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We sampled leaves from 678 individuals in 21 natural populations (30-36 individuals per population), covering the entire distribution of Euptelea pleiospermum in China.Total DNA was isolated from about 50 mg powdered leaf tissue following the protocol of a DNA extraction kit (Tiangen Biotech Co., LTD., Beijing, China). We used seven fluorescence-labeled microsatellite loci (EP036, EP059, EP081, EP087, EP091, EP278 and EP294; Zhang et al., 2008) to genotype our 678 DNA samples.