低温胁迫下水稻苗期低温失绿突变体cisc(ta)的miRNA表达谱分析

Analysis of miRNA Expression Profiles of a Cold-induced Seedling Chlorosis Gene(cisc(ta))in Rice under Cold Stress

[目的]鉴定水稻耐冷相关功能基因、解析其调控耐冷性的分子机制。[方法]以耐冷籼稻材料“P1211”和冷敏型籼稻突变体cisc(ta)为试验材料,进行了低温胁迫处理和水稻幼苗转录组测序分析。[结果]冷敏型突变体cisc(ta)低温胁迫处理后共有4542个差异表达基因,其中有3673个基因上调,869个基因下调。在冷敏突变体cisc(ta)的差异表达基因中筛选到一些与低温胁迫相关的典型基因,如基本的螺旋-环-螺旋(bHLH)、活性氧清除(SOD1)和富亮氨酸重复序列(LRR)等调控基因,此外,还筛选出一些与生长素、脱落酸(ABA)、赤霉酸(GA)和冷应激反应调控(DREB/CBF)等相关的基因。为了确认RNA-seq数据,随机选择差异表达基因进行实时荧光定量PCR(qRT-PCR)表达分析,结果与这些基因的RNA-seq表达测定一致,证实了RNA-seq数据的准确性。[结论]该研究为水稻耐冷机理的研究和筛选水稻遗传改良有用的候选基因奠定基础。

[Objective]To identify genes for cold tolerance and understand its underlying molecular mechanism of its regulation of cold tolerance.[Method]In the present study,a cold tolerance variety‘P1211’and a cold sensitive variety‘cisc(ta)’were used for transcriptomics analysis under low temperature stress.[Result]The results showed that in response to cold,a total of 4542 differentially expressed genes(DEGs)were detected in cisc(ta),including 3673 up-regulated and 869 down-regulated genes.Many common and special DEGs were analyzed in cisc(ta).Some typical genes related to cold stress such as the basic helix-loop-helix(bHLH)gene,superoxide dismutase(SOD1)and leucine-rich repeat(LRR)domain gene etc.In addition,some genes related to several plant hormones such as auxin,abscisic acid(ABA),gibberellic acid(GA)and dehydration responsive element binding protein/C-repeat binding factor(DREB/CBF)were identified.To confirm the RNA-seq data,qRT-PCR were performed on 7 randomly selected DEGs.The expression patterns of RNA-seq on these genes corresponded with the qRT-PCR method,which further confirmed the accuracy of RNA-seq results.[Conclusion]This study provides some insights into the cold tolerance mechanism and digging out useful candidate genes for genetic improvement in rice.