Induces Anthocyanin Biosynthesis Research Articles

Combined transcriptomic, metabolomic and physiological analysis reveals the key role of nitrogen, but not phosphate and potassium in regulating anthocyanin biosynthesis induced by nutrient deficiency in Eucalyptus

Anthocyanin biosynthesis in Eucalyptus plants can be flexibly and rapidly modulated in response to hormones or environmental stimuli, including nutrient deprivation (ND). However, the underlying mechanism of ND in inducing anthocyanin biosynthesis in plants remains largely elusive. In this study, we discovered that anthocyanin levels in leaves and stems could well reflect nitrogen availability in Eucalyptus. Supplementation with nitrogen, but not with phosphate or potassium, effectively inhibited ND-induced anthocyanin biosynthesis. To further investigate how nitrogen regulates this process, comprehensive time-resolved transcriptomic and targeted metabolomic analyses were conducted. The results revealed that 3405, 2706, and 3153 genes were differentially regulated by nitrogen treatment at 1, 2, and 3 d, respectively. Pathway analysis indicated the majority of the enriched KEGG pathways were associated with cellular metabolism, such as amino acid and flavonoid biosynthesis. Moreover, metabolomic analysis showed that the most abundantly accumulated anthocyanins, cyanidin-3-O-galactoside and cyanidin-3-O-glucoside, along with key intermediates in the flavonoid pathway, were downregulated by nitrogen treatment. Furthermore, nitrogen-responsive MYB-bHLH-WDR complex genes were identified, including six EgrMYB113 family members. Overexpression of one of the EgrMYB113-like genes induced anthocyanin biosynthesis in the transgenic hairy roots of Eucalyptus. Interestingly, mutation of AtMYB113 inhibited nitrogen deficiency-induced anthocyanin biosynthesis in Arabidopsis, suggesting that MYB113 is a novel N-responsive MYB transcriptional regulator of anthocyanin biosynthesis. Additionally, nitrogen treatment upregulated a few GA biosynthesis genes while simultaneously downregulated the expression of several GA2ox genes. Exogenous application of GA3 decreased ND-induced anthocyanin biosynthesis. Conclusively, this study provides novel insights into the molecular mechanism of nitrogen in the regulation of anthocyanin biosynthesis in Eucalyptus.

Transcriptomic Analyses Reveal the Mechanism by Which Different Light Qualities and Light Duration Induce Anthocyanin Biosynthesis in ‘Kyoho’ Grapes

Transcriptomic Analyses Reveal the Mechanism by Which Different Light Qualities and Light Duration Induce Anthocyanin Biosynthesis in ‘Kyoho’ Grapes

Reactive oxygen species act as the key signaling molecules mediating light-induced anthocyanin biosynthesis in Eucalyptus

Light plays a pivotal role in regulating anthocyanin biosynthesis in plants, and the early light-responsive signals that initiate anthocyanin biosynthesis remain to be elucidated. In this study, we showed that the anthocyanin biosynthesis in Eucalyptus is hypersensitive to increased light intensity. The combined transcriptomic and metabolomic analyses were conducted on Eucalyptus leaves after moderate (ML; 100 μmol m−2 s−1) and high (HL; 300 μmol m−2 s−1) light intensity treatments. The results identified 1940, 1096, 1173, and 2756 differentially expressed genes at 6, 12, 24, and 36 h after HL treatment, respectively. The metabolomic results revealed the primary anthocyanin types, and other differentially accumulated flavonoids and phenylpropane intermediates that were produced in response to HL, which well aligned with the transcriptome results. Moreover, biochemical analysis showed that HL inhibited peroxidase activity and increased the ROS level in Eucalyptus leaves. ROS depletion through co-application of the antioxidants rutin, uric acid, and melatonin significantly reduced, and even abolished, anthocyanin biosynthesis induced by HL treatment. Additionally, exogenous application of hydrogen peroxide efficiently induced anthocyanin biosynthesis within 24 h, even under ML conditions, suggesting that ROS played a major role in activating anthocyanin biosynthesis. A HL-responsive MYB transcription factor EgrMYB113 was identified to play an important role in regulating anthocyanin biosynthesis by targeting multiple anthocyanin biosynthesis genes. Additionally, the results demonstrated that gibberellic acid and sugar signaling contributed to HL-induced anthocyanin biosynthesis. Conclusively, these results suggested that HL triggers multiple signaling pathways to induce anthocyanin biosynthesis, with ROS acting as indispensable mediators in Eucalyptus.

