中国苦荞SSR分子标记体系构建及其在遗传多样性-NUSCommunity.PDFVIP

2012,45(6):1042-1053 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2012.06.002 1 2 3 030006 100081 100081 · μ· μ· · Construction and Application of SSR Molecular Markers System for Genetic Diversity Analysis of Chinese Tartary Buckwheat Germplasm Resources GAO Fan1,2, ZHANG Zong-wen2,3, WU Bin2 1 2 ( College of Life Science of Shanxi University, Taiyuan 030006; Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081; 3Bioversity International, Office for East Asia, Beijing 100081) Abstract: Objective The SSR molecular markers system was optimized and constructed for genetic diversity analyses of Chinese tartary buckwheat germplasm resources, which is helpful for evaluating Chinese tartary buckwheat collections. Method The SSR-PCR system was optimized by [L 5 16(4 )]orthogonal design, the optimized gel concentration of PAGE was confirmed, and the genetic diversity of 50 tartary buckwheat accessions was analyzed by 19 SSR primer pairs screened from 250 ones of different crops. ResultThe optimized SSR-PCR system was as follows: 30 ng DNA template, 150 μmol·L-1 dNTP, 0.1 μmol·L-1 primer, 2.0 -1 -1 2+ U·L TaqDNA polymerase, 2.0 mmol·L Mg , 1×Taq buffer and ddH O then added up to terminal volume of 25 μL with 6% PAGE 2 for testing. The primers screening efficiency was 7.6%, and the primers from common buckwheat were applicable. A total of 157 alleles were detected by 19 prime