| The phytohormone abscisic acid (ABA) is an important regulatory factor during plant growth and development. It plays an important role in the synthesis of seed storage protein, water-tolerance, seed dormancy and germination, seeding growth and regulations of stress. Plants can synthetize and accumulate ABA when exposed to environmental stress such as draught, high salinity and cold. SnRK protein is a kind of Ser/Thr protein kinase widely in plants and is critical in resistance of abiotic stress by participating in many signaling pathway. So illustrating the function of SnRK in ABA signaling makes much significance.In this study we investigated the level of gene expression and kinase activity of SAPK8-10, homogenous to SnRK2.6in Arabidopsis thaliana, and their subcellular location and relationship with Osrboh. The results as follows:The kinase activity of SAPK8-10increased when leaves were exposed to100μM ABA for60min and10mM H2O2for30min while the gene expressions had no apparent changes. And the function of H2O2is earlier than ABA.The transient expression and interference system and Q-PCR were used to analyze the effect of Osrbohs when SAPK8-10were overexpressed and interfered. But we found that overexpression and RNAi of SAPK8-10had no significance to Osrbohs at transcriptional level.Coding sequence of SAPK8-10were cloned and inserted into sites of pXZP008vector for the study of subcellular location. The constructs were transformed into rice protoplasts by40%PEG and then observed with confocal laser microscope. The results showed that SAPK8-10mainly located at the cytoplasm and nucleus.For further research of the function of SAPK8-10, BiFC was performed to detect interactions between SAPK8-10and Osrbohs. CDS regions of SAPK8-10and Osrbohs were inserted into YFPN and YFPC vectors. The constructs were transformed into onion epidermal cells, and then detected fluorescence by confocal. The results showed that SAPK10and Osrboh B might interact with each other in vivo. |
