Isolation and characterization of microsatellite loci in Rehmannia glutinosa (Scrophulariaceae), a medicinal herb1

Ap Applicati tions ons in in Pl Plant t Scien Sciences ces Rehmannia glutinosa (Gaertn.) Libosch. ex Fisch. & C. A. Mey. (Scrophulariaceae) is a perennial herbaceous species of medicinal value (Zhou, 2002 ; Shao et al., 2008). Its tuberous roots are commonly used in Chinese traditional medicine (Wen et al., 2002). In recent decades, a signifi cant number of chemical and pharmacological studies have been performed on R. glutinosa (Zhang et al., 2008 ; Chang et al., 2011). More than 70 compounds, including iridoids, saccharides, amino acids, inorganic ions, and other trace elements, have been found in the herb (Zhang et al., 2008). Many studies show that some active ingredients in the roots of R. glutinosa possess broad pharmacological actions for protecting gastric mucosa and restraining pulmonary fi brosis (Liu et al., 2009). In addition, the root has been demonstrated to improve hematopoiesis, have anti-infl ammation and antitumor activities, decrease blood sugar, and promote the proliferation of vascular endothelial cells (Liu et al., 2009). Genetic knowledge about R. glutinosa , such as its genetic diversity, genetic structure, and gene fl ow, serves as a foundation for cultivating improved varieties and exploiting and utilizing Chinese traditional medicine resources (Zhang et al., 2012). Simple sequence repeats (SSRs) are highly polymor-phic, multiallelic, reproducible, abundantly distributed in the genome, and easy to interpret (Tanya et al., 2011). They are also codominant inheritance markers and can provide the am-plifi ed result of the heterozygote or the homozygote. In this study, we isolated 16 microsatellite loci from R. glutinosa and used these loci as markers to estimate the genetic diversity of two wild populations. METHODS AND RESULTS Forty-four individual leaves were collected from two wild populations of The voucher specimens (Rehmannia glutinosa HB09001 and Rehmannia glutinosa JZ09001 for the HB and JZ populations, respectively) are deposited in the herbarium of Henan Agricultural University (HEAC). These leaf samples were dried quickly with silica gel and stored at −20 ° C. The total DNA was isolated from the dried leaves using a modifi ed cetyltrimethylam-monium bromide (CTAB) method (Fang et al., 2009) and purifi ed with a Universal DNA Purifi cation Kit (Tiangen Biotech, Beijing, China). The purifi ed DNA was digested with the enzyme Rsa I, and the digested DNA fragments were linked to SuperSNX-24F (5 ′-GTTTAAGGCCTAGCTAGCAGAATC-3 ′) and SuperSNX-24R (5 ′-GATTCTGCTAGCTAGGCCTTAAACAAAA-3 ′) adapters. Using biotinylated (AC) 8 , (AG) 8 , and (ATG) 12 as …