SlGAD2 is the target of SlTHM27, positively regulates cold tolerance by mediating anthocyanin biosynthesis in tomato.

Cold stress significantly limits the yield and quality of tomato. Deciphering the key genes related to cold tolerance is important for selecting and breeding superior cold-tolerant varieties. γ-aminobutyric acid (GABA) responds to various types of stress by rapidly accumulating in plant. In this study, glutamic acid decarboxylase (GAD2) was a positive regulator to enhance cold stress tolerance of tomato. Overexpression of SlGAD2 decreased the extent of cytoplasmic membrane damage and increased the endogenous GABA content, antioxidant enzyme activities, and reactive oxygen species (ROS) scavenging capacity in response to cold stress, whereas Slgad2 mutant plants showed the opposite trend. In addition, SlGAD2 induced anthocyanin biosynthesis in response to cold stress by increasing the content of endogenous GABA. Further study revealed that SlGAD2 expression was negatively regulated by the transcription factor SlTHM27. However, the transcript levels of SlTHM27 were repressed under cold stress. Antioxidant enzyme activities, SlGAD2 transcript levels, GABA and anthocyanin contents were significantly increased in Slthm27 mutant plants. Further, our study demonstrated that SlTHM27 decreases SlGAD2-promoted cold resistance in tomato by repressing SlGAD2 transcription. Overall, our results showed that the SlTHM27-SlGAD2 model regulates the cold tolerance in tomato by regulating GABA and anthocyanin.

ABA functions in low phosphate-induced anthocyanin accumulation through the transcription factor ABI5 in Arabidopsis.

ABI5 functions in ABA-mediated anthocyanin accumulation in plant response to low phosphate. Low phosphate (LP)-induced anthocyanin biosynthesis and accumulation play an important role in plant adaptive response to phosphate starvation conditions. However, whether and how the stress phytohormone abscisic acid (ABA) participates in LP-induced anthocyanin accumulation remain elusive. Here, we report that ABA is required for LP-induced anthocyanin accumulation in Arabidopsis thaliana. Disrupting ABA DEFICIENT2 (ABA2), a key ABA-biosynthetic gene, or BETA-GLUCOSIDASE1 (BG1), a major gene implicated in converting conjugated ABA to active ABA, significantly impairs LP-induced anthocyanin accumulation, as LP-induced expression of the anthocyanin-biosynthetic genes Chalcone Synthase (CHS) is dampened in the aba2 and bg1 mutant. In addition, LP-induced anthocyanin accumulation is defective in the mutants of ABA signaling pathway, including ABA receptors, ABA Insensitive2, and the transcription factors ABA Insensitive5 (ABI5), suggesting a role of ABI5 in ABA-mediated upregulation of anthocyanin-biosynthetic genes in plant response to LP. Indeed, LP-induced expression of CHS is repressed in the abi5-7 mutant but further promoted in the ABI5-overexpressing plants compared to the wild-type. Moreover, ABI5 can bind to and transcriptionally activate CHS, and the defectiveness of LP-induced anthocyanin accumulation in abi5-7 can be restored by overexpressing CHS. Collectively, our findings illustrates that ABI5 functions in ABA-mediated LP-induced anthocyanin accumulation in Arabidopsis.

RNA-sequencing analysis reveals novel genes involved in the different peel color formation in eggplant.

