Detection and separation of Rhizobium and Bradyrhizobium Nod metabolites using thin-layer chromatography.
Detection and separation of Rhizobium and Bradyrhizobium Nod metabolites using thin-layer chromatography.
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DOI：
10.1094/mpmi-5-072
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发表时间：
1992
期刊：
影响因子：
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通讯作者：
H. Spaink;A. Aarts;G. Stacey;G. Bloemberg;B. Lugtenberg;E. P. Kennedy
中科院分区：
文献类型：
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作者：
H. Spaink;A. Aarts;G. Stacey;G. Bloemberg;B. Lugtenberg;E. P. Kennedy
使用放射性乙酸盐作为前体，表明根瘤菌属和慢生根瘤菌属菌株的共同的HLA-ABC基因参与分泌到生长培养基中的一种或多种代谢物的产生。一种快速薄层色谱（TLC）系统已经开发出来，以分离这些所谓的Nod代谢物，然后可以通过放射自显影可视化。交叉接种组的供试菌株结瘤代谢产物的模式不同。豆科植物viceae、河草R. L. BV. trifolii、三叶草R.草木犀和B.山茱萸只有根瘤菌的代谢产物。当培养基中存在放射性硫酸盐时，苜蓿草被标记。本文还讨论了其它促排基因在R. L. BV.进一步详细测试了viceae在检测到的Nod代谢物的产生中的作用。除了常见的NodABC基因，NodFE和NodL基因也参与了Nod代谢产物的产生。相反，染色体背景不影响检测到的Nod代谢物的数量或它们在TLC板上的迁移率。结瘤代谢产物也可以在大肠杆菌细胞中产生和分泌，其中适当的表达基因。
Using radioactive acetate as a precursor, it was shown that the common nodABC genes of Rhizobium and Bradyrhizobium strains are involved in the production of one or more metabolites that are excreted into the growth medium. A rapid thin-layer chromatography (TLC) system has been developed to separate these so-called Nod metabolites that can then be visualized by autoradiography. Different patterns of Nod metabolites were observed in the tested strains of the cross-inoculation groups of R. leguminosarum bv. viceae, R. l. bv. trifolii, R. meliloti, and B. japonicum. Only Nod metabolites of R. meliloti became labeled when radioactive sulphate was present in the medium. The role of the other nodulation genes of R. l. bv. viceae in the production of the detected Nod metabolites was tested in further detail. In addition to the common nodABC genes, the nodFE and nodL genes are involved in the production of Nod metabolites. In contrast, the chromosomal background did not influence the number of detected Nod metabolites or their mobilities on TLC plates. Nod metabolites could also be produced and excreted in Escherichia coli cells in which the appropriate nodulation genes were expressed.