학과)
  • Published : 2013.06.30
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Abstract

Phytoplasmas were detected consistently in 42 mulberry cultivars showing dwarf disease using DNA analysis by amplification with phytoplasma universal primer pairs P1/P7 (about 1.8 kb and R16F2n/R2 (about 1.2 kb). The point mutation from 42 cultivars of mulberry tree was detected by single-strand conformation polymorphism (SSCP) analysis. The SSCP profiles were clearly observed from all of cultivars in 8% polyacrylamide gel, electrophoresizing for and running 8-15 hrs. at 150V, $10^{\circ}C$. The MD and JWB phytoplasma PCR products was mixed and electrophoresis was performed to detect their polymorphism. In this results, the SSCP profiles of all bands of MD and JWB were analyzed on single lane and were distinct in their each of band patterns. The SSCP analysis was possible to detect of 1.8 kb and 1.2 kb nucleotide size and near close band patterns were distinct by mix of two samples. Previously, it was only possible to detect of point mutation under 600 bp nucleotide sequence by SSCP analysis but this modification of SSCP technique was possible to detect clearly SSCP band patterns of about 1.8 kb and 1.2 kb nucleotides.

파이토플라스마 증폭 프라이머, P1/P7 및 R16F2n/R2를 이용하여 뽕나무 품종 42개체에 대하여 뽕나무오갈병 파이토플라스마의 전염여부를 조사한 결과 공시 모두에서 파이토플라스마가 검출되었다. 뽕나무오갈병 파이토플라스마 단일염기변이를 SSCP분석기법을 응용하여 분석조건을 조사한 결과 P1/P7(약 1.8 kb) 및 R16F2n/R2(약 1.2kb)로 증폭한 PCR산물에서는 6% polyacrylamide gel 농도, 150V, $10^{\circ}C$의 전기영동 조건에서 SSCP밴드패턴이 나타났다. 유사한 SSCP밴드 패턴을 보이는 두 시료간의 밴드형태를 뚜렷하게 구별하는 방법을 찾기 위하여 뽕나무 오갈병파이토플라스마와 대추나무 빗자루병 파이토플라스마의 P1/P7 및 R16F2n/R2 프라이머로 증폭한 PCR산물을 혼합한 후 SSCP분석 결과, 전기영동상에서 대추나무 파이토플라스마와 뽕나무 파이토플라스마의 SSCP 밴드패턴 모두를 관찰할 수 있었다. 본 연구 결과, 기존에 약 600 bp 크기로 한정된 것으로 알려진 SSCP 분석법을 응용하여 파이토플라스마 PCR 산물 1.8 kb 또는 1.2 kb 크기에서도 유사한 SSCP 밴드패턴에 의하여 단일염기변이를 검출할 수 있었다.

Keywords

References

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