Cloning Of Tomato Response To Cadmium Stress Related Gene SlJMJ And Functional Verification

Plants are threatened by many abiotic stresses during growth and development,such as heavy metal stress.Heavy metal ions are characterized by strong concealment and toxicity,which seriously affect the growth and development of plants.Solanaceae lycopersicum is one of the important vegetable crops,and its fruit is rich in vitamin C.Tomatoes mainly adopt the facility cultivation method.The growth period is long,the multiple cropping index is high,and the excessive application of fertilizers and pesticides is serious,which leads to serious degradation of soil quality in tomato cultivation areas,and the problems of heavy metal residues are increasingly prominent.Therefore,as the demand for tomatoes increases,it is urgent to cultivate tomatoes that are not polluted by the environment.SlJMJ is a histone demethylase.Many scholars have studied the processes involved in drought,chilling and salt stress in Arabidopsis,but their response to plant tolerance to cadmium stress has rarely been reported.In this paper,"Red Luocheng" tomato was used as experimental material to analyze its function under stress conditions.It is expected to provide a theoretical basis for furt.The main research results obtained are as follows:1.Cloning and sequence analysis of the S1 JMJ geneThe SlJMJ gene was cloned from tomato.Sequence analysis showed that the SlJMJ gene was 1254 bp in length and encoded 417 amino acids,containing the JmjC domain and belonging to the JmjC domain only subfamily.her study of histone demethylase gene in plant abiotic stress.2.SlJMJ gene expression in heavy metal Cd stress induced by EBLTomato seedlings were treated with different concentrations of heavy metals Cd(0?10?20?30 mmol/L),and tomatoes were treated with heavy metal Cd(10 mmol/L)and then sprayed with EBL 0?3?6?12 h taken.The expression pattern of SlJMJ gene was analyzed by qRT-PCR.The results showed that theSlJMJgene was up-regulated at a concentration of 10 mmol/L in Cd treatment and up-regulated at 6 h after E-spray after Cd stress.3.Transcriptional activity analysis of the S1 JMJ geneUsing the yeast one-hybrid system,the recombinant plasmid pGBKT7-SlJMJ was constructed and transferred into yeast AH109.The fusion vector pGBKT7-SlJMJ can be grown on SD/-Trp-His deletion medium and contained in SD/-Trp-His containing X-?-Gal.showed a blue color on the deletion medium,indicating that the SlJMJ protein has transcriptional activation activity.4.Yeast Genetic Transformation and Stress Resistant Analysis of S1 JMJ GeneThe yeast expression vector pYES2-SlJMJ was constructed,transformed into yeast cells,and the transgenic yeast was salt(400?500?600 mmol/L),drought(0.5?1?1.5 mol/L)and heavy metal Cd(5?15?25?mol/L)treatment showed that the transgenic yeast showed some tolerance to salt,drought and Cd stress by observing the growth of yeast.5.SlJMJ gene genetic transformation of Arabidopsis thalianaThe pBI121-SlJMJ plant expression vector was constructed and the model plant Arabidopsis thaliana was genetically transformed by Agrobacterium tumefaciens infection method.Using kanamycin for screening,the green plants were screened and cultured,and the Arabidopsis thaliana plants were identified by molecular identification(PCR).Three Arabidopsis thaliana positive seedlings were obtained and T2 seeds were harvested.Compared with the wild type,the transgenic Arabidopsis thaliana leaves were thick and the fresh weight increased,and the expression of SlJMJ gene was significantly increased.