1 Akdeniz University, Vocational School of Technical Sciences, Antalya
2 Akdeniz University, Faculty of Agriculture, Department of Field Crops, Antalya
Abstract
Cannabis (Cannabis sativa L.) is a valuable crop utilized for the production of food, fiber, oils, and psychotropic compounds. DNA molecular markers are essential in cannabis breeding research, enabling the identification of genes associated with favorable features, evaluation of genetic diversity, clarification of genetic links, genetic mapping, and hybrid detection in germplasm. The utilization of simple sequence repeat (SSR) markers, a molecular marker approach, is constrained by developmental problems and the requirement for specific genomic data. Therefore, fresh approaches or resources are essential for the development of microsatellite markers for cannabis. In this study, comprehensive development of expressed sequence tag simple sequence repeat (EST-SSR) markers was conducted to assess genetic diversity in cannabis plants and provide more valuable genetic markers. This work employed 12,907 transcribed genomic DNA sequences/expressed sequence tags to analyze the frequency of microsatellite patterns, encompassing di- to hexa-nucleotides with four to ten tandem repeats, as well as repeat counts beyond ten. We noted that as the motif lengths increased from di- to hexa-nucleotides and the number of repeats escalated from four to ten and beyond, the contexts of the motifs altered. Among the di-nucleotides, TC repeats exhibited a majority over all repeat motifs, whereas AG repeats were characterized by a higher frequency of 4 to 5 repeats relative to the other motifs. CT repeats exhibited a predominance, excluding 4, 5, and 10. The tri-nucleotide motif context CTT/AAG was the most common variant, especially in repeats ranging from nine to ten. Sixty potential microsatellite markers were constructed and evaluated at the sequence level using mined transcribed data. We examined the biological significance of motif lengths and repeat counts in the cannabis genome. Overall, this study offers a comprehensive EST-SSR resource for the cannabis transcriptome and highlights the predominance of specific microsatellite motifs in transcribed regions. The developed primer pairs represent promising candidates for future validation and application in cannabis breeding and germplasm identification.
İNCE, A. G., & KARACA, M. (2026). EST-SSR Marker Development from Cannabis Sequence Data: A Resource for Genetic Diversity and Breeding Research. ISPEC Journal of Agricultural Sciences, 10(3), 1003–1014. https://doi.org/10.5281/zenodo.22498136
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Mehmet KARACA2