化学杀雄剂CH1诱导的小麦雄性不育花药转录组学及相关基因表达的分析

Analysis of Transcriptome and Related Gene Expression in Male-Sterile Anthers of Wheat Induced by Chemical Hybridizing Agent CH1

为了揭示化学杀雄剂CH1诱导雄性不育的作用机理,以小麦品种普冰151为试验材料,利用测序技术对化杀不育系(FHS)和正常可育系(FZC,对照)的花药RNA进行转录组学分析,筛选差异表达基因,并进行GO、KEGG富集分析及实时荧光定量PCR分析。结果表明,FHS和FZC之间筛选出差异表达基因共3 895个,其中上调表达基因1 334个,下调表达基因2 561个,主要富集在碳水化合物代谢、多糖代谢、外部结构组织、细胞壁组织等;经KEGG分析,差异基因主要集中在戊糖和葡萄糖醛酸转化、淀粉和蔗糖代谢利用等通路中。利用实时定量PCR技术对转录组数据中所筛选出的5个表达差异较大的基因进行验证,在花药发育的单核期,处理与对照相比,这5个基因表现出不同的上下调趋势;而在二核期和三核期,这5个基因都表现出明显的下调趋势,推测这5个基因的下调表达有可能与化杀不育系花粉的败育有关。

In order to reveal the mechanism of male sterility induced by the chemical hybridizing agent CH1,in this study,we used the wheat cultivar Pubing 151 as the test material,and analyzed the anther RNAs of chemical hybridizing sterile lines(FHS)and the control normal fertile lines(FZC)by transcriptome sequencing,screened the differentially expressed genes,analyzed the GO enrichment,KEGG enrichment and the expression pattern of major differentially expressed genes using the real-time fluorescent quantitative PCR.The results showed that,a total of 3895 differentially expressed genes were screened between FHS and FZC,including 1334 up-regulated genes and 2561 down-regulated genes,which were mainly enriched in carbohydrate metabolism,polysaccharide metabolism,external structural organization,and cell wall organization,etc..The results of KEGG analysis showed that the differentially expressed genes were mainly enriched in the pentose and glucuronate interconversions,and the metabolism of starch and sucrose pathways.To validate the five differentially expressed genes screened in the transcriptome data,real-time quantitative PCR was conducted.The five genes performed differently up-and down-regulated trends,between the control group and CH1 treatment at the uninucleate period.But in the binucleate period,the five genes all showed obviously down-regulated trends,in agreement with the results of the transcriptomic analysis.It can be assumed that the down-regulated genes expression may be relative to pollen abortion.This study provides a reference for further revealing the mechanism of male sterility induced by chemical hybridizing agent CH1 in wheat.