Function Analysis Of FAE1 Promoter

Rapeseed oil consists of different plant fatty acid, which with high erucic acid (55-60% usually) represents an important industrial feedstock for the production of lubricants. But rapeseed oil with even higher erucic acid (40-55% usually) than other plant edible oil is not easy to be digested and with low nutrition. Linoleic acid and oleic acid raise remarkably in rapeseed oil with no erucic acid by cultivar developing, oleic acid is not in animal oil, but in plant oil, and it is easy to be digested. Research on FAE1 promoter is important to learn the molecular mechanism of very-long-chain fatty acid biosynthesis and genomic engineering development of rapeseed oil components.The research mainly focus on FAE1 gene promoter deletion experiment; construction of expression vector; Arabidopsis thaliana transformation, obtaining transgenic plants and identification; GUS staining analysis. The main results were as follows:1. Five FAE1 promoter deletion fragments were obtained by deletion experiment, which have not one base fault with the full-length promoter, the full-length promoter has a whole length of 934 bp and more than 99.7% homology to known sequence of FAE1 promoter with Accession No. AF355982 in GenBank.2. A set of binary plant expression vectors with the full-length and deletion promoter driving reports genes GUS was constructed basing on the vector pBI121. They were conformed by PCR and enzyme digestion.3. 156 plants resistant to km were obtained by Arabidopsis thaliana transformation in planta. 102 plants were PCR positive in 107 resistant plants. It indicated basically that those promoter and promoter fragments had been inserted into Arabidopsis thaliana genomes.4. GUS expression identification of the transgenic plants with the full-length promoter found that it does not express in roots, stems, leaves, flowers and siliques, but in seeds, and start expressing after flowering 5 days, reaching its peak 11 days after flowering, weaking at 13 days after flowering.5. Function analysis of each promoter deletion fragment discovered that only the 406bp fragment can express abundantly in seeds.6. GUS staining analysis discovered that FAE1 promoter and all deletion fragment can not express in transgenic plant seedlings, but 35S promoter can express in the different tissues, such as roots, stems and leaves.