摘要
以卷丹、麝香百合品种富田和离体鲜切花的花蕾为材料,比较Trizol法、异硫氰酸胍法、CTAB改进法和SDS改进法提取总RNA的效果,结果表明,SDS改进法能有效去除多糖,提取的RNA中28SrRNA亮度约为18SrRNA的两倍,OD260/OD280值介于1.7～2.2之间,卷丹RNA得率为132.52μg/g,富田RNA得率为186.88μg/g。进一步用SDS改进法分别提取富田外轮花瓣、内轮花瓣、雄蕊、雌蕊、叶和茎的RNA,同样可以获得完整的纯度高的RNA,其中28SrRNA亮度约为18SrRNA的两倍,OD260/OD280值介于1.7～2.2之间,说明此方法适用于百合各个组织RNA的提取。经RT-PCR获得了花发育基因的特异性条带,说明用SDS改进法从百合中提取的RNA质量好、产率高、完整性强,完全适合于百合进一步的分子生物学研究。
The performances of four extraction methods of total RNA： Trizol, guanidinium isothiocyanate extraction, modified CTAB and modified SDS, were compared with in this report. The results showed that the modified SDS extraction was capable of efficiently removing polysaccharides, the brightness 28S rRNA of the RNAs isolated by the modified SDS was two times as much as that of 18S rRNA, OD260/OD280 ranged between 1.7-2.2, and the RNA productivities from Lanceleaf lily and Futian were 132.52μg/g and 186.88μg/g. The modified SDS extraction was also used in the lily tissue of outer petal, inner petal, stamen, carpel, leaves and stems, the RNA performances were the same as that of buds. It was indicated that the modified SDS extraction could be used in other tissue of lily. The specific RNA bands obtained by RT-PCR explained that the modified SDS could isolate high-quality and highly-complete RNA at a higher productivity and thus was worth further biological research.
出处
《分子植物育种》
CAS
CSCD
2006年第6期871-876,共6页
Molecular Plant Breeding
基金
上海科技兴农重点课题“百合种质创新和快繁技术研究”（2005-2007）资助.
