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Tomato R2R3-MYB Proteins SlANT1 and SlAN2: Same Protein Activity, Different Roles.

Kiferle C, Fantini E, Bassolino L, Povero G, Spelt C, Buti S, Giuliano G, Quattrocchio F, Koes R, Perata P, Gonzali S - PLoS ONE (2015)

Bottom Line: Despite the fact that cultivated tomato varieties do not accumulate anthocyanins in the fruit, the biosynthetic pathway can be activated in the vegetative organs by several environmental stimuli.Little is known about the molecular mechanisms regulating anthocyanin synthesis in tomato.Here, we carried out a molecular and functional characterization of two genes, SlAN2 and SlANT1, encoding two R2R3-MYB transcription factors.

View Article: PubMed Central - PubMed

Affiliation: PlantLab, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.

ABSTRACT
Anthocyanins are water-soluble polyphenolic compounds with a high nutraceutical value. Despite the fact that cultivated tomato varieties do not accumulate anthocyanins in the fruit, the biosynthetic pathway can be activated in the vegetative organs by several environmental stimuli. Little is known about the molecular mechanisms regulating anthocyanin synthesis in tomato. Here, we carried out a molecular and functional characterization of two genes, SlAN2 and SlANT1, encoding two R2R3-MYB transcription factors. We show that both can induce ectopic anthocyanin synthesis in transgenic tomato lines, including the fruit. However, only SlAN2 acts as a positive regulator of anthocyanin synthesis in vegetative tissues under high light or low temperature conditions.

No MeSH data available.


Related in: MedlinePlus

Identification of possible tomato R2R3-MYB, bHLH, and WDR regulators of anthocyanin synthesis.Evolutionary relationships of R2R3-MYB (A), bHLH (B) and WDR (C) proteins involved in anthocyanin pigmentation in different plant species. The evolutionary history was inferred using the Neighbor-Joining method [42]. The optimal tree with the sum of branch length (A = 4.04792561, B = 3.00531532, C = 0.83964559) is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches [57]. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the p-distance method [58] and are in the units of the number of amino acid differences per site. The analysis involved 20 (A), 14 (B) and 12 (C) amino acid sequences. All ambiguous positions were removed for each sequence pair. There were a total of 790 (A), 364 (B) and 372 (C) positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [41]. Expression heatmap (Log2 scale of FPKM values) of SlAN2, SlANT1, SlAN1, SlAN11, SlJAF13 and SlDFR genes in different tissues of tomato, analyzed by Illumina RNA-Seq (D). MG: Mature Green fruit; B: Breaker fruit; B+10: ripe fruit 10 days after breaker stage. Subcellular localization of GFP-SlANT1 and GFP-SlAN2 fusion proteins in transiently transformed A. thaliana mesophyll protoplasts (E). Pictures were taken with bright field, green fluorescent protein (GFP) and 4’6-diamidino-2-phenylindole (DAPI) filters.
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pone.0136365.g001: Identification of possible tomato R2R3-MYB, bHLH, and WDR regulators of anthocyanin synthesis.Evolutionary relationships of R2R3-MYB (A), bHLH (B) and WDR (C) proteins involved in anthocyanin pigmentation in different plant species. The evolutionary history was inferred using the Neighbor-Joining method [42]. The optimal tree with the sum of branch length (A = 4.04792561, B = 3.00531532, C = 0.83964559) is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches [57]. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the p-distance method [58] and are in the units of the number of amino acid differences per site. The analysis involved 20 (A), 14 (B) and 12 (C) amino acid sequences. All ambiguous positions were removed for each sequence pair. There were a total of 790 (A), 364 (B) and 372 (C) positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [41]. Expression heatmap (Log2 scale of FPKM values) of SlAN2, SlANT1, SlAN1, SlAN11, SlJAF13 and SlDFR genes in different tissues of tomato, analyzed by Illumina RNA-Seq (D). MG: Mature Green fruit; B: Breaker fruit; B+10: ripe fruit 10 days after breaker stage. Subcellular localization of GFP-SlANT1 and GFP-SlAN2 fusion proteins in transiently transformed A. thaliana mesophyll protoplasts (E). Pictures were taken with bright field, green fluorescent protein (GFP) and 4’6-diamidino-2-phenylindole (DAPI) filters.

Mentions: By analyzing the whole genome of tomato, all the annotated putative MYB proteins were compared with the main MYB regulatory factors involved in anthocyanin synthesis in other species (S1 Fig). Four different tomato MYB proteins, encoded by the genes Solyc10g086250, Solyc10g086260, Solyc10g086270, and Solyc10g086290, corresponding, respectively, to SlAN2 [33–35], SlANT1 [32,36], SlANT1like [35,54] and SlAN2like [35], grouped in one clade with MYB proteins of tobacco (N. tabacum), petunia and potato (S. tuberosum) involved in anthocyanin synthesis [5,55,56] (S1 Fig, Fig 1A). This clade included members from the Solanaceae family and was clearly separated from anthocyanin MYBs from other dicots, such as A. thaliana or A. majus, and monocots (S1 Fig, Fig 1A). This analysis confirms that SlANT1 and SlAN2 are indeed tomato MYB TFs involved in anthocyanin synthesis regulation.


