Isolation and characterization of polymorphic microsatellite markers in the endangered Nothotsuga longibracteata (Pinaceae)

Abstract

Twelve polymorphic microsatellite loci were isolated and characterized from an ACenriched genomic library of Nothotsuga longibracteata . The average allele number of these microsatellites was 8.3 per locus, ranging from two to 13. The ranges of observed and expected heterozygosities were 0.03 -0.97 and 0.09 -0.88, respectively. These polymorphic markers provide useful tools for the study of evolutionary history and conservation genetics of N. longibracteata .

Spatial and temporal population genetic variation and structure of Nothotsuga longibracteata (Pinaceae), a relic conifer species endemic to subtropical China

“…DNA quality and quantity were determined by electrophoresis in 1% agarose gels with λDNA markers. Microsatellite genotyping of the nuclear genome of N. longibracteata was performed according to the methods described by Qiu et al (2007) using the loci NT01, NT02, NT03, NT04, NT06 and NT07. In addition, three cpDNA microsatellite primer pairs (Pt15169, Pt63718, Pt71936) derived from Pinus thunbergii ( Vendramin et al , 1996 ) were used due to their high genetic polymorphism found in a preliminary screen for genetic variation of cpDNA in N. longibracteata.…”

“…Polymerase chain reaction (PCR) assays were done in a volume of 10 μL, containing 10 mM Tris-HCl (pH 8.4), 50 mM (NH 4 ) 2 SO 4 , 1.5 mM MgCl 2 , 0.2 mM dNTPs, 0.2 μM of each primer, 50 ng of genomic DNA, and 1 unit Taq polymerase (Fermentas, Lithuania) for both nSSR and cpSSR. The amplification protocol was essentially similar to that described by Qiu et al (2007) for SSR and by Vendramin et al (1996) for cpSSR. The amplified products were separated in a 6% denaturing polyacrylamide gel and revealed using silver staining.…”