Structure, variation and expression analysis of glutenin gene promoters from Triticum aestivum cultivar Chinese Spring shows the distal region of promoter 1Bx7 is key regulatory sequence

Abstract

In this study, ten glutenin gene promoters were isolated from model wheat (Triticum aestivum L. cv. Chinese Spring) using a genomic PCR strategy with gene-specific primers. Six belonged to high-molecular-weight glutenin subunit (HMW-GS) gene promoters, and four to low-molecular-weight glutenin subunit (LMW-GS). Sequence lengths varied from 1361 to 2,554 bp. We show that the glutenin gene promoter motifs are conserved in diverse sequences in this study, with HMW-GS and LMW-GS gene promoters characterized by distinct conserved motif combinations. Our findings show that HMW-GS promoters contain more functional motifs in the distal region of the glutenin gene promoter (> -700 bp) compared with LMW-GS. The y-type HMW-GS gene promoters possess unique motifs including RY repeat and as-2 box compared to the x-type. We also identified important motifs in the distal region of HMW-GS gene promoters including the 5'-UTR Py-rich stretch motif and the as-2 box motif. We found that cis-acting elements in the distal region of promoter 1Bx7 enhanced the expression of HMW-GS gene 1Bx7. Taken together, these data support efforts in designing molecular breeding strategies aiming to improve wheat quality. Our results offer insight into the regulatory mechanisms of glutenin gene expression.

Keywords: ABRE; BAC; CTAB; DNA; E-box; GA-responsive element; GARE; GCN4-like motif; GUS; HMW-GS; LMW-GS; N-box; P-box; PBF; PCR; Regulatory motif; SDS-PAGE; SSP; The distal region of glutenin gene promoter; Transient expression; Triticum aestivum cv. Chinese Spring; UTR; abscisic acid-responsive element; bacterial artificial chromosome; cetyl trimethyl ammonium bromide; dNTP; deoxynucleotide; deoxyribonucleic acid; endosperm box; high-molecular-weight glutenin subunit; low-molecular-weight glutenin subunit; polymerase chain reaction; prolamin box; prolamin box binding factor; seed-specific protein; sodium dodecyl sulfate polyacrylamide gel electrophoresis; untranslated region; β-glucuronidase.