大豆慢生根瘤菌3种标记载体构建与应用
- Title:
- Construction and Application of Three Labeling Vectors for Bradyrhizobia in Soybean
- 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。
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