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A plasmid-born Rap-Phr system regulates surfactin production, sporulation and genetic competence in the heterologous host, Bacillus subtilis OKB105

  • Applied microbial and cell physiology
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Abstract

According to the change of environment, soil-dwelling Bacillus species differentiate into distinct subpopulations, such as spores and competent cells. Rap-Phr systems have been found to be involved in this differentiation circuit by interacting with major regulatory proteins, such as Spo0A, ComA, and DegU. In this study, we report that the plasmid-born RapQ-PhrQ system found in Bacillus amyloliquefaciens B3 affects three regulatory pathways in the heterologous host Bacillus subtilis. Expression of rapQ in B. subtilis OKB105 strongly suppressed its sporulation efficiency, transformation efficiency, and surfactin production. Co-expression of phrQ or addition of synthesized PhrQ pentapeptide in vitro could compensate for the suppressive effects caused by rapQ. We also found that expression of rapQ decreased the transcriptional level of the sporulation-related gene spoIIE and surfactin synthesis-related gene srfA; meanwhile, the transcriptional levels of these genes could be rescued by co-expression of phrQ and in vitro addition of PhrQ pentapeptide. Electrophoretic mobility shift (EMSA) result also showed that RapQ could bind to ComA without interacting with ComA binding to DNA, and PhrQ pentapeptide antagonized RapQ activity in vitro. These results indicate that this new plasmid-born RapQ-PhrQ system controls sporulation, competent cell formation, and surfactin production in B. subtilis OKB105.

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Acknowledgments

This work was supported by grants from the Agro-scientific Research in the Public Interest (20130315), the Special Fund for the Fundamental Research Funds for the Central Universities (KYZ201404), the National Natural Science Foundation of China (31100056, 31471811), the Doctoral Fund of Ministry of Education of China (20100097120011), and the National High-tech R&D Program of China (2012AA101504).

Ethical statement/conflict of interest

I hereby certify that this paper consists of original, unpublished work which is not under consideration for publication elsewhere and all the authors listed have approved the manuscript that is enclosed.

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Authors and Affiliations

  1. Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Yang Yang, Hui-Jun Wu, Ling Lin, Qing-qing Zhu & Xue-Wen Gao

  2. Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Nanjing, 210095, People’s Republic of China

    Yang Yang, Hui-Jun Wu, Ling Lin, Qing-qing Zhu & Xue-Wen Gao

  3. ABiTEP GmbH, Berlin, Germany

    Rainer Borriss

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  1. Yang Yang
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  2. Hui-Jun Wu
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Correspondence to Xue-Wen Gao.

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Yang Yang and Hui-Jun Wu contributed equally to this work and share the first authorship.

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Yang, Y., Wu, HJ., Lin, L. et al. A plasmid-born Rap-Phr system regulates surfactin production, sporulation and genetic competence in the heterologous host, Bacillus subtilis OKB105. Appl Microbiol Biotechnol 99, 7241–7252 (2015). https://doi.org/10.1007/s00253-015-6604-3

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