Production of Doubled Haploid Lines with High-Molecular-Weight Alleles via Microspore Cultures in Wheat

The objectives of this study were to generate doubled-haploid wheat lines (DH) from microspore cultures, to compare the agronomic performance of DH wheat lines derived via microspore culture against their corresponding parental lines under field conditions and to identify DH wheat lines carrying glutenin allelic combinations which are related with good or poor bread making quality. A double haploid production system from microspore cultures method have been established and subsequently, seeds were obtained from the regenerated haploid wheat plants derived from the F₁ plants of the two wheat populations (Irena × Ksu102 and Klasic × Ksu105). The number of harvested plants with high amount of seeds derived from microspore cultures system for Irena × Ksu102 and Klasic × Ksu105 wheat populations were 49 and 19, respectively. Field trials of wheat doubled haploid lines were conducted over two seasons. Genotypic differences were highly significant for all agronomic traits. These differences are very useful in screening for selecting the high- yielding DH lines as potential sources of new cultivars. The highest- yielding DH lines, for most traits, had similar performance as compared with the parental genotypes and check cultivar. Some of the selected DH lines had similar or higher grain yield than the parental cultivars and the check cultivar Yecora Rojo in both populations. Sets of 6 gene- or allele-specific PCR markers were used for identification of genes encoding HMW glutenin subunits Ax1 and Ax2* at the Glu-A1 locus, Bx7, Bx7*, By8 and By17 at the Glu-B1 locus and Dx2, Dx5, Dx10 and Dx12 at the Glu-D1 locus in wheat inbred lines. The 49 and 19 wheat inbred lines for the populations; Irena × Ksu102 and Klasic × Ksu105, respectively, showed seven and eight different allelic combinations at the Glu A, Glu B and Glu D loci for Irena × Ksu102 and Klasic × Ksu105, respectively. Rapid identification of wheat Glu-1 alleles by molecular markers could be an efficient for early selection of useful doubled-haploid wheat lines with good bread-making quality.