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Protein extraction and database construction in tea rhizosphere soil

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Abstract

Soil protein extraction and database construction are the key points of soil proteomics research. In this paper, tea tree rhizosphere soil was used as material. The soil proteins were extracted three times by citrate, SDS, and mixture of citrate and SDS, respectively. The total proteins were separated by 2-DE electrophoresis and identified by Data Dependent Acquisition (DDA). The DDA data collection was further separated by High-Performance Liquid Chromatography (HPLC) and identified by LC–MS/MS, then to build the database of soil protein and microbial species using fungus and bacteria databases. The result showed soil protein was identified and reached 2741 points, and the molecular weight was mainly distributed in between 2.64 and 338.33 kDa, and Isoelectric point (pI) is mainly distributed in between 3.78 and 12.15. The soil protein was mainly from 138 families, 346 species of microorganisms. This optimization method could obtain more proteins than previous methods, with a wider range of molecular weight and pI. This study lays an important foundation for the research and development of soil metaproteomics.

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Acknowledgements

This work was supported by China Postdoctoral Science Foundation (2016M600493), the National 948 Project (2014-Z36), Natural Science Foundation of Fujian Province (2017J05057), Science and Technology Project of Longyan City (2017LY71), the Project of Scientific Research of Young and Middle-aged teachers, Fujian Province (JAT170573), Fujian Outstanding Research Talent Cultivation Project, National Program for Innovation and Entrepreneurship Training for College Students (201911312001, 202011312004), and Youth Top Talent Training Program of Longyan University (2019ZJ19).

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Authors and Affiliations

  1. College of Life Sciences, Longyan University, Longyan, 364012, China

    Hai-bin Wang, Chun-lian Zhu, Yu-hua Wang, Qing-xu Zhang, Peng Wang & Ding Li

  2. Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring, Fujian Agriculture and Forestry University, Fuzhou, 35002, China

    Hai-bin Wang, Qing-xu Zhang, Peng Wang & Hai-bin He

  3. College of Ecology and Resources Engineering, Wuyi University, Wuyishan, 354300, China

    Xiao-li Jia & Jiang-hua Ye

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  1. Hai-bin Wang
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  2. Chun-lian Zhu
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  9. Hai-bin He
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Correspondence to Hai-bin He.

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Wang, Hb., Zhu, Cl., Wang, Yh. et al. Protein extraction and database construction in tea rhizosphere soil. Acta Physiol Plant 42, 158 (2020). https://doi.org/10.1007/s11738-020-03146-5

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