毛竹Ph-TElncRNA1的鉴定及对靶基因的调控  

Identification of Ph-TElncRNA1 in Phyllostachys edulis and its regulation of target genes

作　　者：赵佳敏 余璐 丁一倩 周明兵[1] ZHAO Jiamin;YU Lu;DING Yiqian;ZHOU Mingbing(State Key Laboratory of Subtropical Silviculture/Institute of Bamboo Research,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]浙江农林大学省部共建亚热带森林培育国家重点实验室/竹子研究院,浙江杭州311300

出　　处：《浙江农林大学学报》2023年第2期314-320,共7页Journal of Zhejiang A&F University

基　　金：国家自然科学基金资助项目(31870656);浙江自然科学基金重点资助项目(LZ19C160001)。

摘　　要：【目的】旨在探究毛竹Phyllostachys edulis Ph-TElncRNA1及靶基因的表达模式,初步分析lncRNA的功能。【方法】基于高温、低温、紫外、高盐胁迫处理毛竹实生苗全转录组测序数据,筛选在胁迫下差异表达的lncRNA。通过CNCI、Pfam、CPC2、PLEK等4个软件对lncRNA的编码特性进行分析,用LncRNATar软件鉴定lncRNA的靶基因,通过实时荧光定量PCR分析lncRNA和靶基因在紫外胁迫下和叶片着色过程中的表达模式。【结果】筛选到1个在紫外胁迫下差异表达的lncRNA,此lncRNA来源于LARD转座子序列(large retrotransposons derivatives),命名为PhTElncRNA1 (Phyllostachys edulis transposable element derived lncRNA1)。Ph-TElncRNA1是一个典型的非编码RNA,全长为342 bp,其中,185个碱基来源于外显子,157个碱基来源于内含子。Ph-TElncRNA1的靶基因为psbA,编码photosystemⅡprotein D1。实时荧光定量结果表明:Ph-TElncRNA1与psbA的相对表达量呈完全相同的趋势,说明在紫外胁迫下Ph-TElncRNA1与psbA呈正相关。通过Ph-TElncRNA1和psbA在不同着色时期的毛竹叶片的相对表达量分析,发现在叶绿体形成期,Ph-TElncRNA1和psbA表达量达到峰值。【结论】Ph-TElncRNA1和psbA在紫外胁迫下协同表达,Ph-TElncRNA1可以通过调控靶基因psbA参与毛竹叶片发育。[Objective]The purpose of this study is to explore the expression patterns of Ph-TElncRNA1 and target genes in Phyllostachys edulis,and to preliminarily analyze the function of lncRNA.[Method]Based on the whole transcriptome sequencing data of Ph.edulis seedlings under high temperature,low temperature,ultraviolet and high salt stress,lncRNAs differentially expressed under stress were screened.The coding characteristics of lncRNA were analyzed by four kinds of software(CNCI,Pfam,CPC2 and PLEK),and the target genes of lncRNA were identified by LncTar software.The expression patterns of lncRNA and target genes under UV stress and leaf coloration were analyzed by real-time fluorescent quantitative PCR.[Result]One differentially expressed lncRNA under UV stress was screened,which was derived from the lncRNA of the Large retrotransposons derivatives and named Ph-TElncRNA1(Ph.edulis transposable element derived lncRNA1).Ph-TElncRNA1 was a typical non-coding RNA with a total length of 342 bp,185 bases from exons and 157 bases from introns.The target gene of Ph-TElncRNA1 was psbA,encoding photosystemⅡprotein D1.Real time fluorescence quantitative results showed that the relative expression of Ph-TElncRNA1 and psbA was the same,indicating that Ph-TElncRNA1 was positively correlated with psbA under UV stress.Through the analysis of the relative expression of Ph-TElncRNA1 and psbA in Ph.edulis leavesat different coloring stages,it was found that the expression of psbA and Ph-TElncRNA1 reached the peak during chloroplast formation stage.[Conclusion]Ph-TElncRNA1 and psbA are co-expressed under UV stress,and Ph-TElncRNA1 can participate in the leaf development of Ph.edulis by regulating the target gene psbA.

关 键 词：Ph-TElncRNA1 叶片着色 D1蛋白 毛竹 

分 类 号：Q943.2[生物学—植物学] S722.3[农业科学—林木遗传育种]