应用RT-PCR技术检测甜菜坏死黄脉病毒  

RT-PCR Detection for Beet Necrotic Yellow Vein Virus (BNYVV)

作　　者：刘大丽[1] 马龙彪[1] 王皙玮[2] 吴玉梅[2] 张福顺[2] 

机构地区：[1]黑龙江省普通高等学校甜菜遗传育种重点实验室,哈尔滨150080 [2]中国农业科学院甜菜研究所

出　　处：《中国糖料》2008年第2期12-14,17,共4页Sugar Crops of China

基　　金：农业部标准项目[农财发(2006)64号]

摘　　要：为了更加准确地检测患有丛根病的甜菜植株中BNYVV病毒的含量,实验采用了RT-PCR的方法分别从26株待测甜菜的根中扩增获得了324bp的BNYVV病毒基因片断。通过调整反转录cDNA模板的浓度,研究发现,当cDNA模板稀释到100倍时,能更加精准地检测到不同丛根病染病植株中BNYVV病毒含量的差异。同时,半定量PCR的统计结果进一步表明.当待测植株中BNYVV病毒的相对含量低于阳性对照的一半时,甜菜丛根病病症并不明显;而当其相对含量超过阳性对照的1.3倍时,甜菜受害严重,有些几乎死亡。本检测方法为更加及时、快捷、准确和系统地检测甜菜植株的染病程度打下了良好的基础。A diagnostic test based on reverse-transcription for the detection of beet necrotic yellow vein virus （BNYVV）. polymerase chain reaction （RT-PCR） was developed RNA was extracted from 26 sugarbeet seedling roots with different symptom of rhizomania. A specific 324bp fragment was amplified based on primers designed according to RNA-5 sequence in BNYVV genome, and the different seedlings exhibited different contents of BNYVV virus with the exact cDNA concentrations in dilutions of 1×10^-2. Meanwhile, semi-quantitative PCR analysis further indicated that the symptom BNYVV in sugarbeet below 0.5-fold of that of rhizomania was no obvious when the comparative content of in the positive control, and the sugarbeet seedlings were damaged severely, and some almost dead when the content of BNYVV exceeded 1.3-fold of that in the positive control. The RT-PCR technology supplied a useful tool to study the rhizomania of sugarbeet exactly and comprehensively.

关 键 词：甜菜 RT-PCR 丛根病 甜菜坏死黄脉病毒 

分 类 号：S435.663[农业科学—农业昆虫与害虫防治]