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. 2020 Oct 14;10:17219. doi: 10.1038/s41598-020-73709-6

Figure 1.

Figure 1

Schematic overview of stable combinatorial-transformation of tomato plants to simultaneously introduce multiple genes under different promoters to confer appropriate tissue specificity. Transgenes are involved in three different processes of carbon and nitrogen fluxes. (i) assimilation ([1] SlmMDH, Solanum lycopersicum mitochondrial malate dehydrogenase; [2] AtSBP, Arabidopsis thaliana sedoheptulose 1,7-bisphosphatase; [3] SlSPA, Solanum lycopersicum sugar partitioning affected; [4] EcPP, Escherichia coli pyrophosphatase; [5] NtGS2, Nicotiana tabacum chloroplast glutamine synthetase 2; [6] FpGLDH, Flaveria pringlei H-protein of glycine decarboxylase); (ii) transport ([7] AtSWEET11, Arabidopsis thaliana sugar efflux transporter 11; [8] AtSUC2, Arabidopsis thaliana sucrose transporter 2; [9] AtAAP1, Arabidopsis thaliana amino acid permease 1); and (iii) sink metabolism ([10,11] AtSUC2/9, Arabidopsis thaliana sucrose transporter 2/9; [12, 13] AtSTP3/6, Arabidopsis thaliana sugar transporter 3/6; [14] SpLIN5, Solanum pennellii tomato apoplastic invertase 5; [15] AtSUS1, Arabidopsis thaliana sucrose synthase 1; [16] ShAgpL1, Solanum habrochaites Large subunit of ADPglucose pyrophosphorylase 1; [17] AtTMT1, Arabidopsis thaliana tonoplast monosaccharide transporter 1; [18] AtAAP6, Arabidopsis thaliana amino acid permease 6; [19] SlINVINH, Solanum lycopersicum apoplastic invertase inhibitor; [20] SlCAT9, Solanum lycopersicum cationic amino acid transporter 9). Overexpression (showed as red color) or silencing (showed as blue color) of these genes were achieved using different tissue-specific promoters; (i) leaf- and mesophyll-specific, ribulose-bisphosphate carboxylase (RbcS), and fructose-1,6-bisphosphate (cyFBP); (ii) constitutive, 35S-cauliflower mosaic virus (35S); (iii) companion cell-specific, commelina yellow mottle virus (CoYMV); (iv) fruit specific, patatin B33 (B33), and ripening-specific ethylene-inducible E8 (E8); and (v) native promoter of S. habrochaites Large subunit of ADPglucose pyrophosphorylase 1 (ShAgpL1). Transgenic lines were grown under glasshouse and polytunnel conditions. SlSPA resides in the plastid but is not known to catalyze an enzymatic reaction, GLDH is associated to the inner mitochondrial membrane where it catalyzes the terminal reaction of ascorbate biosynthesis.