Abstract
Type IV pilus (T4P) is widespread in bacteria, yet its biogenesis mechanism and functionality is only partially elucidated in a limited number of bacterial species. Here, by using strain OH11 as the model organism, we reported the identification of 26 T4P structural or functional component (SFC) proteins in the Gram-negative Lysobacter enzymogenes, which is a biocontrol agent potentially exploiting T4P-mediated twitching motility for antifungal activity. Twenty such SFC coding genes were individually knocked-out in-frame to create a T4P SFC deletion library. By using combined phenotypic and genetic approaches, we found that 14 such SFCs, which were expressed from four operons, were essential for twitching motility. These SFCs included the minor pilins (PilEi, PilXi, PilVi, and FimTi), the anti-retraction protein PilY1i, the platform protein PilC, the extension/extraction ATPases (PilB, PilT, and PilU), and the PilMNOPQ complex. Among these, mutation of pilT or pilU caused a hyper piliation, while the remaining 12 SFCs were indispensable for pilus formation. Ten (FimTi, PilY1i, PilB, PilT, PilU, and the PilMNOPQ complex) of the 14 SFC proteins, as well as PilA, were further shown to play a key role in L. enzymogenes biofilm formation. Overall, our results provide the first report to dissect the genetic basis of T4P biogenesis and its role in biofilm formation in L. enzymogenes in detail, which can serve as an alternative platform for studying T4P biogenesis and its antifungal function.
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Acknowledgments
We thank Prof. Shan-Ho Chou (National Chung Hsing University) and Dr. Benard Omondi Odhiambo in our laboratory for critical reviewing and revising this manuscript.
Funding
This study was supported by the Fundamental Research Funds for the Central Universities (Y0201600126 and KYTZ201403 to G. Qian), National Basic Research (973) program of China (2015CB150600 to G. Qian), National Key Research and Development Program (2017YFD020110), Special Fund for Agro-Scientific Research in the Public Interest (No. 201303015 to G. Qian and F. Liu), National Natural Science Foundation of China (31371981 and 31572046 to G. Qian), Innovation Team Program for Jiangsu Universities (2017), the Jiangsu Provincial Key Technology Support Program (BE2015354 and BE2015354 to F. Liu), “948” Project of the Ministry of Agriculture (2014-Z24 to F. Liu), and National pear industry technology system (CARS-29-09 to F. Liu and G. Qian).
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G. Q., and F. L. conceived and designed experiments. J. X., J. C., and Y. C. carried out experiments. J. X., J. C., Y. C., G. Q., and F. L. analyzed data and prepared figures. G. Q. wrote the manuscript. F. L reviewed and revised the manuscript. All authors reviewed the manuscript.
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Xia, J., Chen, J., Chen, Y. et al. Type IV pilus biogenesis genes and their roles in biofilm formation in the biological control agent Lysobacter enzymogenes OH11. Appl Microbiol Biotechnol 102, 833–846 (2018). https://doi.org/10.1007/s00253-017-8619-4
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DOI: https://doi.org/10.1007/s00253-017-8619-4