百脉根Rboh基因家族在共生固氮中的功能研究
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国家重点研发计划项目(2016YFD0100700);国家自然科学基金项目(31370278)

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    摘要:

    为探索植物呼吸爆发氧化酶(respiratory burst oxidase homologue,Rboh)基因家族在共生固氮中的功能和作用机制,利用分子生物学、细胞生物学和遗传学等方法,围绕百脉根Rboh基因家族中在根瘤中特异性表达的基因LjRbohA(Lj5g3v1497840)以及在根瘤中高表达的基因LjRbohB(Lj6g3v1549190)和LjRbohC(Lj5g3v1497820),分别对其蛋白的亚细胞定位、基因的表达模式、基因的启动子活性及蛋白的功能进行了研究。结果显示:LjRbohA、LjRbohB和LjRbohC蛋白都定位在植物的细胞膜上;LjRbohA、LjRbohB和LjRbohC基因在接种根瘤菌后表达上调;LjRobhA和LjRobhB基因在根瘤、维管束和侧根原基表达;利用CRISPRCas9技术对LjRbohA、LjRbohB和LjRbohC 3个基因分别进行敲除导致结瘤数目减少。这些研究结果表明,LjRbohA、LjRbohB和LjRbohC这 3个基因可能都参与调控百脉根与根瘤菌的共生结瘤过程。

    Abstract:

    genes including RbohA(Lj5g3v1497840),RbohB(Lj6g3v1549190) and RbohC (Lj5g3v1497820) were found to be primarily expressed in nodules based on the transcriptions of Rboh genes in symbiosis in Lotus japonicus. Their function involved in symbiosis were further studied in Lotus japonicus.The subcellular location of proteins,the expression pattern of Rboh genes,the proteins function and the promoter activity of Rboh genes in Lotus japonicas were studied with methods of molecular biology,cell biology and genetics. The results showed that LjRbohA,LjRbohB and LjRbohC proteins were localized at the plasma membrane.The transcript of LjRbohA,LjRbohB and LjRbohC accumulated abundantly in roots and nodules. The activity of LjRbohA and LjRbohB promoters were detected at vascular tissue,nodule,and the lateral root primordia.The numbers of nodules were reduced in LjRbohB,LjRbohALjRbohC  and LjRbohALjRbohBLjRbohC knockout transgenic roots compared with control transgenic roots. It is indicated that these three Rbohs might have an overlapping function in regulating nitrogen fixation symbiosis in L. japonicus.

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凌颖,张婷婷,端木德强,张忠明,曹扬荣.百脉根Rboh基因家族在共生固氮中的功能研究[J].华中农业大学学报,2019,38(1):1-9

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  • 收稿日期:2018-05-08
  • 在线发布日期: 2019-01-03
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