Fig 1.
Actinidia chinensis TPS1 genes.
A. A. chinensis TPS1 gene structure. B. A. chinensis (Ac) TPS1 predicted protein structure compared with TREHALOSE-6-PHOSPHATE SYNTHASE proteins from Arabidopsis thaliana (At), Escherichia coli (Ec) and Saccharomyces cerevisiae (Sc). The TPS and TPP domains are highlighted black and grey, respectively. C. Cladogram of Class I TPS proteins. A. chinensis (Ac) TPS1 predicted protein sequences (indicated by arrowheads) were aligned with Class I TPS proteins from Arabidopsis thaliana (At), Oryza sativa (Os), Populus trichocarpa (Pt), Solanum lycopersicum (Solyc), and Solanum tuberosum (Sotub) and a tree was constructed using the Neighbour-joining method and 1000 bootstrap replicates, with Class II AtTPS5 as an outgroup.
Fig 2.
Relative expression of Actinidia chinensis TPS1 in kiwifruit tissues, normalized to kiwifruit PP2A.
Error bars represent standard errors (SE) for three biological replicates. DAA, days after anthesis.
Fig 3.
Yeast mutant complementation assays.
A. Complementation assay of the yeast tps1Δ tps2Δ mutant. B. Complementation assay of the single tps1Δ mutant. W303-1A is the wild-type control which can grow on bot galactose and glucose. The mutation results in impaired growth on glucose. Columns represent ten-fold serial dilutions.
Fig 4.
Phenotypes of transgenic Arabidopsis constitutively expressing TPS1 genes.
A. Early growth on ½ MS medium supplemented with sucrose and kanamycin. B. Plants grown in soil.
Fig 5.
Phenotypes of transgenic Nicotiana benthamiana constitutively expressing TPS1 genes.
A. Branching and flowering of representative plants. B. Early growth on ½ MS medium supplemented with sucrose and kanamycin.
Fig 6.
The response of Nicotiana benthamiana to transiently elevated kiwifruit TPS1.2a transcript one day (T1) and three days (T3) after infiltration.
A. Agrobacterium tumefaciens carrying GFP under control of 35S promoter was used as control. Exposure to UV light confirmed uniform GFP activity, easily detected in all infiltrated leaves after three days. B. Increased GFP transcription after infiltration. C. Increased TPS1 transcription after infiltration. RPKM, frequency counts normalized to number of reads and length of gene. D. Venn diagram showing the number of differentially expressed genes at two sampling times. E. GO term categories of genes with differential expression in response to elevated TPS1.2a. The graph presents the number of differentially expressed genes between TPS1.2a and control GFP samples identified as significant at T1, T3 and both time points (T1T3).
Table 1.
Differentially expressed genes between TPS1.2a and control GFP samples identified as significant at both time points (T1 and T3).
Fig 7.
A model for TPS1 action in Nicotiana benthamiana leaf.
T6P signalling modulates transcription, hormone signals and cell wall biogenesis to coordinate growth, development and stress responses with carbon availability.