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Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (07): 1114-1125.doi: 10.3724/SP.J.1006.2010.01114

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Molecular Markers for Estimating Distinctness, Uniformity, and Stability of Wheat Lines in Regional Trials

WANG Li-Xin,CHANG Li-Fang**,LI Hong-Bo,JI Wei,LIU Li-Hua,ZHAO Chang-Ping*   

  1. Beijing Engineering and Technique Research Center for Hybrid Wheat / Beijign Academy of Agricultural and Forest Sciences, Beijing 100097, China
  • Received:2010-02-03 Revised:2010-04-12 Online:2010-07-12 Published:2010-04-28
  • Contact: ZHAO Chang-Ping,E-mail: bjhwc2003@yahoo.com.cn, Tel: 010-51503968

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Abstract:

To determine the molecular markers for testing the distinctness, uniformity，and stability (DUS) of wheat (Triticum aestivum L.) lines in regional trials, 145 wheat cultivars were used to screen 1 334 pairs of primers of SSR, EST-SSR, and AFLP-SCAR markers. According to the even distribution of molecular markers on 21 wheat chromosomes, the higher polymorphism information content (PIC), the clear PCR bands, the easy discrimination of different alleles, and the stable PCR amplification, 105 pairs of primers, including 63 pairs of SSR primers, 21 pairs of EST-SSR primers, and 21 pairs of AFLP-SCAR primers, were selected for wheat cultivar (lines) DUS testing. A total of 754 alleles at 122 loci can be identified with 105 pairs of primers, with 5.8 loci per chromosome and 7.2 alleles per locus on average. The three kinds of molecular markers can amplify the DNA fragments that are inside/outside of genes and repeative/non-repetitive sequences. The 105 pairs of primers were classified into 21 pairs of core primers and 84 pairs of standby primers based on their pattern definition, distribution on 21 chromosoms, and PIC value. The 21 core primers distributing on 21 chromosoms could determine the distinctness test for about 80% of total cultivars (lines), the seed purity test for 95%, and the uniformity and stability test for 60%. The 84 standby primers were used as the first grade (29 pairs) and the second grade (55 pairs) to the homozygous DNA locus ratio of lines with no-homozygous loci of coer markers and the genetic similarities or the distinctness of similar cultivars (lines) pairs revealed by the core primers. At the same time, we determined the seed purity for the other 5% of lines. These 105 pairs of primers could finish DUS test for more than 90% of lines in the winter wheat regional trials in 3 cropping seasons from 2006 to 2009. Although less than 10% of lines should be evaluated in combination with phenotypic identification in the field, the effectiveness and efficiency of the 105 pairs of primers are satisfactory in bulk screening for wheat DUS test.

Key words: Wheat, DUS, Molecular markers, DNA fingerprint


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