大豆高盐响应蛋白GmOSM的调控机理研究

Osmotins and OLPs (osmotin like proteins）are kinds of proteins which were produced during plant adapting to the environmental stress. These proteins were closely related to osmotic regulation and resistance stress. The expression of these genes of these protein was induced by high salt stress, which played an important role in the process of salt stress in plants. Our previous study showed that GmOSM (Glycine max osmotin protein) located on the cell membrane was induced expressed by high salt stress. Overexpressing GmOSM1 can significantly improve resistant to high salt capacity in yeast. Further study in this research, we attempt to study the function of GmOSM by determine the salt tolerance in over expression and knochout expression soybean using high salt treatment; to reveal the high salt tolerance physiological mechanism of GmOSM by microscopic observation, determination of physiological index and gene expression related proline synthesis and degradation, and hydrogen peroxide synthesis; improve the molecular mechanism of GmOSM resistant to high salt and involved in the metabolic pathway by Yeast two hybrid, BiFC (bimolecular fluorescence complementation), Co-IP (Co-Immunoprecipitation), RNA-seq and transcription factor - promoter - GUS co-transformation. This research will provide important gene resources for soybean molecular breeding and theoretic support for studying plant molecular mechanism of signal transduction network system under osmotic stress.

渗调蛋白(Osmotins)是由外界胁迫诱导植物形成的一类逆境蛋白，与植物细胞的渗透调节、抗逆境等重要生理过程密切相关。该类蛋白基因的表达受高盐胁迫诱导，在植物应答盐胁迫过程中起重要作用。我们前期研究发现大豆一个渗调蛋白基因GmOSM受高盐胁迫诱导表达，异源表达能显著提高酿酒酵母耐高盐能力，且定位于植物细胞膜上。本项目拟进一步通过研究大豆过表达和敲除GmOSM后的耐盐能力，解析其在大豆耐盐中的作用；通过显微观察、生理指标测定、脯氨酸合成和降解以及过氧化氢合成相关基因表达来揭示其耐高盐的生理机制；利用酵母双杂交、BiFC、Co-IP、 RNA-seq、转录因子-启动子-GUS共表达技术来完善GmOSM耐高盐的分子机理和参与的代谢通路。该项目对深入探讨植物渗透胁迫信号转导的网络体系的构建具有重要的理论意义，同时将为大豆抗逆育种提供理论基础和耐高盐种质。

渗调蛋白(Osmotins)是由外界胁迫诱导植物形成的一类逆境蛋白，与植物细胞的渗透调节、抗逆境等重要生理过程密切相关。该类基因的表达受高盐胁迫诱导，通过脯氨酸积累和抑制活性氧和自由基的产生在植物应答盐胁迫过程中起重要作用。我们研究发现大豆一个渗调蛋白基因GmOSM (Glycine max osmotin) 受高盐胁迫诱导表达，异源表达能显著提高酿酒酵母耐高盐能力，且定位于植物细胞膜上。本项目明确了过表达GmOSM在大豆中的耐盐性，通过生理指标测定、脯氨酸合成和降解以及过氧化氢合成相关基因表达来揭示其通过增加脯氨酸积累和抑制活性氧和自由基的产生耐高盐的生理机制；利用酵母单、双杂交RNA-seq等技术来完善了GmOSM可能通过与GmMYB、GmHMGB1、GmSIP1;3及GmRboh等蛋白相互作用而耐高盐的分子机理。该项目对深入探讨植物渗透胁迫信号转导的网络体系的构建具有重要的理论意义，同时将为大豆抗逆育种提供理论基础和耐高盐种质。