Establishment and Application of Multiplex PCR Systems Based on Molecular Markers for HMW-GSs in Wheat
Chuxuan Yao,
Cuimian Zhang,
Caili Bi,
Shuo Zhou,
Fushuang Dong,
Yongwei Liu,
Fan Yang,
Bo Jiao,
He Zhao,
Mengyu Lyu,
Haibo Wang,
Jianfang Chai
Affiliations
Chuxuan Yao
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Cuimian Zhang
Institute of Agricultural Resources and Environment, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050051, China
Caili Bi
Life Science College, Hebei Normal University, Shijiazhuang 050024, China
Shuo Zhou
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Fushuang Dong
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Yongwei Liu
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Fan Yang
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Bo Jiao
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
He Zhao
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Mengyu Lyu
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Haibo Wang
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
Jianfang Chai
Institute of Biotechnology and Food Science, Hebei Academy of Agriculture and Forestry Sciences/Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China
High-molecular-weight glutenin subunits (HMW-GSs) encoded by alleles at the Glu-A1, Glu-B1, and Glu-D1 loci confer unique end-use quality properties of common wheat (Triticum aestivum L.). Wheat accessions with the high-quality HMW-GSs combination of Ax2*/Bx7OE/Dx5 usually exhibit strong gluten characteristics. In order to stack these three high-quality subunit genes by molecular markers in strong gluten wheat breeding, an agarose gel-based multiplex PCR marker for these high-quality HMW-GSs and two agarose gel-based multiplex PCR markers detecting the homozygosity of Ax2* and Bx7OE subunits were developed. These markers were verified in an F2 segregating population from a cross between a medium-gluten winter wheat cultivar with the HMW-GSs combination of Ax null/Bx7 + By8/Dx4 + Dy12 and a strong-gluten spring wheat cultivar with the HMW-GSs combination of Ax2*/Bx7OE + By8*/Dx5 + Dy10. By integrating the newly established multiplex PCR markers and a published co-dominant PCR marker of the Dx5 subunit, a complete molecular marker selection system was established. After multiple rounds of molecular marker-assisted selection with the system, 17 homozygous winter wheat lines that stacked the three high-quality HMW-GSs were generated. The gluten strength of these homozygous lines was comparable to their strong-gluten parent, but significantly higher than that of their medium-gluten parent by measuring their lactic acid-sodium dodecyl sulfate solvent retention capacities of whole wheat meal. The multiplex PCR systems established in the present study can be used for molecular marker-assisted selection of strong gluten wheats.
