1江苏省农业科学院经济作物研究所,南京 210014,2 农业农村部油料作物生物学与遗传育种重点实验室,武汉 430000
国家现代农业产业技术体系建设项目(CARS-13);江苏省农业科技自主创新资金项目(cx(18)2015);农业农村部油料作物生物学与遗传育种重点实验室开放课题项目(KF2019011)
1 Jiangsu Academy of Agricultural Sciences,Nanjing 210014;2 Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture and Rural Affairs,Wuhan 430000
Special Fund for Modern Agroindustry Technology Research System of China (CARS-13), Independent Innovation Fund of Agricultural Science and Technology of Jiangsu Province(cx(18)2015), Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, P.R. China(KF2019011)
前期根据花生干旱转录组测序结果克隆了一个胁迫诱导表达基因 AhMYB44 的全长序列。为进一步研究基因的功能,本 研究利用酵母双杂交系统进行其互作蛋白的筛选。结果显示:根据基因序列构建的诱饵载体 pGBKT7-AhMYB44 本身无毒性,且 无自激活活性;经与花生干旱及高盐胁迫相关的 cDNA 文库杂交、筛选、鉴定出 16 个插入序列,它们对应的同源蛋白功能涉及植物 转录调控、生长发育、信号转导、生物及非生物胁迫响应等多个方面;其中 46 号候选互作蛋白与非生物胁迫密切相关,结构域分析 属于钙依赖性磷脂结合蛋白(膜联蛋白,ANNs),植物膜联蛋白已被证实参与多种胁迫响应过程。因此推测 AhMYB44 可能通过胞 内钙信号调控花生胁迫响应过程。这也为今后深入开展 AhMYB44 的功能及作用机制研究提供了参考依据。
We previously cloned the full-length coding sequence of the gene AhMYB44,which showed a stress-induced expression from peanut drought transcriptome sequencing datasets. In order to investigate the function of AhMYB44,a yeast two-hybrid system was deployed for identifying its interacting proteins. The results showed that the bait vector pGBKT7-AhMYB44 was not toxic and showed no self-activation activity. Sixteen coding sequences were identified by screening of cDNA libraries which were constructed from a drought and high-salt stress of peanut. We further conducted an in-silico analysis of their homologous proteins,such as plant transcriptional regulation,growth and development,signal transduction,biotic and abiotic stress responses. Out of that,the candidate gene(No.46)was annotated with the abiotic stress,and encoded for a calcium-dependent phospholipid binding protein(ANNEXIN). Therefore,we speculated that AhMYB44 maybe participate into regulation of peanut stress response process through intracellular calcium signal. This result might provide a reference for further research on the function and mechanism of AhMYB44.
刘永惠,沈一,沈悦,等.利用酵母双杂交系统筛选花生AhMYB44互作蛋白[J].植物遗传资源学报,2020,21(1):201-207.
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