Clone, Expression And Analysis Of Wheat Abiostress Resistance Gene Wcs120

Plant growth and development need suitable environment, under optimum conditions, it will get the maximum biomass and highest growth efficiency. Abiotic stress always cause great threat on plant growth and huge yield loss when climate is serious. So its very important to reveal the mechanism of plant resistant or tolerance to salt stress. Identify and isolate salt tolerant genes by modern genetic techniques and made some necessary genetic modification to salt sensitive plant will be greatly benefit to breeder for salt tolerance improvement of plant.Wheat is one of the important food crops in the world, to increase salt and drought tolerance of it is very crucial to its yield and quality. Shanrong No.3, the first somatic hybrid introgression line from common wheat JN177 and Agropyron elongatum, had been generated in our laboratory and with high salt and drought tolerance. To understand the mechanism of plant response to abiostress resistance, we need to study some stress-relative genes extensively, this may be very important to breeder for improve wheat abiostress resistance.Suo Li et al., obtained a large number of ESTs which related to salt stress from SR3 through the suppressive subtractive hybridization (SSH). In this paper we got a sequence from those ESTs that is greatly induced by salt stress, after clone, sequence and blast NCBI, the result shows that this gene is a cold induced protein (WCS120) gene of wheat. WCS120 is mainly induced and accumulated during the vernalization of wheat. There are some difference in wcs120 gene between Shanrong No.3 and Chinese spring, for example:3 SNPs in ORF,21 bp INDEL in promoter and there may be a alternative splicing of this gene (due to different poly A site of its mRNA). We study the expression patten of this gene in various abiostress condition (such as cold, salt and PEG) between SR3 and JN177, the result show that wcs120 gene had similar expression pattern between JN177 and SR3, while it's expression more early, strong and durable in SR3 than JN177. The OFR of wcs120 gene were inserted into pET24a+, after transformed E.coli BL21 and induced with IPTG, we got a 50 kDa protein that is heat stable (up to 100℃), the bacteria with this protein is also cold tolerance.In order to study the function of wcs120 gene in eukaryote, we constructed two eukaryotic expression vectors, one of this is fused wcsl20 gene to green fluorescent protein (GFP), after transformation of Arabidopsis thaliana protoplast, WCS120 fusion protein is targeted to nucleus and cytoplasm; another one is wcs120 gene overexpressing vector, we transfer it to Arabidopsis thaliana (Col-0) and Nicotiana tabacum (NC89), pure lines of the transgenic plants were obtained. Transgenic plants show significant tolerance to salt, drought, cold and other stress, this result confirmed that WCS120 protein does protect plants from various abiotic stresses. This work provides some clues to investigate the mechanism of plant response to abiotic stress and must be benefit to breeder for new stress resistance varieties cultivatation.