| Wheat (Triticum aestivum L.) is an important crops grown worldwide. However, due to frequent low temperature stress, the production of wheat is severely affected. Therefore, it is particularly important to breed low temperature resistant varieties of wheat. Cloning of new genes responsible to low temperature from wheat and their expressing analysis is helpful for understanding the mechanism of plant resistant to low-temperature and can provide candidate genes for molecular breeding of low-temperature resistant wheat variety.In the present study, separate new genes from low temperature-induced wheat species by means of cDNA-AFLP and verify the function and expression analysis , to provide certain theoretical foundation for breeding new wheat varieties with resistant to low temperature. The results are as follows:(1) Four gene fragments induced by low temperature were isolated, and the semi-RT-PCR results showed that expression of two genes was enhanced, and expression of other two genes was decreased when treated with low temperature.(2) Our target is to clone two genes which were decreased at transcriptional level, We directly cloned the two full-length genes from Arabidopsis, named AtADK and AtPBI whose full length is 1767 bp and 591 bp.(3) We studied the expression of AtADK and AtPBI by using the semi-RT-PCR method, and results showed that the expression of AtADK and AtPBI was relevant to the environmental stress.The result showed that both genes were responsible to not only temperature and salt, but also the plant hormones.(4) Genes were overexpressed in Arabidopsis under the control of Cauliflower Mosaic Virus 35S promoter with a binary expression vector pSN1301 in order to study gene function in plant growth and metabolism. These two genes were transferred into Arabidopsis by using the floral-dip method. Presently, we have got the T0 generation seeds, which would help us to continue further studies to affirm the function of the identified genes. |
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