Tomato R2R3-MYB Proteins SlANT1 and SlAN2: Same Protein Activity, Different Roles.

Kiferle C, Fantini E, Bassolino L, Povero G, Spelt C, Buti S, Giuliano G, Quattrocchio F, Koes R, Perata P, Gonzali S - PLoS ONE (2015)

Identification of possible tomato R2R3-MYB, bHLH, and WDR regulators of anthocyanin synthesis.Evolutionary relationships of R2R3-MYB (A), bHLH (B) and WDR (C) proteins involved in anthocyanin pigmentation in different plant species. The evolutionary history was inferred using the Neighbor-Joining method [42]. The optimal tree with the sum of branch length (A = 4.04792561, B = 3.00531532, C = 0.83964559) is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches [57]. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the p-distance method [58] and are in the units of the number of amino acid differences per site. The analysis involved 20 (A), 14 (B) and 12 (C) amino acid sequences. All ambiguous positions were removed for each sequence pair. There were a total of 790 (A), 364 (B) and 372 (C) positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [41]. Expression heatmap (Log2 scale of FPKM values) of SlAN2, SlANT1, SlAN1, SlAN11, SlJAF13 and SlDFR genes in different tissues of tomato, analyzed by Illumina RNA-Seq (D). MG: Mature Green fruit; B: Breaker fruit; B+10: ripe fruit 10 days after breaker stage. Subcellular localization of GFP-SlANT1 and GFP-SlAN2 fusion proteins in transiently transformed A. thaliana mesophyll protoplasts (E). Pictures were taken with bright field, green fluorescent protein (GFP) and 4’6-diamidino-2-phenylindole (DAPI) filters.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556288&req=5

pone.0136365.g001: Identification of possible tomato R2R3-MYB, bHLH, and WDR regulators of anthocyanin synthesis.Evolutionary relationships of R2R3-MYB (A), bHLH (B) and WDR (C) proteins involved in anthocyanin pigmentation in different plant species. The evolutionary history was inferred using the Neighbor-Joining method [42]. The optimal tree with the sum of branch length (A = 4.04792561, B = 3.00531532, C = 0.83964559) is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches [57]. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the p-distance method [58] and are in the units of the number of amino acid differences per site. The analysis involved 20 (A), 14 (B) and 12 (C) amino acid sequences. All ambiguous positions were removed for each sequence pair. There were a total of 790 (A), 364 (B) and 372 (C) positions in the final dataset. Evolutionary analyses were conducted in MEGA6 [41]. Expression heatmap (Log2 scale of FPKM values) of SlAN2, SlANT1, SlAN1, SlAN11, SlJAF13 and SlDFR genes in different tissues of tomato, analyzed by Illumina RNA-Seq (D). MG: Mature Green fruit; B: Breaker fruit; B+10: ripe fruit 10 days after breaker stage. Subcellular localization of GFP-SlANT1 and GFP-SlAN2 fusion proteins in transiently transformed A. thaliana mesophyll protoplasts (E). Pictures were taken with bright field, green fluorescent protein (GFP) and 4’6-diamidino-2-phenylindole (DAPI) filters.
Mentions: By analyzing the whole genome of tomato, all the annotated putative MYB proteins were compared with the main MYB regulatory factors involved in anthocyanin synthesis in other species (S1 Fig). Four different tomato MYB proteins, encoded by the genes Solyc10g086250, Solyc10g086260, Solyc10g086270, and Solyc10g086290, corresponding, respectively, to SlAN2 [33–35], SlANT1 [32,36], SlANT1like [35,54] and SlAN2like [35], grouped in one clade with MYB proteins of tobacco (N. tabacum), petunia and potato (S. tuberosum) involved in anthocyanin synthesis [5,55,56] (S1 Fig, Fig 1A). This clade included members from the Solanaceae family and was clearly separated from anthocyanin MYBs from other dicots, such as A. thaliana or A. majus, and monocots (S1 Fig, Fig 1A). This analysis confirms that SlANT1 and SlAN2 are indeed tomato MYB TFs involved in anthocyanin synthesis regulation.

Bottom Line: Despite the fact that cultivated tomato varieties do not accumulate anthocyanins in the fruit, the biosynthetic pathway can be activated in the vegetative organs by several environmental stimuli.Little is known about the molecular mechanisms regulating anthocyanin synthesis in tomato.Here, we carried out a molecular and functional characterization of two genes, SlAN2 and SlANT1, encoding two R2R3-MYB transcription factors.

View Article: PubMed Central - PubMed

Affiliation: PlantLab, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.

ABSTRACT
Anthocyanins are water-soluble polyphenolic compounds with a high nutraceutical value. Despite the fact that cultivated tomato varieties do not accumulate anthocyanins in the fruit, the biosynthetic pathway can be activated in the vegetative organs by several environmental stimuli. Little is known about the molecular mechanisms regulating anthocyanin synthesis in tomato. Here, we carried out a molecular and functional characterization of two genes, SlAN2 and SlANT1, encoding two R2R3-MYB transcription factors. We show that both can induce ectopic anthocyanin synthesis in transgenic tomato lines, including the fruit. However, only SlAN2 acts as a positive regulator of anthocyanin synthesis in vegetative tissues under high light or low temperature conditions.

No MeSH data available.


Related in: MedlinePlus