Abstract
Premise of the Study Ephedra sinica (Ephedraceae) is a gymnosperm shrub with a wide distribution across Central and Eastern Asia. It is widely cultivated as a medicinal plant, but its wild populations are monitored to determine whether protection is needed.Methods and ResultsThirty‐six microsatellite markers, including 11 polymorphic markers, were developed from E. distachya RNA‐Seq data deposited in the National Center for Biotechology Information dbEST database. Among 100 genotyped E. sinica individuals originating from five different population groups, the allele number ranged from three to 22 per locus. Levels of observed and expected heterozygosity ranged from 0 to 0.866 (average 0.176) and 0 to 0.876 (average 0.491), respectively. Allelic polymorphism information content ranged from 0.000 to 0.847 (average 0.333). Cross‐species amplifications were successfully conducted with two related Ephedra species for all 11 di‐ or trinucleotide simple sequence repeats.ConclusionsThis study provides the first set of microsatellite markers for genetic monitoring and surveying of this medicinal plant.
Highlights
Si-Qian Jiao1, Yan-Qiang Sun1, Dong-Xu Zhang2, Qiong Gao1, Yuqing Jin1, Hui Liu1, Yongpeng Ma3, Yong Yang4, Ilga Porth5,6, and Jian-Feng Mao1,7
This study provides the first set of microsatellite markers for genetic monitoring and surveying of this medicinal plant
Ephedra sinica is recorded on the International Union for Conservation of Nature (IUCN) Red List of Threatened Species (Bell and Bachman, 2011)
Summary
A total of 4981 ESTs generated from mRNA sequencing of E. distachya were retrieved from the National Center for Biotechnology Information (NCBI) Expressed Sequence Tags database (dbEST) (accessed by searching with “(Ephedra) AND “Ephedra distachya”[porgn:__txid3389]”). In order to test for successful amplification of the 88 EST-S SR loci selected, we conducted PCR analysis using eight individual plants of E. sinica. After final capillary electrophoresis analysis on an ABI 3730 sequencer (Applied Biosystems, Waltham, Massachusetts, USA), SSR alleles were called with GeneMarker version 2.20 (SoftGenetics, State College, Pennsylvania, USA) Of these 38 loci, 36 showed clear, single peaks for each allele as essential for confident scoring, and 11 of these loci were polymorphic among the initially screened eight individuals. Note: A = number of alleles; He = expected heterozygosity; Ho = observed heterozygosity; n = number of individuals sampled; N = number of successfully amplified individuals; PIC = polymorphism information content. Except for primers at the E-4 9 locus, the remaining primer pairs showed moderate polymorphism in E. equisetina, possibly due to the small sample size
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