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Arsenic methylation by a genetically engineered Rhizobium-legume symbiont

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Abstract

Background and aims

Arsenic (As) is one of the most widespread environmental contaminants. The aim of our study was to test a novel bioremediation system based on the symbiosis between leguminous plant and genetically engineered rhizobia.

Methods

The arsenite [As(III)] S-adenosylmethionine methyltransferase gene (CrarsM) from the alga Chlamydomonas reinhardtii was inserted into the chromosome of Rhizobium leguminosarum bv. trifolii strain R3. The As methylation ability of the recombinant Rhizobium was tested under free living conditions and in symbiosis with red clover plants. Arsenic speciation was determined using high-performance liquid chromatography-inductively coupled plasma mass spectrometry.

Results

Under free-living conditions, CrarsM-recombinant R. leguminosarum gained the ability to methylate As(III) to methylated arsenicals, including methylarsenate [MAs(V)], dimethylarsenate [DMAs(V)] and trimethylarsine oxide [TMAs(V)O]. Red clover plants were inoculated with either control (non-recombinant) or CrarsM-recombinant R. leguminosarum and exposed to 5 or 10 μM arsenite. No methylated As species were detected in red clover plants inoculated with control R. leguminosarum. In contrast, all three methylated species were detected in both the nodules and the shoots when the recombinant Rhizobium established symbiosis with red clover, accounting for 74.7–75.1% and 29.1–42.4% of the total As in the two plant tissues, respectively. The recombinant symbiont also volatilized small amounts of As.

Conclusions

The present study demonstrates that engineered rhizobia expressing an algal arsM gene can methylate and volatilize As, providing a proof of concept for potential future use of legume-rhizobia symbionts for As bioremediation.

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Acknowledgements

The study was supported by the Natural Science Foundation of China (grant No. 41330853 and 41571312), the Innovative Research Team Development Plan of the Ministry of Education of China (grant no. IRT1256), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) and NIH grant R37 GM55425 to B.P.R.

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Authors and Affiliations

  1. Jiangsu Provincial Key Laboratory for Organic Waste Utilization, Jiangsu Collaborative Innovation Center for Solid Organic Waste Resource Utilization, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095, China

    Jun Zhang, Yan Xu, Tingting Cao & Fang-Jie Zhao

  2. Department of Cellular Biology and Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, FL, 33199, USA

    Jian Chen & Barry P. Rosen

  3. Sustainable Soils and Grassland Systems Department, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK

    Fang-Jie Zhao

Authors
  1. Jun Zhang
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  2. Yan Xu
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  3. Tingting Cao
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  4. Jian Chen
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  5. Barry P. Rosen
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  6. Fang-Jie Zhao
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Corresponding author

Correspondence to Fang-Jie Zhao.

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Responsible Editor: Juan Barcelo.

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Zhang, J., Xu, Y., Cao, T. et al. Arsenic methylation by a genetically engineered Rhizobium-legume symbiont. Plant Soil 416, 259–269 (2017). https://doi.org/10.1007/s11104-017-3207-z

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