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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(1College of Life Science of Shanxi University,Taiyuan 030006;2Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081;3Bioversity International,Office for East Asia,Beijing 100081)  
【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 [L16(45)]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.【Result】The optimized SSR-PCR system was as follows: 30 ng DNA template,150 μmol?L-1 dNTP,0.1 μmol?L-1 primer,2.0 U?L-1 TaqDNA polymerase,2.0 mmol?L-1 Mg2+,1×Taq buffer and ddH2O then added up to terminal volume of 25 μL with 6% PAGE 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 primers,with 2-11 alleles for each primer pair,and the average was 7.42.Moreover,the averaged PIC and DP values were 0.8881 and 5.684,respectively.Using Popgen Ver.1.31,50 accessions were clustered into 5 groups at GS 0.578.The clustering results revealed that the genetic diversity of accessions of tartary buckwheat was not correlated to their geographic origins.The genetic diversity of tartary buckwheat from Sichuan was very rich as genetic parameters were the highest.The core primers could be used to identify the similar accessions.【Conclusion】 The SSR molecular markers system was effective for assessment of genetic diversity of Chinese tartary buckwheat germplasm resources.SSR primers of common buckwheat could be used in tartary buckwheat.TBP5 and Fes2695 were SSR core primers.It showed a high genetic diversity in 50 Chinese tartary buckwheat accessions which could be classified into 5 groups.
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