红地球葡萄GLRaV-3的茎尖培养脱毒及RT-PCR检测.PDFVIP

中国农学通报 2011,27(04): 198-202 Chinese Agricultural Science Bulletin 红地球葡萄GLRaV-3的茎尖培养脱毒及RT-PCR检测 1 1 1 2 1 1 1 韩艳婷 ，石雪晖 ，杨国顺 ，王先荣 ，刘昆玉 ，徐 丰 ，倪建军 1 2 （湖南农业大学园艺园林学院，长沙410128 ；湖南神州庄园葡萄酒业有限公司，湖南澧县415500 ） 摘 要：笔者以带GLRaV-3 病毒的红地球为试材，研究了6-BA、NAA 、IBA、KT 不同配组及茎尖大小对茎 尖培养的影响，并利用RT-PCR 技术对部分红地球组培苗进行了GLRaV-3 病毒检测。研究结果表明：红 地球葡萄茎尖培养脱毒最佳茎尖初始培养基为：1/2MS+6-BA 0.5 mg/L+KT 0.5 mg/L+NAA 0.2 mg/L ，最 佳茎尖分化培养基为：B5+KT 1.0 mg/L+NAA 0.2 mg/L ；茎尖大小与成活率成正比，与脱毒率成反比；运 用RT-PCR 检测技术进行了脱毒后组培苗的检测，得到了GLRaV-3 的特异片段为300 bp ，获得了28 株脱 毒苗，平均脱毒率为43%。 关键词：红地球葡萄；茎尖培养；脱毒；葡萄卷叶病；RT-PCR 中图分类号：S663.1 文献标志码：A 论文编号：2010-2514 ShootTipTissueVirus-freeCultureandRT-PCRDetectionofGLRaV-3inRedGlobeGrape 1 1 1 2 1 1 1 Han Yanting , Shi Xuehui , Yang Guoshun , Wang Xianrong , Liu Kunyu , Xu Feng , Ni Jianjun 1 ( CollegeofHorticultureandLandscape, HunanAgriculturalUniverdity, Changsha 410128; 2HunanShenzhouZhuangyuanGrapeWineIndustryLimitedCompany, Li County Hunan 415500) Abstract: Effects of the stem-apex size and different combinations among 6-BA, NAA, IBA and KT on shoot tip culture invitrowere studied by using red globe grape infected with GLRaV-3 virus. GLRaV-3 was detected early in some of red globe grape of plantlets in vitro with RT-PCR technique. The results showed that the best suitable initial medium was 1/2MS+6-BA 0.5 mg/L+ KT 0.5 mg/L+NAA 0.2 mg/L. The optimum medium for shoot differentiation was B5 + KT 1.0 mg/L + NAA0.2 mg/L. Plants free of GLRaV-3 were found. The size of excise