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Cited by 38Plant tissue culture and regenerationFlowering Plant Growth and CultivationGrowth and nutrition in plants

Effect of Explant Source on Phenotypic Changes of In Vitro Grown Cannabis Plantlets over Multiple Subcultures

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Hesami, Mohsen, Adamek, Kristian, Pepe, Marco, Jones, Andrew Maxwell Phineas·2023·Biology
DOI 10.3390/biology12030443PMC PMC10044989
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Abstract

Drug-type cannabis is often multiplied using micropropagation methods to produce genetically uniform and disease/insect-free crops. However, micropropagated plantlets often exhibit phenotypic variation, leading to culture decline over time. In cannabis, the source of these changes remains unknown, though several factors (e.g., explant’s sources and prolonged in vitro culture) can result in such phenotypical variations. The study presented herein evaluates the effects of explant sources (i.e., nodal segments derived from the basal, near-basal, middle, and apical parts of the greenhouse-grown mother plant) over multiple subcultures (4 subcultures during 235 days) on multiplication parameters and leaf morphological traits of in vitro cannabis plantlets. While initial in vitro responses were similar among explants sourced from different regions of the plant, there were significant differences in performance over the course of multiple subcultures. Specifically, explant source and/or the number of subcultures significantly impacted plantlet height, number of nodes, and canopy surface area. The explants derived from the basal and near-basal parts of the plant resulted in the tallest shoots with the greatest number of nodes, while the explants derived from the middle and apical regions led to shorter shoots with fewer nodes. Moreover, the basal-derived explants produced cannabis plantlets with shorter but wider leaves which demonstrated the potential of such explants for in vitro rejuvenation practices with minimal culture decline. This study provides new evidence into the long-term impacts of explant source in cannabis micropropagation.

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  • In Cannabis sativa, somaclonal variation was more pronounced in meristematic regions compared to differentiated tissues, indicating higher susceptibility of actively dividing cells to in vitro-induced genetic and epigenetic changes.

    “However, Hesami et al. reported that in Cannabis sativa, somaclonal variation was more pronounced in meristematic regions compared to differentiated tissues, indicating a higher susceptibility of actively dividing cells to in vitro-induced genetic and epigenetic changes [72].”

    From Somaclonal Variation and Clonal Fidelity in Commercial Micropropagation: Challenges and Perspectives (2025) · cites this paper as [72]

    The Discussion reports that reduced node numbers after repeated subcultures—a possible morphological sign of somaclonal variation—were especially evident in cannabis explants from the middle and apical parts of the mother plant. It does not compare meristematic regions with differentiated tissues or establish that actively dividing cells are more susceptible to genetic and epigenetic changes. The Conclusions say molecular aspects of the variation still need investigation. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Culture conditions profoundly impact somaclonal variation, as in vitro environments differ from natural conditions, subjecting plant cells to stresses that alter metabolism and trigger somaclonal variation.

    “Culture conditions also profoundly impact, as in vitro environments differ significantly from natural conditions, subjecting plant cells to stresses that can alter their metabolism and trigger somaclonal variation [72,73].”

    From Somaclonal Variation and Clonal Fidelity in Commercial Micropropagation: Challenges and Perspectives (2025) · cites this paper as [72]

    The Discussion (page unavailable) links prolonged in vitro culture to DNA hypermethylation and describes a change in cannabis plantlets that may be a sign of somaclonal variation. It does not say that differences from natural conditions stress plant cells, alter their metabolism, and thereby trigger that variation. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

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References 71 · Cited by 38

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Authors
Hesami, M., Adamek, K., Pepe, M., & Jones, A. M. P.
Affiliations
University of Guelph (CA)
Corresponding author
Jones, Andrew Maxwell Phineas
ORCID
Venue
Biology
Publisher
MDPI AG
Published
2023-03-13
Volume
12
Issue
3
Pages
443
ISSN
2079-7737 (ISSN-L)
DOI
10.3390/biology12030443
PMC
PMC10044989
OpenAlex
W4324157363
Cited by
38
References
71
Citations by year
2026: 9 · 2025: 10 · 2024: 13 · 2023: 6
FWCI
3.46
Type
journal article
Language
en
Open access
Licence
Locations
  • Biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Brantmed Inc. (ALLRP555969−20)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Flowering Plant Growth and Cultivation
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Growth and nutrition in plants
Keywords
cannabis, micropropagation, explant source, leaf morphology, shoot length, nodal explants, mother plants, in vitro plantlets

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