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Cannabis sativa L. photoautotrophic micropropagation: a powerful tool for industrial scale in vitro propagation

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Abstract

Global demands for an in vitro culture of cannabis have never been more sought after as countries shift their paradigm towards legalization. Cannabis conventional (photomixotrophic) micropropagation has not been suitable enough for large-scale propagation due to a high degree of plant hyperhydricity, low growth rate, poor rooting, and acclimation efficiency. In the present study, cannabis photoautotrophic micropropagation method is introduced with the purpose of overcoming the difficulties that conventional micropropagation entails when conducted at a large scale. The roles of rockwool medium pH and moisture content, cutting length, basal wounding methods, light intensity, and culture vessel gas exchange capacity were assessed with the intention of increasing productivity of micropropagation method. The results showed 300 mL per vessel of fertilizer solution containing 5-mM MES buffer stabilized medium pH and increased rooting success. Both 5- and 7-cm cutting lengths significantly increased the percent of rooted plants compared to 3-cm cutting length. However, the basal wounding methods did not significantly improve or impede the rooting success. The highest rooting success was also obtained with 150 μmol m−2 s−1 of photosynthetic photon flux density compared to 50 and 100 μmol m−2 s−1. Increasing gas exchange rates either using more permeable vessels or aeration practices significantly improved the rooting success. Overall, more than 90% of cannabis plantlets grown in photoautotrophic micropropagation are rooted in 2 wk of culture followed by 4 d ex vitro acclimation period, which was remarkably shorter than any other available method in cannabis micropropagation. This study not only optimizes a method for cannabis photoautotrophic micropropagation using passive ventilation for the first time but also scales up in vitro clonal propagation for in vitro commercial production.

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Acknowledgements

The authors gratefully thank two NRC Industrial Technology Advisors Mr. Hugh Scholaert and Mr. Ron Evans for their constructive advices, connections, and supports.

Funding

Financial support for this research was provided in part by National Research Council of Canada (NRC), Industrial Research Assistance Program (NRC_IRAP, Project No. 944131).

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Authors and Affiliations

  1. Safari Flower Co., 2818 House Road, Stevensville, Ontario, L0S 1S0, Canada

    Adel Zarei, Behrang Behdarvandi, Elham Tavakouli Dinani & Jennifer Maccarone

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  1. Adel Zarei
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  2. Behrang Behdarvandi
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  3. Elham Tavakouli Dinani
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Correspondence to Adel Zarei.

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Zarei, A., Behdarvandi, B., Tavakouli Dinani, E. et al. Cannabis sativa L. photoautotrophic micropropagation: a powerful tool for industrial scale in vitro propagation. In Vitro Cell.Dev.Biol.-Plant 57, 932–941 (2021). https://doi.org/10.1007/s11627-021-10167-3

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