Cloning, identification and expression analysis of Triticum aestivum 4-hydroxyphenylpyruvic acid dioxygenase gene

摘要

Tocopherols, the lipid-soluble antioxidants known collectively as vitamin E, are produced by photosynthetic organisms, which play important roles not only in human and animals but plants as well. 4-hydroxyphenylpyruvic acid dioxygenase is the first committed enzyme of this biosynthesis pathway. To better understand the biosynthesis pathway and mechanism in the one of the most important staple crop of common wheat, a cDNA sequence of 4-hydroxyphenylpyruvic acid dioxygenase gene was cloned by using in silico cloning combined with experimental RT-PCR with a GenBank accession number DQ139267. Nucleotide sequence analysis showed that the cDNA has an intact open reading frame (ORF) encoding 436 amino acids. Three tailing signals and seven in frame terminal codons were found in 3′ region. Protein multiple alignment and phylogenetic analysis suggested that common wheat HPPD was high similarity among different plant species. In silico expression analysis results showed that its expression was high correlated with photosynthesis organs. Differential expression results revealed by semi-quantitative RT-PCR were consistent with the in silico results and it was upregulated by high light stress to relieve oxidative damage, which is as the same in Arabidopsis. These results provide fundamental and important information for wheat quality improvement by using metabolic pathway engineering.