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Selection and characterization of in vitro–induced mutants of Dendrobium ‘Earsakul’ resistant to black rot

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Abstract

The development of Dendrobium ‘Earsakul’ with improved resistance to black rot is essential for sustainable orchid production. In this study, in vitro mutagenesis and selection was used to breed D. ‘Earsakul’ for black rot resistance, evaluated black rot resistance levels in mutants and controls using detached leaf assay, and characterized putative resistant mutants using molecular markers, cytology, and morphological traits. Mutagenized protocorm-like bodies (PLBs) obtained by 1.4% (LD30) and 1.8% (LD50) ethyl methanesulfonate (EMS) concentrations and non-mutagenized PLBs were selected on a pea sucrose broth (PSB) medium supplemented with 0, 30, and 50% (first 2 cycles) and 0, 40, and 60% (third cycle) of Phytophthora palmivora culture filtrates (CFs). Fifty putative resistant mutants were obtained, and 42 of these were used for resistance level evaluation by detached leaf assay using P. palmivora isolate NK-53–9. The results revealed 13 resistant putative mutants, including 4 highly resistant putative mutants and 9 resistant putative mutants. Ten surviving resistant putative mutants were genetically different from the non-mutagenized controls and were confirmed as mutants after inter-simple sequence repeat (ISSR) analysis. Based on flow cytometry, the resistant mutants and non-mutagenized controls possessed the same chromosome number of 2n + 4n + 8n. Moreover, a mutant, SUT17E18316, exhibiting maximum DNA content and genome size was identified. Morphological characterization revealed that most of the black rot–resistant mutants were morphologically different on some characters from non-mutagenized lines, such as plant height and number of roots. Particularly, the highly resistant mutant SUT13E18305 which possessed outstanding characters may be useful for future commercialization.

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Acknowledgements

This work was supported by Suranaree University of Technology (SUT) and by Thailand Science Research and Innovation (TSRI). The authors would also like to acknowledge Ministry of Education and grants from National Research Council of Thailand (NRCT). We are very grateful to Peter C. Bint for editing the manuscript.

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Authors and Affiliations

  1. Institute of Agricultural Technology, School of Crop Production Technology, Suranaree University of Technology, 111 University Avenue, Muang District, Nakhon Ratchasima, Thailand

    Apinya Khairum, Sasitorn Hualsawat, Witsarut Chueakhunthod, Akkawat Tharapreuksapong, Benjawan Tomsantia, Pitchakon Papan & Piyada A. Tantasawat

  2. Faculty of Natural Resources, Agricultural Innovation and Management Division, Prince of Songkla University, Hat Yai, Songkhla, Thailand

    Sureerat Yenchon

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  1. Apinya Khairum
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  8. Piyada A. Tantasawat
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Contributions

PAT designed and guided the experiments. AK, PAT, WC, SH, AT, BT, SY, and PP performed the experiments, analyzed the data, wrote the article, and revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Piyada A. Tantasawat.

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Khairum, A., Hualsawat, S., Chueakhunthod, W. et al. Selection and characterization of in vitro–induced mutants of Dendrobium ‘Earsakul’ resistant to black rot. In Vitro Cell.Dev.Biol.-Plant 58, 577–592 (2022). https://doi.org/10.1007/s11627-022-10261-0

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