Deletion analysis of the Brassica napus cruciferin gene cru 1 promoter in transformed tobacco: promoter activity during early and late stages of embryogenesis is influenced by cis-acting elements in partially separate regions

Abstract

To define sequences in the cruciferin gene cru1 promoter of importance for expression, tobacco (Nicotina tabacum L.) plants were transformed with constructs in which the cru1 promoter, in front of the intact cru1 structural gene, was truncated at -1216, -974, -736, -515, -306, -46 and -17 bp relative to the cap-site. Cru1 expression in tobacco seeds was studied by Northern analysis, Western analysis and in-situ hybridizations. Comparisons of the Northern analysis of RNA from tobacco seeds harvested at 18 d after pollination with the Western analysis of protein from mature seeds showed that the regions between -974 to -736 and -306 to -46 were important for the expression of cru1 at an early developmental stage, whereas the regions -736 to -515 and -515 to -306 were important for expression throughout embryogenesis. By investigating the mRNA levels in transgenic seeds at different stages of development, indications were obtained that the two latter regions exerted their effects during the later stages. The in-situ hybridization showed that cru1 mRNA was distributed in parenchyma cells throughout the embryo in seeds expressing constructs -974 and -736. Constructs -515 and -306 showed an expression restricted to the axis or axis and parts of the cotyledons. Sequence comparisons of the cru1 promoter with other storage-protein gene promoters, identified several motifs implicated in gene regulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Abscisic acid regulates pinoresinol–lariciresinol reductase gene expression and secoisolariciresinol accumulation in developing flax (Linum usitatissimum L.) seeds

“…In this region, SEF3 and SEF4 motifs are good candidates because these motifs are important binding sites for transcription factors that enhance gene expression of several storage proteins in soybean seed (for example, b-conglycinin, Lessard et al 1991;or 7S globulin, Hayashi et al 1997). These motifs, observed in various species such as in the promoter of the cruciferin CRU1 gene of Brassica napus (Sjödahl et al 1995), were also observed in the promoter region of the chalcone synthase genes CHS7 and CHS8 which are expressed in the seed coat in soybean (Yi et al 2010). …”

Quality Evaluation of Plant-Derived Foods Ⅱ

“…Conserved motifs exist in the promoter regions of seed storage protein genes across diverse plant species [157] suggesting that the regulation of seed storage protein biosynthesis is governed by transcriptional mechanisms that evolved early in the evolution of the plant kingdom. The disruption of some of these conserved motifs in the canola cruciferin-encoding cru1 gene [158] and the napin-encoding napA gene [159] led to reduced accumulation of the respective protein product. Multiple transcription factors are known to regulate the expression of seed storage protein genes in Arabidopsis: ABSCISIC ACID INSENSITIVE 3 (ABI3) [160][161][162][163], FUSCA 3 (FUS3) [161,164], LEAFY COTYLEDON 1 (LEC1) [164][165][166], LEC2 [161], and multiple MYC transcription factors [167].…”

Breeding Canola (Brassica napus L.) for Protein in Feed and Food

“…Conserved motifs exist in the promoter regions of seed storage protein genes across diverse plant species [ 157 ] suggesting that the regulation of seed storage protein biosynthesis is governed by transcriptional mechanisms that evolved early in the evolution of the plant kingdom. The disruption of some of these conserved motifs in the canola cruciferin-encoding cru1 gene [ 158 ] and the napin-encoding napA gene [ 159 ] led to reduced accumulation of the respective protein product. Multiple transcription factors are known to regulate the expression of seed storage protein genes in Arabidopsis : ABSCISIC ACID INSENSITIVE 3 ( ABI3 ) [ 160 , 161 , 162 , 163 ], FUSCA 3 ( FUS3 ) [ 161 , 164 ], LEAFY COTYLEDON 1 ( LEC1) [ 164 , 165 , 166 ], LEC2 [ 161 ], and multiple MYC transcription factors [ 167 ].…”