甜瓜种子中种传病毒的RT-PCR检测  

Detection of seed- transmitted viruses from dry melon seeds using RT-PCR

作　　者：彭斌[1] 汪丽刚[1] 古勤生[1] 

机构地区：[1]河南省果树瓜类生物学重点实验室.中国农业科学院郑州果树研究所,郑州450009

出　　处：《中国瓜菜》2015年第6期10-15,共6页China Cucurbits And Vegetables

基　　金：国家西甜瓜产业技术体系项目(CARS-26-13);中国农业科学院科技创新工程专项经费项目(CAAS-ASTIP-2015-ZFRI);河南省基础与前沿技术研究项目(152300410138)

摘　　要：为了建立葫芦科作物干种子主要种传病毒检测方法,在现有的TRIZOL法、CTAB法等提取植物总RNA方法的基础上进行改良,从富含有多糖的甜瓜干种子中提取高质量的总RNA。建立了18 S r RNA为内参基因的侵染葫芦科作物4种主要种传病毒的两重RT-PCR检测方法。应用改良TRIZOL法分别提取单粒和0.1 g(约20粒)带毒甜瓜种子的总RNA作为检测模板,分别对4种种传病毒进行RT-PCR检测,结果表明可分别从携带4种种传病毒的甜瓜种子上检测到对应的病毒,18 S r RNA基因能够很好的作为内参基因对病毒检测结果进行评估。In order to establish a method of detection of the major seed-transmitted virus in dry cucurbit seeds, we modified the total RNA extract methods based on TRIZOL and CTAB, and then the high quality of total RNA was obtained from dry melon seeds rich in polysaccharide. A duplex RT-PCR method was developed for four kinds of seed-transmitted viruses in- fecting cucurbits respectively, including an internal amplification control 18 S rRNA gene of host. The total RNA extracted from single and mixture （about 20 seeds） infected melon seeds by modified TRIZOL method were used as templates of dul- plex RT-PCR detection for the four kinds of viruses respectively. The results indicated that RT-PCR detection system was capable of detecting four different viruses pathogens in samples, and the 18 S rRNA gene as internal control was a useful in- dicator of the quality of plant total RNA extraction effect and the effectiveness of RT-PCR.

关 键 词：葫芦科作物 干种子 多糖 种传病毒 反转录聚合酶链式反应 

分 类 号：S436.5[农业科学—农业昆虫与害虫防治]