Functional Metagenomics to Mine Soil Microbiome for Novel Cadmium Resistance Genetic Determinants

ZHENG Xin; CHEN Liang; CHEN Miaomiao; CHEN Jinghao; LI Xiaofang

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Functional Metagenomics to Mine Soil Microbiome for Novel Cadmium Resistance Genetic Determinants

机译：新型土壤抗镉遗传决定因素的矿土壤微生物组功能元基因组学。

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Metal resistance genes are valuable resources for genetic engineering of bioremediation tools.In this study,novel genetic determinants involved in cadmium (Cd) resistance were identified using a small-insert metagenomic DNA library constructed from an arable soil microbiome.A total of 16 recombinant plasmids harboring 49 putative open reading frames (ORFs) were found to be associated with enhanced Cd tolerance.In addition to several ORFs for ion transport/chelation and stress response,most ORFs were assumed to be associated with non-direct metal resistance mechanisms such as energy metabolism,protein/amino acid metabolism,carbohydrate/fatty acid metabolism,and signal transduction.Furthermore,13 ORFs from five clones selected at random were cloned and subject to Cd resistance assay.Eight of these ORFs were positive for Cd resistance when expressed in Escherichia coli,among which four ORFs significantly reduced Cd accumulation and one increased Cd enrichment of the host cells.Notably,C1-ORF1,potentially encoding a histidine kinase-like adenosine triphosphatase,was the most effective Cd resistance determinant and reduced host Cd accumulation by 33.9％.These findings highlight the vast capacity of soil microbiome as a source of gene pool for bioengineering.The novel genetic determinants for Cd resistance identified in this study merit further systematic explorations into their molecular mechanisms.

机译：金属抗性基因是生物修复工具基因工程的宝贵资源。在这项研究中，使用由可耕土壤微生物组构建的小插入宏基因组DNA库，鉴定了与镉（Cd）抗性有关的新遗传决定因素。带有49个推定的开放阅读框（ORF）被发现与增强的Cd耐受性有关。除了几个用于离子迁移/螯合和应力响应的ORF之外，大多数ORF还被认为与非直接的金属电阻机制有关，例如能量代谢，蛋白质/氨基酸代谢，碳水化合物/脂肪酸代谢和信号转导。此外，从随机选择的五个克隆中克隆了13个ORF，并对其进行Cd抗性测定。这些ORF中有8个在大肠杆菌中表达时对Cd抗性呈阳性。大肠杆菌中，有四个ORF显着降低了Cd的积累，而一个增加了Cd对宿主细胞的富集。 ORF1可能编码组氨酸激酶样腺苷三磷酸酶，是最有效的Cd抗性决定簇，使宿主Cd积累减少了33.9％。这些发现突显了土壤微生物组作为生物工程基因库来源的巨大能力。在这项研究中确定的对Cd耐药性的研究值得对其分子机制进行进一步的系统探索。

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《土壤圈（英文版）》 |2019年第3期|298-310|共13页
作者
ZHENG Xin; CHEN Liang; CHEN Miaomiao; CHEN Jinghao; LI Xiaofang;
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Key Laboratory for Agricultural Water Resources, Hebei Key Laboratory of Soil Ecology, Center for Agricultural Resources Research,Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang 050021 China;

Key Laboratory for Agricultural Water Resources, Hebei Key Laboratory of Soil Ecology, Center for Agricultural Resources Research,Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang 050021 China;

Key Laboratory for Agricultural Water Resources, Hebei Key Laboratory of Soil Ecology, Center for Agricultural Resources Research,Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang 050021 China;

Key Laboratory for Agricultural Water Resources, Hebei Key Laboratory of Soil Ecology, Center for Agricultural Resources Research,Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang 050021 China;

University of Chinese Academy of Sciences, Beijing 100049 China;

Key Laboratory for Agricultural Water Resources, Hebei Key Laboratory of Soil Ecology, Center for Agricultural Resources Research,Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang 050021 China;

Environment Center, Sustainable Minerals Institute, The University of Queensland, Brisbane 4072 Australia;

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Functional Metagenomics to Mine Soil Microbiome for Novel Cadmium Resistance Genetic Determinants

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