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Cannabis sativa L. (cannabis) is one of the earliest domesticated crops cultivated for medicinal, food, industrial, and recreational use throughout the world. A growing number of jurisdictions have lifted the prohibition on medical and recreational cannabis with legislation that allows researchers to conduct important and urgently needed research, such as understanding the molecular basis of agronomically important traits. However, a dearth of suitable molecular genetic techniques for the analysis of cannabis species has hampered the elucidation of gene function. Here, we developed a robust protoplast transient transformation system by optimizing conditions of protoplast isolation and polyethylene glycol-mediated transformation using cannabis cotyledons. A protoplast isolation yield of up to 1.15 x [10.sup.7] cells per gram of fresh material and 98.5% viability and a transformation efficiency of 75.4% were achieved. This optimized protocol for protoplast isolation and transformation was successfully applied to eight representative cannabis varieties, which highlight the versatility and robustness of this technique. Using this protocol, the nuclear localization of a basic helix-loop-helix transcription factor (CsMYC2) was visualized, and the transcriptional activity of CsMYC2 was assessed. These results demonstrate the effectiveness of our cannabis protoplast isolation and transient transformation method that should serve as a powerful molecular research tool in a wide range of molecular studies. Key words: cannabis, protoplast, isolation, transformation, transient expression Cannabis sativa L. (cannabis) est l'une des premières plantes domestiquées cultivées à des fins médicinales, alimentaires, industrielles et récréatives dans le monde entier. Un nombre croissant de juridictions ont levé l'interdiction du cannabis à des fins médicales et récréatives au moyen d'une législation qui permet aux chercheurs de mener des recherches importantes et urgentes, telles que la compréhension de la base moléculaire de caractéristiques agronomiques importantes. Cependant, le manque de techniques de génétique moléculaire adaptées à l'analyse des espèces de cannabis a entravé l'élucidation de la fonction des gènes. Les auteurs ont développé ici un système robuste de transformation transitoire de protoplastes en optimisant les conditions d'isolement des protoplastes et de transformation médiée par le PEG en utilisant des cotylédons de cannabis. Ils ont obtenu un rendement d'isolement de protoplastes allant jusqu'à 1,15 x 107 cellules par gramme de matériel frais, une viabilité de 98,5% et une efficacité de transformation de 75,4%. Ce protocole optimisé d'isolement et de transformation des protoplastes a été appliqué avec succès à huit variétés représentatives de cannabis, ce qui souligne la polyvalence et la robustesse de cette technique. Ce protocole a permis de visualiser la localisation nucléaire d'un facteur de transcription héliceboucle-hélice basique (CsMYC2) et d'évaluer l'activité transcriptionnelle de CsMYC2. Ces résultats démontrent l'efficacité de cette méthode d'isolement et de transformation transitoire de protoplastes de cannabis qui devrait servir d'outil de recherche moléculaire puissant dans un large éventail d'études moléculaires. [Traduit par la Rédaction] Mots-clés : cannabis, protoplaste, isolement, transformation, expression transitoire, Introduction Cannabis sativa L. (cannabis) is an annual, predominantly dioecious, multi-purpose plant that has been cultivated for millennia as a source of fiber, food, and medicine (Bonini et al. 2018). [...]

SUMMARY: Cannabis glandular trichomes (GTs) are economically and biotechnologically important structures that have a remarkable morphology and capacity to produce, store, and secrete diverse classes of secondary metabolites. However, our understanding of the developmental changes and the underlying molecular processes involved in cannabis GT development is limited. In this study, we developed Cannabis Glandular Trichome Detection Model (CGTDM), a deep learning‐based model capable of differentiating and quantifying three types of cannabis GTs with a high degree of efficiency and accuracy. By profiling at eight different time points, we captured dynamic changes in gene expression, phenotypes, and metabolic processes associated with GT development. By integrating weighted gene co‐expression network analysis with CGTDM measurements, we established correlations between phenotypic variations in GT traits and the global transcriptome profiles across the developmental gradient. Notably, we identified a module containing methyl jasmonate (MeJA)‐responsive genes that significantly correlated with stalked GT density and cannabinoid content during development, suggesting the existence of a MeJA‐mediated GT formation pathway. Our findings were further supported by the successful promotion of GT development in cannabis through exogenous MeJA treatment. Importantly, we have identified CsMYC4 as a key transcription factor that positively regulates GT formation via MeJA signaling in cannabis. These findings provide novel tools for GT detection and counting, as well as valuable information for understanding the molecular regulatory mechanism of GT formation, which has the potential to facilitate the molecular breeding, targeted engineering, informed harvest timing, and manipulation of cannabinoid production. [ABSTRACT FROM AUTHOR]

