RESUMEN
Increasing the yield of plant oil is an important objective to meet the demand for sustainable resources and energy. Some attempts to enhance the expression of genes involved in oil synthesis in seeds have succeeded in increasing oil content. In many cases, the promoters of seed-storage protein genes have been used as seed-specific promoters. However, conventional promoters are developmentally regulated and their expression periods are limited. We constructed a chimeric promoter that starts to express in the early stage of seed development, and high-level expression is retained until the later stage by connecting the promoters of the biotin carboxyl carrier protein 2 (BCCP2) gene encoding the BCCP2 subunit of acetyl-CoA carboxylase and the fatty acid elongase 1 (FAE1) gene from Arabidopsis. The constructed promoter was ligated upstream of the TAG1 gene encoding diacylglycerol acyltransferase 1 and introduced into Arabidopsis. Seeds from transgenic plants carrying AtTAG1 under the control of the chimeric promoter showed increased oil content (up by 18â»73%) compared with wild-type seeds. The novel expression profile of the chimeric promoter showed that this could be a promising strategy to manipulate the content of seed-storage oils and other compounds.
Asunto(s)
Acetil-CoA Carboxilasa/genética , Acetiltransferasas/genética , Proteínas de Arabidopsis/genética , Diacilglicerol O-Acetiltransferasa/genética , Aceites de Plantas/metabolismo , Semillas/genética , Arabidopsis/genética , Elongasas de Ácidos Grasos , Ácidos Grasos/genética , Ácidos Grasos/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Aceites de Plantas/química , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Regiones Promotoras Genéticas/genética , Proteínas de Almacenamiento de Semillas/biosíntesis , Proteínas de Almacenamiento de Semillas/química , Proteínas de Almacenamiento de Semillas/genética , Semillas/químicaRESUMEN
Plastidic acetyl-CoA carboxylase (ACCase) regulates the rate of fatty acid synthesis. This enzyme is composed of biotin carboxyl carrier protein (BCCP), biotin carboxylase (BC), and carboxyltransferase (CT), which consists of α and ß subunits. Among these components, CTß is encoded by the plastidic genome. In Arabidopsis, BC and CTα are each encoded by a single gene, and there are two genes for BCCP, BCCP1 and BCCP2. Promoter analysis revealed that the 5'-UTR containing the AW box is necessary for the expression of these genes in seeds and seedlings. The results indicated that there are other transcription factors besides WRI1 that bind to the AW box and regulate these genes in organs other than seeds. Although the AW boxes at 748 and 532 bp upstream from the transcription start sites (TSSs) of the BC and CTα genes, respectively, were not functional in seeds, the latter was functional in seedlings. In addition, when these AW boxes were moved to approximately 200 bp upstream from the TSS, they became active in seeds but not in seedlings. These results suggest that the distance from the TSS affects the function of the AW box, and the AW box alone is not sufficient for expression in seedlings. A comparison of the protein levels of BC, BCCP1, BCCP2 and CTß between a wri1 mutant, a WRI1-overexpressing line and control plants showed that protein levels of BCCP2 and BC but not BCCP1 and CTß are affected by WRI1. The results suggest that ACCase subunits are differentially regulated by WRI1.