Eggplant (Solanum melongena L.) is a highly nutritious vegetable. Here, the molecular mechanism of color formation in eggplants was determined using six eggplant cultivars with different peel colors and two SmMYB113-overexpressing transgenic eggplants with a purple peel and pulp. Significant differentially expressed genes (DEGs) were identified by RNA-sequencing analysis using the following criteria: log2(sample1/sample2)≥ 0.75 and q-value ≤ 0.05. Two analytical strategies were used to identify genes related to the different peel color according to the peel color, flavonoids content, delphinidins/flavonoids ratio, and the content of anthocyanins. Finally, 27 novel genes were identified to be related to the color difference among eggplant peels and 32 novel genes were identified to be related to anthocyanin biosynthesis and regulated by SmMYB113. Venn analysis revealed that SmCytb5, SmGST, SmMATE, SmASAT3, and SmF3'5'M were shared among both sets of novel genes. Transient expression assay in tobacco suggested that these five genes were not sufficient for inducing anthocyanin biosynthesis alone, but they play important roles in anthocyanin accumulation in eggplant peels. Yeast one-hybrid, electrophoretic mobility shift assay and dual-luciferase assays indicated that the expression of the five genes could be directly activated by SmMYB113 protein. Finally, a regulatory model for the mechanism of color formation in eggplant was proposed. Overall, the results of this study provide useful information that enhances our understanding of the molecular mechanism underlying the different color formation in eggplant.

The ethylene-responsive transcription factor PpERF9 represses PpRAP2.4 and PpMYB114 via histone deacetylation to inhibit anthocyanin biosynthesis in pear.

Ethylene induces anthocyanin biosynthesis in most fruits, including apple (Malus domestica) and plum (Prunus spp.). By contrast, ethylene inhibits anthocyanin biosynthesis in pear (Pyrus spp.), but the underlying molecular mechanism remains unclear. In this study, we identified and characterized an ethylene-induced ETHYLENE RESPONSE FACTOR (ERF) transcription factor, PpETHYLENE RESPONSE FACTOR9 (PpERF9), which functions as a transcriptional repressor. Our analyses indicated PpERF9 can directly inhibit expression of the MYB transcription factor gene PpMYB114 by binding to its promoter. Additionally, PpERF9 inhibits the expression of the transcription factor gene PpRELATED TO APETALA2.4 (PpRAP2.4), which activates PpMYB114 expression, by binding to its promoter, thus forming a PpERF9-PpRAP2.4-PpMYB114 regulatory circuit. Furthermore, PpERF9 interacts with the co-repressor PpTOPLESS1 (PpTPL1) via EAR motifs to form a complex that removes the acetyl group on histone H3 and maintains low levels of acetylated H3 in the PpMYB114 and PpRAP2.4 promoter regions. The resulting suppressed expression of these 2 genes leads to decreased anthocyanin biosynthesis in pear. Collectively, these results indicate that ethylene inhibits anthocyanin biosynthesis by a mechanism that involves PpERF9-PpTPL1 complex-mediated histone deacetylation of PpMYB114 and PpRAP2.4. The data presented herein will be useful for clarifying the relationship between chromatin status and hormone signaling, with implications for plant biology research.

The long noncoding RNA LINC15957 regulates anthocyanin accumulation in radish.

