香蕉不同组织中总RNA提取方法的研究

A Study of the Methods for Total RNA Extraction from Different Tissues of Banana

针对香蕉组织中多糖、多酚等次生物质含量高的特点,以香蕉根、茎、叶、果实为试材,对其总RNA提取进行研究．比较了SDS法、改良SDS法、改良CTAB法和Trizol试剂盒法提取总RNA的效果,结果分析表明:改良SDS法能从香蕉根、茎、叶中获得质量高、完整性较好的总RNA,而改良CTAB法对果实的提取效果较佳,所得A 260/A 280均高于1．8,A 260/A 230大于2．0,28 S rRNA带的亮度约是18 SrRNA的2倍．其产率在根、茎、叶中均在400μg·g^(-1)以上,果实中可达49．34μg·g^(-1)．以上2种方法所得的总RNA均能满足RT-PCR、Northern blot和cDNA文库建立等分子生物学研究,且具有快速、简便易行、成本较低的特点,适合于富含多糖、多酚的植物材料．

SDS, modified SDS, modified CTAB and Trizol Kit were employed to extract total RNA from the tissues of banana root, stem, leaf and fruit. Modified SDS was found to be the best procedure for RNA extraction from banana root, stem and leaf, while modified CTAB was the best for RNA extraction from the fruit. The ratios of A260/A280 and A260/A230 were up to 1.8 and 2.0, respectively, for both methods. The band 28S was twice as bright as the band 18 S by visualizing the ribosomal RNA of the samples on agarose gel electrophoresis. The yield was above 400 μg/g in the root, stem and leaf samples and 49.34 μg/g in the fruit samples. Therefore, the extracted RNA should be suitable for application in RT-PCR, Northern blot, cNDA library construction and other biotechnological researches, and the two methods are believed to be especially useful for total RNA extraction of plant materials which have plenty of polysaccharides and hydroxybezene.