Genetic Analysis And Study On The Complementation Function Of OsELF3

Rice(Oryza sativa L.)is one of the most important food crops.Advantages such as smaller genome,well developed genetic maps and being transferred easily,made rice become a model plant of functional genomics research.After the completion of sequencing of the rice genome,the post-genome era comes.Post-genome,namely,is functional genome and proteome.In order to research the functional genome of rice,researchers created a mass of rice mutants by various methods,especially created many T-DNA inserted mutants,which contained the specific T-DNA as a tag of isolating the specific gene.In this study,the mutants with late heading date obtained by screeing of the mutants of T-DNA inserted ZH11.The flanking sequence of this mutant had been obtained by adaptor-mediated nest PCR.By sequencing and aligment,it is found that T-DNA inserted into the forth exon of the OsELF3.In this study,we investigated the agronomic characters between ZH11 and Oself3,then analysed the number of the T-DNA insertion.In order to test the function of OsELF3,we constructed the function complement vector: pZH01-OsELF3.Through agrobactetium-mediated transformation with the callus from immature embryos of Oself3 mutant,the function complement vector were introduced into rice.The main research results as follows:1 Co-segregation examining the F₂ populations of zhonghua11×Oself3 mutant and T-DNA insertion label demenstrated that the T-DNA inserted into Oself3 mutant is single copy and changes of Oself3 mutant characters were caused by T-DNA label inserted into OsELF3 gene.2 The results of agronomic characters surveying between Oself3 mutant and CK(zhonghua11) indicated Oself3 mutant has a longer heading date,narrower grain, shorter panicle and lighter kilo-grain weight than CK.Furthermore,statistical analysis showed significant difference.3 The surfaces of mature glumes of Oself3 mutant and ZH11 were observed by scanning electron microscope(SEM).There was an evident difference in the size of glumes surface cell.ZH11 has wider cells than Oself3 mutant on the surface of glumes.4 The fragment of the OsELF3 gene was cloned into the pMD20-T vector and sequenced using M13F/R primers.Then the target gene of OsELF3 was inserted into the pZH01 expression vector and the vector with OsELF3 was named pZH01-OsELF3.5 The complement expression vector pZH01-OsELF3 was transferred into callus of Oself3 mutant through Agrobactetium-mediated transformation.After tissue culture,antibiotics resistance gene screening and callus differentiation,the regeneration plants were gained. Screening these regeneration plants by PCR,11 positive plants are obtained.