The scientific literature contains evidence that the prevalence of celiac disease (CD) and non-celiac gluten sensitivity (NCGS) has increased in the population during the second half of the 20th century. There are numerous factors which can be attributed to the increased prevalence. Besides new wheat processing techniques, agricultural practices in wheat cultivation, changes in the dietary habits, improved diagnostic techniques, modern wheat breeding practices might have contributed to the increase in the prevalence, essentially due to changes of the protein content and composition of wheat cultivars over the past 120 years. The aim of this work was an in-depth investigation of 60 hexaploid German winter wheat cultivars from the past 120 years to form the basis for developing new wheat cultivars, which cause less harmful sensitivities in the population. In the first part all cultivars were comprehensively analyzed for their quantitative protein composition and distribution. The second part emphasized the necessity of a new RP-HPLC pattern integration method for an in-depth elucidation of protein fingerprints. In the third part the focus was on the distribution of four CD-active peptides in all wheat cultivars. Thus, the main hypothesis of this work was that the selection criteria for modern breeding may have led to an altered composition of wheat proteins and possibly to a higher immunostimulatory potential of modern cultivars (registered after the 1950s) compared to older cultivars.
The quantitation of the crude protein contents showed highly variable contents depending on the cultivar and harvest year with a slightly decreasing trend based on the median of five cultivars per decade. Wheat proteins were extracted stepwise by means of the modified Osborne fractionation and measured by RP-HPLC. While the proportions in the total RP-HPLC content of albumins/globulins remained unaffected, the percentages of gliadins showed a clear decreasing trend and those of glutenins an increasing trend. Concerning the protein types ω5- and ω1,2-gliadins, no trends were revealed for their percentages, whereas the proportions of α- and γ-gliadins showed a significant decreasing trend. In contrast, the proportions of the glutenin subunits showed an increasing trend. Because of the high variability of the protein contents, a new approach of RP-HPLC profile integration was developed, in order to enable a better differentiation between wheat cultivars. For the novel approach, the integration was carried out every 20 s for all protein fractions and the fractions were no longer subdivided into protein types. This way of integration allowed us to consider the distribution of individual peaks without information loss and to identify cultivars with a unique protein composition. Further, all wheat cultivars were analyzed for their contents of four immunodominant peptides, namely the 33-mer and peptides containing the DQ2.5-glia-α1a/DQ2.5-glia-α2 (P1), DQ2.5-glia-α3 (P2) and DQ2.5-glia-γ1 (P3) epitopes by means of SIDA combined with targeted LC-MS/MS in order to identify the influence of breeding on the contents.
The values for the 33-mer and P1 proportions in α-gliadins showed a slightly increasing trend from old to modern cultivars. Concerning the P2 and P3 peptides present in α- and γ-gliadins, no trends, either in relative or absolute values were observed. This means therefore, that old and modern cultivars appear to have largely the same immunoreactive potential. What is more important is the effect of the harvest year, which influenced the contents of the CD-active peptides more strongly than the wheat cultivars.
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The scientific literature contains evidence that the prevalence of celiac disease (CD) and non-celiac gluten sensitivity (NCGS) has increased in the population during the second half of the 20th century. There are numerous factors which can be attributed to the increased prevalence. Besides new wheat processing techniques, agricultural practices in wheat cultivation, changes in the dietary habits, improved diagnostic techniques, modern wheat breeding practices might have contributed to the incre...
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