Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (11): 293-302.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1494

Previous Articles     Next Articles

Establishment of a Method Identifying the Varieties of Hulless Barley Based on Fluorescence Detection Technology

GUAN Jun-jiao1(), YANG Xiao-hong1, ZHANG Peng1, HUANG Qing-mei1, ZHANG Jian-hua2()   

  1. 1. Quanlity Standard and Testing Technology Research Indtitute,Yunnan Academy of Agriculture Sciences,Kunming 650205
    2. Food Crops Research Institute,Yunnan Academy of Agriculture Sciences,Kunming 650205
  • Received:2020-12-09 Online:2021-11-26 Published:2021-12-03
  • Contact: ZHANG Jian-hua E-mail:guanjunjiao@163.com;zhjhua748@163.com

Abstract:

A series of SSR core primers were screened to provide a DNA fingerprinting of hulless barley varieties,which may provide high-throughput detection method for the breeding and variety identification of hulless barley. First of all,24 hulless barley germplasms with large morphological differences were used,42 primers were screened from the original 198 SSR primers,which had clear amplification bands,rich polymorphism and stable amplification. These screened primers were labeled with fluorescent,and used for the PCR amplification of the total 226 cultivars. The amplified products were detected by capillary electrophoresis. Finally,28 pairs of SSR fluorescent primers were screened to establish an identification system for hulless barley variety based on the high-throughput fluorescent SSR marker. And the selected 28 primers were used to construct DNA fingerprinting for identifying the 226 cultivars of hulless barley. A total of 252 alleles were detected by 28 pairs of SSR primers,the effective number of alleles ranged from 5 to 16,and the average number of alleles was 9. The genetic diversity ranged from 0.48 to 0.86 and the average value was 0.68. The polymorphism information content(PIC)changed from 0.44 to 0.84 and the average value of PIC was 0.65. The genetic distance of 226 materials was 0-0.99. This set of primers distinguished the most of the test materials. Seven SSR primers(Bmag0211,Scssr07759,HVM62,EBmac0679,Scssr03907,Bmag0870,and EBmac0827)with good amplification and detection effect as well as distributed in different chromosomes were chosen from the 28 SSR primers,and they were used for constructing DNA fingerprinting spectrum. The corresponding reference cultivars to major allelic variations of 7 pairs of primers were selected to eliminate the errors caused by different platforms and experimental batches. These reference cultivars can be used as the basis for reading bands of ordinary polyacrylamide gel electrophoresis. In this study,a SSR high-throughput identification system was constructed in two detection platforms(fluorescent capillary electrophoresis platform and polyacrylamide gel electrophoresis platform)based on 28 pairs of primers.

Key words: hulless barley, SSR marker, core primers, fingerprinting, variety identification