Genotype-independent de novo regeneration protocol in Cannabis sativa L. through direct organogenesis from cotyledonary nodes.

ABSTRACT: Efficient regeneration protocols are essential for large-scale propagation and genetic manipulation of recalcitrant medicinal species such as Cannabis sativa. Existing direct and indirect regeneration methods are highly genotype and explant-dependent, limiting broader applicability. Here, we report a five-stage (S₀-S₄) optimised protocol that is reproducible and achieves high-efficiency direct de novo regeneration using cotyledonary node explants from both hemp and medicinal cannabis genotypes. A 1% (v/v) H₂O₂-based sterilisation method significantly improved seed germination and reduced endophyte contamination. Among embryo-derived explants, the cotyledonary node attached to the cotyledon showed superior regeneration efficiency through two distinct pathways: axillary

SUBMITTER: Bennur PL 

PROVIDER: S-EPMC12595704 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature