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Rhizobium Inoculation Enhances the Resistance of Alfalfa and Microbial Characteristics in Copper-Contaminated Soil.

ABSTRACT: Some studies have reported the importance of rhizobium in mitigating heavy metal toxicity, however, the regulatory mechanism of the alfalfa-rhizobium symbiosis to resist copper (Cu) stress in the plant-soil system through biochemical reactions is still unclear. This study assessed the effects of rhizobium (Sinorhizobium meliloti CCNWSX0020) inoculation on the growth of alfalfa and soil microbial characteristics under Cu-stress. Further, we determined the regulatory mechanism of rhizobium inoculation to alleviate Cu-stress in alfalfa through plant-soil system. The results showed that rhizobium inoculation markedly alleviated Cu-induced growth inhibition in alfalfa by increasing the chlorophyll content, height, and biomass, in addition to nitrogen and phosphorus contents. Furthermore, rhizobium application alleviated Cu-induced phytotoxicity by increasing the antioxidant enzyme activities and soluble protein content in tissues, and inhibiting the lipid peroxidation levels (i.e., malondialdehyde content). In addition, rhizobium inoculation improved soil nutrient cycling, which increased soil enzyme activities (i.e., β-glucosidase activity and alkaline phosphatase) and microbial biomass nitrogen. Both Pearson correlation coefficient analysis and partial least squares path modeling (PLS-PM) identified that the interactions between soil nutrient content, enzyme activity, microbial biomass, plant antioxidant enzymes, and oxidative damage could jointly regulate plant growth. This study provides comprehensive insights into the mechanism of action of the legume-rhizobium symbiotic system to mitigate Cu stress and provide an efficient strategy for phytoremediation of Cu-contaminated soils.

SUBMITTER: Duan C

PROVIDER: S-EPMC8791600 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description: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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Rhizobium Inoculation Enhances the Resistance of Alfalfa and Microbial Characteristics in Copper-Contaminated Soil.

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