红地球葡萄GLRaV-3的茎尖培养脱毒及RT-PCR检测  

Shoot Tip Tissue Virus-free Culture and RT-PCR Detection of GLRaV-3 in Red Globe Grape

作　　者：韩艳婷[1] 石雪晖[1] 杨国顺[1] 王先荣 刘昆玉[1] 徐丰[1] 倪建军[1] 

机构地区：[1]湖南农业大学园艺园林学院,长沙410128 [2]湖南神州庄园葡萄酒业有限公司,湖南澧县415500

出　　处：《中国农学通报》2011年第4期198-202,共5页Chinese Agricultural Science Bulletin

基　　金：国家农业部农业公益性行业科研项目"葡萄现代产业技术体系研究与建立"(nyhyzx07-027);国家葡萄产业技术体系专项资金资助;湖南省科技厅重点科研项目"酿酒剌葡萄产业化技术示范"(2009CK-2010);湖南省教育厅省级科技计划(成果推广计划)项目"酿酒剌葡萄产业化示范"(09CY008)

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-BA0.5mg/L+KT0.5mg/L+NAA0.2mg/L,最佳茎尖分化培养基为:B5+KT1.0mg/L+NAA0.2mg/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300bp,获得了28株脱毒苗,平均脱毒率为43%。Effects of the stem-apex size and different combinations among 6-BA,NAA,IBA and KT on shoot tip culture in vitro were 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 excised shoot tip was proportional to the survival sate and inverse ratio to the virus elimination rate.TheRT-PCR detection technology was used for the GLRaV-3 detection and a 300 bp specific fragment was effectively amplified.The virology detection showed that virus-free red globe grape were obtained in 28 and the average rate of virus elimination was 43%.

关 键 词：红地球葡萄 茎尖培养 脱毒 葡萄卷叶病 RT-PCR 

分 类 号：S663.1[农业科学—果树学]