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Development of T3SS Mutants (hrpB− and hrcV−) of Ralstonia solanacearum, Evaluation of Virulence Attenuation in Brinjal and Tomato—A Pre-requisite to Validate T3Es of R. solanacearum

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Abstract

Ralstonia solanacearum is an important plant pathogen which infects a large number of agriculturally important crops. The Type Three Secretion System (T3SS) plays a major role in its pathogenicity by secreting type III effectors (T3Es) which overthrow the host defence mechanism. The secretion of T3Es is transcriptionally regulated by hrpB and its secretion is dependent on the pili formed by hrcV gene. In this study, two T3SS mutants of R. solanacearum strain Rs-09-161 viz. Rs-HrpB− and Rs-HrcV− were developed through insertional mutagenesis. The method of development of insertional mutant is quite simple and reliable. The plasmid integrates through homologous recombination and in vitro studies have proved that the integration was stable for several generations. The mutants are non-pathogenic on its highly susceptible hosts, brinjal and tomato inoculated by soil drench method and by petiole inoculation directly into the vascular system. Further it was observed that the colonisation ability of the mutants was also highly reduced in the susceptible host. These mutants will be useful in validating putative T3E through translocation studies.

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Acknowledgements

We gratefully acknowledge the financial support by Indian Council of Agricultural Research, New Delhi, India through “Outreach project on Phytophthora, Fusarium and Ralstonia diseases of horticultural and field crops”—(PhytoFuRa). We are grateful to Director, ICAR- Central Coastal Agricultural Research Institute, Old Goa for providing other necessary facilities.

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  1. ICAR-Central Coastal Agricultural Research Institute, Old Goa, Goa, 403402, India

    Trupti Asolkar & Raman Ramesh

  2. Department of Microbiology, Goa University, Taleigao Plateau, Goa, 403206, India

    Trupti Asolkar

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  1. Trupti Asolkar
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Correspondence to Raman Ramesh.

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Asolkar, T., Ramesh, R. Development of T3SS Mutants (hrpB− and hrcV−) of Ralstonia solanacearum, Evaluation of Virulence Attenuation in Brinjal and Tomato—A Pre-requisite to Validate T3Es of R. solanacearum. Indian J Microbiol 58, 372–380 (2018). https://doi.org/10.1007/s12088-018-0736-y

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