大豆慢生根瘤菌3种标记载体构建与应用  

Construction and Application of Three Labeling Vectors for Bradyrhizobia in Soybean

作　　者：王小倩 钟永嘉[2] 廖红[2] WANG Xiao-qian;ZHONG Yong-jia;LIAO Hong(College of Life Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Root Biology Center,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学生命科学学院,福建福州350002 [2]福建农林大学根系生物学研究中心,福建福州350002

出　　处：《大豆科学》2023年第3期276-283,共8页Soybean Science

基　　金：国家重点研发计划项目子课题“玉米、大豆互作调控养分协同高效的机制”(2021YFF1000504)。

摘　　要：大豆与根瘤菌的共生固氮是农业生态系统中主要的氮来源之一,研究其互作机制意义重大,但是由于根瘤菌个体小,转化难,目前尚缺乏高效、稳定标记根瘤菌的方法,限制了其在互利共生方面的研究。为开发用于大豆与慢生根瘤菌互作研究的根瘤菌标记方法,本研究通过改造原始载体pMG103,构建高效表达tdTomato、GUS、LUC的标记载体,以高效固氮的慢生根瘤菌株系Bradyrhizobium elkanii BXYD3为试验菌株,采用电击转化法进行标记。利用显微镜、GUS染液、化学成像仪观察标记的B.elkanii BXYD3根瘤菌在大豆不同时期不同部位的定殖情况。结果表明:成功获得了tdTomato、GUS及LUC标记的慢生根瘤菌转化菌株,携带不同标记的慢生根瘤菌株B.elkanii BXYD3能在大豆根表及根瘤内稳定定殖,并且可满足不同试验需求。本研究建立了较为稳定的表达多种标记蛋白的载体,且能较好地应用于慢生根瘤菌株系中,为直观研究大豆与慢生根瘤菌的互作提供有效的方法参考。Symbiotic nitrogen fixation between soybean and rhizobia is one of the most important sources of nitrogen for agricultural ecosystems.And it is of great significance to study its interaction mechanism.However,rhizobia are small and difficult to be labeled by transformation and lack of efficient and stable method for rhizobial labeling.This study aimed to provide a method for rhizobial labeling to investigate the interaction between soybean and Bradyrhizobia.In this study,the original vector pMG103 was modified to construct a labeled vector that efficiently expressed tdTomato,GUS,and LUC.Bradyrhizobium elkanii BXYD3,a slow-growing rhizobia strain with high nitrogen fixation,was used as the experimental strain and labeled by electric shock conversion.Microscope,GUS staining and chemoluminescence were used for BXYD3 colonization visualization.Our results suggested BXYD3 rhizobia was successfully labeled with different tags and labeling signaling were stable in different compartments of roots for different intentions.In summary,this study established a relatively stable vector expressing a variety of marker proteins,which should be well applied to Bradyrhizobia strains,providing effective method for studying the interaction between soybean and Bradyrhizobia.

关 键 词：慢生根瘤菌 大豆 标记 共生固氮 tdTomato GUS LUC 

分 类 号：S565.1[农业科学—作物学]