Using The Microarray Technology To Study On Differential Expressed Genes Annotation And Clone Of Swainsonine (Swainsona Salsula Taub.) Roots Under Salinity Stress

Swainsonine is one of the most important drought resistant and salt tolerant legumes in Xinjiang, Salt stress is one of the most important factors limiting plant cultivation. Seed germination and seedling are markedly harmed when it is cultivated in soil salinity. The effects of different concentrations of NaCl on the germinating swainsonine seeds were studied. It was shown that: (1) there was less germination in 400 mmol/L NaCl solution than solutions, and seedling are markedly harmed in 300 mmol/L NaCl solution (2) it is implied that the inhibition of membrane restoration might be one of the important reasons for the postponed germination and declined germination vigor under salt stress. In addition, the content of proline might be related to the salt tolerance of Swainsonine.We confirmed that Swainsonine is a diploid legume through the chromosome pressing technology, and then we used Affymetrix Medicago Genome GeneChip containing 61,278 probes to identify genes whose expression changed in relative abundance without salt treatment and exposure to 180 mmol/L NaCl following 24 hours in roots. The results shown that: (1) 4,057 genes were identified to be significant differential expressed between the treatments, and we found those 155 upwards genes and 105 downwards genes; (2) We used Gene Ontology annotation to analysis those differential expressed genes at molecular function, cell components,biological process level, respectively, we implement sequence similarity search against NCBI database, and some of these genes do have similarity to known genes related to Medicago truncatula and Arabidopsis thaliana.The experimental results indicated that some of differential expressed genes are known genes related to stress-inducible gene,signal transduction,electron transport,protein metabolism and so on, and these are some of salt-stress related gene in plant root; (3) We found upwards gene from Affymetrix Medicago Genome GeneChip, according to the conserved regions of genes from other plant in Genbank, by homologous based cloning, we isolated a cDNA sequence in Swainsonine with 957 bp in length, which encoding 319 amino acid. The cDNA fragment including ORF is obtained by RT-PCR, The gene is named IFR, and the change of IFR's expression can be detected by RT-PCR.