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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (7): 155-159.

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

一种适合葡萄多种组织的总RNA提取方法

李红熙 徐美隆 杨智   

  • 收稿日期:2011-11-15 修回日期:2012-01-13 出版日期:2012-03-05 发布日期:2012-03-05

A Method Suited to Extract Total RNA from Various Tissues of Grapevine

  • Received:2011-11-15 Revised:2012-01-13 Online:2012-03-05 Published:2012-03-05

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摘要：

为了解决葡萄组织中总RNA提取难度大的问题，以感染葡萄卷叶病毒-3的葡萄品种‘蛇龙珠’为试验材料，采用CTAB结合SDS法对总RNA提取方法进行了研究，建立了一套适合葡萄叶片、叶柄及韧皮部等多种组织总RNA提取的方法。结果表明，该方法在多种组织中均能获得高质量的RNA，OD260/OD280=1.8~1.9，电泳结果表明，所得RNA完整性好，无降解。同时用所得RNA进行了GLRaV-3的检测，电泳结果表明该法所得RNA可满足葡萄病毒病RT-PCR检测的质量要求。该方法操作简单，结果稳定，对其他富含多酚、多糖类植物组织总RNA的提取具有借鉴意义。

关键词: 油茶, 油茶, 硬脂酰-酰基载体蛋白脱饱和酶, 原核表达载体构建

Abstract:

In order to solve the problem of extracting total RNA from grape tissues, the author took the leaf-roll associated virus 3 infected grapes varieties ‘Gernischt’ as materials, the method of CTAB combined with SDS was studied in this paper, a suitable method for total RNA extraction from leaves, petioles and phloem of grapevine was established. The results indicated that high-quality RNA could be obtained by this method, with OD260/OD280=1.8-1.9, without degradation. The results showed that the RNA obtained could be used for RT-PCR to detect grapevine viruses. The method was simple, the results were stable. It also gave better suggestions to extract total RNA of tissues which rich in polyphenol and polysaccharides.