应用RT-PCR检测温州蜜柑萎缩病毒资料.PDFVIP

33 2 植 物 保 护 学 报 Vo.l 33 No. 2 2006 6 ACTA PHYTOPHYLACICA SINICA June 2006 RT-PCR 1, 2 1, 2* 2 2 2 高艳玲 周常勇 王雪峰 周 彦 刘 英 (1. , 400712; 2. , 400716) : 温州蜜柑萎缩病是以日本为主的少数亚洲国家柑橘上的重要病毒病害, 多数柑橘品种隐症 毒, 不易发现, 对其早期准确快速检测尤为重要以毒源植株的叶枝皮为材料, 对提取的总 RNA和总核酸进行反转录和 PCR扩增,通过 SDV特异引物的设计与筛选,反应体系与反应程序的 建立与优化,扩增得到 一个251bp的特异片段,测序结果与日本 Iwanami报道的 SDV序列同源性 为 99. 6 RT-PCR检测体系的灵敏度为 100ng, 同时应用该检测体系可以全年检测到毒源植株 嫩叶嫩皮老叶老皮中的 SDV : 温州蜜柑萎缩病; RT-PCR; 检测 RT-PCR detection of Satsum a dwarf virus 1, 2 1, 2* 2 2 2 GAO Yan-ling ZHOU Chang-yong WANG Xue-feng ZHOU-Yan LIU-Ying ( 1. Citrus Research Institute, ChineseAcademy ofAgriculture Sciences, Chongqing 400712, China; 2. Plant Protection College, SouthwestUniversity, Chongqing 400716, China) Abstract: Satsumadwarf virus( SDV) is an mi portant citrus virusmainly occurred in Japan and other Asian countries. Generallymost hostscarry the viruswithout symptom. To facilitate efficiently detecting SDV, especially from the symptomlesshosts, new diagnosticmethods are in demand. In this study, the totalRNAsor totalnucleic acidsextracted from the tissue of citrus infectedwith SDV were used as tem- plate. The prmi erswere specifically designed and selected. The reaction system of reverse transcription- polymerase chain reaction (RT-PCR) hasbeen set up and optmi ized. A target band of 251bpwas ob- tained, which has99. 6 of sequence homology comparedwith the sequence reported by Iwanami in Ja- pan. The sensitivity ofRT-PCR system forSDV in