Abstract：Rhododendron henanense subsp. lingbaoense belongs to Ericaceae, Rhododendron, and is a rare and endemic species in Henan Province. Because of its narrow distribution and poor natural regeneration, the population size has been shrinking, and systematic research and protection has become increasingly urgent. In order to develop SSR (simple sequence repeat) molecular markers for conservation genetics research on R. henanense subsp. lingbaoense, simplified genome sequencing by restriction-site associated DNA sequencing (RAD-seq), SSR search and primer design were performed. A total of 6 107 SSR loci were detected in 120 970 assembled sequences. Among them, the proportion of di-nucleotide motifs was the highest, accounting for 63.01% of the total loci; followed by tri-nucleotides with 765, accounting for 12.53%. 3 843 pairs of primers were successfully designed for these SSR loci using software Primer Premier 3.0. A total of 100 pairs of primers were randomly selected and verified with 20 individuals of R. henanense subsp. lingbaoense, and 87 were able to amplify the target bands. Fifteen pairs of polymorphic SSR loci were further analyzed using Cervus 3.0, and the polymorphic information content ranged from 0.463 to 0.893 with an average of 0.649. Among them, there were 13 highly polymorphic loci, no deviation from Hardy-Weinberg equilibrium was detected in the 15 loci. Above results could provide data support for the study of SSR markers in R. henanense subsp. lingbaoense and Rhododendron.
周晓君, 王海亮, 李方玲, 张凯, 王俞娜, 押辉远. 基于RAD-seq技术开发灵宝杜鹃多态性SSR标记[J]. 农业生物技术学报, 2019, 27(1): 55-62.
ZHOU Xiao-Jun, WANG Hai-Liang, LI Fang-Ling, ZHANG Kai, WANG Yu-Na, YA Hui-Yuan. Development of Polymorphic SSR Markers in Rhododendron henanense subsp. lingbaoense Based on RAD-seq. 农业生物技术学报, 2019, 27(1): 55-62.
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