Cloning Of Chorispora Bungeana Gene Cb4773 And Analyzing Of Its Functions In Abiotic Stress

Chorispora bungeana is a typical perennial herb that belongs to Brassicaceae and it could survive in rapid changing freezing temperature from an ice-free cirque in Tianshan Mountains, Xinjiang. The survival ability of Chorispora bungeana in cold environment is of great values, They may be associated with particular genes to adapt to harsh environment. According to the some related articles and gene Cb4773 increased expressed in response to low temperature. We cloned the Chorispora bungeana gene Cb4773 and its Arabidopsis thaliana homologous gene At5g62360,overexpression of Arabidopsis thaliana study its biological function and characteristics, The main research contents and results are as follows:1.By the RACE and Tail-PCR, obtain Cb4773 gene sequences and promoter sequences, coding region sequence contains the 591 bp,the promoter sequence contains1320 bp. By BLAST alignment obtains similarity was 88% in the evolutionary relationships between coding sequence and At5g62360. Analyzing their sequences and protein structure characteristics found they contain PMEI domain structure with the same function and the Cb4773 contains transmembrance domains.2. Cb4773 and At5g62360 promoter sequences connect with GUS reporter gene and analyse tissue orientation. By GUS staining,Cb4773 and At5g62360 appears to mainly localize in microtubules of root the respectively, leaf, pods, filaments and stima.By construct a Cb4773 and At5g62360 coding area linked with YFP found the subcellular localization is in the plasma membrane.3. The obtainment of Cb4773 overexpressed genes and At5g62360 and abiotic stress treatment, found that the transgenic plants frost resistance and salt resistance improved. After cold acclimation can increase the content of proline and total sugar. The overexpression analysis of Cb4773 expressed genes and At5g62360 found could inhibit the PME activity, Atpme41 mutants can inhibit PME activity. Inhibition of PME activity may cause transgenic Cb4773 and At5g62360 plant mature early in advance. Inhibition of PME activity may cause mutant BR deletion mutant det2 could restore some partial phenotypes, such as the growth of root and stem branch and so on.These conclusions in the experiment have important significance in reponse to the abiotic stress, RT-PCR studies showed that they were expressed in response to low temperature,The research of this gene plays significant roles in agricultural production. The determination of LT50 conclude the lethal temperature lower than that of WT after plants overexpressed and they have more frost resistance. The phenotype of these overexpressed genes have great significances in later crops maturing early and new varieties cultivating.