首页|期刊导航|中国农业科学(英文版)|Dot-Blot Hybridization for Detection of Five Cucurbit Viruses by Digoxigenin-Labelled cDNA Probes
中国农业科学(英文版)2007，Vol.6Issue(12)：1450-1455,6.
Dot-Blot Hybridization for Detection of Five Cucurbit Viruses by Digoxigenin-Labelled cDNA Probes
Dot-Blot Hybridization for Detection of Five Cucurbit Viruses by Digoxigenin-Labelled cDNA Probes
摘要
Abstract
Dot-blot hybridization was applied in this paper to detect five viruses infecting cucurbitaceous crops,Zuccini yellow mosaic virus(ZYMV),Watermelon mosaic virus(WMV),Cucumber mosaic virus(CMV),Papaya ringspot virus watermelon strain(PRSV-W)and Squash mosaic virus(SqMV),as a good alternative assay in seed health test and epidemiological and transgenic research.Digoxigenin-labelled cDNA probes of the five viruses were synthesized by PCR with the specific primers and applied in dot-blot hybridization to detect five viruses in crude extraction of the infected leaves.And three SqMV probes of different lengths(0.55,1.6,and 2.7 kb,respectively)were designed to investigate the effect of hybridization.The results showed that the sensitivity for detecting the crude extraction of infected leaves by ZYMV,WMV,CMV,PRSV-W,and SqMV was down to 1:160,1:160,1:320,1:160,and 1:320,respectively.Three SqMV probes of different length showed no differences on the sensitivity and specificity.The digoxigenin-labelled probes prepared by PCR could be used for accurate and rapid identification of 5 viruses infecting cucurbitaceous crops with good stabilities,sensitivities,specificity,and reproducibilities.关键词
PCR/digoxigenin-labelled cDNA probe/dot-blot hybridization/ZYMV/WMV CMV/PRSV-W/SqMVKey words
PCR/digoxigenin-labelled cDNA probe/dot-blot hybridization/ZYMV/WMV CMV/PRSV-W/SqMV分类
农业科技引用本文复制引用
MENG Juan,GU Qin-sheng,LIN Shi-ming,PENG Bin,LIU Li-feng,TIAN Yan-ping,LI Li..Dot-Blot Hybridization for Detection of Five Cucurbit Viruses by Digoxigenin-Labelled cDNA Probes[J].中国农业科学(英文版),2007,6(12):1450-1455,6.基金项目
This study was supported bv the Special Program for Key Basic Research of the Ministry of Science and Technology,China(2005CCA01700) （2005CCA01700）
and the fund of General Administration of Quality Supervision,Inspection,and Quarantine of China (2005IK0061). （2005IK0061）
