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Genetic modification can be an effective strategy for improving the agronomic traits of tomato (Solanum lycopersicum) to meet demands for yield, quality, functional components, and stress tolerance. However, limited numbers of available tissue-specific promoters represent a bottleneck for the design and production of transgenic plants. In the current study, a total of 25 unigenes were collected from an RNA-sequence dataset based on their annotation as being exclusively expressed in five type of tissues of tomato pericarp (outer and inner epidermis, collenchyma, parenchyma, and vascular tissues), and every five unigenes, was respectively selected from each tissue based on transcription expression. The 3-kb 5′ upstream region of each unigene was identified from the tomato genome sequence (SL2.50) using annotated unigene sequences, and the promoter sequences were further analyzed. The results showed an enrichment in T/A (T/A > 70%) in the promoter regions. A total of 15 putative tissue-/organ-specific promoters were identified and analyzed by real-time (RT) quantitative (q) PCR analysis, of which six demonstrated stronger activity than widely used tissue-specific tomato promoters. These results demonstrate how high spatiotemporal and high throughput gene expression data can provide a powerful means of identifying spatially targeted promoters in plants.

Found a trans-splicing of PHYTOENE SYNTHASE 1 alters tomato fruit color by map-based cloning, functional complementation and RACE providing an insight into fruit color development. Color is an important fruit quality trait and a major determinant of the economic value of tomato (Solanum lycopersicum). Fruit color inheritance in a yellow-fruited cherry tomato (cv. No. 22), named yellow-fruited tomato 2 (yft2), was shown to be controlled by a single recessive gene, YFT2. The YFT2 gene was mapped in a 95.7 kb region on chromosome 3, and the candidate gene, PHYTOENE SYNTHASE 1 (PSY1), was confirmed by functional complementation analysis. Constitutive over expression of PSY1 in yft2 increased the accumulation of carotenoids and resulted in a red fruit color, while no causal mutation was detected in the YFT2 allele of yft2, compared with red-fruited SL1995 cherry tomato or cultivated variety (cv. M82). Expression of YFT2 3' region in yft2 was significantly lower than in SL1995, and further studies revealed a difference in YFT2 post-transcriptional processing in yft2 compared with SL1995 and cv. M82, resulting in a longer YFT2 transcript. The alternatively trans-spliced allele of YFT2 in yft2 is predicted to encode a novel LT-YFT2 protein of 432 amino acid (AA) residues, compared to the 412 AA YFT2 protein of SL1995. The trans-spliced event also resulted in significantly down regulated expression of YFT2 in yft2 tomato, and the YFT2 allele suppressed expression of the downstream genes involved in the carotenoid biosynthesis pathway and carotenoids synthesis by a mechanism of the feed-forward regulation. In conclusion, we found that trans-splicing of YFT2 alters tomato fruit color, providing new insights into fruit color development.

Fruit firmness is an important trait in tomato ( Solanum lycopersicum), associated with shelf life and economic value; however, the precise mechanism determining fruit softening remains elusive. A yellow fruit tomato 1 ( yft1) mutant harbors a genetic lesion in the YFT1 gene and has significantly firmer fruit than those of the cv. M82 wild type at a red ripe stage, 54 days post-anthesis (dpa). When softening was further dissected, it was found that the yft1 firm fruit phenotype correlated with a difference in cellulose, hemicellulose, and pectin deposition in the primary cell wall (PCW) compared to cv. M82. Alterations in the structure of the pericarp cells, chemical components, hydrolase activities, and expression of genes encoding these hydrolases were all hypothesized to be a result of the loss of YFT1 function. This was further affirmed by RNA-seq analysis, where a total of 183 differentially expressed genes (DEGs, 50/133 down-/upregulated) were identified between yft1 and cv. M82. These DEGs were mainly annotated as participating in ethylene- and auxin-related signal transduction, sugar metabolism, and photosynthesis. This study provides new insights into the mechanism underlying the control of fruit softening.

Additional file 1: Fig. S1. Results from RNA-seq and Protein array. a Heatmap from RNA-seq representing expression levels for differentially expressed genes (DEGs) between hBMSCs and hEMSCs. Red: upregulation; Blue: downregulation. Expression intensity was based on R software analysis of gene expression levels. b Twenty categories with significant enrichment were screened for functional enrichment analysis of gene ontology (GO). Yellow line: Number of genes. Bars: Significance level. c Heatmap showing up- and down-regulation for DEGs in hBMSCs and hEMSCs related to angiogenesis. d Venn diagram and protein array analysis between hBMSCs and hEMSCs for DEGs. Black circle in Venn diagram represents DEGs related to angiogenesis between hEMSCs and hBMSCs by RNA-seq, while gray circle represents DEGs identified through protein arrays. The intersection of the two circles represents overlapping DEGs between RNA-seq and protein array for angiogenesis, where five angiogenesis-related factors were found, of which PGF and ANG4 were up-regulated in hBMSCs, while FLT-1, TEK (also known as TIE2), and FGF9 were upregulated in hEMSCs. FGF9: Fibroblast growth factor 9; FLT-1: Fms-like tyrosine kinase 1; PGF: Placental growth factor; ANG4: Angiopoietin 4. Table S1. Formulation of Differentiation Media. Table S2. Real-time qPCR primer sets.