Abstract
Cannabis sativa L. is an important economic plant for the production of food, fiber, oils, and intoxicants. However, lack of sufficient simple sequence repeat (SSR) markers has limited the development of cannabis genetic research. Here, large-scale development of expressed sequence tag simple sequence repeat (EST-SSR) markers was performed to obtain more informative genetic markers, and to assess genetic diversity in cannabis (Cannabis sativa L.). Based on the cannabis transcriptome, 4,577 SSRs were identified from 3,624 ESTs. From there, a total of 3,442 complementary primer pairs were designed as SSR markers. Among these markers, trinucleotide repeat motifs (50.99%) were the most abundant, followed by hexanucleotide (25.13%), dinucleotide (16.34%), tetranucloetide (3.8%), and pentanucleotide (3.74%) repeat motifs, respectively. The AAG/CTT trinucleotide repeat (17.96%) was the most abundant motif detected in the SSRs. One hundred and seventeen EST-SSR markers were randomly selected to evaluate primer quality in 24 cannabis varieties. Among these 117 markers, 108 (92.31%) were successfully amplified and 87 (74.36%) were polymorphic. Forty-five polymorphic primer pairs were selected to evaluate genetic diversity and relatedness among the 115 cannabis genotypes. The results showed that 115 varieties could be divided into 4 groups primarily based on geography: Northern China, Europe, Central China, and Southern China. Moreover, the coefficient of similarity when comparing cannabis from Northern China with the European group cannabis was higher than that when comparing with cannabis from the other two groups, owing to a similar climate. This study outlines the first large-scale development of SSR markers for cannabis. These data may serve as a foundation for the development of genetic linkage, quantitative trait loci mapping, and marker-assisted breeding of cannabis.
Highlights
Cannabis sativa L., a member of the Cannabinaceae, is a diploid (2n = 20) monocotyledon, and one of the oldest cultivated plants
100 varieties were from 18 provinces of China, Longdama 1 (LDM), Jinma 1 (JM1), Wandama 1 (WDM), Yunma 1 (YM1), Yunma 4 (YM4), Yunma 5 (YM5) and Yunwan 6 (YW6) were cultivated varieties and the others were all wild varieties in China
Lack of sufficient simple sequence repeat (SSR) markers has limited the development of cannabis molecular genetics
Summary
Cannabis sativa L., a member of the Cannabinaceae, is a diploid (2n = 20) monocotyledon, and one of the oldest cultivated plants. It has been cultivated or grows wild around the world, and is used in diverse applications [1]. Cannabis is a botanical genus of flowering plants that is divided into two species. Hemp, which has low tetrahydrocannabinol (THC) content, is mainly used to cultivate for fiber and seed production, whereas marijuana, with its relatively higher THC content, is used for its psychoactive potency [2]. Hemp was used for textile production in China more than 6,000 years ago, and is still widely grown in China
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