期刊
MOLECULAR BREEDING
卷 12, 期 1, 页码 51-59出版社
KLUWER ACADEMIC PUBL
DOI: 10.1023/A:1025484523771
关键词
bread wheat (Triticum aestivum L.); doubled haploids; gene-assisted selection; HMW glutenin genes; PCR markers
Molecular markers based on DNA sequence variations of the coding and/or promoter regions of the wheat ( Triticum aestivum L.) HMW glutenin genes located at the Glu-1 loci were developed. Markers characteristic of alleles Glu-A1-1a ( encoding Ax1 subunit) and Glu-A1-1c ( encoding Ax2* subunit) at the Glu-A1 locus, alleles Glu-B1ak ( encoding Bx7* subunit) and Glu-B1al for overexpressed Bx7 subunit at the Glu-B1 locus and alleles Glu-D1-1a ( encoding Dx2 subunit) and Glu-D1-1d ( encoding Dx5 subunit) at the Glu-D1 locus were tested using genomic DNA of haploid leaf tissue. A method for simultaneously extracting DNA from 96 haploid leaf tissue pieces is described. Two of the developed markers were dominant and two were co-dominant. A F-1-derived population segregating for all HMW glutenin genes was used to test the validity of the markers and their usefulness in doubled haploid breeding programs. SDS-PAGE analysis of seed storage protein was performed on seeds from the doubled haploid lines. A total of 299 lines were tested with the DNA markers on the haploid tissue and validated by protein analysis of the corresponding DH seeds. PCR markers and SDS-PAGE analysis showed between 2 and 8.5% discrepancies depending on the marker. Applications of DNA markers for gene-assisted-selection of haploid tissue and use in breeding programs are discussed. Advantages and disadvantages of dominant and co-dominant markers are outlined.
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