Radish (Raphanus sativus L.) is an important root vegetable crop belonging to the Brassicaceae family. Anthocyanin rich radish varieties are popular among consumers because of their bright color and high nutritional value. However, the underlying molecular mechanism responsible for skin and flesh induce anthocyanin biosynthesis in transient overexpression, gene silencing and transcriptome sequencing were used to verify its function in radish anthocyanin accumulation, radish remains unclear. Here, we identified a long noncoding RNA LINC15957, overexpression of LINC15957 was significantly increased anthocyanin accumulation in radish leaves, and the expression levels of structural genes related to anthocyanin biosynthesis were also significantly increased. Anthocyanin accumulation and expression levels of anthocyanin biosynthesis genes were significantly reduced in silenced LINC15957 flesh when compared with control. By the transcriptome sequencing of the overexpressed LINC15957 plants and the control, 5,772 differentially expressed genes were identified. A total of 3,849 differentially expressed transcription factors were identified, of which MYB, bHLH, WD40, bZIP, ERF, WRKY and MATE were detected and differentially expressed in the overexpressed LINC15957 plants. KEGG enrichment analysis revealed the genes were significant enriched in tyrosine, L-Phenylalanine, tryptophan, phenylpropanol, and flavonoid biosynthesis. RT-qPCR analysis showed that 8 differentially expressed genes (DEGs) were differentially expressed in LINC15957-overexpressed plants. These results suggested that LINC15957 involved in regulate anthocyanin accumulation and provide abundant data to investigate the genes regulate anthocyanin biosynthesis in radish.

Melatonin alleviates arsenic (As) toxicity in rice plants via modulating antioxidant defense system and secondary metabolites and reducing oxidative stress

The Arsenic (As) load on the environment has increased immensely due to large-scale industrial and agricultural uses of As in several synthetic products, such as fertilizers, herbicides, and pesticides. Melatonin is a plant hormone that has a key role in abiotic stress inhibition, but the mechanism of resilience to As stress remains unexplored in rice plants. In this study, we determined how As affects rice plant and how melatonin facilitate As stress tolerance in rice. Here we investigated that, exogenous melatonin reduced As stress by inducing anthocyanin biosynthesis. Melatonin induced the expression of anthocyanin biosynthesis genes such as PAL, CHS, CHI, F3H, DFR, and ANS, which resulted in 1659% and 389% increases in cyanidin and delphinidin, respectively. Similarly, melatonin application significantly induced SA and ABA accumulation in response to As stress in rice plant. Application of melatonin also significantly reduced expression of PT-2 and PT-8 (transporter genes) and reduced uptake of As and its translocation to other compartments. Melatonin and As analysis revealed that melatonin application significantly reduced As contents in the melatonin-supplemented plants, suggesting that As uptake is largely dependent on either the melatonin basal level or anthocyanin in rice plants. In this study, we investigated new symptoms on leaves, which can severely damage leaves and impair photosynthesis. However, anthocyanin as a chelating agent, detoxifies As in vacuole and reduces oxidative stress induced by As.

Biotic stress-induced and ripening-related anthocyanin biosynthesis are regulated by alternate phytohormone signals in blueberries

During ripening blueberries accumulate high amounts of flavonoid-derived anthocyanin pigments in fruit skin, but not in flesh. However, we found that anthocyanin biosynthesis was induced in fruit flesh in response to infiltration with Agrobacterium and even with sterile water. We hypothesised that this stress-response was mediated by phytohormones, hence studied their role and associated gene expression patterns in comparison with skin-specific anthocyanin biosynthesis during fruit ripening. In ripening berries, active abscisic acid (ABA) concentrations were highest at the onset of anthocyanin biosynthesis, suggesting a role for ABA in regulating anthocyanins during at the onset of fruit ripening. Concentrations of active ABA, however, were comparable between fruit tissues during berry development, while genes and metabolites associated with ABA catabolism and reversible conjugation were up to 10-fold higher in fruit skin and correlated strongly with anthocyanins. Thus, increased amounts of ABA were processed in fruit skin, likely impacting on tissue-specific pigmentation during fruit development. In response to biotic stress , however, salicylic acid (SA) concentrations increased in fruit flesh and correlated strongly with anthocyanin biosynthesis, while ABA was actively catabolised and concentrations reduced, indicating a role for SA in activating stress-anthocyanin production. Application of SA and ABA showed that both phytohormones increased flavonoid gene expression and anthocyanin accumulation, thus indicating that anthocyanin biosynthesis can be regulated by alternate signals. • Biotic stress induced anthocyanin biosynthesis in non-pigmented blueberry flesh. • In stressed fruit, abscisic acid was catabolised and salicylic acid content increased. • In ripening fruit, abscisic acid metabolism was higher in fruit skin than flesh. • Alternate signals induced stress-related and ripening-related anthocyanins.

