Abstract
PremiseThe natural population size of Huperzia serrata (Lycopodiaceae) has dramatically decreased and the species has become endangered due to overexploitation. Here, we developed simple sequence repeat (SSR) markers for H. serrata to survey both its genetic diversity and population structure.Methods and ResultsBased on 177 individuals, 120 SSR primer pairs were developed and optimized from five regions of the H. serrata transcriptomic data. Of these primer pairs, 20 were successfully amplified and 10 showed obvious polymorphism. These polymorphic loci were investigated to study the genetic diversity of H. serrata. Two to 11 alleles per locus were identified, the level of observed heterozygosity ranged from 0.00 to 1.00, and the level of expected heterozygosity ranged from 0.19 to 0.79. All loci were successfully amplified in H. crispata, H. sutchueniana, and H. selago.ConclusionsThe 10 polymorphic primer pairs developed here will be valuable for studies of the endangered H. serrata and other related species.
Highlights
PREMISE: The natural population size of Huperzia serrata (Lycopodiaceae) has dramatically decreased and the species has become endangered due to overexploitation
20 were successfully amplified and 10 showed obvious polymorphism. These polymorphic loci were investigated to study the genetic diversity of H. serrata
All loci were successfully amplified in H. crispata, H. sutchueniana, and H. selago
Summary
177 H. serrata individuals were collected from the Chinese localities of Luan, Enshi, Jizhou, Hanzhong, and Jinping (Appendix 1). Total RNA was extracted from 100 mg of fresh leaves using TRIzol following the instructions of the manufacturer (TIANGEN, Beijing, China). To eliminate potential DNA contamination, we used DNase to purify total RNA following the manufacturer instructions (QIAGEN, Hilden, Germany). RNA purity and concentration were determined by NanoDrop Spectrophotometer (Qubit2.0, Agilent 2100; Shimadzu, Kyoto, Japan). MRNA was isolated using magnetic oligo (dT) beads, and cut into short fragments using NEBNext Poly(A) mRNA Magnetic Isolation Module according to the manufacturer’s instructions Locus c52026.graph_c0 c63431.graph_c0 c51797.graph_c0 c59934.graph_c0 c52211.graph_c0 c65171.graph_c0 c50318.graph_c0. C52257.graph_c0 c66382.graph_c0 c60778.graph_c0 c38689.graph_c0 c56357.graph_c0 c59441.graph_c0 c60018.graph_c0 c61426.graph_c0 c62215.graph_c0 c62350.graph_c0 c62412.graph_c0 c63947.graph_c0 c64437.graph_c0 Mean
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