冻融法纯化水稻胚乳RNA及稻米品质重要相关基因表达谱研究

Isolation of Total RNA from Rice Endosperm and Expression of Important Genes Related to Grain Quality in Rice (Oryza sativa)

提出了一种从水稻胚乳组织中快速分离总RNA的方法,可以简便地去除RNA中存在的淀粉等多糖类杂质,所获的胚乳总RNA其质量可满足Northern印迹、微阵列制备等分子生物学实验要求。将从966个水稻品种的胚乳中分离所得的RNA制备RNA阵列,用于稻米品质重要基因Wx基因的表达谱分析,结果证实:水稻Wx基因表达丰度总体上与水稻胚乳的直链淀粉含量呈正相关;应用于Northern印迹实验,表明在水稻胚乳组织中,淀粉分支酶Rbe1具有两个长度不同的同源基因,其中一个分子量大的为胚乳特异表达,分子量小的则在水稻各组织中共表达。

A simple method was advanced to wipe starch off RNA and obtain purer total RNA.This method was successfully applied in Northern blot and RNA array experiments to study the expression of starch branching enzyme gene (Rbe1) and Wx gene, which were important for starch synthesis. And the results indicate that on the whole, the expression of Wx gene was linear with amylose content of rice endosperm, and there were two Rbe1 homogenes in rice endosperm, the longer expressed only in endosperm and the shorter in all tissues.