Function Analysis Of Solanum Lycopersicum SYTA And Analyzing The Interaction Between Solanum Lycopersicum SYTA And Nicotiana Benthamiana FerredoxinⅠ

S.lycopersicum as a typical representative plant of Solanaceae,occupies an important position in agricultural production.The growth of S.lycopersicum is affected by abiotic/biotic stresses such as drought,salt,heat,and low temperature,viruses and fungi,causing damage to the plants,which severely limits the yield of the S.lycopersicum.Synaptotagmin,one kind of putative Ca²⁺sensors,widely present in endocrine and neuronal cells,and can regulated the fusion process between the vesicle and the plasma membrane,playing an important role in the transport of proteins and membranes.Previous studies have reported relevant studies on Arabidopsis SYTA in plant disease resistance.In this paper,the real-time fluorescent quantitative PCR（qRT-PCR）was used to analyze the expression of S.l SYTA in different tissue and the change of its expression level by the abiotic/biotic stress in S.lycopersicum,exploring the biological function of Sl SYTA.The results showed that the expression level of S.l SYTA in different tissue of S.lycopersicum was highest in the root,followed by the leaf,and the lowest in the stem.After 12 h of cold stress,the S.l SYTA was highly expressed and up-regulated.In the first 3 h of heat stress,S.l SYTA was suppressed,although returned to normal level after12 h and then decreased.The expression of S.l SYTA did not change significantly within3 h and decreased after 12 h under the salt stress.The expression of S.l SYTA in S.lycopersicum obviously increased on the 1st day and decreased to the normal level on the 7th day after inoculation of green fluorescence protein（GFP）labled Tobacco mosaic virus（TMV）.For further investigation of the S.l SYTA influence on the process of the infection and movement of the plant viruses.The S.l SYTA was transiently expressed inthe Nicotiana benthamiana by agroinfiltration,and then ELISA was used to detection the accumulation and movement of TMV-GFP Immune adsorption assay（ELISA）was used to detect the accumulation and migration of TMV-GFP when transiently expressing Sl SYTA in N.benthamiana.The results showed that the transient expression of S.lSYTA in N.benthamiana,TMV-GFP was inoculated.After 5 days,TMV-GFP had moved to the new leaf of the N.benthamiana.But no TMV-GFP was found in the new leaf of the control N.benthamiana in which the empty vectors were transiently expressed.ELISA assay results showed that the accumulation of the TMV-GFP in the samples under treatment is obviously higher than that in the control group after 5 days of TMV-GFP inoculation.In order to completely analyze the molecular biological functions of Sl SYTA involved in plant antiviral,seven strains of S.l SYTA gene over-expressing transgenic N.benthamiana were obtained by Agrobacterium-mediated leaf disc transformation method.The qRT-PCR and Western Blot were used to verify the expression of Sl SYTA in transgenic tobacco.It was found that the expression of Sl SYTA in transgenic N.benthamiana was over-expressed,and the expression of Fd I decreased,and the expression of DCL increased.Transcriptome sequencing technology（RNA-Seq）analysis between transgenic N.benthamiana T0 and non-transgenic N.benthamiana showed that there were 4160 differentially expression gene（DEGs）,containing 1603up-regulated genes and 2557 down-regulated genes,compared to wild type.The mRNA of TMV resistance gene N-like was significantly lower in the transgenic N.benthamiana than this in the wild type,and the mRNA of PERK3 receptor protein related gene was significantly higher than that of the wild type.The qRT-PCR was used to detection the migration of TMV-GFP in transgenic N.benthamiana.The results showed that TMV-GFP had moved to the new leaf of the transgenic N.benthamiana after 72 h.However,no TMV-GFP was found in the new leaf of the wild type.The movement speed of TMV was significantly increased.To clarify of the S.l SYTA influence on infection of plant virus,yeast two-hybrid（Y2H）and bimolecular fluorescence complementation（BiFC）were used to verify the interaction between Sl SYTA and FdI protein.It was shown that yeast strains which co-transformed with Sl SYTA and FdI can normally grow on auxotrophic medium and turn blue colonies in the presence of X-α-gal,and yellow fluorescent proteins are observed under the confocal microscope.And the interaction of Sl SYTA and FdI,the96-130 amino acids of the C-terminal helical region of Fd I are required.The above indicated that S.l SYTA is highly up-regulated by stress,and plays an important role in S.lycopersicum growth and response stress.The overexpression of S.l SYTA gene can reduce the resistance of transgenic plants by regulating the expression of stress-resistance related genes.The S.l SYTA promotes the movement of plant viruses in plant cells through the interaction of the protein networks of the CP _TMV-Fd I-S.l SYTA-MP _TMV.