Isolation and characterization of polymorphic microsatellite markers in Schisandra chinensis (Turcz.) Baill. (Schisandraceae)

Abstract

Fourteen polymorphic microsatellite markers were developed and characterized in Schisandra chinensis, a widely used traditional Chinese herbal medicine. The average number of alleles of these microsatellites was 6.071, ranging from 2 to 12 per locus. The effective number of alleles was from 1.645 to 7.119 per locus with an average of 3.916. The observed and expected heterozygosity were 0.107-0.871 and 0.399-0.874, respectively. Three loci significantly (P \ 0.01) deviated from Hardy-Weinberg equilibrium. No significant (P \ 0.01) linkage disequilibrium was detected for any pair of loci. Eleven out of 14 SSRs are also polymorphic in S. sphenanthera, a closely related medicinal species of S. chinensis. This set of microsatellite markers has provided a useful tool for the ongoing efforts in studying the population genetic variation of S. chinensis and S. sphenanthera, which will facilitate formulation of appropriate strategies for conservation and sustainable utilization of the two important officinal species.Keywords SSR Á Schisandra chinensis Á Schisandra sphenanthera Á Herbal medicine Á Schisandraceae Schisandra chinensis (Turcz.) Baill is well-known Chinese medicinal plant, belonging to the family Schisandraceae. The fruits of S. chinensis are commonly used in the treatment of cough and asthma, spermatorrhea, enuresis, arrhea, night sweating, diabetes symptoms, palpitation and insomnia (Chinese Pharmacopoeia Commission 2005). Schisandra chinensis are scandent woody vines, occurring over a wide geographical range across north-eastern China and the adjacent regions of Russia, Korea, and Japan. They are often found in deciduous, coniferous and mixed forests (Saunders 2000). The resources of S. chinensis are dramatically diminishing especially in the recent decades because of overexploitation and habitat deterioration (Yu et al. 2005;Zhang et al. 2007). The population genetics information is prerequisite for formulation of appropriate strategies of conservation and management for rare and endangered species. However, the genetic diversity and genetic structure of the natural populations of S. chinensis are little known due to the lack of highly informative molecular marker. Microsatellite markers were developed in the present study to facilitate studying the population genetic variation of S. chinensis.Genomic DNA was isolated using a modified procedure of Doyle and Doyle (1987). Washing buffer (2% PVP 40 000 MW, 100 mM Tris-HCl pH 8.0, 20 mM EDTA pH 8.0, 1.4 M NaCl, 0.2% mercaptoethanol) was used to rinse the leaf powder at 65°C to remove most of polysaccharides, polyphenols and secondary metabolites. Microsatellite isolation were performed according to the procedure of Fast Isolation by AFLP of Sequences Containing repeats (FIASCO) (Zane et al. 2002).

EST-SSR markers development and application in an important medicinal plant, Schisandra chinensis (Schisandraceae)

“…More than two times higher frequency of the dinucleotides compared to the trinucleotides conformed to the trend as observed in peanut (Arachis hypogaea) [24], precocious trifoliate orange (Citrus reticulata) [29], Korean pine (Picea neoveitchii) [30] and litchi (Litchi chinensis) [31], despite some evidence suggesting that trinucleotides are the most common SSR type [17,19,22,32,33]. This difference is potentially attributed to the characteristics and quantity of EST-SSR and the EST resources in plants.…”

Ten polymorphic microsatellite markers in Michelia maudiae (Magnoliaceae)

“…Genomic DNA was isolated from a single individual using a modified CTAB method described in Sun et al (2009). Microsatellites were isolated following the procedure of FIASCO (Fast Isolation by AFLP of Sequences Containing repeats) (Zane et al, 2002).…”

Genetic diversity and population structure of two important medicinal plant species Schisandra chinensis and Schisandra sphenanthera revealed by nuclear microsatellites

“…1. DNA isolation and SSR genotyping were carried out as described in Sun et al (2009). Seven polymorphic and transferable microsatellite markers were used to genotype these two species.…”

“…A set of polymorphic microsatellite markers has been developed recently from S. chinensis, most of them are transferable and successfully cross-amplified in S. sphenanthera (Sun et al 2009). In the present study, the population structure of genetic diversity of S. chinensis and S. sphenanthera are investigated using this same set of microsatellite markers to further our understanding of their population genetics.…”