野大豆天然无序蛋白TIFY10提高植物耐碳酸盐的蛋白调控机制

Intrinsically disordered protein (IDP), interacting with chaperone protein by its structure change, play important role in several physiological and pathological processes. Carbonate stress, including saline and alkaline stresses, are major factors limiting plant growth and development, eliciting adverse effects on the productivity and quality of crops. However, the information about IDP molecular mechanism in plant abiotic stress resistant is limit. We previously identified a nucleus located IDP GsTIFY10, from wild soybean (Glycine soja). GsTIFY10 can positively regulate plant carbonate stress response, but the regulation mechanism is still not clear. In this research, we identified seven GsTIFY10 interacted proteins using the wild soybean cDNA library for yeast two hybrid screening. We will verify GsTIFY10 interaction proteins by Y2H, BiFc and pull down. Then analyze GsTIFY10 protein interaction network using Bayesian modeling and functional annotation. We next aim at uncovering the biological functions of the key GsTIFY10 interaction protein as well as exploring how the protein regulates GsTIFY10 function in plant carbonate stress tolerance. We will combine bioinformatics and proteomics to analyze GsTIFY10 protein regulation mechanism in plant carbonate stress resistant. This study will not only greatly enhance our understanding of the important roles of IDP structure change in plant abiotic stress response, but also lay theoretical foundation for molecular breeding of crop with superior carbonate tolerance and developing and utilizing the saline-alkali land.

天然无序蛋白通过与伴侣蛋白结合行使特定功能，对于许多生物过程的调节非常重要。盐碱等逆境胁迫严重制约我国农业生产，植物中有关天然无序蛋白在耐逆中的作用机制研究还很少。我们在前期研究中发现野大豆天然无序核定位蛋白GsTIFY10能正调控植物对碳酸盐胁迫的耐性，但是其调控机制并不清楚。以野大豆酵母双杂交cDNA文库为材料，筛选到7个与GsTIFY10互作的功能未知的靶蛋白，本项目拟采用Y2H、BiFc 和pull down 技术验证蛋白互作的准确性；贝叶斯方法建模和功能注释构建GsTIFY10蛋白互作调控网络；通过研究GsTIFY10关键互作调控蛋白的生物学功能和对GsTIFY10调控植物胁迫耐性的影响，阐明GsTIFY10参与碳酸盐胁迫蛋白调控机制。本研究的结果将有助于人们从天然无序蛋白与靶蛋白互作网络途径这一角度来研究植物抵御碳酸盐胁迫机制，为作物耐逆分子育种和盐碱地开发利用奠定理论基础。

盐碱等逆境胁迫严重制约我国农业生产，我们在前期研究中发现野大豆天然无序核定位蛋白GsTIFY10能正调控植物对碳酸盐胁迫的耐性，但是其调控机制并不清楚。本研究以野大豆酵母双杂交cDNA文库为材料，通过两次筛选获得17个可能与GsTIFY10互作的蛋白，克隆获得了9个蛋白的基因全长序列；采用Y2H、BiFC和pull-down技术验证确定出4个GsTIFY10关键互作蛋白；初步构建了蛋白互作调控网络和蛋白的功能注释；后期通过拟南芥和大豆转基因材料和Crispr-Cas9基因沉默材料证明了关键互作蛋白的亚细胞定位、胁迫表达特性和耐碳酸盐的生物学功能，初步阐明了GsTIFY10参与碳酸盐胁迫的蛋白调控机制。本研究为揭示植物耐逆分子机理提供了实验证据和奠定理论基础，将有助于人们利用转基因等分子生物学技术来提高植物自身对胁迫逆境的耐性，开发利用盐碱地资源，提高农作物产量，解决粮食安全等问题，促进我国农业经济的可持续发展。