Cloning And Expressing Of ELO Homologue Genes In Arabidopsis Thaliana And Phenotypical Characteriazation Of CERA Mutant Deficient In Stem Cuticular Wax

Very long chain fatty acids play important roles in plant growth and development. In order to make foundation to investigate whether ELO homologue genes are involved in very long chain fatty acid biosynthesis, The cDNA sequences of At1g75000, At4g36830 and At3g06470 were amplified by RT-PCR and fused in the C-terminal of eGFP to form plant transient expression vectors. After transforming these vectors into onion epidermal cells by gene gun, we tested the expression result of vectors by fluorescence microscope. We have successfully cloned cDNA of target genes: At1g75000, At4g36830 and At3g06470, made constructs fused with eGFP in plant transient expression vectors At1g75000::eGFP::NOS/pPZP21, At4g36830::eGFP::NOS/pPZP211 and At1g06470::eGFP ::NOS/pPZ P211 respectively, transformed above vectors in onion epidermal cells and examined green fluorescence. These plant transient expression vectors fusing At1g75000, At4g36830 and At3g06470 with eGFP were successfully constructed and would make a foundation to investigate the function of AtELO proteins.The surface of terrestrial plants is covered by a layer of wax that plays important roles in plant growth and plant-environment interaction. Wax consists of a mixture of very long chain fatty acids (VLCFAs) and other hydrophobic components derived from VLCFAs. By chemically mutagenizing seeds of Arabidopsis thaliana with ethyl-methylsulfonate (EMS), a mutant expressing light green stems and shorter siliques was obtained and named cera. Genetic analysis revealed that the phenotype of cera was led by mutation of a single recessive gene. Scanning electron microscopy on cera stem displayed a severe wax deficient phenotype. Further analysis of chemical composition and content of epicuticular wax on cera stem surface suggested that CERA played an important role in elongating more than twenty four carbons very long chain fatty acids in Arabidopsis thaliana.