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Improving callus regeneration of Miscanthus × giganteus J.M.Greef, Deuter ex Hodk., Renvoize ‘M161’ callus by inhibition of the phenylpropanoid biosynthetic pathway

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Abstract

In previous studies, the regeneration rates of Miscanthus × giganteus J.M.Greef, Deuter ex Hodk., Renvoize from callus tissue cultured on semi-solid media significantly declined after 4 mo of culture, which presents problems with germplasm conservation and use as an alternative propagation system. Due to the species’ lignocellulosic nature, it was hypothesized that the accumulation of phenolic compounds in the callus may be responsible for inhibiting regeneration. The current study aimed to optimize regeneration of M. × giganteus callus by culturing the callus tissue in the presence of 2-aminoindan-2-phosphonic acid (AIP), a competitive inhibitor of phenylalanine ammonia lyase (PAL), to reduce the biosynthesis of phenolics. Embryogenic callus was cultured on media supplemented with 9.0- or 11.3-μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0-, 1-, 10-, 100-, or 1000-μM AIP. Every 28 d for 7 mo, the callus tissue was visually classified based on morphology and regeneration rate. Over the duration of the study, regeneration of shoots was consistently highest in callus cultured on 11.3-μM 2,4-D supplemented with 10- and 100-μM AIP (13–58.3%), and in vitro plantlet development from callus cultured on all concentrations of AIP demonstrated tillering and rooting. Total soluble phenolic content of the callus decreased in a dose-dependent manner from 2242.34-μg g−1 dry weight in the control to 1569.71-μg g−1 dry weight in AIP-treated callus. These data indicate that inhibiting PAL in M. × giganteus cultures increased the percentage of calluses exhibiting regeneration over time.

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Acknowledgements

The authors wish to thank Abhishek Chattopadhyay for training and assistance with determining the soluble phenolic content of the samples, All Weather Farming Inc. for supplying explant material, and Dr. Michelle Edwards for aiding with statistical analyses.

Funding

The authors are also grateful for the funding partner BioFuelNet for financially supporting this research. The funders had no role in the design of the study, data collection and analysis, decision to publish, or preparation of the manuscript.

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  1. Gosling Research Institute for Plant Preservation, Department of Plant Agriculture, 50 Stone Road East, University of Guelph, Guelph, ON, N1G 2W1, Canada

    Cassandra Doll Downey & Andrew Maxwell Phineas Jones

  2. Department of Thermodynamics, Theory of Machine and Thermal Systems, Faculty of Mechanical and Power Engineering, Wrocław University of Science and Technology, 27 Wybrzeże Wyspiańskiego St., 50-370, Wrocław, Poland

    Jerzy Zoń

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  1. Cassandra Doll Downey
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Correspondence to Andrew Maxwell Phineas Jones.

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Downey, C.D., Zoń, J. & Jones, A.M.P. Improving callus regeneration of Miscanthus × giganteus J.M.Greef, Deuter ex Hodk., Renvoize ‘M161’ callus by inhibition of the phenylpropanoid biosynthetic pathway. In Vitro Cell.Dev.Biol.-Plant 55, 109–120 (2019). https://doi.org/10.1007/s11627-018-09957-z

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