三倍体刺参生长优势的分子机理研究

The artificial aquatic animals of polyploidy usually has the characteristics of fast growth, individual large, strong resistance, and has no pollution in germplasm resources, which are widely used, but the molecular genetic expression and traits of analytical research is relatively weak. Application research with independent intellectual property rights "Sea cucumber polyploidy induction of hydrostatic pressure" patent technology, cultivating issue a clear growth advantage of triploid sea cucumber and its diploid control system. This research is based on the calculation of the triploid sea cucumber growth, gene expression and regulation (epigenetics) from two visual systems and comprehensive analysis the growth advantage of the triploid sea cucumber molecular mechanisms. In terms of gene expression level, RNA-Seq transcriptome information analysis is used to clear whether triploid sea cucumber gene expression "dose-effect"? What about the triploid sea cucumber gene expression patterns are? And get the growth advantage related gene of sea cucumber and SNP marker; In terms of genetic manipulation, the triploid sea cucumber gene methylation mapping is built and characteristics of methylation are analyzed with the latest sea cucumber genome-wide data of 2017. Tthe methylation of the promoter and the coding region are examined with BSP technology. Finally, the gene expression and apparent regulation of triploid sea cucumber are analyzed comprehensively ,the result will help to understanding the molecular mechanism of its growth dominant .The research would enrich the application of polyploidy in aquatic animals and provide information for improvement the character of echinoderms.

三倍体水产动物生长快、个体大、抗逆性强，养殖效益显著，但其性状解析和遗传调控研究较薄弱。项目组应用自主知识产权“静水压诱导刺参多倍体”专利技术，已培育出生长优势明显的三倍体刺参，在此基础上，项目从基因表达和表观遗传学两个视域，全面、系统地研究三倍体刺参生长优势的分子机制。基因表达层面，应用RNA-Seq结合表达谱分析，明确三倍体刺参基因表达是否存在“剂量效应”？表达模式如何？并获得三倍体刺参生长优势相关基因和SNP标记；表观调控层面，利用2017年公布的刺参全基因组数据，methylC-Seq和BSP技术，绘制三倍体刺参全基因甲基化图谱，筛选生长优势甲基化差异基因并验检测其启动子和编码区甲基化变化，阐明三倍体刺参表观调控特征；最后综合分析三倍体刺参基因表达和表观调控的关联性，解析其生长优势基因的分子机制，以期丰富水产动物多倍体应用基础研究，为棘皮动物性状改良提供资料。

国家自然科学基金“三倍体刺参生长优势的分子机理研究”按计划完成全部研究内容。研究以三倍体刺参和二倍体刺参为实验材料，实现了静水压诱导刺参多倍体条件的优化，三倍体刺参群体倍化率、存活率、生长、夏眠的检测；并运用分子生物学、生物信息学理论与实验技术，完成了三倍体刺参生长优势相关基因、代谢物及蛋白质的筛选与表达研究。主要内容包括：（1）三倍体刺参诱导技术优化与胚胎发育观察；（2）三倍体刺参群体倍化率、存活率与生长特征测定；（3）三倍体刺参非特异性免疫酶活性特征研究；（4）三倍体刺参基因转录表达模式分析；（5）三倍体刺参DNA甲基化特征研究；（6）基于转录组与代谢组的三倍体刺参生长优势分析；（7）基于蛋白质组学的三倍体刺参生长优势特征研究。重点开展了“三倍体刺参诱导技术优化”、“基于转录组与甲基化联合分析的三倍体刺参生长优势特征分析研究”，“基于转录组和代谢组的三倍体刺参生长优势特征分析”等研究。研究构建了三倍体刺参体壁组织的基因表达谱、DNA甲基化图谱及代谢谱，从生长、生理和分子机制不同水平，“基因表达-调控-代谢”不同层面揭示了刺参多倍体生长优势的分子机制，丰富了水产动物多倍体化的基础理论研究，为刺参健康养殖提供理论和技术支撑，促进刺参种业和养殖业提质增效和健康可持续发展。项目共发表SCI科研论文9篇；获批发明专利2项，其中国外发明专利1项；培养博士生1名，硕士研究生10名，毕业生5名。项目进展顺利，全面完成研究内容和考核指标。