Comparative restriction enzyme analysis of methylation (CREAM) reveals methylome variability within a clonal in vitro cannabis population

被引:0
|
作者
Boissinot, Justin [1 ,2 ,3 ,4 ]
Adamek, Kristian [5 ]
Jones, Andrew Maxwell Phineas [5 ]
Normandeau, Eric [2 ]
Boyle, Brian [2 ]
Torkamaneh, Davoud [1 ,2 ,3 ,4 ]
机构
[1] Univ Laval, Dept Phytol, Quebec City, PQ, Canada
[2] Univ Laval, Inst Biol Integrat & Syst IBIS, Quebec City, PQ, Canada
[3] Univ Laval, Ctr Rech & Innovat Vegetaux CRIV, Quebec City, PQ, Canada
[4] Univ Laval, Inst Intelligence & Donnees IID, Quebec City, PQ, Canada
[5] Univ Guelph, Dept Plant Agr, Guelph, ON, Canada
来源
基金
加拿大自然科学与工程研究理事会;
关键词
Cannabis sativa; DNA methylation; clonal propagation; in vitro tissue culture; epigenetics; methylotyping; CREAM; ELAEIS-GUINEENSIS JACQ; DNA-METHYLATION; TISSUE-CULTURE; SOMACLONAL VARIATION; EPIGENETIC CHANGES; PLANT; PROFILES;
D O I
10.3389/fpls.2024.1381154
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
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.
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页数:13
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