Uniform ripening Encodes a Golden 2-like Transcription Factor Regulating Tomato Fruit Chloroplast Development

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Science  29 Jun 2012:
Vol. 336, Issue 6089, pp. 1711-1715
DOI: 10.1126/science.1222218

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  1. Fig. 1

    Fruit phenotypes. (A) Immature green fruit (IM, 15 dpa) from the S. lycopersicum “M82” x S. pennellii introgression population and S. lycopersicum “Moneymaker” x S. pimpinellifolium backcross lines. (B) IM fruit from S. lycopersicum “T63” containing the CaMV35S promoter (Tcontrol) or AtGLK1 or AtGLK2 with the Ca35S (p35S), RbcS (pRbcS), LTP (pLTP), or PDS (pPDS) promoter. (C) Mature green (32 dpa) fruit from “Ailsa Craig” U/U and “Craigella” u/u. (D) Mature green fruit from Ailsa Craig U/U containing p35S::SlGLK2 with overexpression (OE) or cosuppression (CS) of SlGLK2 and from M82 u/u overexpressing p35S::SlGLK2.

  2. Fig. 2

    Map position and sequence of SlGLK2 alleles. (A) Left to right, the classical morphological map (26), IL map (27), maps derived from the IL 10-1 F2 and the MM x “T0937” populations, candidate genes in the 60,507-bp region defined by markers, and gene model of SlGLK2 with the additional A underlined and the resulting stop codon in Slglk2/u allele circled. (B) SlGLK2/U and Slglk2/u coding sequences. Nucleic acid calls between SL2.40ch10:2292143-2295824 (SGN tomato release 2.4) from cDNA and genomic sequencing of the u/u S. lycopersicum varieties “Heinz 1706” (HEI), M82, T63, Moneymaker (MM), “Castlemart” (CSM), “E6203” (E6), “Fireball” (FB), “Long Red” (uLR), “N93,” S. lycopersicum var. cerasiforme PI114490 (Cer), and Craigella (CRG) and the U/U S. pimpinellifolium (Spi), S. pennellii (Spe), and S. lycopersicoides (Sha) wild tomato relatives and S. lycopersicum varieties Ailsa Craig (AC), “73X,” “T91,” and a “Cuatomate” (CAU) landrace. Translated reverse transcription polymerase chain reaction (RT-PCR) products and Basic Local Alignment Search Tool (BLAST) searches predicted the start codon at 2292050; no differences were detected until position 2292143. The sequence differences at 2292260-2292267 of all u/u varieties compared with all U/U varieties is boxed in orange.

  3. Fig. 3

    GLK expression and chlorophyll accumulation. (A) SlGLK1 and SlGLK2 RT-PCR products from Ailsa Craig U/U and Craigella u/u cotyledons (C), young leaves (YL), developed leaves (OL), flower petals (P), stamens (S), immature fruit (IM, 10-15 dpa), and the pedicel shoulders (Pd) or the stylar ends (St) of mature green (MG, 32 dpa) fruit and ripe fruit (RR, 46 dpa). Results are typical of replicated RNA samples. (B) Chlorophyll in the outer pericarp and epidermis of IM fruit from AtGLK1- or AtGLK2-expressing lines. Statistical significance by means of general linear model (GLM) and Bonferroni multiple comparison test (MCT) at P < 0.05 are indicated. (C) Quantitative RT-PCR of AtGLK1 and AtGLK2 expression in IM fruit. Statistical significance by means of GLM and Tukey’s honestly significant difference (HSD) test at P < 0.05 are indicated by roman (AtGLK1) and italic (AtGLK2) letters, respectively. (D) Chlorophyll in leaves from AtGLK-expressing lines. Statistical significance determined by means of GLM and Bonferroni MCT at P < 0.05 are indicated.

  4. Fig. 4

    AtGLK-expressing fruit characteristics. (A) TEM of IM (15 dpa) green fruit (top row) and leaf (bottom row) chloroplasts, 0.5 μm scale. (B) Cellular components Gene Ontology (GO) terms for significantly (P < 0.05, fold change >2) overexpressed genes in IM fruit identified in microarray hybridizations. The total number of genes with known GO terms is shown below bars. (C) GO categories of overrepresented genes (P < 0.05, n > 3 genes) whose transcript abundance were different (P < 0.05, fold change >2). *P < 0.0001; **0.0001 < P < 0.001; ***0.001 < P <0.01. (D) Starch in IM fruit pericarp and epidermis. (E) Glucose (dark bar) and fructose (light bar) in red ripe (RR, 42 dpa) fruit. (F) Total soluble solids in RR fruit. Statistical significance determined by means of GLM and Tukey’s HSD at P < 0.05 are indicated.