PIP5K基因家族调控蓖麻Lm型雌性系后代花序发育分子机制的研究

Castor (Ricinus communis L.) is one of the most important oilseed crops in the world. However, three-line breeding has not been achieved in castor. In this study, we used Lm type female line progeny of castor as material to analyze the molecular mechanism of inflorescence that is a hot and difficult point. The research group has castor plant breeding parent material and it has independent intellectual property rights. Therefore, the study group is the research object of the Lm type female line with marked trait progeny of castor. Based on the previous studies, we found that six genes of PIP5K family involved in the development of inflorescence in Lm type female line progeny of castor and related to regulation of three types of inflorescence including to isofemale lines, markered female lines and monoecious lines. Here, we detected the six gene functions in the process of inflorescence development in Lm type female line progeny of castor by bioinformatics, gene overexpression and knock-down technologies. Then we also analyzed the related genes and proteins using transcriptomics and proteomics to study its function in the metabolic pathways. And how six genes regulated inflorescence development by synergistic/complementary effects. This study will reveal that the molecular mechanism on regulation of the PIP5K family to the development of inflorescence in Lm type female line progeny of castor, lay the foundation for the study of the molecular mechanism on other types of castor inflorescence development.

蓖麻是世界上重要的油料作物，尚未实现三系配套育种，花序发育分子机制是其研究的热点与难点。课题组以具有自主知识产权的蓖麻育种母本材料——带有标志性状的Lm型雌性系后代为研究对象，在前期工作基础上——发现PIP5K家族6个基因与Lm型雌性系后代花序发育直接相关，提出假说：PIP5K家族6个基因在蓖麻Lm型雌性系后代花序发育中存在功能，协同/递补调控其发育出单雌、标雌、两性系三种花序类型。本课题拟采用生物信息学、同源过表达和敲减表达等技术，明确PIP5K家族6个基因在花序发育过程中的功能；采用转录组学和蛋白质组学等手段，明确6个基因过表达和敲减表达时有哪些基因和蛋白质发生改变及其参与的代谢通路，6个基因间如何通过协同/递补作用完成对花序发育的调控，揭示PIP5K基因家族调控蓖麻Lm型雌性系后代花序发育的分子机制，也为其他类型蓖麻花序发育分子机制研究奠定基础。

蓖麻是重要的油料作物，尚未实现三系配套育种，花序发育分子机制是其研究的热点与难点。课题组以具有自主知识产权的蓖麻育种母本材料——带有标志性状的Lm型雌性系后代为研究对象，采用生物信息学、同源过表达和敲减表达等技术，明确PIP5K家族6个基因在花序发育过程中的功能；采用转录组学和蛋白质组学等手段，明确6个基因过表达和敲减表达时有哪些基因和蛋白质发生改变及其参与的代谢通路，6个基因间如何通过协同/递补作用完成对花序发育的调控，揭示PIP5K基因家族调控蓖麻Lm型雌性系后代花序发育的分子机制，也为其他类型蓖麻花序发育分子机制研究奠定基础。实验结果如下：克隆了蓖麻PIP5K家族6个基因的全长，完成其生物信息学分析；对PIP5K家族6个基因进行亚细胞定位，证明6个基因对应的蛋白均位于细胞膜表面；PIP5K家族6个基因在拟南芥中异源过表达和敲减表达分析，证明基因对拟南芥的发育有影响；同源过表达和敲减表达分析，获得了PIP5K家族基因过表达和敲减表达转基因突变型蓖麻；对6个基因的过表达和敲减表达转基因突变型与野生型蓖麻相同花序类型植株差异组织部位的转录组学、蛋白质组学检测分析，表明6个基因分别在蓖麻中过表达和敲减表达后，会引起下游基因和蛋白质表达量变化，这些基因和蛋白质与植物物质代谢、能量代谢、氨基酸代谢、信号转导、光合作用、发育、运输等相关，完成了对50个差异基因和40个蛋白质的RT-qPCR、Western Blot检测验证；通过WGCNA分析和基因、蛋白质模块功能分析，确定蓖麻Lm型雌性系中PIP5K家族6个基因从RNA到蛋白质所引起的基因之间、蛋白质之间存在调控关系；在蓖麻花序中，PIP5K1、PIP5K6、PIP5K8、PIP5K9、PIP5K11基因表达为协同关系，PIP5K2基因的表达与另外5个基因为递补关系。