Faculty of Life and Environmental Science, Shimane University 1060 Nishikawatsu, Matsue, Shimane 690-8504, Japan.
Faculty of Life and Environmental Science, Shimane University 1060 Nishikawatsu, Matsue, Shimane 690-8504, Japan.
Microbiol Res. 2016 Jul-Aug;188-189:62-71. doi: 10.1016/j.micres.2016.04.014. Epub 2016 May 4.
Herbicides 2,4-dichlorophenoxyacetic acid (2,4-D)- and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T)-degrading Bradyrhizobium strains possess tfdAα and/or cadABC as degrading genes. It has been reported that root-nodulating bacteria belonging to Bradyrhizobium elkanii also have tfdAα and cadA like genes but lack the ability to degrade these herbicides and that the cadA genes in 2,4-D-degrading and non-degrading Bradyrhizobium are phylogenetically different. In this study, we identified cadRABCK in the genome of a type strain of soybean root-nodulating B. elkanii USDA94 and demonstrated that the strain could degrade the herbicides when cadABCK was forcibly expressed. cadABCK-cloned Escherichia coli also showed the degrading ability. Because co-spiked phenoxyacetic acid (PAA) could induce the degradation of 2,4-D in B. elkanii USDA94, the lack of degrading ability in this strain was supposed to be due to the low inducing potential of the herbicides for the degrading gene cluster. On the other hand, tfdAα from B. elkanii USDA94 showed little potential to degrade the herbicides, but it did for 4-chlorophenoxyacetic acid and PAA. The 2,4-D-degrading ability of the cad cluster and the inducing ability of PAA were confirmed by preparing cadA deletion mutant. This is the first study to demonstrate that the cad cluster in the typical root-nodulating bacterium indeed have the potential to degrade the herbicides, suggesting that degrading genes for anthropogenic compounds could be found in ordinary non-degrading bacteria.
除草剂 2,4-二氯苯氧乙酸(2,4-D)和 2,4,5-三氯苯氧乙酸(2,4,5-T)降解菌 Bradyrhizobium 菌株拥有 tfdAα 和/或 cadABC 作为降解基因。据报道,属于根瘤菌属的结瘤细菌也拥有 tfdAα 和 cadA 类似基因,但缺乏降解这些除草剂的能力,并且 2,4-D 降解菌和非降解菌的 cadA 基因在系统发育上存在差异。在本研究中,我们在大豆根瘤菌属 B. elkanii USDA94 的模式菌株基因组中鉴定出了 cadRABCK,并证实当 cadABCK 被强制表达时,该菌株能够降解这些除草剂。cadABCK 克隆的大肠杆菌也表现出了降解能力。由于共添加的苯氧乙酸(PAA)能够诱导 B. elkanii USDA94 降解 2,4-D,因此该菌株缺乏降解能力可能是由于这些除草剂对降解基因簇的诱导潜力较低。另一方面,来自 B. elkanii USDA94 的 tfdAα 对除草剂的降解潜力较小,但对 4-氯苯氧乙酸和 PAA 有一定的降解潜力。通过制备 cadA 缺失突变体,证实了 cad 簇的 2,4-D 降解能力和 PAA 的诱导能力。这是首次证明典型结瘤菌中的 cad 簇确实具有降解这些除草剂的潜力的研究,这表明在普通非降解菌中可能存在用于降解人为化合物的降解基因。