野生稻颖果总RNA提取方法研究

Method for Total RNA Isolation from Wild Rice Caryopses

目的：比较以确定适用于野生稻总RNA提取的方法。方法：用改良的异硫氰酸胍法、TR／ZOL法、SDS法和CTAB法分别提取云南3种野生稻和栽培稻滇超6号在灌浆期颖果的总RNA。结果：SDS法能够提取的普通野生稻和疣粒野生稻的总RNA的纯度高，其A260/A280介于1．9938～1．9267；而TRIZOL法只适用于栽培稻滇超6号，CTAB法对于疣粒野生稻和药用野生稻提取效果一般，电泳条带不很清晰，其A260/A280介于1．8243～1．6928；而新建立的改良的异硫氰酸胍法都能提取完整性好、高得率（浓度介于0．9358～1．2008μ/μl）和高纯度的RnA（A260/A280大于1．8），电泳28SrRNA和18SrRNA条带清晰，且该方法操作简单、耗时少、效率高。结论：改良的异硫氰酸胍法适合于野生稻颖果总RNA的提取，提取的总RNA完整性好、得率高。

Objective and Methods：In order to find the best method to isolate the RNA from the caryopses of three species of wild rice and cultivated rice （dian-chao-six）, the improved Isothiocyanate method, the Trizol method, the SDS method and the CTAB method were utilited and compared in this paper. Results： The purity of the RNA isolated by the SDS method from Oryza. rufipogon and. O. granu/ata was high and the ratios of A260/A280 were between 1.9938 and 1.9267; while TRIZOL method was only applied to dian- chao- six, the extraction effect of the RNA which isolated from O. granulata and O. officinalis by CTAB method is moderate, and the bands was obscure and the ratios of A260/A280 were between 1.8243 and 1.6928; The RNA isolated by the improved Isothiocyanate method produced two clear bands of 28S rRNA and 18S rRNA ,the integrity and purity was good（the ratios of A260/A280 were above 1.8）,the output rate, too（concentration ranged from 0.9358 to 1.2008μg/μl）. and this method was simple and less time- consuming, high efficiency. Conclusion：The improved Isuthiocyanate method suited to the total RNA extraction from the earyopsis of wild rice species, and its good integrity and high output rate.