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Half-high blueberry plants from bioreactor culture display elevated levels of DNA methylation polymorphism

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Abstract

Blueberry (Vaccinium spp. L.) plants exhibit high potential of regeneration via adventitious shoot formation on a semi-solid medium followed by shoot elongation in a liquid medium under bioreactor systems. To find out whether DNA methylation plays a role during shoot elongation, we compared DNA methylation level in the regenerants of two in vitro-grown half-high blueberry (V. corymbosum L. × V. angustifolium Ait.) cultivars Patriot and Chippewa on a semi-solid medium (SSM) in glass bottles and in a liquid medium in temporary immersion bioreactors (TIB), via methylation-sensitive amplification polymorphism (MSAP) technique. The SSM was separately fortified with various combinations of two plant growth regulators, zeatin and thidiazuron for shoot regeneration but elongation was carried out using the same medium under both SSM and TIB systems with only zeatin. Zeatin at 9.2 µM produced the maximum shoots in SSM and TIB for both cultivars, which varied from 30 to 33 in SSM and from 28 to 29 in TIB for Patriot and Chippewa, respectively. However, shoots proliferated in TIB were more vigorous. Noticeable changes in the methylation profiles were detected using MSAP in the regenerants grown in each type of culture system. The TIB system exhibited a significant increase in total methylation percentage (47–50%) in comparison to SSM (25–32%) in both cultivars. Similarly, the plants regenerated from TIB system were more polymorphic than those from SSM system. We describe here the effects of culture in vitro on DNA methylation that induced during the process of shoot elongation in two half-high blueberry cultivars.

Key message

In vitro shoot culture was achieved in half-high blueberry cultivars using a semi-solid and a liquid medium; plantlets developed from liquid medium showed higher level of DNA methylation alterations.

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Acknowledgments

This is an Agriculture and Agri-Food Canada (AAFC) St. John’s Research and Development Centre Contribution No. 244. The authors are thankful to Darryl Martin and Glenn Chubs for their excellent technical help. All the experiments have been done in St. John’s Research and Development Centre, Agriculture and Agri-Food Canada.

Funding

Agriculture and Agri-Food Canada St. John’s Research and Development Centre Contribution Number 244.

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Authors and Affiliations

  1. Department of Biology, Memorial University of Newfoundland, 232 Elizabeth Avenue, St. John’s, Newfoundland and Labrador, A1B 3X9, Canada

    Amrita Ghosh & Abir U. Igamberdiev

  2. St. John’s Research and Development Centre, Agriculture and Agri-Food Canada, Bldg. 25, 204 Brookfield Road, St. John’s, Newfoundland and Labrador, A1E 6J5, Canada

    Amrita Ghosh & Samir C. Debnath

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  1. Amrita Ghosh
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  2. Abir U. Igamberdiev
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Contributions

AG conceived the idea, designed the study, performed the experiments, analyzed the data, prepared the tables and figures, and wrote the original manuscript. AUI participated in study design and revised the manuscript. SCD conceptualized, designed, managed and supervised the study, and reviewed and edited the manuscript. All authors read and approved the final manuscript.

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Correspondence to Samir C. Debnath.

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Communicated by Pamela J. Weathers.

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Ghosh, A., Igamberdiev, A.U. & Debnath, S.C. Half-high blueberry plants from bioreactor culture display elevated levels of DNA methylation polymorphism. Plant Cell Tiss Organ Cult 146, 269–284 (2021). https://doi.org/10.1007/s11240-021-02067-6

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  • DOI: https://doi.org/10.1007/s11240-021-02067-6

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