Shah Ritu, Emerich David W
Department of Biochemistry, University of Missouri, Columbia, 65211, USA.
J Bacteriol. 2006 Nov;188(21):7600-8. doi: 10.1128/JB.00671-06. Epub 2006 Aug 25.
A mutant strain of Bradyrhizobium japonicum USDA110 lacking isocitrate dehydrogenase activity was created to determine whether this enzyme was required for symbiotic nitrogen fixation with soybean (Glycine max cv. Williams 82). The isocitrate dehydrogenase mutant, strain 5051, was constructed by insertion of a streptomycin resistance gene cassette. The mutant was devoid of isocitrate dehydrogenase activity and of immunologically detectable protein, indicating there is only one copy in the genome. Strain 5051 grew well on a variety of carbon sources, including arabinose, pyruvate, succinate, and malate, but, unlike many microorganisms, was a glutamate auxotroph. Although the formation of nodules was slightly delayed, the mutant was able to form nodules on soybean and reduce atmospheric dinitrogen as well as the wild type, indicating that the plant was able to supply sufficient glutamate to permit infection. Combined with the results of other citric acid cycle mutants, these results suggest a role for the citric acid cycle in the infection and colonization stage of nodule development but not in the actual fixation of atmospheric dinitrogen.
构建了一株缺乏异柠檬酸脱氢酶活性的日本慢生根瘤菌USDA110突变株,以确定该酶对于与大豆(Glycine max cv. Williams 82)共生固氮是否必需。异柠檬酸脱氢酶突变株5051是通过插入一个链霉素抗性基因盒构建而成的。该突变株缺乏异柠檬酸脱氢酶活性且无免疫可检测蛋白,表明基因组中只有一个拷贝。5051菌株能在多种碳源上良好生长,包括阿拉伯糖、丙酮酸、琥珀酸和苹果酸,但与许多微生物不同,它是谷氨酸营养缺陷型。尽管结瘤形成稍有延迟,但该突变株能够在大豆上结瘤并像野生型一样还原大气中的氮气,这表明植物能够提供足够的谷氨酸以允许感染。结合其他柠檬酸循环突变体的结果,这些结果表明柠檬酸循环在根瘤发育的感染和定殖阶段起作用,但在实际固定大气中的氮气方面不起作用。