Cloning And Expression Of The Related Stomatal Movement Genes In Brassica Campestris Ssp. Chinensis

Chinese cabbage (Brassica campestris ssp. chinensis), which originated from China, is one of the widely cultivated vegetables grown in southern China and has an important position in the "vegetable basket" project occupy.Cold stress is an important environmental factor to limit plant growth and production, abscisic acid (ABA) regulate stomatal opening through a complex signal transduction network and it is one of the important mechanisms to obtain the resistance.H2O2 as an important signal substances involve in the ABA-induced stomatal movement. SLAC1 was firstly cloned by Negi in Arabidopsis, it is one member of the SLAHs gene family, its protein involves in the environmental stimuli and stress, salt stress, low temperature response.The studies have shown that SLAC1 related ion channels in guard cells. Under stress conditions, the protein may be produced by responding to ABA and participating in the control of ion channels to regulate stomatal closure. Research to clarify the abiotic stress responses molecular mechanism in chinese cabbage has an important practical significance. In this study, based on previous studies, by using stomata and fluorescence microscopy analysis of stomatal movement records ABA signaling pathway.Through RT-PCR technology,the gene family's cDNA sequences which control the stomatal opening are cloned from non-heading chinese cabbage.The aim of our study is to analyse molecular biology of the stomatal momvent mechanism signaling pathway in non-heading chinese cabbage.The results are as follows:1. Using stomata measurements and fluorescence microscopy analysis:Under stress stimulation, exogenous ABA inhibite stomatal opening with a dose effect. In the ABA signaling way,the H2O2 as a downstream signaling molecule produce and involve in the process of stomatal closure. Under cold stress, using RT-PCR, BcSLAC1 gene is cloned in chinese cabbage leaves,the phylogenetic analysis showes that:BcSLACl has closer genetic distance with SLAC1 genes in Arabidopsis. Real-time quantitative RT-PCR analysis showes that BcSLACl increases gene expression under the cold stress, The BcSLAC1 gene expression significantly increases more than Vc while with the treatment of ABA, indicating that the function of ABA induced BcSLACl is related with H2O2 that induced by ABA.2. On the basis of RT-PCR method and specific primers, the BcSLAHs gene family's cDNA open reading frame are cloned, they are named as:BcSLAHl, BcSLAH2, BcSLAH3. The prediction and analysis of cabbage BcSLAHs sequences:the nucleotide sequence full length of BcSLAHl is 1 146 bp, BcSLAH2:1917 bp, BcSLAH3:1614 bp. The followed amino acid composition are 381,638,557. Phylogenetic analysis showes that:compared to other BcSLAHs genes,family, BcSLAC1 has a closer genetic distance with BcSLAHl gene.