This article discusses the concept of knowledge co-creation for community resilience to sustainability challenges. It emphasizes the importance of engaging diverse actors, including scientists, practitioners, and community members, in the research process to collectively generate actionable insights and solutions. The article highlights various case studies that demonstrate the value of knowledge co-production in addressing sustainability issues, such as youth engagement in territorial governance, advancing agroecological transitions, and citizen contributions to environmental science. The authors emphasize the need for building trust, sharing power, and facilitating learning among all research participants to achieve meaningful and impactful results. [Extracted from the article]

Gene expression analysis provides important clues regarding gene functions, and quantitative real-time PCR (qRT-PCR) is a widely used method in gene expression studies. Reference genes are essential for normalizing and accurately assessing gene expression. In the present study, 16 candidate reference genes (ACTB, CyPA, EF1-[alpha], GAPDH, HSP90, NDPk, RPL13a, RPL18, RPL19, RPL32, RPL4, RPL8, RPS13, RPS4, [alpha]-TUB, and [beta]-TUB) from Plutella xylostella were selected to evaluate gene expression stability across different experimental conditions using five statistical algorithms (geNorm, NormFinder, Delta Ct, BestKeeper, and RefFinder). The results suggest that different reference genes or combinations of reference genes are suitable for normalization in gene expression studies of P. xylostella according to the different developmental stages, strains, tissues, and insecticide trea[TM]ents. Based on the given experimental sets, the most stable reference genes were RPS4 across different developmental stages, RPL8 across different strains and tissues, and EF1-[alpha] across different insecticide trea[TM]ents. A comprehensive and systematic assessment of potential reference genes for gene expression normalization is essential for post-genomic functional research in P. xylostella, a notorious pest with worldwide distribution and a high capacity to adapt and develop resistance to insecticides. Key words: reference genes, diamondback moth, expression pattern, normalization, qRT-PCR. L'analyse de l'expression génique fournit des informations importantes sur la fonction des gènes et la PCR quantitative en temps réel (qRT-PCR) est une méthode largement employée pour l'étude de l'expression des gènes. Des gènes de référence sont essentiels pour normaliser et mesurer précisément l'expression génique. Dans le présent travail, les auteurs ont évalué 16 gènes du Plutella xylostella (ACTB, CyPA, EF1-[alpha], GAPDH, HSP90, NDPk, RPL13a, RPL18, RPL19, RPL32, RPL4, RPL8, RPS13, RPS4, [alpha]-TUB, and [beta]-TUB) pour mesurer la stabilité de leur expression dans différentes conditions expérimentales au moyen de cinq algorithmes statistiques (geNorm, NormFinder, Delta Ct, BestKeeper et RefFinder). Les résultats suggèrent que différents gènes ou combinaisons de gènes de référence sont adéquats pour normaliser l'expression génique chez le P. xylostella selon différents stades de développement, souches, tissus et traitements insecticides. Basé sur les ensembles expérimentaux donnés, les gènes de référence les plus stables étaient RPS4 à différents stades de développement, RPL8 à travers différentes souches et tissus, et EF1-[alpha] à travers différents traitements insecticides. Une évaluation exhaustive et systématique des gènes de référence potentiels en vue de la normalisation de l'expression est essentielle dans le cadre des recherches fonctionnelles dans l'ère post-génomique chez le P. xylostella, un ravageur important présent partout dans le monde et qui est doté d'une grande capacité d'adaptation et de développement de résistance aux insecticides. [Traduit par la Rédaction] Mots-clés : gènes de référence, teigne des crucifères, profil d'expression, normalisation, qRT-PCR., Introduction An increasing number of large genomes have been published in the past few years and studies are now focusing on post-genomic functional research. Gene expression analysis is increasingly important [...]