Interspecific plasmid and genomic DNA sequence homologies and localization of nif genes in effective and ineffective strains of Rhizobium japonicum.

R Haugland, D P Verma
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Abstract

Three strains of Rhizobium japonicum were examined for the presence of interspecific conserved plasmid-borne DNA sequences and the location of their nif DNA sequences. Strains 61A76 and 110, which both form effective (nitrogen fixing) nodules on soybeans show very low (24%) total DNA sequence homology with each other; strain 61A76 contains one plasmid, and strain 110 contains no identifiable plasmids. Strain 61A24 which forms ineffective nodules on soybeans shows relatively high (50%) sequence homology to strain 110 and contains two plasmids. While the total sequence homology between purified plasmid DNA isolated from strains 61A76 and 61A24 appears limited, heterologous hybridizations between plasmids and total DNAs from these strains and DNA from strain 110 suggest that several sequences which are plasmid-borne in one strain can occur in the chromosome or in very large plasmid(s) in the other strains. The plasmid isolated from strain 61A24 exhibits some homology with several SmaI fragments of an octopine Ti plasmid from Agrobacterium tumifaciens, whereas the 61A76 plasmid shows no homology to this Ti plasmid. DNA sequences from five strains of Rhizobium japonicum were found to hybridize with pSA30, a plasmid carrying the Klebsiella pneumoniae nif-KDH genes, but these sequences were not located on the isolated plasmids. The organization of these genes appears to be similar in strains 110 and 61A24, but is different in strain 61A76.

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日本根瘤菌有效和无效菌株种间质粒和基因组DNA序列同源性及nif基因定位。
对3株日本根瘤菌进行了种间保守性质粒携带DNA序列的检测和nif序列的定位。在大豆上形成有效固氮根瘤的菌株61A76和110的DNA序列同源性极低(24%);菌株61A76含有1个质粒,菌株110不含可识别的质粒。在大豆上形成无效根瘤的菌株61A24与菌株110序列同源性较高(50%),含有两个质粒。虽然从菌株61A76和61A24中分离的纯化质粒DNA的总序列同源性有限,但这些菌株的质粒和总DNA与菌株110的DNA之间的异源杂交表明,一个菌株的质粒携带的几个序列可能出现在染色体上或在其他菌株的非常大的质粒中。从菌株61A24中分离出的质粒与农杆菌中章鱼钛质粒的几个smi片段具有一定的同源性,而从菌株61A76中分离出的质粒与该钛质粒没有同源性。发现5株日本根瘤菌的DNA序列与携带肺炎克雷伯菌nif-KDH基因的质粒pSA30杂交,但这些序列不在分离的质粒上。这些基因的组织在菌株110和61A24中相似,但在菌株61A76中不同。
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