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Selection of available suicide vectors for gene mutagenesis using chiA (a chitinase encoding gene) as a new reporter and primary functional analysis of chiA in Lysobacter enzymogenes strain OH11

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Abstract

Here, three different suicide vectors were evaluated for the possibility of performing gene mutagenesis in strain OH11 using the chiA gene (accession number: DQ888611) as a new reporter. Suicide vector pEX18GM was selected, and it was successfully applied for disruption and in-frame deletions in the chiA gene in strain OH11, which was confirmed by PCR amplification and Southern hybridization. The chiA-deletion mutant OH11-3 did not have the ability to produce chitinase on chitine selection medium. Interestingly, the chiA-deletion mutants displayed wild-type antimicrobial activity against Saccharomyces cerevisiae, Magnaporthe grisea, Phytophthora capsici, Rhizoctonia solani, Sclerotinia sclerotiorum and Pythium ultimum. Our data suggest that chitinase might not be a unique lytic enzyme in controlling S. cerevisiae, M. grisea, P. capsici, and P. ultimum. R. solani, S. sclerotiorum. Also, suicide vector pEX18GM might be explored as a potential tool for gene deletions in L. enzymogenes, which will facilitate the molecular study of mechanisms of biological control in L. enzymogenes.

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Acknowledgments

We are very grateful to Prof. Jun Zhu (Nanjing Agricultural University) for kindly providing suicide vector pEX18GM. We also thank Prof. Gary Y Yuen (Department of Plant pathology, University of Nebrask-Lincoln) for kindly providing strains (Saccharomyces cerevisiae and Pythium ultimum) as gifts. This work was supported by Special Fund for Agro-scientific Research in the Public Interest (No. 200903052; No. 201003004), Natural Science Foundation of Jiangsu Province (No. BK2010445), Doctoral Fund of Ministry of Education of China (No. 20100097120013), the National High Technology Research and Development Program of China (No. 2006AA10A211) and A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China

    Guoliang Qian, Yansheng Wang, Dongyu Qian, Jiaqin Fan, Baishi Hu & Fengquan Liu

  2. Key Laboratory of Integrated Management of Crop Diseases and Pests, Nanjing Agricultural University, Ministry of Education, Nanjing, China

    Guoliang Qian, Yansheng Wang, Jiaqin Fan, Baishi Hu & Fengquan Liu

  3. Bureau of Agriculture and Forestry of Xishan District, Wuxi, 214100, China

    Dongyu Qian

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  1. Guoliang Qian
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  2. Yansheng Wang
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  3. Dongyu Qian
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  4. Jiaqin Fan
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  5. Baishi Hu
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Correspondence to Fengquan Liu.

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Guoliang Qian and Yansheng Wang contributed equally to this work.

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Qian, G., Wang, Y., Qian, D. et al. Selection of available suicide vectors for gene mutagenesis using chiA (a chitinase encoding gene) as a new reporter and primary functional analysis of chiA in Lysobacter enzymogenes strain OH11. World J Microbiol Biotechnol 28, 549–557 (2012). https://doi.org/10.1007/s11274-011-0846-8

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  • DOI: https://doi.org/10.1007/s11274-011-0846-8

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