Identification and characterization of high-molecular-weight glutenin genes in Polish triticale cultivars by PCR-based DNA markers

Abstract

Molecular markers were used to identify the allele/gene composition of complex loci Glu-A1 and Glu-B1 of high-molecular-weight (HMW) glutenin subunits in triticale cultivars. Forty-six Polish cultivars of both winter and spring triticale were analysed with 7 PCR-based markers. Amplified DNA fragments of HMW glutenin Glu-1 genes were separated by agarose slab-gel electrophoresis. Differences between all 3 alleles at the locus Glu-A1 [Glu-A1a (encoding Ax1), 1b (Ax2*), and 1c (AxNull)], 4 alleles at Glu-B1-1 [Glu-B1-1a (Bx7), 1b (Bx7*), 1d (Bx6), 1ac (Bx6.8)], and 5 alleles at Glu-B1-2 [Glu-B1-2a (By8), 2b (By9), 2o (By8*), 2s (By18*), and 2z (By20*)] were revealed. In total, 16 allele combinations were observed. Molecular markers are particularly helpful in distinguishing the wheat Glu-A1a and Glu-A1b alleles from the rye Glu-R1a and Glu-R1b alleles in triticale genotypes, respectively, as well as subunits Bx7 from Bx7* and By8 from By8*, which could not be distinguished by SDS-PAGE. Novel glutenin subunits By18* and By20* (unique to triticale) were identified. HMW glutenin subunit combinations of Polish triticale cultivars, earlier identified by SDS-PAGE analyses, were verified by PCR-based DNA markers. Rapid identification of wheat Glu-1 alleles by molecular markers can be an efficient alternative to the standard separation procedure for early selection of useful triticale genotypes with good bread-making quality.

Survey and new insights in the application of PCR‐based molecular markers for identification of HMW‐GS at the Glu‐B1 locus in durum and bread wheat

“…Ofanto and Capeiti, carrying the Bx20+By20 combination. Similar results were reported for triticale, where a 750-bp band was observed in cultivars carrying the By20* gene (Salmanowicz & Dylewicz, 2007). Moreover, cv.…”

“…David (and confirmed in cv. Carme) as previously detected also in triticale (Salmanowicz & Dylewicz, 2007). The same profile was also observed in cv.…”

“…Notwithstanding the large number of markers developed, the allelic specificity has been verified on specific and limited numbers of test cultivars (cvs) selected on a case by case basis and representative of the expected HMW‐GS subunits at the Glu‐B1 locus (Lei et al., ; Ma et al., ). When applied to different species such as triticale (x Tricosecale) , a wider discrimination of HMW‐GS alleles at the Glu‐B1 locus was observed (Salmanowicz & Dylewicz, ).…”

Molecular, Physicochemical and Rheological Characteristics of Introgressive Triticale/Triticum monococcum ssp. monococcum Lines with Wheat 1D/1A Chromosome Substitution

“…The presence of the GS III primer pair, which is an allele-specific marker for the triticale subunit By20* [30,31], in the reaction mixture, gave rise to one band of 750 bp (Figure 2c) in all triticale samples. The allelic composition encoding subunit pair 6.8 + 20* frequently appears among European triticale cultivars [27,32].…”

“…Genomic DNA was extracted from the leaves of single plants, according to Salmanowicz and Dylewicz [31]. Two pairs of primers were used for the amplification of DNA fragments corresponding to the x-type genes present at the Glu-A1 and Glu-D1 loci.…”