Comparative transcriptome analysis between wild-type and gpa1 mutant in response to ABA

Mutations in the heterotrimeric G-protein a-subunit of Arabidopsis, GPA1, leads to deficiency in ABA-induced stomatal closure (Wang et al., 2001). To further investigate whether GPA1 is involved in the regulation of gene expression in response to ABA, we examined the induction of known ABA-inducible genes in the gpa1 mutant and compared it to wild-type. We found significant differences in levels of ABA-induced expression between wild-type and gpa1 mutant. In order to systematically investigate GPA1 involvement in ABA signalling leading to gene expression, we are requesting the transcriptome analysis of the gpa1 mutant in response to ABA.In detail, 2 week old wild-type and gpa1 plants grown in the 16/8 hrs light and dark cycle will be treated with either ABA or with a control solution for 3 hours. 10 plants will be used per sample to produce RNA, to give 4 samples, one for each chip: gpa1-1 mutant in the presence (chip1) or absence of ABA (chip2) and wild-type (WS2) in the presence (chip3) or absence of ABA (chip4). Keywords: strain_or_line_design Number of plants pooled:10 plants

拟南芥异三聚体G蛋白（heterotrimeric G-protein）α亚基GPA1发生突变，会导致脱落酸（ABA）诱导的气孔关闭功能缺陷（Wang等人，2001年）。为进一步探究GPA1是否参与脱落酸（ABA）应答过程中的基因表达调控，我们检测了gpa1突变体中已知ABA诱导基因的诱导表达情况，并与野生型植株进行对比。结果发现，野生型与gpa1突变体在ABA诱导的基因表达水平上存在显著差异。为系统探究GPA1在介导ABA信号通路调控基因表达过程中的作用，我们申请对ABA处理后的gpa1突变体进行转录组分析。具体实验方案如下：将在16小时光照/8小时黑暗周期下培养的2周龄野生型与gpa1植株，分别用ABA溶液或对照溶液处理3小时。每份样本选取10株植株提取RNA，共设置4个样本对应4张芯片：分别为经ABA处理的gpa1-1突变体（chip1）、未用ABA处理的gpa1-1突变体（chip2）、经ABA处理的野生型(WS2)（chip3）以及未用ABA处理的野生型(WS2)（chip4）。关键词：株系设计；混合植株数量：10株