Identification of Candidate HY5-Dependent and -Independent Regulators of Anthocyanin Biosynthesis in Tomato

Qiu, Zhengkun; Wang, Haijing; Li, Dongjing; Yu, Bingwei; Hui, Qiu-Ling; Yan, Shuangshuang; Huang, Zejun; Cui, Xia; Cao, Bihao

doi:10.1093/pcp/pcy236

Cited by 96 publications

(84 citation statements)

References 79 publications

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“…The extracted DNA was then used as template to amplify the desired fragments using primers flanking the targeted sites (Table S2). The PCR conditions followed those described in a previous study (Qiu et al , ). PCR products were directly sequenced to identify mutations by the Sangon Biotech Company using the Sanger methods.…”

Section: Methodsmentioning

confidence: 99%

“…Samples were collected following the procedure described in a previous study (Qiu et al , ). Each tissue sample was composed of at least three biological replicates; for replicates of leaf samples, leaves were collected from three different plants and fruit for fruit peel replicates, peels were collected from at least five fruits from three different plants.…”

Section: Methodsmentioning

confidence: 99%

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Anthocyanin Fruit encodes an R2R3‐MYB transcription factor, SlAN2‐like, activating the transcription of SlMYBATV to fine‐tune anthocyanin content in tomato fruit

et al. 2019

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Anthocyanin fruit (Aft) and atroviolacea (atv) were characterized in wild tomato and can enhance anthocyanin content in tomato fruit. However, the gene underlying the Aft locus and the mechanism by which Aft and atv act remain largely unknown.In this study, the Aft locus was fine-mapped to an approximately 145-kb interval on chromosome 10, excluding SlAN2 (Solyc10g086250), SlANT1 (Solyc10g086260) and SlANT1like (Solyc10g086270), which have previously been suggested as candidates. Thus, the R2R3-MYB transcription factor SlAN2-like (Solyc10g086290) was considered the best candidate gene for Aft.The CRISPR/Cas9-mediated SlAN2-like mutants show a much lower accumulation of anthocyanins associated with the downregulation of multiple anthocyanin-related genes compared to the wild-type tomato, indicating that SlAN2-like is responsible for the Aft phenotype. The repressive function of SlMYBATV also was confirmed through the CRISPR/Cas9 approach. A yeast-two-hybrid assay revealed that SlMYBATV interacts with the bHLH protein SlJAF13. Furthermore, yeast-one-hybrid and dual-luciferase transient expression assays showed that Aft directly binds to the SlMYBATV promoter and activates its expression.The results herein provide candidate genes to enhance anthocyanin content in tomato fruit. This research also provides insight into a mechanism involving the Aft-SlMYBATV pathway that fine-tunes anthocyanin accumulation in tomato fruit.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Anthocyanin Fruit encodes an R2R3‐MYB transcription factor, SlAN2‐like, activating the transcription of SlMYBATV to fine‐tune anthocyanin content in tomato fruit

et al. 2019

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“…Several flavonol and anthocyanin biosynthetic genes can be activated by HY5 and R2R3-MYBs [33]. However, a small amount of anthocyanin could still be detected in the tomato hy5 mutant, indicating the existence of some HY5-independent transcription factors that mediate anthocyanin metabolism [34].…”

Section: The Accumulation Of Flavonoids and Anthocyanins Under Uv-b Rmentioning

confidence: 99%

Quantitative Proteomic Analyses Identify STO/BBX24 -Related Proteins Induced by UV-B

Lyu

Xiong

et al. 2020

IJMS

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Plants use solar radiation for photosynthesis and are inevitably exposed to UV-B. To adapt to UV-B radiation, plants have evolved a sophisticated strategy, but the mechanism is not well understood. We have previously reported that STO (salt tolerance)/BBX24 is a negative regulator of UV-B-induced photomorphogenesis. However, there is limited knowledge of the regulatory network of STO in UV-B signaling. Here, we report the identification of proteins differentially expressed in the wild type (WT) and sto mutant after UV-B radiation by iTRAQ (isobaric tags for relative and absolute quantitation)-based proteomic analysis to explore differential proteins that depend on STO and UV-B signaling. A total of 8212 proteins were successfully identified, 221 of them were STO-dependent proteins in UV-B irradiated plants. The abundances of STO-dependent PSB and LHC (light-harvesting complex) proteins in sto mutants decreased under UV-B radiation, suggesting that STO is necessary to maintain the normal accumulation of photosynthetic system complex under UV-B radiation to facilitate photosynthesis photon capture. The abundance of phenylalanine lyase-1 (PAL1), chalcone synthetase (CHS), and flavonoid synthetase (FLS) increased significantly after UV-B irradiation, suggesting that the accumulation of flavonoids do not require STO, but UV-B is needed. Under UV-B radiation, STO stabilizes the structure of antenna protein complex by maintaining the accumulation of PSBs and LHCs, thereby enhancing the non-photochemical quenching (NPQ) ability, releasing extra energy, protecting photosynthesis, and ultimately promoting the elongation of hypocotyl. The accumulation of flavonoid synthesis key proteins is independent of STO under UV-B radiation. Overall, our results provide a comprehensive regulatory network of STO in UV-B signaling.

