木本植物根系总RNA提取方法的比较和分析

Comparison with methods for total RNA extracting from roots of woody plants

以杂种马褂木不定根和杨树、柳树、柽柳根系组织为材料,比较异硫氰酸胍法、Tiangen试剂盒、CTAB改良法、Trizol法和改良Trizol法提取总RNA的效果。结果表明,改良Trizol法通过在离心后的匀浆上清液中加入低浓度的无水乙醇以及结合后续的高盐溶液,能有效去除多糖,提取的RNA中28S rRNA亮度约为18S rRNA的两倍,OD260/OD280值介于1.7～2.1之间,各林木树种根系组织RNA得率均大于150μg/g,而且整个提取时间只要2～3 h。表明改良Trizol法方便、快捷、高效,完全适用于林木树种根系组织RNA的提取。

The performances of five extraction methods of total RNA： guanidinium isothiocyanate extraction,Tiangen RNA plant Reagent,modified CTAB,Trizol and modified Trizol,were compared using the roots of four woody plants,such as Tamarix chinensis,Salix babylonica,Populus deltoids and Liriodendron hybrid,as the materials in this report.The results showed that the modified Trizol extraction was capable of efficiently removing polysaccharides by putting low concentration absolute alcohol into cleared homogenate solution and followed by adding 0.25 mL of a high salt precipitation solution.The brightness of 28S rRNA was two times as much as that of 18S rRNA in the total RNAs isolated by the modified Trizol.OD260/OD280 was ranged between 1.7～2.2,and the RNA productivities from roots of four woody plants were more than 150 μg/g.Meantime,all time of extracting RNA was only 2～3 h.Therefore,the modified Trizol method was convenient and efficient and could be used for extracting RNA from root of woody plants.