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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (4): 198-202.

所属专题： 园艺

• 林学 园艺 园林 • 上一篇    下一篇

红地球葡萄GLRaV-3的茎尖培养脱毒及RT-PCR检测

韩艳婷 石雪晖 杨国顺 王先荣 刘昆玉 徐 丰 倪建军   

  • 收稿日期:2010-08-23 修回日期:2010-10-26 出版日期:2011-02-10 发布日期:2011-02-10
  • 基金资助:

    国家农业部农业公益性行业科研项目;湖南省科技厅重点科研项目

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

  • Received:2010-08-23 Revised:2010-10-26 Online:2011-02-10 Published:2011-02-10

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摘要：

本文以带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: 链霉菌702, 链霉菌702, 红谷霉素, 抑菌活性, 毒力方程

Abstract:

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. The RT-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%.