The low-molecular-weight glutenin subunits (LMW-GS) with extra cysteine numbers have attracted great research interest for their potential quality value. In this study, 14 LMW-i type genes (YD1-YD14) were isolated from three types of Chinese wheat landraces; and nine of 14 genes (YD1-YD9) had nine cysteines, and the other five genes contained eight cysteines. Phylogenic analysis suggested that all 14 LMW-i genes were related to Glu-A3-1 variants Glu-A3-17/FJ 549934 and Glu-A3-15/FJ 549932. Six randomly selected genes, five genes including YD1 with nine cysteines and the remaining one with eight cysteines, were successfully expressed in bacteria as mature proteins with a molecular mass of ~ 46 kDa. These proteins were traced to corresponding seed storage proteins for having similar elution times in reverse phase high-performance liquid chromatography (RP-HPLC) profiles. Mass spectrometry verified that bacterial expressed protein pET-30a-YD1 was LMW-i. Dough mixing experiments for incorporation of 50 mg pET-30a-YD1 proteins into the base flour of weak gluten wheat cv. "Chuannong 16" indicated that the dough strength of mixing flours was noticeably weaker than that of the control, which was reflected by mixing parameters in 8-min curve width, peak width, peak height, mixing time, and right of peak slope. The results suggested that the LMW-i genes with nine cysteine residues in the present study contributed to inferior quality properties for wheat flour.

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