Development of EST-SSR Molecular Markers in Dendrobium officinale
To develop EST-SSR molecular markers for Dendrobium officinale,15 383 ESTs from Dendrobium nobile (Related Species of Dendrobium officinale) in NCBI were screened using SSRIT software.The primers were designed and evaluated by using Primer 5 and Oligo 7 software,followed by agarose gel electrophoresis for preliminary screening after PCR amplification,and reusing polyacrylamide gel electrophoresis to detect the effectiveness of the selected primers.The results showed that 379 SSRs were mined from 370 ESTs with a frequency of 2.46％.Dinucleotide,Trinucleotide and Hexanucleotide nucleotide repeats EST-SSRs were dominant,which account for 41.42％,26.91％and 27.44％,respectively.Among them,AG/CT (20.84％) and GA/TC (18.73％) were highly abundant in Dinucleotide nucleotide repeats,TCT/AGA (4.22％) and GA/TC (3.96％) were highly abundant in Trinucleotide nucleotide repeats,and Hexanucleotide repeat motifs were higher.The average length of EST-SSR was 24bp,and every 8.5 EST sequences contained a SSR site.Among 106 designed primer pairs,56 primer pairs showed ideal amplifications and the effective amplification rate was 47.17％.Moreover,43 primer pairs displayed polymorphic,which accounts for 86％ of the effective amplification.Taken together,43 primer pairs developed in this study could be used in genetic diversity analysis,germplasm identification,genetic map construction and functional gene study of Dendrobium officinale to provide an important reference value.
Dendrobium officinaleDendrobium nobileEST-SSR,molecular markersprimer development
国家科技期刊平台
NSTL
万方数据
维普
