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Bulletin of Botanical Research ›› 2017, Vol. 37 ›› Issue (3): 447-452.doi: 10.7525/j.issn.1673-5102.2017.03.016

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Analysis on SSR in Sinoswertia tetraptera Base on RAD-seq

WANG Jiu-Li1,2, ZHU Ming-Xing3, XU Ming-Hang3, CHEN Shi-Long1, ZHANG Fa-Qi1   

  1. 1. Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Xining 810008;
    2. University of the Chinese Academy of Sciences, Beijing 100039;
    3. College of Ecology-Environment Engineering, Qinghai University, Xining 810016
  • Received:2016-12-26 Online:2017-05-15 Published:2017-06-03
  • Supported by:
    National Natural Science Foundation of China(31400322);Applied Basic Research Programs of Qinghai Province(2016-ZJ-761)

Abstract: We used the restriction-site associated DNA sequencing(RAD-seq) technology to analyze simple sequence repeats(SSR) information of Sinoswertia tetraptera(Maxiowicz) T.N.Ho, S.W.Liu & J.Q.Liu. The 5844 SSR loci, with at least 100 bp at two ends, were identified using SR search software. The 5339(91.38%) loci's primers were designed successfully. Among which, amount of tri-nucleotide SSR loci is the most(3323); repeat sequence length type number is 17, while repeat motif type number is 227, and type number of penta-nucleotide motif is the most(106). We employed 32 individuals from 4 natural populations of S.tetraptera to estimate usability and polymorphism of 10 pairs of SSR primers selected randomly from the 5 339 pairs of primers. According to the result of PCR and Polyacrylamide gel electrophoresis, four pairs of primers(ST2, ST3, ST6 and ST10) amplified favorably and showed polymorphism. By the GENEPOP 4.4, the mean number of alleles of the four loci is 6; these loci do not link to each other(P<0.01); these loci deviate from HWE(P<0.01) in most populations and have many homozygotes(observed heterozygosity mean 0.023), which due to the cleistogamous pollination mode of S.tetraptera. ST2 and ST6 were successfully amplified in Halenia elliptica. Our study will offer a SSR dataset in the future based on SSR markers of S.tetraptera.

Key words: RAD-seq, reduced-representation sequencing, Sinoswertia tetraptera, SSR, subtribe Swertiinae

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