日本血吸虫正常发育和发育阻遏雌虫的转录组差异分析

Analysis of Different Transcriptome between Normally Developed and Undeveloped Female Schistosoma japonicum

为探讨日本血吸虫雌虫生长发育、性成熟及产卵的调控机制,本研究采用RNA-seq 高通量技术对 25日龄两性感染而来的正常发育雌虫(MF)和单性感染而来的发育阻遏雌虫(SF)的转录组进行测序、定量及差异基因的筛选。从 MF 和 SF 中分别获得 3 696 376 条和 3 662109 条 clean reads,将 clean reads 与日本血吸虫基因组数据库进行匹配发现 2 组样本的匹配率皆达 79%以上。以| Log2 Ratio |≥1,p-Value<0.001 为筛选条件,得到 775 个在 MF 和 SF 间差异表达的基因,其中有401 个基因在 MF 中高表达,374 个基因在 SF 中高表达。利用荧光实时定量RT-PCR对差异基因进行验证,结果与RNA-seq相符。生物信息学分析表明,在 MF 中高表达的基因多与代谢和生物合成过程相关,这些过程对于促进和维持雌性性成熟和产卵至关重要;而在 SF 中高表达的基因多与运动和细胞通讯等功能相关。这些发现可为深入研究日本血吸虫雌虫生殖发育以及产卵的生物学机理提供一定的理论依据。

In order to investigate the regulation mechanism of growth, sexual maturity and oviposition of Schistosoma japonicum females, RNA-seq high-throughput technology was used to analyze the differential transcriptome of 25 d normally developed mature females come from bisexual infected(MF) and undeveloped females come from single-sex infected(SF). A total of 3 696 376 and 3 662 109 clean reads were obtained separately form the MF and SF of 25 d Schistosoma japonicum. Clean reads were matched with the Schistosoma japonicum genomic database, the result found that the matching rates of the two groups were over 79%, respectively. A total of 775 differentially expressed genes between MF and SF were identified by│log2 Ratio│≥1, p-Value<0.001 as screening condition. Among them, 401 genes were highly expressed in MF, 374 genes were highly expressed in SF. For these differentially expressed genes, their expression was verified by quantitative RT-PCR, the results showed that they were consistent with the results of RNA-seq. Bioinformatics analysis showed that on the biological process, MF express more genes related to metabolic processes and biosynthesis, these processes are essential for promoting and maintaining female sexual maturity and spawning, whereas more genes related to locomotion and cellular communication were expressed in SF. These findings provide a theoretical basis for study of the biological mechanisms of reproductive development and spawning of Schistosoma japonicum females.