| Soluble sugars,organic acids,carotenoids,flavonoids,alkaloids and volatiles are contributed to the quality of citrus,apples,tomatoes and so on,which were influenced by various factors of developmental stages of fruit,and environment.Among them,the effects of hormones are especially complex.Tomatoes are excellent research material in effects of hormone response genes on the primary and secondary metabolism.We found that in citrus callus carotenoid content greatly changed by hormone treatmented,and hormone response genes caused quality related metabolites qualitative significantly changed in tomato by silence treatmented.In this study,we chose Micro-Tom tomato as the research material,techniques of real-time fluorescent quantitative PCR(q RT-PCR),high performance liquid chromatography(HPLC),liquid chromatography mass spectrometry(LC-MS)and gas chromatography mass spectrometry(GC-MS)were applied techniques to analysis the function of three hormone response genes ACO1,ABAR and GARP.The results were showed below:We constructed over-expressed OE-ACO1,OE-ABAR,and OE-GARP vectors successfully using gateway system.All of those three vectors were transformed into agrobacterium tumefaciens and then infected bud and root of tomato.Positive transgenic lines were obtained.Gene expression in each transgenic line is much higher than in wild type,which indicated that genes transformations were fulfilled.For carotenoids: The contents of total carotenoids in over-expression lines were up-regulated,expression levels in OE-GARP and OE-ACO1 were higher than in the OE-ABAR lines,while neo-xanthin,lutein of OE-GARP and OE-ACO1 have shown significant difference in comparison with the control.The composition of carotenoids in the fruit was more abundant than in the leaves,and the total content of carotenoids was significantly higher than the control.Among them,the OE-ACO1 changed significantly in the most lines,the content of neo-neoxanthin,δ-carotene and γ-carotene raised nearly as2 folds of the control,while that of lycopene up-regulated 7 folds of the higher than control,up to 147.97 ±14.6(μg/g,FW)(P < 0.05);The content of lycopene was also increased in OE-ABAR and OE-GARP plants,and the content of lutein and other metabolites was not significantly changed.For volatiles: the volatile compounds were consisted of aldehydes,alcohols,terpenoids,esters,alkanes and acids.Terpenoids were limited to be detected and quantified,so as to furans and phenolic.The proportion of 3-hexanal in the total volatiles was the highest,while the pentaerythritol and linoleic acid had significantly changed after each treatment.For other metabolites: In leaves of transgenic lines,the highest compounds isα-tomatidine,reaching 18.10±1.10(μg/g,DW).Such as petunidin-3-(trans-coumaroyl)-rutinoside-5-glucoside and naringenin chalcone compounds up or down regulated significantly.Meanwhile,four amino acids were detected in the fruit,with the highest one was phenylalanine,reaching 41.20 ± 2.90(μg/g,DW).While 26 phenolic acids and flavonoids were determined,among them,contents of 3-caffeinic acid,pomelanin,7-o-glucoside were significantly changed.Taken together,those three hormones response related gene,ACO1 、 ABAR and GARP,were of great significance in regulating primary and secondary metabolic biosynthesis of tomatoes. |
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