Journal of Plant Biochemistry and Biotechnology
  • Year: 2007
  • Volume: 16
  • Issue: 2

Detection of Cymbidium mosaic virus in Protocorm-Like Bodies in Dendrobium Sonia using One-Step RT-PCR

  • Author:
  • Yuphin Khentry1, Srimek Chowpongpang1, Ampaiwan Paradornuwat1, Sureeya Tantiwiwat2, Niphone Thaveechai1,
  • Total Page Count: 3
  • Page Number: 123 to 125

1Department of Plant Pathology, Faculty of Agriculture, Kasetsart University, Bangkok 10900, Thailand

2Department of Botany, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand

*Corresponding author. E-mail: agrnpt@ku.ac.th

AbbreviationsCymMV

Cymbidium mosaic virus;

ELISA

enzymelinked immunosorbent assay;

PLB

protocorm - like body;

RT-PCR

reverse transcription-polymerase chian reaction;

OD

optical density.

Abstract

Dendrobium orchids are clonally mass propagated by tissue culture techniques. However, Cymbidium mosaic virus is prevalent in vegetatively propagated Dendrobium spp. Therefore, a sensitive virus detection method is an important requirement for the production of CymMV-free orchid plants and control of virus spreading in the orchid industry. The presence of CymMV was screened in in vitro PLBs of Dendrobium Sonia using a one-step RT-PCR with specific primers to amplify a 858 bp fragment of a CymMV coat protein gene and flanking regions. Results were compared with those obtained from a conventional indirect ELISA. Only 22% samples showed the presence of CymMV in indirect-ELISA as compared to 78% in RT-PCR. Thus, the one-step RT-PCR appears to be a more sensitive method than indirect-ELISA for detecting CymMV in PLBs.

Keywords

CymMV, Dendrobium orchids, indirect ELISA, RT-PCR, PLBs