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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 method for rapid isolation of total RNA from rice endosperm tissue is proposed, which can easily remove polysaccharides such as starch present in RNA. The obtained endosperm total RNA can meet the requirements of Northern blotting and microarray preparation of molecular biology Learn experiment requirements. The obtained RNA was isolated from the endosperm of 966 rice cultivars and was used for RNAi analysis of Wx gene. The results showed that the abundance of Wx gene in rice was overall positive with the amylose content of rice endosperm The results showed that in rice endosperm tissue, the starch branching enzyme Rbe1 has two homologous genes with different length, one of which is specific for endosperm and one with low molecular weight, which is co-expressed in all tissues of rice.