广聚萤叶甲雄虫生殖系统特异性精液蛋白基因的筛选及表达分析

Screening and expression analysis of seminal fluid protein genes specific to the male reproductive tract in ragweed leaf beetle Ophraella communa

为明确广聚萤叶甲Ophraella communa雄虫精液蛋白基因在性别间的表达差异并筛选出在雄虫生殖系统特异性表达的基因,从转录组及蛋白组数据库选取13个精液蛋白基因,采用实时荧光定量PCR技术检测其在广聚萤叶甲不同性别、组织及发育阶段的表达特征。结果显示,在广聚萤叶甲雄虫13个精液蛋白基因中,有9个精液蛋白的mRNA表达水平在雄虫中更高,有5个精液蛋白基因具有专一的雄虫生殖系统特异性,分别为PGK、sucA、ENO、SDH和PLA2b基因,在雄虫生殖系统中的表达量分别较其在头部的表达量高273.01倍、401.48倍、11.21倍、64.27倍和1.86倍。PGK、sucA和ENO基因均在广聚萤叶甲老熟蛹期急剧上调表达,在性成熟成虫中的表达量最高,分别是卵期表达量的43.48倍、31.03倍和11.89倍,三者具有明显的组织和发育阶段时空特异性,推测PGK、sucA和ENO基因参与广聚萤叶甲雄虫生殖的可能性较其余基因更高。

To clarify the expression difference of seminal fluid protein(SFP)genes between sexes,and obtain the specific expression genes in the male reproductive tract(MRT)in ragweed leaf beetle Ophraella communa,13 SFP genes were selected the O.communa transcriptome and proteome databases based on bioinformatics analysis.Quantitative real-time PCR(qPCR)was used to determine the expression patterns of these genes in different genders,tissues and developmental stages.The results showed that the mRNA expression levels of nine SFP genes were significantly higher in males than in females,and five SFP genes(i.e.PGK,sucA,ENO,SDH and PLA2 b)were specifically expressed in MRT,and their expression levels in MRT were 273.01,401.48,11.21,64.27,and 1.86 times higher than those in the head,respectively.The expression levels of PGK,sucA and ENO genes were rapidly up-regulated in the mature pupal stage,reaching the highest in mature adults,which increased by 43.48,31.03,and 11.89 times,respectively,compared to those at egg stage.In addition,the expression of PGK,sucA and ENO genes had spatial-temporal specificity in tissue and developmental stage.Therefore,it was suggested that PGK,sucA,and ENO genes were more likely to be involved in male reproductive regulation compared to any other genes in O.communa.