Construction Of Coexpression Vectors For Tomato Carbon Assimilation Related Genes And Their Genetic Transformation

Shanxi Province is a major province for the production of vegetables for plants,of which tomato?Solanum lycopersicum.Mill.?is one of the most widely grown species and has a considerable economic value.The output of fruits and vegetables depends on photosynthesis to a certain extent.When light and temperature are appropriate,CO₂ is one of the key factors restricting the quality of tomatoes in the facility.However,the CO₂ in the facility is far from meeting the demand of tomato for CO₂.Therefore,in this experiment,the Agrobacterium-mediated multi-gene co-transformation technology was used to clone the target gene.Each of the two genes was connected in tandem with the connecting peptide 2A and the guiding peptide SSU to construct a multi-gene co-expression vector and transformed into tomato.In cooperation with 908 plants,high-efficiency tomato plants were obtained through the verification of regenerated seedlings.The results of the study are as follows:1.?CA1,SBP,and FBA were cloned as target genes,and three gene fragments were ligated into the polygene ?CA1-2A-SSU-SBP-2A-SSU-FBA via the linker peptide 2A and the leader peptide SSU using overlap extension PCR technology.In the expression vector pCAMBIA1301,a multi-gene co-expression vector pCAMBI A1301-?CAI+SBP+FBA was successfully constructed.2.By constructing a multi-gene co-expression vector for tomato,C58C1 Agrobacterium fluid with pCAMBIA1301-?CA1+SBP+FBA was used to infect the cotyledon of Tomato Collaborative 908 for genetic transformation.Through the screening of anti-browning agents on the browning of the cotyledon during the induction of tomato buds,the genetic transformation system of Tomato Cooperative 908 was optimized and established.Finally,the bud induction medium was MS+0.5 mg/L6-BA+0.5 mg/LIAA+500 mg/LCef+400 mg/mL PVP/40,rooting medium MS+0.1 mg/L IAA+250 mg/L Cef,and the transgene was successfully obtained.Tomato plants.3.PCR was used to verify the DNA extracted from the cotyledons of tomato regenerated plants.Fluorescent quantitative qRT-PCR further verified that the transgenic tomato with increased expression levels of ?CA1,SBP and FBA genes was successfully obtained.