Articles
Optimization of culture conditions and analysis of DNA methylation changes in tissue cultures of Cannabis sativa L.
Article number
1454_11
Pages
77 – 84
Language
English
Abstract
Micropropagation serves as a key method for the mass production of Cannabis sativa L. plants characterized by genetic uniformity and the absence of pathogens.
Nevertheless, its effectiveness is frequently affected by genotype-specific responses and low multiplication rates.
This research assessed how different basal media and plant growth regulators influence shoot multiplication and rooting in different medical cannabis genotypes during in vitro micropropagation.
The findings demonstrated that shoot multiplication was significantly influenced by the interaction between genotype and culture medium.
Shoot numbers were comparable between MS and DKW media, but DKW resulted in longer shoots, a higher node number per culture, and increased biomass.
TDZ enhanced the rate of shoot proliferation, but also caused a reduction in shoot length and induced callus formation.
Successive subcultures led to decreased shoot length and node number, particularly when using DKW medium, with MS medium showing a milder decline.
GA3 efficiently stimulated shoot elongation and aided in subsequent root formation and acclimatization.
Given the limited rooting efficiency under in vitro conditions, a new ex vitro protocol was developed that integrated rooting and acclimatization, achieving high rooting success within three weeks and thereby considerably shortening the propagation cycle.
Epigenetic evaluation was also conducted to compare plant methylation profiles over the course of micropropagation process, where it proved to be a useful tool for sequence-specific detection of differentially methylated genomic regions.
Nevertheless, its effectiveness is frequently affected by genotype-specific responses and low multiplication rates.
This research assessed how different basal media and plant growth regulators influence shoot multiplication and rooting in different medical cannabis genotypes during in vitro micropropagation.
The findings demonstrated that shoot multiplication was significantly influenced by the interaction between genotype and culture medium.
Shoot numbers were comparable between MS and DKW media, but DKW resulted in longer shoots, a higher node number per culture, and increased biomass.
TDZ enhanced the rate of shoot proliferation, but also caused a reduction in shoot length and induced callus formation.
Successive subcultures led to decreased shoot length and node number, particularly when using DKW medium, with MS medium showing a milder decline.
GA3 efficiently stimulated shoot elongation and aided in subsequent root formation and acclimatization.
Given the limited rooting efficiency under in vitro conditions, a new ex vitro protocol was developed that integrated rooting and acclimatization, achieving high rooting success within three weeks and thereby considerably shortening the propagation cycle.
Epigenetic evaluation was also conducted to compare plant methylation profiles over the course of micropropagation process, where it proved to be a useful tool for sequence-specific detection of differentially methylated genomic regions.
Authors
S. Troha, D. Kastelec, S. Svetik, T. Trafela, J. Murovec
Keywords
in vitro plant tissue culture, ex vitro rooting, epigenetics, DNA methylation, acclimatization
Groups involved
- Division Plant Genetic Resources, Breeding and Biotechnology
- Working Group Genetic Transformation and Gene Editing
- Working Group Horticultural Biotechnology and Breeding
- Division Ornamental Plants
- Division Vegetables, Roots and Tubers
- Division Temperate Tree Fruits
- Division Vine and Berry Fruits
- Division Horticulture for Development
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