Recalcitrance of Cannabis sativa to de novo regeneration; a multi-genotype replication study
| Autor: | Sean T. Kyne, Andrew Maxwell Phineas Jones, Adrian S. Monthony, Christopher M. Grainger |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Leaves
Somatic embryogenesis Organogenesis Artificial Gene Amplification and Extension Plant Science Polymerase Chain Reaction Plant Roots Genotype Medicine and Health Sciences Morphogenesis Cultivar Flowering Plants Cannabis Sativa Multidisciplinary Plant Anatomy Eukaryota food and beverages Plants Behavioral Pharmacology Research Design Shoot Medicine Plant Shoots Research Article Science Replication Studies Biology Research and Analysis Methods Recreational Drug Use Genetics Regeneration Molecular Biology Techniques Molecular Biology Cannabis Pharmacology business.industry Regeneration (biology) fungi Organisms Biology and Life Sciences Reproducibility of Results Biotechnology Plant Leaves Callus business Organism Development Developmental Biology Explant culture |
| Zdroj: | PLoS ONE, Vol 16, Iss 8, p e0235525 (2021) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Cannabis sativais relatively recalcitrant tode novoregeneration, but several studies have reported shoot organogenesis or somatic embryogenesis from non-meristematic tissues. Most report infrequent regeneration rates from these tissues, but a landmark publication from 2010 achieved regeneration from leaf explants with a 96% response rate, producing an average of 12.3 shoots per explant in a single drug-type accession. Despite the importance regeneration plays in plant biotechnology and the renewed interest in this crop the aforementioned protocol has not been used in subsequent papers in the decade since it was published, raising concerns over its reproducibility. Here we attempted to replicate this importantCannabisregeneration study and expand the original scope of the study by testing it across 10 drug-typeC.sativagenotypes to assess genotypic variation. In our study, callus was induced in all 10 genotypes but callus growth and appearance substantially differed among cultivars, with the most responsive genotype producing 6-fold more callus than the least responsive. The shoot induction medium failed to induce shoot organogenesis in any of the 10 cultivars tested, instead resulting in necrosis of the calli. The findings of this replication study raise concerns about the replicability of existing methods. However, some details of the protocol could not be replicated due to missing details in the original paper and regulatory issues, which could have impacted the outcome. These results highlight the importance of using multiple genotypes in such studies and providing detailed methods to facilitate replication. |
| Databáze: | OpenAIRE |
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