基于转录组分析不同温度下池蝶蚌性别相关基因的表达

Transcriptome analysis of sex related gene expression in Hyriopsis schlegelii at different temperatures

采用Pacbio三代和Illumina二代测序技术对不同温度(15℃,20℃,25℃,30℃,33℃)条件下养殖的30月龄池蝶蚌性腺组织进行转录组分析。通过差异分析获得各个温度下雌雄对比组的差异基因,数量分别为6311,3013,6673,6798和5685个。其中在20℃和33℃下差异基因数量最少。差异表达基KEGG富集显示,25℃条件下出现卵母细胞的有丝分裂,细胞周期和notch信号通路等雌性相关活动。对五个温度处理组之间的差异表达基因进行组间比较,发现含有802个共同表达差异的基因,其中鉴定到wnt4,rspo1,fem1和tra1等4个性别决定相关基因。在qRT-PCR和转录组数据中,各温度下wnt4,rspo1的表达水平雌性均显著(P<0.01)高于雄性;而fem1,tra1相反。根据wnt4,rspo1,fem1和tra1在不同温度下的相对表达情况和差异基因的KEGG富集结果表明,25℃以上更利于雌性发育,其发育机制还有待深入研究。

The 30-month-old Hyriopsis schlegelii were cultured at 15℃,20℃,25℃,30℃and 33℃.The Pacbio third-generation sequencing technology and the Illumina second-generation sequencing technology were used to acquire the full-length transcripts of Hyriopsis schlegelii’s gonad.Through the difference analysis,the differential genes of the male and female comparison groups at each temperature were obtained,and the numbers were 6311,3013,6673,6798 and 5685,respectively.Among them,the number of differential genes was the least at 20°C and 33°C.KEGG enrichment showed that the oocyte meiosis appeared in the Hyriopsis schlegelii at 20℃and 25℃,and the progesterone-mediated oocyte maturation and cell cycle pathways appeared at 30℃.The differentially expressed genes between the five temperature treatment groups were compared,and it was found that there were 802 co-expressed differential genes,among which four sex-determining genes such as wnt4,rspo1,fem1 and tra1 were identified.In the qRT-PCR and transcriptome data,the expression levels of wnt4 and rspo1 in females were higher than those in males at each temperature;while fem1,and tra1 were the opposite.According to the relative expression of wnt4,rsop1,fem1 and tra1 at different temperatures and KEGG enrichment of different genes,25℃may be more conducive to female development,while the mechanism remained to be further studied.