Rossbach S, Rasul G, Schneider M, Eardly B, de Bruijn F J
NSF-Center for Microbial Ecology, Michigan State University, East Lansing 48824, USA.
Mol Plant Microbe Interact. 1995 Jul-Aug;8(4):549-59. doi: 10.1094/mpmi-8-0549.
Rhizopine (L-3-O-methyl-scyllo-inosamine; 3-O-MSI) synthesis (mos) and catabolism (moc) genes were originally isolated from Rhizobium meliloti strain L5-30 (Murphy et al., Proc. Natl. Acad. Sci. U.S.A., 84:493, 1987). These genes have been postulated to give a competitive advantage to this strain in the rhizosphere, since the ability to utilize the unusual nutritional mediator rhizopine as nitrogen and carbon source appears to be correlated with the ability of Moc+ bacteria to efficiently infect alfalfa plants. This study examines the distribution of rhizopine catabolism (moc) genes among different soil bacteria. By using oligonucleotide primers homologous to the moc genes and the polymerase chain reaction (PCR), moc genes were shown to be absent from a random collection of 100 different soil isolates. However, screening 50 different electrophoretic type strains of a worldwide R. meliloti collection (Eardly et al., Appl. Environ. Microbiol. 56:187, 1990) revealed the presence of moc genes in three additional strains, S33, 102F51, and 74B3. These three strains were found to be able to synthesize rhizopine in planta (Mos+) and to catabolize it (Moc+). To determine the relatedness of the Mos+/Moc+ strains to each other and to other R. meliloti strains, we used the rep-PCR method to generate genomic fingerprints, and to create a phylogenetic tree with the help of an optical imaging system and data analysis program (AMBIS). Because of the apparent infrequent occurrence of moc genes among soil bacteria, we suggest that the use of moc genes as a selectable marker trait for tracking genetically manipulated organisms is feasible.
根瘤碱(L-3-O-甲基-scyllo-肌醇胺;3-O-MSI)合成(mos)和分解代谢(moc)基因最初是从苜蓿根瘤菌L5-30菌株中分离出来的(Murphy等人,《美国国家科学院院刊》,84:493,1987)。据推测,这些基因赋予该菌株在根际环境中的竞争优势,因为利用这种不寻常的营养介质根瘤碱作为氮源和碳源的能力似乎与Moc+细菌有效感染苜蓿植物的能力相关。本研究考察了根瘤碱分解代谢(moc)基因在不同土壤细菌中的分布情况。通过使用与moc基因同源的寡核苷酸引物和聚合酶链反应(PCR),在随机收集的100种不同土壤分离物中未检测到moc基因。然而,对来自世界各地的苜蓿根瘤菌集合的50种不同电泳类型菌株进行筛选(Eardly等人,《应用与环境微生物学》,56:187,1990),结果显示在另外三个菌株S33、102F51和74B3中存在moc基因。发现这三个菌株能够在植物体内合成根瘤碱(Mos+)并对其进行分解代谢(Moc+)。为了确定Mos+/Moc+菌株彼此之间以及与其他苜蓿根瘤菌菌株的亲缘关系,我们使用rep-PCR方法生成基因组指纹图谱,并借助光学成像系统和数据分析程序(AMBIS)构建系统发育树。由于moc基因在土壤细菌中明显不常见,我们认为将moc基因用作追踪基因改造生物的选择标记性状是可行的。