Duan Jin, Müller Kirsten M, Charles Trevor C, Vesely Susanne, Glick Bernard R
Department of Biology, University of Waterloo, Waterloo, ON, Canada.
Microb Ecol. 2009 Apr;57(3):423-36. doi: 10.1007/s00248-008-9407-6. Epub 2008 Jun 12.
A collection of 233 rhizobia strains from 30 different sites across Saskatchewan, Canada was assayed for 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, with 27 of the strains displaying activity. When all 27 strains were characterized based on 16S rRNA gene sequences, it was noted that 26 strains are close to Rhizobium leguminosarum and one strain is close to Rhizobium gallicum. Polymerase chain reaction (PCR) was used to rapidly isolate ACC deaminase structural genes from the above-mentioned 27 strains; 17 of them have 99% identities with the previously characterized ACC deaminase structural gene (acdS) from R. leguminosarum bv. viciae 128C53K, whereas the other ten strains are 84% identical (864-866/1,020 bp) compared to the acdS from strain 128C53K. Southern hybridization showed that each strain has only one ACC deaminase gene. Using inverse PCR, the region upstream of the ACC deaminase structural genes was characterized for all 27 strains, and 17 of these strains were shown to encode a leucine-responsive regulatory protein. The results are discussed in the context of a previously proposed model for the regulation of bacterial ACC deaminase in R. leguminosarum 128C53K.
对从加拿大萨斯喀彻温省30个不同地点收集的233株根瘤菌菌株进行了1-氨基环丙烷-1-羧酸(ACC)脱氨酶活性检测,其中27株显示出活性。当根据16S rRNA基因序列对所有27株菌株进行鉴定时,发现26株与豌豆根瘤菌相近,1株与法国根瘤菌相近。采用聚合酶链反应(PCR)从上述27株菌株中快速分离ACC脱氨酶结构基因;其中17株与先前鉴定的豌豆根瘤菌蚕豆生物型128C53K的ACC脱氨酶结构基因(acdS)具有99%的同一性,而其他10株与128C53K菌株的acdS相比同一性为84%(864 - 866/1,020 bp)。Southern杂交表明每个菌株只有一个ACC脱氨酶基因。使用反向PCR对所有27株菌株ACC脱氨酶结构基因的上游区域进行了鉴定,其中17株被证明编码一种亮氨酸响应调节蛋白。在先前提出的豌豆根瘤菌128C53K中细菌ACC脱氨酶调控模型的背景下对结果进行了讨论。