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“…Accumulation of anthocyanins in fruits in response to light is mediated by a R2R3 MYB transcription factor in many plants including apple, litchi, pear, Chinese bayberry, and eggplant (Bai et al, ; M. Jiang, Ren, Lian, Liu, & Chen, ; Lai et al, ; Niu et al, ; Takos et al, ). HY5 is a basic leucine zipper (bZIP) transcription factor involved in light‐induced anthocyanin biosynthesis by directly binding to the G‐box motif within the promoter of R2R3 MYB regulators of anthocyanin biosynthesis and promoting anthocyanin accumulation (An et al, ; M. Jiang et al, ; Qiu et al, ; Shin et al, ). In combination with HY5 protein, B‐box containing zinc‐finger proteins were shown to activate anthocyanin genes in Arabidopsis and red pear (Bai et al, ; D. Xu et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…Jiang, Ren, Lian, Liu, & Chen, 2016;Lai et al, 2014;Niu et al, 2010;Takos et al, 2006). HY5 is a basic leucine zipper (bZIP) transcription factor involved in light-induced anthocyanin biosynthesis by directly binding to the G-box motif within the promoter of R2R3 MYB regulators of anthocyanin biosynthesis and promoting anthocyanin accumulation Qiu et al, 2018;Shin et al, 2013). In combination with HY5 protein, B-box containing zinc-finger proteins were shown to activate anthocyanin genes in Arabidopsis and red pear (Bai et al, 2019;D.…”

mentioning

confidence: 99%

Retrotransposon promoter of Ruby1 controls both light‐ and cold‐induced accumulation of anthocyanins in blood orange

Ding

Yuan

Tang

et al. 2019

Plant Cell & Environment

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Blood orange is generally recognized to accumulate anthocyanins in its fruit pulp in a cold‐inducible manner. We observed that the fruit peel of blood orange can also accumulate anthocyanins under ample light conditions. Interestingly, purple pummelo can accumulate anthocyanins only in its fruit peel but not in its pulp. The mechanism underlying the tissue specificity of anthocyanin accumulation in citrus is unknown. Here, we show that the active promoter of Ruby1, a key activator of anthocyanin biosynthesis, is also light inducible in addition to its already known cold inducibility in blood orange. Electrophoretic mobility shift assays and transient expression assays showed that HY5 positively regulated the transcription of Ruby1 by binding to the G‐box motif (CACGTC). The tissue specificity of anthocyanin accumulation in the peel of purple pummelo may be due to the lack of a low temperature responsive element and a MYC binding site, which were shown to be involved in cold inducibility of CsRuby1 in blood orange by insertion of a long terminal repeat type retrotransposon in the promoter. These results bring new insights into the regulatory mechanism of anthocyanin biosynthesis in response to environmental stimuli and provide cis‐elements for genetic improvement of anthocyanin‐stable fruits rich in antioxidant metabolites.

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Identification of Candidate HY5-Dependent and -Independent Regulators of Anthocyanin Biosynthesis in Tomato

Cited by 96 publications

References 79 publications

Anthocyanin Fruit encodes an R2R3‐MYB transcription factor, SlAN2‐like, activating the transcription of SlMYBATV to fine‐tune anthocyanin content in tomato fruit

Anthocyanin Fruit encodes an R2R3‐MYB transcription factor, SlAN2‐like, activating the transcription of SlMYBATV to fine‐tune anthocyanin content in tomato fruit

Quantitative Proteomic Analyses Identify STO/BBX24 -Related Proteins Induced by UV-B

Retrotransposon promoter of Ruby1 controls both light‐ and cold‐induced accumulation of anthocyanins in blood orange

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