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基于慢生根瘤菌甘氨酸氧化酶基因(thiO)的土壤中草甘膦生物降解潜力

Glyphosate Biodegradation Potential in Soil Based on Glycine Oxidase Gene (thiO) from Bradyrhizobium.

作者信息

Hernández Guijarro Keren, De Gerónimo Eduardo, Erijman Leonardo

机构信息

National Institute of Agricultural Technology (INTA), Balcarce Experimental Station, Ruta Nac, 226, Km 73,5, CP 7620 Balcarce, Buenos Aires, Argentina.

National Scientific and Technical Research Council (CONICET), Buenos Aires, Argentina.

出版信息

Curr Microbiol. 2021 May;78(5):1991-2000. doi: 10.1007/s00284-021-02467-z. Epub 2021 Apr 2.

Abstract

Despite the intensive use of glyphosate (GP) and its ubiquitous presence in the environment, studies addressing the presence of microbial genes involved in glyphosate degradation in natural conditions are scarce. Based on the agronomical importance of Bradyrhizobium genus and its metabolic versatility, we tested the hypothesis that species or genotypes of Bradyrhizobium could be a proxy for GP degrader potential in soil. A quantitative PCR assay was designed to target a specific region of the glycine oxidase gene (thiO), involved in the oxidation of glyphosate to AMPA, from known sequences of Bradyrhizobium species. The abundance of the thiO gene was determined in response to herbicide application in soils with different GP exposure history both under field and microcosm conditions. The gene coding for RNA polymerase subunitB (rpoB) was used as a reference for the abundance of total Bradyrhizobia. The assay using the designed primers was linear over a very large concentration range of the target and showed high efficiency and specificity. In a field experiment, there was a differential response related to the history of glyphosate use and the native Bradyrhizobium genotypes. In a soil without previous exposure to herbicides, thiO gene increased over time after glyphosate application with most genotypes belonging to the B. jicamae and B. elkanni supergroups. Conversely, in an agricultural soil with more than 10 years of continuous glyphosate application, the abundance of thiO gene decreased and most genotypes belonged to B. japonicum supergroup. In a microcosm assay, the amount of herbicide degraded after a single application was positively correlated to the number of thiO copies in different agricultural soils from the Pampean Region. Our results suggest that Bradyrhizobium species are differently involved in glyphosate degradation, denoting the existence of metabolically versatile microorganisms which can be explored for sustainable agriculture practices. The relationship between the abundance of thiO gene and the GP degraded in soil point to the use of thiO gene as a proxy for GP degradation in soil.

摘要

尽管草甘膦(GP)被大量使用且在环境中普遍存在,但针对自然条件下参与草甘膦降解的微生物基因存在情况的研究却很少。基于慢生根瘤菌属在农学上的重要性及其代谢多样性,我们检验了这样一个假设,即慢生根瘤菌的物种或基因型可以作为土壤中草甘膦降解潜力的指标。设计了一种定量PCR测定法,以已知的慢生根瘤菌物种序列中参与草甘膦氧化为氨甲基膦酸的甘氨酸氧化酶基因(thiO)的特定区域为靶点。在田间和微观试验条件下,针对不同草甘膦暴露历史的土壤施用除草剂后,测定了thiO基因的丰度。编码RNA聚合酶亚基B(rpoB)的基因被用作总慢生根瘤菌丰度的参照。使用设计引物进行的测定在目标物的非常大的浓度范围内呈线性,且显示出高效率和特异性。在田间试验中,与草甘膦使用历史和本地慢生根瘤菌基因型存在差异响应。在未曾接触过除草剂的土壤中,施用草甘膦后,thiO基因随时间增加,大多数基因型属于菜豆慢生根瘤菌和埃尔坎尼慢生根瘤菌超群。相反,在连续施用草甘膦超过10年的农业土壤中,thiO基因丰度下降,大多数基因型属于日本慢生根瘤菌超群。在微观试验中,单次施用后降解的除草剂量与来自潘帕斯地区不同农业土壤中thiO拷贝数呈正相关。我们的结果表明,不同的慢生根瘤菌物种参与草甘膦降解的程度不同,这表明存在代谢多样的微生物,可用于可持续农业实践。thiO基因丰度与土壤中降解的草甘膦之间的关系表明,可将thiO基因用作土壤中草甘膦降解的指标。

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