Development of a Direct in vitro Plant Regeneration Protocol From Cannabis sativa L. Seedling Explants: Developmental Morphology of Shoot Regeneration and Ploidy Level of Regenerated Plants

Frontiers in Plant Science
Alberto Galán-ÁvilaFrancisco Javier Herraiz

Abstract

In vitro shoot regeneration can efficiently contribute to the improvement of recalcitrant Cannabis sativa L. We aimed at developing a highly efficient protocol for in vitro direct regeneration of C. sativa plants from different explants (cotyledon, hypocotyl, and true leaf) from seedlings of monoecious C. sativa short-day varieties Ferimon, Felina32, Fedora17, and USO31, together with dioecious neutral-day variety Finola. Ten regeneration media, including already published protocols, and self-designed combinations of plant growth regulators were tested. The developmental morphology since germination of seeds to the development of rooted plantlets was followed. Additionally, the ploidy level of explants and in vitro regenerants was analyzed. We concluded that hypocotyl is the best explant for in vitro direct regeneration of C. sativa plants with 49.45% of responding explants, while cotyledon and true leaf had a poor response with, respectively, 4.70 and 0.42% of explants developing plantlets. In terms of shoot regeneration, we found significant differences among the culture media evaluated and the varieties studied. Overall, the best regeneration media were ZEARIB 2.0 (mg/L) and ZEARIB 1.0 (mg/L) + NAA 0.02 (mg/L) with 66.67% of...Continue Reading

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Citations

Jan 23, 2021·Plants·Adrian S MonthonyAndrew Maxwell P Jones
Mar 23, 2021·Frontiers in Plant Science·Dinesh AdhikaryPankaj Bhowmik
May 1, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Mohsen HesamiAndrew Maxwell Phineas Jones
Jun 3, 2021·International Journal of Molecular Sciences·Mohsen HesamiAndrew Maxwell Phineas Jones
Aug 14, 2021·PloS One·Adrian S MonthonyAndrew Maxwell P Jones

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Methods Mentioned

BETA
flow cytometry
dissection

Software Mentioned

R

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