The Cloning And Expression Analysis Of The ACS Family Gene Of Vitis Vinifera

During the growth and development the plant will be subjected to various biotic or abiotic stress factors, for example, drought, O3, SO2, low temperature, high salt and pathogenic bacteria etc. These stress factors will seriously affect plant growth ability, which will lead to reducing the economic benefits of the crop (reduced crop yield and quality or shortened florescence). With the arable land area decreased sharply, destruction of the ozone layer, acid rain, global warming and other effects, grape cultivation is facing severe challenge, which is currently the world’s important economic and horticultural crop. Because most of the grape cultivars are susceptible to viral, bacterial and fungal diseases, what’s more, the resistance and tolerance ability to a variety of adverse environment are poorer; The cultivation of new stress-resistant grape has become one of the effective ways of solving the above problems.1-aminocyclopropane-l-carboxylic acid synthase is the critical rate-limiting enzyme during ethylene formation process. Catalyze S-adenosylmethionine into ethylene precursor1-aminocyclopropane-l-carboxylic acid. And ethylene plays an important role in seed germination, leaf and flower abscission, fruit ripening, cell elongation, trauma and pathogen infection, responses from seed germination to fruit riping until the plant senescence in the whole life process. So the researching of1-aminocyclopropane-l-carboxylic acid synthase gene and1-aminocyclopropane-1-carboxylic acid synthase has a great significance.We got eight amino acid sequences translated from ACS genes of V. Vinifera in the NCBI database and analyse their conserved domain. The results showed that the eight ACS genes all have AAT-like conserved domains. Comparing the eight VvACS genes with the identified nine tomato ACS genes and eleven Arabidopsis ACS genes, the phylogenetic tree was established. The results showed that the eight VvACS genes have different degrees of similarity with the identified nine tomato ACS genes and eleven Arabidopsis ACS genes, and this laid a foundation for further researching VvACS genes. Phosphorylation analysis showed that the above eight VvACSs are all have multiple of serine, threonine and tyrosine phosphorylation sites, multiple of phosphorylation sites are also explained that the VvACSs are easily to be phosphorylation in post-translational modification stage, and complete the folding to form an effective three-dimensional conformation, and then transport to the specific part to play its role. Then the total RNA of grape was extracted by improved SDS method. And design primers useing the mentioned nucleotide sequenceses, and get the VvACS1, VvACS5, VvACS7, VvACS8genes, for the followed experiment laid the foundation. To investigate the VvAVS5and VvACS8genes relative expression induced by exogenous ethylene, salicylic acid and wounding, we used real-time fluorescence quantitative PCR. The results showed that ethylene can inhibit the expression of VvACS5and VvACS8, while salicylic acid and wounding treatment promote VvACS5and VvACS8expression in different degrees. This lay a theoretical basis for the construction of gene over expression vector in the followed experiment.