Abstract
Ralstonia solanacearum causes a bacterial wilt of ginger (Zingiber officinale) that hampers ginger production. Because pathogenicity tests using rhizomes are laborious and time-consuming, here we developed a new test using ginger plants that were aseptically regenerated in vitro from shoot tips using 6-benzyl adenine and 1-naphthalene acid (NAA) in Murashige–Skoog (MS) agar. The regenerated plants were then cultured in MS broth with NAA to evaluate pathogenicity after the roots were dipped into a bacterial solution. The wild-type strain (MAFF 211479) caused wilt symptoms such as leaf yellowing within 15 days after inoculation (dpi), and plants had died by 28 dpi. MAFF 301069, an avirulent strain of ginger, did not cause any symptoms. Because method was suitable for assessing virulence of R. solanacearum, we then used to test the pathogenicity of MAFF 211479 mutants that we constructed and were defective in the type III secretion system. None of the mutants were pathogenic in the regenerated ginger plants or in eggplant. The mutant cells proliferated less efficiently than the wild type in the inoculated ginger plants. We thus conclude that aseptically regenerated ginger plants can be used to elucidate the infection mechanism of R. solanacearum in ginger.
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Acknowledgements
We thank Yuzhu Cao and Rana Lokendra for caring for plants.
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This work was supported by JSPS KAKENHI Grant-in-Aid for Scientific Research (C) Grant Number 20K06054 (to KO) and by a Cabinet Office grant in aid, the Advanced Next-Generation Greenhouse Horticulture by IoP (Internet of Plants), Japan.
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All authors contributed to study conception and design. Material preparation, data collection and analysis were performed by MH, AK, MT, and KO. The first draft of the manuscript was written by MH, YH, and KO, and all authors commented on previous and read and approved the final manuscript.
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Hasan, M., Kiba, A., Hikichi, Y. et al. Method for aseptically regenerating ginger plants and studying infection by Ralstonia solanacearum using hrp mutants. J Gen Plant Pathol 89, 185–191 (2023). https://doi.org/10.1007/s10327-023-01121-9
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DOI: https://doi.org/10.1007/s10327-023-01121-9