Expression Vector Construction Of Plant Cuticle Wax-related Transcription Factor Gene SHN1/WIN1

During their life activities, plants are subjected to a variety of environment stresses, including pathogen and insect attack, exposure to UV light. The plant cuticular forms a layer between the plant and the environment, covering aerial plant organs, and plays an important role in protecting aerial organs from damage caused by biotic and abiotic stresses. Cuticular compounds form a layer that is deposited on the outer epidermal cell wall of plant aerial organs, which is composed of cutin and waxes. Wax components are deposited both within and on the surface of the cutin polymer. This layered, semirigid hydrophobic structure makes the cuticular an efficient barrier that waterproofs and protects internal tissues. Cuticle composition and permeability affect the plant resistance on various biological and abiotic stress. This may be an efficient method in stress resistance gene engineering through the transcription factor to enhance the plant stress resistance. WIN1 influences wax accumulation through the direct or indirect regulation of metabolic pathway genes. Analysis of 35S:WIN1 overexpressors suggest that the phenotype of transgenic plants may result from a more permeable cuticular structure, and found that epidermal wax accumulation and displayed significant drought tolerance.In this paper, the leaves of Euonymus japonicus and the fruits of Capsicum frutescens was used as the experimental materials. intact cuticular leaves of Euonymus japonicus and the fruits of Capsicum frutescens were gained through method of enzymatic isolation,then trend of the cuticle ornamentation, the ultrastructure of wax and cutin were observed by scanning electron microscopy. The results show that Euonymus japonicus wax is thick, dense and lamellar; but Capsicum frutescens wax is thin and wax crystal is uneven distribution, Euonymus japonicus cutin shows network structure, which is smaller and regularer aperture to compared Capsicum frutescens cutin.An SHN1/WIN1 gene was amplified by Reverse Transcription Polymerse Chain Reaction(RT-PCR) from Wild-type Arabidopsis flower bud. A 600bp PCR product that regulates wax and cutin biosynthesis was cloned. Sequence analysis showed that SHN1/WIN1 (SHN1/WAX INDUCER1) nucleotide sequence was 100% identity with SHN1/WIN1 in GenBank. The plant express vector PBI121 was digested and ligated with the cloned cDNA fragment and got the SHN1/WIN1 gene express vector PBI121-SHN1/WIN1 regulated by CaMV35S promoter, then which was transferred into agrobacterium tumefaciens LBA4404 by the liquid nitrogen freezing thaw method. This expression vecotor provides material basis for changing cuticular structure and composition, improving plant resistance, especially drought resistance.