Construction And Analysis Of Cotton Promoter-trapping Population

Cotton is an important economic crop in China and in the world. Cotton fiber is considered as one of the best nature resource for its cellulose purity, and has an important position in textile spin industry. Cotton's functional genomics research lags far relatively. Gene-trapping mediated by T-DNA is used in identification and isolation novel gene.Now, this technique plays an important role in the plant functional genomics.In order to discover new information in cotton genome , assort with the strategies of the high-throughout plant functional genome research and provide new gene resource to the cotton biotechniques and cotton general breeding, we construct a cotton promoter-trapping system. This system uses a non-promoter GUS gene as the report gene and the NPT-II gene as the selectable gene. If the reporter gene inserted into the down stream of a promoter, it may be drived by cotton endogenesis promoter, so the expression pattern of GUS is the one of targeted gene expression pattern.Using agrobacterium-mediated transformation method, the promoter-trapping element was insert into cotton genome randomly, and its feasibility in the cotton functional genomic research was discussed. The progress we gained as follows:1) We discussed some factor's effection of the transformation efficiency of agrobacteria transforming cotton's hypocotyl, these factors include transformation time, co-culture temperature, and co-culture time. We find the best infection time, co-culture time and co-culture temperature is 5min, 60h and 21℃respectively.2) Using agrobacterum-mediated transformation method, we obtained 212 transformants and 172 plants.After PCR test, 78% transformed plants was positive transgenic plants.3) We identified these mutant plants and found some of them have significant different characterization between normal plants.4) Using histochemical localization we detected GUS gene's expression pattern and expression frequency in calli, somatic embryo, seedling stem and young leave, we found the diversity of GUS gene's expression pattern.