ABA signaling mediates 5-aminolevulinic acid-induced anthocyanin biosynthesis in red pear fruits

Peel color is an important quality trait of pears and directly affects its commercial value. Although several studies have reported that exogenous 5-aminolevulinic acid (ALA) induces anthocyanin biosynthesis, the mechanisms involved remain unclear. The purpose of this study was to confirm the effects of ALA on anthocyanin biosynthesis in the red pear ‘Nanhong’ and elucidate the effect of abscisic acid (ABA) on ALA-induced anthocyanin accumulation. Application of ALA solution (100 or 300 mg L−1) promoted arbutin, flavonol, and anthocyanin accumulation, but decreased chlorogenic acid and flavanol content, especially at 300 mg L−1. The structural gene transcriptional levels and transcription factors (PuMYB10 and PubHLH33) associated with anthocyanin synthesis were also enhanced by ALA treatment. Furthermore, ALA treatment increased the endogenous ABA content and upregulated the expression of ABA biosynthesis-related genes. Additionally, the influence of ALA on anthocyanin biosynthesis was removed by the ABA biosynthesis inhibitor fluridone, while the expression of PuMYB10, PuUFGT, and PuGSTF12, which were induced by exogenous ABA and ALA treatments, was suppressed. These results suggest that ABA signaling plays a crucial role in ALA-induced anthocyanin accumulation in red pear fruits. The results can provide a theoretical basis for improving the color and fruit quality of red pears.

De novo assembly of a fruit transcriptome set identifies AmMYB10 as a key regulator of anthocyanin biosynthesis in Aronia melanocarpa

Aronia is a group of deciduous fruiting shrubs, of the Rosaceae family, native to eastern North America. Interest in Aronia has increased because of the high levels of dietary antioxidants in Aronia fruits. Using Illumina RNA-seq transcriptome analysis, this study investigates the molecular mechanisms of polyphenol biosynthesis during Aronia fruit development. Six A. melanocarpa (diploid) accessions were collected at four fruit developmental stages. De novo assembly was performed with 341 million clean reads from 24 samples and assembled into 90,008 transcripts with an average length of 801 bp. The transcriptome had 96.1% complete according to Benchmarking Universal Single-Copy Orthologs (BUSCOs). The differentially expressed genes (DEGs) were identified in flavonoid biosynthetic and metabolic processes, pigment biosynthesis, carbohydrate metabolic processes, and polysaccharide metabolic processes based on significant Gene Ontology (GO) biological terms. The expression of ten anthocyanin biosynthetic genes showed significant up-regulation during fruit development according to the transcriptomic data, which was further confirmed using qRT-PCR expression analyses. Additionally, transcription factor genes were identified among the DEGs. Using a transient expression assay, we confirmed that AmMYB10 induces anthocyanin biosynthesis. The de novo transcriptome data provides a valuable resource for the understanding the molecular mechanisms of fruit anthocyanin biosynthesis in Aronia and species of the Rosaceae family.

Two MYB and Three bHLH Family Genes Participate in Anthocyanin Accumulation in the Flesh of Peach Fruit Treated with Glucose, Sucrose, Sorbitol, and Fructose In Vitro.

