《植物生理学报》 2014, 50(4): 558-562

两种适用于葡萄不同组织RNA提取方法的比较

初雪鹏, 佟少明, 侯和胜*

辽宁师范大学生命科学学院, 辽宁省植物生物工程重点实验室, 辽宁大连116082

通信作者：侯和胜；E-mail: hesheng_hou@126.com；Tel: 0411-82159112

摘　要：

以不同葡萄组织为材料对目前常用的2种总RNA提取方法——改良SDS法和CTAB-LiCl法进行研究。2种RNA提取方法中均不使用酚。采用这两种方法从葡萄不同组织中均成功地提取到RNA, 琼脂糖凝胶电泳结果显示28S和18S rRNA条带完整清晰。检测A₂₆₀/A₂₈₀值分布在1.7~2.0之间, A₂₆₀/A₂₃₀值分布于1.9~2.3之间, 说明RNA质量较高。管家基因Actin和ACS5基因的检测表明2种方法所得RNA能够满足RT-PCR和基因克隆等研究需要。改良SDS法的RNA得率是CTAB-LiCl法的RNA得率的2~3倍, 而CTAB-LiCl法获得的RNA纯度高。可以根据原料的数量和对RNA质量的要求来选取最佳提取方法。

关键词：葡萄; 改良SDS法; CTAB-LiCl法; RNA提取

收稿：2013-11-26   修定：2014-02-25

资助：国家自然科学基金(30671439)和教育部博士点基金(2009-2136110001)。

Comparison of Two Methods for RNA Extraction from Different Tissues of Grape (Vitis vinifera L.)

CHU Xue-Peng, TONG Shao-Ming, HOU He-Sheng*

Liaoning Key Laboratory of Plant Biotechnology, College of Life Sciences, Liaoning Normal University, Dalian, Liaoning 116082, China

Corresponding author: HOU He-Sheng; E-mail: hesheng_hou@126.com; Tel: 0411-82159112

Abstract:

Two total RNA extraction methods commonly used at present, modified SDS and CTAB-LiCl methods, were studied with different grape tissues. Phenol wasn’t used in the two methods. Higher quality total RNAs from different grape tissues were obtained with both of two methods. Agrose gel electrophoresis showed that 28S and 18S bands were clear and bright. The ratio of A260/A280 was from 1.7 to 2.0 while that of A260/A230 was from 1.9 to 2.3 which demonstrated the high quality of the isolated RNAs. The results of amplification of Actin, a housekeeping gene in plant, and ACS5 gene showed that the RNAs obtained could meet the requirements of RTPCR and gene cloning. The yield of RNA obtained by modified SDS method was 2−3 times that by CTAB-LiCl method. The total RNA isolated by the CTAB-LiCl method was highly integrate and pure. A preferable method could be selected according to the material quantity and the requirements of the RNA quality.

Key words: grape (Vitis vinifera); modified SDS method; CTAB-LiCl method; RNA extraction