The Expression Of Calmodulin-binding Transcription Factor SR2L In Tomato Flower Pedicel Abscission And Analysis Of Transgenic Plant

Plant organ abscission is a part of the plant body droped from the mother plant.It is a common phenomenon that the plant could adapt to the environment.In the crop production process,the abnormal abscission is an important factor affecting crop yield.Protein phosphorylation is a kind of post-translational modification,plays an important role in the process of plant growth and development,and the process of plant organ abscission.Thisresearch based on the identification of phosphorylated proteins in the process of abscission,the calmodulin-binding transcription factor family was selected as the research object.The expression level of different calmodulin-binding transcription factors in abscission was analyzed.The selection was closely related to the abscission process.This gene was transferred into cultivated tomato in order to clarify the regulation of the process in the tomatoflower pedicel abscission.The main results of the paper are as follows:?1?According to treated tomato flower pedicel with ethylene,1-MCP,Ca²⁺ EGTA.The abscission rate and the expression of SR1L,SR2,SR2L,SR3.SR3L,SR4 were analyzed at different time.The results showed that only SR2L had no significant change in gene level.Combined with the results of phosphorylated peptide analysis of different flowerabscission time in ethy'lene treatment.The phosphorylation level was significantly different,but the level of protein transcription was not obvious,that is,SR2L in the phosphorylation andabscission process is closely related.Therefore,from the selection of calcium phosphate transcription factor SR2L to shed the process of phosphorylated protein research.?2?The expression vector pB7YWG2-SR2L,pB7YWG2-SR2Ls9'4A was constructed by ligation the SR2L and SR2LS914A genes with the plasmid,by TA cloning vector pMD18-T.Agrobacterium tumefaciens-mediated method was used to transferr into tomato cotyledons and screened to obtain resistant tomato plant.Finally,3 TO generation pB7YWG2-SR2L and 4 TO generationtransgenic tomato plantswere obtained,The rate of TO plant were 2.9%,4.1%?3?Select the excitation wavelength of 515nm,laser confocal detection of the obtained plant roots.The results showed that compared with the wild type "Zhongshu No.6 tomato"there was a significant yellow signal in the nucleus in the lateral root of the transgenic plants.These results suggest that SR2L is located in the nucleus of the tomato epidermis.PCR detection,qRT-PCR and laser confocal detection of TO and T1 generation plants,the result showed that the target gene had been successfully integrated into the tomato genome and transcribed at the RNA level?4?By analysis the changes of abscission rate of pedicels in phosphatase inhibitors treatment and the abscission rate of T1 tomatotransgenicplants,two speculations were obtained:SR2L was localized to the nucleus,SR2L might bind to EIN2 in the nucleus and undergo a dephosphorylation to control the response of EIN3.SR2L may also be directly E1N3 promoter region is used to participate in the ethylene-controlled abscission reaction.