摘要
赤霉素(GA)广泛参与植物生命周期的各种生长过程,DELLA蛋白作为GA信号通路中的抑制子,对植物的生长发育起抑制作用。铁皮石斛(Dendrobium officinale Kimura et Migo)种子中高表达的DELLA家族基因可能与其种子自然萌发困难有关。为筛选铁皮石斛DoDELLA蛋白的互作蛋白,解析DoDELLA在种子萌发过程中的作用机制,本研究利用Gateway重组技术构建了铁皮石斛种子cDNA文库,采用同源重组的方法构建DoDELLA4重组诱饵载体,从cDNA文库中筛选DoDELLA4的互作蛋白。结果显示,铁皮石斛种子cDNA文库的库容量为1.20×10^(7)CFU,平均插入片段大于1 000 bp,重组率100%。在SD/-Trp/-Leu/-His/-Ade培养基中添加20 mmol/L的3-AT可抑制重组诱饵载体pGBKT7-DoDELLA4的自激活。从文库中筛选获得了70个潜在的互作蛋白,通过酵母双杂交实验,验证了DoELIP1、5807、DoTZF、DoUBQ、DoGRPA3与DoDELLA4蛋白存在互作关系。研究结果为进一步探究DoDELLA基因调控铁皮石斛种子萌发的分子机制提供了基础。
Gibberellins(GAs)regulate diverse growth processes throughout the plant life cycle.DELLA proteins function as key repressors in the GA signaling pathway,inhibiting plant growth and development.The high expression of DELLA family genes in Dendrobium officinale Kimura et Migo seeds may be related to their poor natural germination.To identify interacting proteins of DoDELLA and elucidate its role in seed germination,a cDNA library was constructed from D.officinale seeds using Gateway recombination technology.A recombinant bait vector for DoDELLA4 was generated using homologous recombination and used to screen interacting proteins from the cDNA library.The constructed library exhibited a capacity of 1.20×10^(7) CFU,with an average insert size exceeding 1000 bp and a recombination efficiency of 100%.The recombinant bait vector pGBKT7-DoDELLA4 effectively suppressed self-activation in SD/-Trp/-Leu/-His/-Ade medium supplemented with 20 mmol/L 3-AT.Screening identified 70 potential interacting proteins,and yeast two-hybrid assays confirmed interactions between DoDELLA4 and DoELIP1,5807,DoTZF,DoUBQ,and DoGRPA3.These findings establish a foundation for further investigation into the molecular mechanisms by which DoDELLA regulates seed germination in D.officinale.
作者
李琳
付强
杨涛
莫小连
陈晓龙
赵杰宏
邹颉
Li Lin;Fu Qiang;Yang Tao;Mo Xiaolian;Chen Xiaolong;Zhao Jiehong;Zou Jie(School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)
出处
《植物科学学报》
北大核心
2025年第2期221-229,共9页
Plant Science Journal
基金
国家自然科学基金项目(82160720)。