Anthocyanins are important pigments in peach fruit and are beneficial to human health. Sugars are both energy-storing and signaling molecules and their roles in inducing anthocyanin biosynthesis have received a great deal of research attention. However, the mechanism by which sugars induce anthocyanin biosynthesis in peach fruit is unknown. In order to understand this induction mechanism, comprehensive transcriptome and metabolome were performed in fruit flesh treated with four different sugars for 12 and 24 h, respectively. Here, we found that cyanidin-3-O-(6-O-p-coumaroyl) glucosides accumulated in fruit flesh treated with glucose, sucrose, sorbitol, and fructose in vitro. Two key structural genes of the anthocyanin biosynthesis pathway, namely, PpDFR and PpUFGT, were upregulated in the flesh of sugar-treated peach fruit. By contrast, the two main transcription factors (TFs) PpMYB10.1 and PpBL regulating anthocyanin biosynthetic genes in peach fruit were not upregulated accordingly. Interestingly, two MYB family genes (PpMYB6 and PpMYB44-like) and three bHLH family genes (PpbHLH35, PpbHLH51, and PpbHLH36-like) were upregulated. A dual luciferase assay revealed that PpMYB6 strongly activated the PpUFGT promoter when it was co-infiltrated with PpbHLH35, PpbHLH51, and PpbHLH36-like. When PpMYB44-like was co-infiltrated with PpbHLH35, it also potently activated the PpUFGT promoter. The results of this study help clarify the molecular mechanisms by which glucose, sucrose, sorbitol, and fructose regulate anthocyanin accumulation in peach fruit.

RBOH-dependent hydrogen peroxide signaling mediates melatonin-induced anthocyanin biosynthesis in red pear fruit

Although several studies have confirmed that exogenous melatonin promotes anthocyanin accumulation, the molecular mechanism of this remains elusive. Here, the signaling cross-talk between melatonin and NADPH oxidase (RBOH) -mediated ROS during anthocyanin biosynthesis were investigated. We found that application of exogenous melatonin not only induced anthocyanin biosynthesis, but also increased endogenous H2O2 and O2‾ content in pear fruits. The effect of melatonin on anthocyanin biosynthesis was abolished by inhibitors of RBOH. We also observed that genes encoding RBOH (PuRBOHF) were ubiquitously and highly expressed after melatonin treatment. Transient PuRBOHF overexpression significantly enhanced anthocyanin accumulation and activated transcription of anthocyanin biosynthesis genes, whereas PuRBOHF silencing repressed melatonin-promoted anthocyanin accumulation and H2O2 production. Moreover, RBOH-derived H2O2 induced PuMYB10 transcription, and PuRBOHF enhanced the PuMYB10-induced activation of the PuUFGT promoter. PuMYB10, in turn, activated PuRBOHF transcription, revealing a positive feedback loop. These results provide molecular evidence supporting the essential roles of PuRBOHF-dependent H2O2 in melatonin-induced anthocyanin accumulation in pears.

Ectopic Expressed LcMYB1 Induced Anthocyanin Biosynthesis in Petunia and Tomato

Ectopic Expressed LcMYB1 Induced Anthocyanin Biosynthesis in Petunia and Tomato

Fungal Infection Induces Anthocyanin Biosynthesis and Changes in DNA Methylation Configuration of Blood Orange [Citrus sinensis L. (Osbeck)

The biosynthesis of sweet orange anthocyanins is triggered by several environmental factors such as low temperature. Much less is known about the effect of biotic stress on anthocyanin production in sweet orange, although in other species anthocyanins are often indicated as “defense molecules”. In this work, citrus fruits were inoculated with Penicillium digitatum, the causal agent of green mold, and the amount of anthocyanins and the expression of genes related to their biosynthesis was monitored by RT-real time PCR after 3 and 5 days from inoculation (DPI). Moreover, the status of cytosine methylation of DFR and RUBY promoter regions was investigated by McrBC digestion followed in real-time. Our results highlight that fungal infection induces anthocyanin production by activating the expression of several genes in the biosynthetic pathway. The induction of gene expression is accompanied by maintenance of high levels of methylation at the DFR and RUBY promoters in the inoculated fruits, thus suggesting that DNA methylation is not a repressive mark of anthocyanin related gene expression in sweet orange subjected to biotic stress. Finally, by measuring the expression levels of the Citrus DNA demethylase genes, we found that none of them is up-regulated in response to fungal infection, this result being in accordance with the observed maintenance of high-level DFR and Ruby promoter regions methylation.

