Arabidopsis Thaliana Screening, Body Swallowed The Family Atatg8 Cloning And Transgenic Seedlings

Abiotic stresses such as salinity, drought and low temperature affect plant growth and crop production extensively. Using gene engineering method to cultivate resistant plant is the important approach for resistant biology research. The regulation of gene expression plays an important role in plant in response to stresses. There are many stress-related genes in plant. Plant can regulates stress-related genes expression and improve the tolerance under environmental stresses. Now so many stress-related genes have been cloned and used for the resistance-engineering research, and some transgenic plants have been got which could tolerate the stresses to some extent.Cell death has always been divided into necrosis and programed cell death(PCD).And the later is a active cell death process in which the caspase plays an determined role. Most higher plants have to cope with a lack of carbohydrates at some time during their life history. Carbohydrate deprivation may occur in young seed- lings during the critical transition to autotrophy or during environmental changes leading to a decrease in photosynthesis. To sustain respiratory activity and thus survive the lack of carbohydrates, plant cells must rapidly replace carbohydrate metabolism with both protein and lipid catabolism. Protein breakdown in plant cells is mainly mediated by two different proteolytic systems, namely the selective ubiquitin proteasome-dependent proteolysis and the non-selective vacuolar proteolysis known as selfcannibalism or autophagy. The central plant vacuole that contains hydrolytic enzymes, like its animal equivalent, the lysosome, is involved in autophagy that is critical for survival during stress condition.The plant coerces in the stress circumstances usually can't maintain normal physiology function because of absorbing not enough nourishment material.Autophagy (eating oneself) is the lysosomal degradation of cytosolic components, usually the long-lived proteins and organelles, and recycle the digested food for cellular metabolism during starvation. Hence, autophagy is functionally involved in cell development, immunity, tissue remodeling and cell adaptation to the adversary circumstances. ATG8 protein is the most important component of autophagic lysosomal degradation pathway. Through ATP-dependent conjugation cascades analogous to ubiquitination, ATG8 become activated by the ATG7, and then are transferred to ATG3, which are specific for ATG8. Finally,ATG8 are covalently attached to their respective targets, the lipid phosphatidylethanolamine(PE). Linkage is via an amide bond between the C-terminal Gly carboxyl group of the tags and the free amino group in PE.The ATG8 gene was isolated from Arabidopsis by RT-PCR strategy and confirmed by sequencing. And they are completely identical to the previous reports. Then, a integrated vector of ATG8(e,f,h) and 3301 plasmid was constructed and introduced into Arabidopsis thaliana by in planta transformation method mediated by Agrobacterium tumefaciens. Transformants were screened by their ability of growing with basta. The gene transformation was confirmed by PCR analysis.