Function of MCHK_7135 gene in symbiotic nitrogen fixation of Mesorhizobium huakuii
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1.College of Life Science and Technology,Huazhong Agricultural University,Wuhan 430070,China;2.Hubei Provincial Plant Protection General Station,Wuhan 430070,China

Clc Number:

S182

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    Abstract:

    In the previous studies,the global transcriptome databases of Mesorhizobium huakuii 7653R in free living and in symbiotic condition were constructed respectively. Some differential expression genes in free living and symbiotic condition were identified with RNA-Seq and Microarray. Among these genes,MCHK_7135 belongs to the peroxiredoxin (Prxs) family. Its expression was significantly up-regulated at the stage of symbiotic nitrogen fixation.In this article,the replacement mutants and complementary strains of MCHK_7135 were constructed by homologous recombination to study the function of MCHK_7135 gene in symbiotic nitrogen fixation between rhizobia and host plants. The phenotypes of mutant and complementary strains were investigated under free living and symbiotic condition with Astragalus sinicus. Results showed that the mutant strain of MCHK_7135 gene was more sensitive to H2O2 stress compare with the wild-type strain. The growth of A. sinicus plant inoculated with the mutant was inhibited,the number of infect thread and nodule primordia decreased,the number of nodule was small,and the activity of nitrogenase was low. However,when the mutant strain was complemented by complete MCHK_7135 gene,some symbiotic phenotypes were restored to different degree. It is indicated that the peroxiredoxin gene MCHK_7135 of M. huakuii plays an important role in the symbiotic nitrogen fixation between rhizobia and host plants.

    Table 1 PCR primers used in this study
    Fig.1 Scheme of SOE-PCR
    Fig.2 DNA and amino acid sequence of MCHK_7135 in rhizobium
    Fig.3 Conserved domains analysis of MCHK_7135 peptide
    Fig.4 The phylogenetic tree of MCHK_7135 gene
    Fig.5 Growth curve of each rhizobium strains in liquid medium
    Fig.6 The colony number of each strains on TY plate after H2O2 stress treatment
    Fig.7 Infect thread and nodules primordium labeled by GFP(bar=100 μm)
    Fig.8 Data of early infect event by each rhizobium strains
    Fig.9 Whole plant and root phenotypes of A. sinicus inoculated by vary rhizobium strains for 30 days
    Fig.10 Paraffin section of nodule after inoculated with rhizobium for 30 days
    Table 2 Symbiotic phenotypes of A. sinicus inoculated by vary rhizobium strains
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胡珍珍,谢华伦,彭杰丽,李友国,陈大松. Function of MCHK_7135 gene in symbiotic nitrogen fixation of Mesorhizobium huakuii[J]. Jorunal of Huazhong Agricultural University,2023,42(2):123-130.

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History
  • Received:August 12,2022
  • Online: March 31,2023
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