Genomics and Applied Biology 2024, Vol.15, No.5, 223-234 http://bioscipublisher.com/index.php/gab 228 5.3 Applications in improving yield, cannabinoid profiles, and environmental adaptability The application of genomic tools, including CRISPR-Cas9, has significant implications for improving yield, cannabinoid profiles, and environmental adaptability in Cannabis. By enabling precise modifications at the genetic level, these tools can enhance the expression of genes associated with higher yield and better cannabinoid profiles, such as THC and CBD content (Ahmar et al., 2020; Rao and Wang, 2021). Moreover, CRISPR-Cas9 has been used to develop Cannabis varieties with improved resistance to biotic and abiotic stresses, such as pests, diseases, and environmental extremes, thereby increasing the plant's adaptability and overall productivity (Zhang et al., 2017; Jaganathan et al., 2018; Nascimento et al., 2023). The ability to fine-tune gene regulation and create high-throughput mutant libraries further supports the development of Cannabis strains that meet specific agricultural and medicinal needs (Chen et al., 2019; Thomson et al., 2022). 6 Epigenetic Studies in Cannabis 6.1 Understanding epigenetic regulation in Cannabis gene expression Epigenetic regulation plays a crucial role in the gene expression of Cannabis, influencing various biological processes without altering the DNA sequence. Epigenetic mechanisms such as DNA methylation, histone modifications, and RNA-associated alterations are pivotal in modulating gene expression. For instance, the endocannabinoid system (ECS), which includes cannabinoid receptors and their endogenous ligands, is subject to epigenetic regulation. This regulation can affect the expression of genes involved in neurotransmitter signaling and other critical functions (Basavarajappa and Subbanna, 2022; Bunsick et al., 2023). Additionally, the spatial organization of the cell nucleus and the three-dimensional chromatin architecture are essential for the precise control of gene expression, which can be epigenetically coordinated (Reece and Hulse, 2023). 6.2 Role of epigenetics in phenotype expression, including cannabinoid production Epigenetic modifications significantly impact phenotype expression in Cannabis, including the production of cannabinoids. These modifications can lead to long-term changes in gene expression that influence the plant's metabolic pathways. For example, DNA methylation and histone modifications can alter the expression of genes involved in cannabinoid biosynthesis, affecting the levels of compounds such as THC and CBD (Wu et al., 2021; Bunsick et al., 2023). Moreover, epigenetic changes can be heritable, potentially influencing the phenotype across generations. This transgenerational inheritance can result from environmental factors, such as exposure to cannabinoids, which can induce epigenetic reprogramming and affect the plant's metabolic phenotype (Bunsick et al., 2023). 6.3 Emerging research in Cannabis epigenomics Recent advancements in next-generation sequencing technologies have enabled more detailed studies of the Cannabis epigenome. These studies have revealed complex networks of gene regulation involving alternative splicing, microRNAs (miRNAs), and long non-coding RNAs (lncRNAs). For instance, comprehensive transcriptome analyses have identified numerous transcripts encoding key enzymes in cannabinoid biosynthesis, with many of these transcripts undergoing alternative splicing. Additionally, miRNAs and lncRNAs have been shown to target transcripts involved in cannabinoid production, further highlighting the intricate regulatory mechanisms at play (Wu et al., 2021). Emerging single-cell epigenomic methods also hold promise for transforming our understanding of gene regulation and cell identity in Cannabis, offering insights into how epigenetic information is integrated with genomic and transcriptional data (Clark et al., 2016). 7 Case Study: Cannabis sativa Genome 7.1 In-depth analysis of Cannabis sativagenomic structure The genomic structure of Cannabis sativa has been extensively studied, revealing significant insights into its complexity and diversity. Initial genome assemblies were found to be incomplete, with approximately 10% of the genome missing and 10%-25% unmapped, including critical regions such as ribosomal DNA clusters and centromeres (Kovalchuk et al., 2020). Recent advancements have led to the development of a high-quality reference genome using PacBio single-molecule sequencing and Hi-C technology, resulting in an assembled genome of approximately 808 Mb with a high level of heterozygosity (Gao et al., 2020). This comprehensive genome provides a valuable resource for further genetic and molecular studies.
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