The characterization and expression analysis under stress conditions of PCST1 in Arabidopsis

Analysis of PCST1 expression characteristics and the role of PCST1 in response to osmotic stress in Arabidopsis thaliana. The structure of PCST1 was analyzed using Bioinformatics method. Real-time PCR, GUS tissue localization and subcellular localization were adopted to analyze the expression pattern of PCST1 in Arabidopsis. To validate the transgenic positive strain of PCST1 using Real-time PCR, overexpression experiments were performed in wild type. Full-length cDNA was cloned and connected into a binary vector with 35S promoter, and the construction was transformed into wild type. With NaCl and mannitol treatments, the germination rate, green leaves rate, physiological indexes were carried out and counted in Arabidopsis with overexpression of PCST1 and T-DNA insertion mutants. The molecular mechanism of PCST1 in response to osmotic stress in Arabidopsis was analyzed. Based on the bioinformatic analysis, PCST1 is a hydrophobin with 403 amino acids, and the molecular weight is 45.3236 KDa. It contains only the START (the lipid/sterol - binding StAR - related lipid transfer protein domains) conservative domain. PCST1 possesses phosphatidylcholine binding sites and transmembrane region. Expression pattern analysis showed that expression of PCST1 increased with time. The PCST1 widely expressed in Arabidopsis, including roots, axils of stem leaves, flowers (sepal, conductive tissue of the petal, thrum, anther and stigmas), and the top and basal parts of the siliquas. It mainly localized in cell membrane. The overexpression of PCST1 enhanced the sensitivity to osmotic stress in Arabidopsis based on the germination rate. While expression of PCST1 decreased, and the sensitivity to osmotic stress had no obvious change in Arabidopsis. Its molecular mechanism study showed, that PCST1 response to osmotic stress resistance by regulating the proline, betaine synthesis, as well as the expression of key genes SOS, NCED, CIPK. PCST1 is composed of 403 amino acids. The START conservative domain, a transmembrane structure, the phosphatidyl choline binding sites are contained in PCST1. It is localized in cytoplasmic membrane. The PCST1 widely expressed in the root, leaf, flower and siliquas. NaCl and mannitol suppressed the expression of PCST1 and PCST1 can negatively control action of Arabidopsis in the osmotic stress. PCST1 regulates the synthetic pathway of proline, betaine and the expression of SOS, NCED and CIPK in response to the osmotic stress resistance.