Genes Conferring Copper Resistance in Sinorhizobium meliloti CCNWSX0020 Also Promote the Growth of Medicago lupulina in Copper-Contaminated Soil
Genes Conferring Copper Resistance in Sinorhizobium meliloti CCNWSX0020 Also Promote the Growth of Medicago lupulina in Copper-Contaminated Soil
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苜蓿中华根瘤菌CCNWSX0020赋予铜抗性的基因也促进铜污染土壤中羽扇豆苜蓿的生长
DOI：
10.1128/aem.03381-13
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发表时间：
2014-01
影响因子：
4.4
通讯作者：
Wei, Gehong
中科院分区：
文献类型：
--
作者：
Ma, Zhanqiang;Hao, Xiuli;Rensing, Christopher;Wei, Gehong
摘要苜蓿中华根瘤菌（Sinorhizobium meliloti）CCNWSX 0020是从我国西北地区金矿尾矿中的苜蓿（Medicago lupulina）根瘤中分离得到的一株具有抗铜和促进豆科植物在铜污染土壤中生长的双重功能的菌株。尽管如此，S.苜蓿CCNWSX 0020仍然未表征。为了研究与铜抗性相关的基因，我们用一个S。创建了14，000个突变体的苜蓿CCNWSX 0020 Tn 5插入文库。分离到5个铜敏感突变体，命名为SXa-1，SXa-2，SXC-1，SXC-2和SXn，并通过反向PCR和随后的测序鉴定所涉及的破坏区域。SXa-1和SXa-2均在lpxXL（SM0020_18047）中携带转座子插入，编码LpxXL C-28酰基转移酶; SXc-1和SXc-2在merR（SM0020_29390）中携带转座子插入，编码调节激活因子; SXn在omp（SM0020_18792）中包含转座子插入，编码假设的外膜蛋白。反转录PCR（RT-PCR）结合转座子基因破坏的结果显示，SM0020_05862编码一种不寻常的P型ATP酶，受MerR蛋白的调控。基因组序列分析表明，这种P型ATP酶不包含N-末端金属结合结构域或CPC基序，而是TPCP相比，CopA从大肠杆菌。通过盆栽试验，研究了铜胁迫对寄主植物的生长和铜积累的影响。在野生型或不同突变体的存在下，啤酒花属受到影响。土壤样品进行了三个水平的铜污染，即，未污染的控制和47.36和142.08 mg/kg，和三个重复进行每个处理。结果表明，野生型S.苜蓿CCNWSX 0020能使寄主植物生长更好，并积累铜离子。植株干重和铜含量均高于对照。在CuSO_4存在下，用5个铜敏感突变体接种的啤酒花显著降低。
ABSTRACT Sinorhizobium meliloti CCNWSX0020, isolated from root nodules of Medicago lupulina growing in gold mine tailings in the northwest of China, displayed both copper resistance and growth promotion of leguminous plants in copper-contaminated soil. Nevertheless, the genetic and biochemical mechanisms responsible for copper resistance in S. meliloti CCNWSX0020 remained uncharacterized. To investigate genes involved in copper resistance, an S. meliloti CCNWSX0020 Tn5 insertion library of 14,000 mutants was created. Five copper-sensitive mutants, named SXa-1, SXa-2, SXc-1, SXc-2, and SXn, were isolated, and the disrupted regions involved were identified by inverse PCR and subsequent sequencing. Both SXa-1 and SXa-2 carried a transposon insertion in lpxXL (SM0020_18047), encoding the LpxXL C-28 acyltransferase; SXc-1 and SXc-2 carried a transposon insertion in merR (SM0020_29390), encoding the regulatory activator; SXn contained a transposon insertion in omp (SM0020_18792), encoding a hypothetical outer membrane protein. The results of reverse transcriptase PCR (RT-PCR) combined with transposon gene disruptions revealed that SM0020_05862, encoding an unusual P-type ATPase, was regulated by the MerR protein. Analysis of the genome sequence showed that this P-type ATPase did not contain an N-terminal metal-binding domain or a CPC motif but rather TPCP compared with CopA from Escherichia coli. Pot experiments were carried out to determine whether growth and copper accumulation of the host plant M. lupulina were affected in the presence of the wild type or the different mutants. Soil samples were subjected to three levels of copper contamination, namely, the uncontaminated control and 47.36 and 142.08 mg/kg, and three replicates were conducted for each treatment. The results showed that the wild-type S. meliloti CCNWSX0020 enabled the host plant to grow better and accumulate copper ions. The plant dry weight and copper content of M. lupulina inoculated with the 5 copper-sensitive mutants significantly decreased in the presence of CuSO4.
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影响因子：
64.8
作者：
David R. Brown;K. Qin;J. Herms;A. Madlung;J. Manson;R. Strome;P. Fraser;T. Kruck;A. Bohlen
通讯作者：
David R. Brown;K. Qin;J. Herms;A. Madlung;J. Manson;R. Strome;P. Fraser;T. Kruck;A. Bohlen
DOI：
10.1038/nbt1183-784
发表时间：
1983-01-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
SIMON, R;PRIEFER, U;PUHLER, A
通讯作者：
PUHLER, A
影响因子：
4.4
作者：
Fraser, JA;Davis, MA;Hynes, MJ
通讯作者：
Hynes, MJ
影响因子：
4.4
作者：
S. Rossbach;Danielle J. Mai;E. L. Carter;L. Sauviac;D. Capela;C. Bruand;F. D. de Bruijn
通讯作者：
S. Rossbach;Danielle J. Mai;E. L. Carter;L. Sauviac;D. Capela;C. Bruand;F. D. de Bruijn
影响因子：
3.2
作者：
Ardissone, Silvia;Kobayashi, Hajime;Deakin, William J.
通讯作者：
Deakin, William J.