小麦lncRNA27195及其靶基因TaRTS克隆及表达分析

Cloning and expression analysis of lncRNA27195 and its target gene TaRTS in wheat(Triticum aestivum L.)

长链非编码RNA(long non-coding RNA,lncRNA)是一种大于200 bp的非编码RNA,大量存在于植物体中,其可通过调节基因表达或蛋白功能,在植物生长发育与胁迫应答中发挥重要作用。前期在研究光照和温度对小麦光温敏核雄性不育系BS366育性诱导中,利用转录组测序筛选获得与育性相关的lncRNA(lncRNA2719)。为研究小麦lncRNA27195的功能,本研究在BS366中克隆lncRNA27195及其靶基因TaRTS,并对TaRTS进行生物信息学分析,结果显示,TaRTS基因全长315 bp,编码104个氨基酸,且发现该RTS蛋白仅存在于禾本科植物中。通过实时荧光定量PCR,对lncRNA27195及TaRTS基因在不同组织不同光温处理及茉莉酸甲酯处理下进行表达模式分析,发现lncRNA27195和TaRTS均在雄蕊中高表达,呈显著的正相关,且两者在不同光温条件下呈现出不同的表达模式。光照和温度均对lncRNA27195和TaRTS有调控作用,适当浓度的MeJA促进两者的表达,SA抑制两者表达。以上结果表明,在光周期、温度和植物激素的诱导下,lncRNA27195正向调节TaRTS基因表达,进而影响花粉育性,本研究结果有助于促进对小麦光温敏核型雄性不育系的机理研究和生产应用。

Long non-coding RNA(lncRNA)is a non-coding RNA length over 200 bp,which is abundant in plants.It plays important roles in plant growth,development,and stress response by regulating gene expression or protein function.In the previous study,a fertility-related lncRNA named lncRNA27195 was screened and obtained by transcriptome sequencing from the anther of wheat Photoperiod-thermo Sensitive Genic Male Sterility(PTGMS)line BS366.To investigate the function of lncRNA27195 in wheat,the lncRNA27195 gene and its target gene TaRTS were cloned from BS366.Bioinformatics analysis were performed on TaRTS.The expressions of lncRNA27195 and TaRTS in different tissues and their expression correlation between them were analyzed by qRT-PCR.Meanwhile,the expression patterns of lncRNA27195 and TaRTS under different light and temperature treatments,and methyl jasmonate(MeJA)treatments were investigated.The results showed that the TaRTS gene with 315 bp length,encoded 104 amino acids.Additionally,RTS proteins were only found as anther-specific proteins in gramineae plants.Both lncRNA27195 and TaRTS with a significantly positive correlation were highly expressed in stamens,and revealed different expression patterns in different fertility environments.The results demonstrated that the expression of lncRNA27195 and TaRTS were also regulated by light and temperature.In addition,we found that the appropriate concentration of MeJA could promote the expression of lncRNA27195 and TaRTS while SA could inhibit the expression.The results indicated that under the induction of photoperiod,temperature,and plant hormones,IncRNA27195 positively regulated TaRTS gene expression,resulting in affecting pollen development and male fertility.This study contributed to the mechanism research and production application of PTGMS wheat.