水稻籼爪重组自交系PEG胁迫早期基因差异表达分析

Differentially expressed gene of indica/javanica recombinant inbred line in rice at early stage of PEG stress

把从籼爪重组自交系中筛选出的最耐旱材料(47-274)和最敏感材料(47-299)在五叶一心时用20%PEG处理6h,取叶片进行mRNA差异显示分析,获得10个差异表达的cDNA片段。利用反式Northern斑点杂交法去除6个假阳性片段,对4个阳性片段(DT-DF1、DT-DF2、DT-DF3、DT-DF4)测序后进行同源性分析,发现DT-DF3的蛋白产物与核酸结合位点和富亮氨酸重复结构域类蛋白B高度同源,DT-DF2与L-乳酸氧化酶部分序列有同源性,DT-DF1没有找到同源蛋白质序列,DT-DF4与一个假设蛋白高度同源。

The drought tolerant line and drought sensitive line screened from the population of indica/javanica recombinant inbred line were stressed with 20% PEG（Polyethyleneglycol） for 6 h at five-leaf stage of rice. Ten of cDNA fragments were isolated by mRNA differentially display reverse transcription PCR（DDRT-PCR）,but only four fragments were approved truth by reverse Northern dot blotting. These fragments named as DT-DF1,DT-DF2,DT-DF3 and DT-DF4 were sequenced and then analyzed by Blast program. The putative protein of DT-DF3 was highly homologous with NBS-LRR-like protein B and that of DT-DF2 was homologous with L-lactate oxidase. The putative protein of DT-DF1 was not found homologous protein,and that of DT-DF4 was highly homologous with a hypothetical protein.