Analysis Of Differentiated Expression Of Salt Stress Related Genes By DDRT-PCR

Soil salinization is one of the major problems that can affect crop yield, which is becoming more and more serious and wide-spreaded. Beets, grown in many parts of the world, is an important economic sugar crops. Compared to other crops, sugar beet has abetter resistance salinity.That salt stress-tolerance as well as the underlying the moleculemechanisms and improving and getting more salt stress-tolerance genes of sugar beetin study is beneficial for improving the cultivation of sugar beet and other plants saltstress tolerance and of significance to enrich the resource of salt stress-tolerance.Salt stress-tolerant sugarbeet lines "O"68was used as our material in this researchand after high salt treatment (300mM NaCl), improved DDRT-PCR technology aswell as Ag staining method was used to get mRNA differential display analysis of beetleaves with different processing conditions under normal conditions as well as high saltin order to find valuable salt stress-tolerance related genes and these differentiallyexpressed cDNAfragments were cloned, sequenced and functional identification.In order to obtain beet gene fragments induced by high salt, RNA extraction kitwas used to extract treatment group and the control group total leaf RNA. Agarose gelelectrophoresis was used to detect integrity; after spectrophotometer being used todetect the purity of the RNA sample, then reverse transcription synthesizes cDNA.cDNA as a template,26random primers with three anchor primers Oligo (dT)10G (Cor A) was used to combine into78pairs of primer for PCR amplification. PCR productswere detected by polyacrylamide gel electrophoresis and silver staining was took forcoloration.More than one thousand amplified bands were obtained, differential bands50,of which the raised15, the down35. Differential fragments were selected and thesecondary amplification was in practice; recovery fragments were verified byNorthern hybridization, and the positive results are for cloning,sequencing andmatching as well as homology analysis in the NCBI.12salt stress-tolerant relatedcDNA fragments were obtained in this study, involving chloroplast mRNA and tRNA,SOS signaling pathway protein cDNA, NADH dehydrogenase cDNA, iron sulfurprotein cDNA and so on.These results suggested that high salt stress enhancedtranscription of beet chloroplast with related proteins expression; the expression ofsalt-sensitive signaling pathways (such as SOS) and related proteins under high saltconditions were altered along with it.