Detection of Potato Spindle Tuber Viroid Using RT-PCR Technique

RT-PCR 기법을 이용한 감자 걀쭉 바이로이드 (Potato Spindle Tuber Viroid)의 검정

  • Joung, Young-Hee (Plant Tissue Culture Research Unit, Korea Research Institute of Bioscience & Biotechnology(KRIBB)) ;
  • Jeon, Jae-Heung (Plant Tissue Culture Research Unit, Korea Research Institute of Bioscience & Biotechnology(KRIBB)) ;
  • Choi, Kyung-Hwa (Plant Tissue Culture Research Unit, Korea Research Institute of Bioscience & Biotechnology(KRIBB)) ;
  • Kim, Hyun-Soon (Plant Tissue Culture Research Unit, Korea Research Institute of Bioscience & Biotechnology(KRIBB)) ;
  • Joung, Hyouk (Plant Tissue Culture Research Unit, Korea Research Institute of Bioscience & Biotechnology(KRIBB))
  • 정영희 (생명공학연구소 식물조직배양연구 Unit) ;
  • 전재홍 (생명공학연구소 식물조직배양연구 Unit) ;
  • 최경화 (생명공학연구소 식물조직배양연구 Unit) ;
  • 김현순 (생명공학연구소 식물조직배양연구 Unit) ;
  • 정혁 (생명공학연구소 식물조직배양연구 Unit)
  • Published : 1997.08.01

Abstract

Potato spindle tuber viroid(PSTVd) RNAs were isolated from PSTVd-inoculated potato cv. Superioc and carried out RT-RCR with reverse transcriptase and PSTVd specifie primer pair desigened to amplify the 356 nucleotides of PSTVd genome. As a result, 356 nucleotides PCR products were amplified from PSTVd-inoculated potato cv. Superior. The 356 nucleotides DNA fragment was indeed the PSTVd geneby sequencing analysis. PSTVd could be successfully detected from infected leaf and tuber tissue of potato by using RT-PCR technique. Especially PSTVd was more effectively detected when both downstream and upstream primer were used than only downstream primer was used in RT reaction.

PSTVd 를 접종시킨 감자의 Superior 품종으로부터 PSTVd RNA를 분리하여 역전사효소와 PSTVd genome 중 356 uncleotides를 증폭할 수 있는 PSTVd 특이적 primer 쌍을 사용하여 RT-PCR를 수행한 결과 356 nuclcotides의 DNA 절편이 증폭되었고 이 절편을 sequencing인 결과 PSTVd 의 유전자임을 확인하였다. 감염된 잎과 괴경에수 모두 RT-PCR 기법을 이용한 PSTVd 의 검정이 가능하였고 특히 RT 반응시 downstream primer만을 이용할 때보다 downstream과 upstream primer를 동시에 사용할 때가 PSTVd 의 검정에 더 효과적이었다.

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