AN AZORHIZOBIUM-CAULINODANS ORS571 LOCUS INVOLVED IN LIPOPOLYSACCHARIDE PRODUCTION AND NODULE FORMATION ON SESBANIA-ROSTRATA STEMS AND ROOTS
AN AZORHIZOBIUM-CAULINODANS ORS571 LOCUS INVOLVED IN LIPOPOLYSACCHARIDE PRODUCTION AND NODULE FORMATION ON SESBANIA-ROSTRATA STEMS AND ROOTS
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DOI：
10.1128/jb.176.1.92-99.1994
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发表时间：
1994-01-01
影响因子：
3.2
通讯作者：
HOLSTERS, M
中科院分区：
文献类型：
--
作者：
GOETHALS, K;LEYMAN, B;HOLSTERS, M
Azorhizobium caulinodans ORS571 is able to nodulate roots and stems of the tropical legume Sesbania rostrata. An ORS571 Tn5 insertion mutant, strain ORS571-X15, had a rough colony morphology, was nonmotile, and showed clumping behavior on various media. When this pleiotropic mutant was inoculated on roots or stems of the host, no nodules developed (Nod(-)). Compared with the wild type, strain ORS571-X15 produced lipopolysaccharides (LPS) with an altered ladder pattern on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, suggestive of a different O-antigen structure with a lower degree of polymerization. A cosmid clone, pRG20, that fully complemented all phenotypes of ORS571-X15 was isolated. With a 6-kb EcoRI subfragment of pRG20, clumping was relieved and nodulation was almost completely restored, but the strain was still nonmotile. LPS preparations from these complemented strains resembled the wild-type LPS, although minor quantitative and qualitative differences were evident. The sequence of the locus hit by the Tn5 in ORS571-X15 (the oac locus) revealed a striking homology,vith the rfb locus of Salmonella typhimurium, which is involved in O-antigen biosynthesis. The Tn5 insertion position was mapped to the oac3 gene, homologous to rfbA, encoding dTDP-D-glucose synthase. Biochemical assaying showed that ORS571-X15 is indeed defective in dTDP-D-glucose synthase activity, essential for the production of particular deoxyhexoses. Therefore, it was proposed that the O antigen of the mutant strain is devoid of such sugars.
