Proceedings of the Korean Society of Plant Pathology Conference (한국식물병리학회:학술대회논문집)

The Korean Society of Plant Pathology (한국식물병리학회)
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A simple method for detection of CMV viral RNAs and satellite RNAs in Korean pepper.

  • J.H. Sung (Dept. Biology, Yeungnam University) ;
  • Park, J.H. (Dept. Biology, Yeungnam University) ;
  • H.Y. Shin (Dept. Biology, Yeungnam University) ;
  • M.U. Chang (Dept. Biology, Yeungnam University) ;
  • H. Sayama (Nippon Del Monte Corporation, R＆D) ;
  • H. Atarashi (Nippon Del Monte Corporation, R＆D)
  • Published : 2003.10.01
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Abstract

To analyze the genome of Cucumber mosaic virus(CMV) in pepper, we developed a new extraction method for double-stranded RNA(dsRNA). To isolate the dsRNA, 0.1g of pepper leaves homogenized with 1ml of 5${\times}$EXB extraction buffer[0.5M glycin, 0.5M NaCl, 5mM EDTA(pH9.0/NaOH), 10％ Sodium N-lauryl salcosinate(NLS), 10％ Sodium dodecylsulfate(SDS)] and purified with the 1/4 volume of phenol： chloroform： isoamylalcohol(25：24：1). dsRNAs from the aqueous phase was precipitated with isopropanol. This procedure was able to detect a minimal amount of dsRNA from CMV infected plant tissue and to distinguish different CMV satellite RNAs by polyacrylamide gel electrophoresis(PAGE). Moreover, this method can be applied CMV infected in pepper or Rice dwarf virus (RDV) infected rice.

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