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To better understand the transcriptional regulation of high molecular weight glutenin subunit (HMW-GS) expression, we isolated four  Glu-1Bx  promoters from six wheat cultivars exhibiting diverse protein expression levels. The activities of the diverse  Glu-1Bx  promoters were tested and compared with β-glucuronidase ( GUS ) reporter fusions. Although all the full-length  Glu-1Bx  promoters showed endosperm-specific activities, the strongest GUS activity was observed with the  1Bx7 OE   promoter in both transient expression assays and stable transgenic rice lines. A 43 bp insertion in the  1Bx7 OE   promoter, which is absent in the  1Bx7  promoter, led to enhanced expression. Analysis of promoter deletion constructs confirmed that a 185 bp MITE (miniature inverted-repeat transposable element) in the  1Bx14  promoter had a weak positive effect on  Glu-1Bx  expression, and a 54 bp deletion in the  1Bx13  promoter reduced endosperm-specific activity. To investigate the effect of the 43 bp insertion in the  1Bx7 OE   promoter, a functional marker was developed to screen 505 Chinese varieties and 160 European varieties, and only 1Bx7-type varieties harboring the 43 bp insertion in their promoters showed similar overexpression patterns. Hence, the  1Bx7 OE   promoter should be important tool in crop genetic engineering as well as in molecular assisted breeding.

plos one

Expression of Wheat High Molecular Weight Glutenin Subunit 1Bx Is Affected by Large Insertions and Deletions Located in the Upstream Flanking Sequences