Evidence map›Paper›PMID 41286625›Full record

ArticleBMC plant biology2025

A novel protocol for protoplast isolation, transfection, and culture in Cannabis sativa L.

Katarzyna Stelmach-Wityk, Kamil Szymonik, Andrew Maxwell Phineas Jones, Ewa Grzebelus

Abstract read
In one paragraph

Article in BMC plant biology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

4 citing papers in PubMed.

  1. From Model Plants toPlants (Basel, Switzerland) · 2026
    Review
  2. Review
  3. Review
  4. Genetic Advances inPlants (Basel, Switzerland) · 2026
    Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

4 authors.

Katarzyna Stelmach-WitykDepartment of Plant Biology and Biotechnology, Faculty of Biotechnology and Horticulture, University of Agriculture in Krakow, Mickiewicza 21, Krakow, 31-120, Poland. k.stelmach@urk.edu.pl.ORCID http://orcid.org/0000-0002-9435-8753
Kamil SzymonikDepartment of Plant Biology and Biotechnology, Faculty of Biotechnology and Horticulture, University of Agriculture in Krakow, Mickiewicza 21, Krakow, 31-120, Poland.
Andrew Maxwell Phineas JonesDepartment of Plant Agriculture, Ontario Agricultural College, University of Guelph, Guelph, ON, N1G 2W1, Canada.
Ewa GrzebelusDepartment of Plant Biology and Biotechnology, Faculty of Biotechnology and Horticulture, University of Agriculture in Krakow, Mickiewicza 21, Krakow, 31-120, Poland.ORCID http://orcid.org/0000-0003-2067-585X

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

backgroundProtoplasts are a valuable tool for studying gene expression and applying genome editing techniques. Given the high medicinal and industrial potential of Cannabis sativa L., developing an efficient protoplast-to-plant regeneration protocol is highly desirable. Due to its recalcitrant nature, a complete plant regeneration from cannabis protoplasts has not yet been achieved.

resultsThis study details a robust protocol for cannabis protoplast isolation, purification, transient transfection, and culture, additionally reporting somatic embryo-like structures derived from protoplast-derived callus. We demonstrated that the age of donor material, the composition of the enzyme solution, and the duration of enzymolysis are crucial for efficient protoplast isolation. Protoplast embedding, coupled with a rich culture medium and plant growth regulators, proved critical for initiating cell wall re-synthesis, cell division, and microcallus formation. Protoplasts isolated using the reported protocol were abundant (2.2 × 10

conclusionsThis system provides a reliable protocol for isolation, transfection and culture of cannabis protoplasts. It also offers a framework for investigating gene function, as well as advancing protoplast fusion and genome editing technologies for this species.

Indexed as

CannabisCell Culture TechniquesProtoplastsTransfection2-aminoindane-2-phosphonic acidHempPhytosulfokineProtoplast embeddingTissue cultures

Identifiers

PMID41286625
PMCPMC12642208

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.