Boissinot Justin, Adamek Kristian, Jones Andrew Maxwell Phineas, Normandeau Eric, Boyle Brian, Torkamaneh Davoud
Département de phytologie, Université Laval, Québec, QC, Canada.
Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, Québec, QC, Canada.
Front Plant Sci. 2024 May 30;15:1381154. doi: 10.3389/fpls.2024.1381154. eCollection 2024.
The primary focus of medicinal cannabis research is to ensure the stability of cannabis lines for consistent administration of chemically uniform products to patients. In recent years, tissue culture has emerged as a valuable technique for genetic preservation and rapid multiplication of cannabis clones. However, there is concern that the physical and chemical conditions of the growing media can induce somaclonal variation, potentially impacting the viability and uniformity of clones. To address this concern, we developed Comparative Restriction Enzyme Analysis of Methylation (CREAM), a novel method to assess DNA methylation patterns and used it to study a population of 78 cannabis clones maintained in tissue culture. Through bioinformatics analysis of the methylome, we successfully detected 2,272 polymorphic methylated regions among the clones. Remarkably, our results demonstrated that DNA methylation patterns were preserved across subcultures within the clonal population, allowing us to distinguish between two subsets of clonal lines used in this study. These findings significantly contribute to our understanding of the epigenetic variability within clonal lines in medicinal cannabis produced through tissue culture techniques. This knowledge is crucial for understanding the effects of tissue culture on DNA methylation and ensuring the consistency and reliability of medicinal cannabis products with therapeutic properties. Additionally, the CREAM method is a fast and affordable technology to get a first glimpse at methylation in a biological system. It offers a valuable tool for studying epigenetic variation in other plant species, thereby facilitating broader applications in plant biotechnology and crop improvement.
药用大麻研究的主要重点是确保大麻品系的稳定性，以便向患者持续提供化学性质均一的产品。近年来，组织培养已成为一种用于大麻克隆体遗传保存和快速繁殖的重要技术。然而，人们担心生长培养基的物理和化学条件会诱发体细胞克隆变异，这可能会影响克隆体的活力和一致性。为了解决这一问题，我们开发了甲基化比较限制性酶切分析（CREAM），这是一种评估DNA甲基化模式的新方法，并将其用于研究在组织培养中保存的78个大麻克隆体群体。通过对甲基化组的生物信息学分析，我们在这些克隆体中成功检测到2272个多态性甲基化区域。值得注意的是，我们的结果表明，克隆群体中传代培养后的DNA甲基化模式得以保留，这使我们能够区分本研究中使用的两个克隆系子集。这些发现极大地增进了我们对通过组织培养技术生产的药用大麻克隆系内表观遗传变异性的理解。这些知识对于理解组织培养对DNA甲基化的影响以及确保具有治疗特性的药用大麻产品的一致性和可靠性至关重要。此外，CREAM方法是一种快速且经济实惠的技术，可让我们初步了解生物系统中的甲基化情况。它为研究其他植物物种的表观遗传变异提供了一个有价值的工具，从而促进其在植物生物技术和作物改良中的更广泛应用。