A Citrus Phosphate Starvation Response Factor CsPHL3 Negatively Regulates Carotenoid Metabolism.

Carotenoids provide precursors for the biosynthesis of strigolactones, which are a new class of hormones that are essential in phosphate (Pi) signaling during plant development. Carotenoid metabolism is a finely tuned pathway, but our understanding of the regulation mechanisms is still limited. In this study, we isolated a protein designated as CsPHL3 from citrus. CsPHL3 belonged to the Pi starvation response factor (PHR)-like subclade and was upregulated by low Pi. Acting as a nucleus-localized protein with transactivation activity, CsPHL3 bound directly to activate the promoter of a key metabolic gene, lycopene β-cyclase1 (LCYb1). Transgenic analysis revealed that the CsPHL3-overexpressing tomato plants exhibited abnormal growth, like the plants grew under limited Pi conditions. The transgenic lines showed reduced carotenoid contents and elevated expression of LCYb genes but downregulation of other key carotenogenic genes, including phytoene synthase (PSY). Moreover, CsPHL3 induced anthocyanin biosynthesis and affected Pi signaling in the transgenic plants. We further demonstrated that the expression of PSY was negatively regulated by CsPHL3 and high Pi. It is concluded that CsPHL3 is a Pi starvation response factor that negatively regulates carotenoid metabolism by modulating the expression of carotenogenic genes. Establishment of the CsPHL3-CsLCYb1 network provides new valuable knowledge of the function and underlying mechanism of PHR transcription factors and expands our understanding of the complex regulation mechanisms of carotenoid biosynthesis.

Ethylene-activated PpERF105 induces the expression of the repressor-type R2R3-MYB gene PpMYB140 to inhibit anthocyanin biosynthesis in red pear fruit.

Ethylene induces anthocyanin biosynthesis in most fruits, including apple (Malus domestica), strawberry (Fragaria × ananassa), and plum (Prunus spp.). However, ethylene inhibits anthocyanin biosynthesis in pear (Pyrus spp.), but the underlying molecular mechanism has not been characterized. In this study, ethylene induced the expression of PpERF105, which encodes a transcription factor. PpERF105 functioned as a transcriptional activator, but it inhibited anthocyanin biosynthesis in pear. A transcriptome analysis revealed that PpERF105 activated the expression of PpMYB140, which encodes an R2R3-MYB transcriptional repressor. Moreover, PpMYB140 directly inhibited the expression of anthocyanin-related structural genes. It also competed with PpMYB114 for the binding to bHLH3, ultimately resulting in the formation of the MYB140-bHLH-WDR complex rather than the conventional MBW complex, thereby further inhibiting anthocyanin biosynthesis. Furthermore, PpMYB140 prevented the overaccumulation of anthocyanins in the absence of ethylene. Collectively, our study data indicate that ethylene-induced PpERF105 inhibits anthocyanin biosynthesis by upregulating PpMYB140 expression. Our findings may be useful for elucidating the molecular basis of the ethylene-mediated inhibition of anthocyanin biosynthesis in fruit.

ABI5 regulates ABA-induced anthocyanin biosynthesis by modulating the MYB1-bHLH3 complex in apple.

Abscisic acid (ABA) induces anthocyanin biosynthesis in many plant species. However, the molecular mechanism of ABA-regulated anthocyanin biosynthesis remains unclear. As a crucial regulator of ABA signaling, ABSCISIC ACID-INSENSITIVE5 (ABI5) is involved in many aspects of plant growth and development, yet its regulation of anthocyanin biosynthesis has not been elucidated. In this study, we found that MdABI5, the apple homolog of Arabidopsis ABI5, positively regulated ABA-induced anthocyanin biosynthesis. A series of biochemical tests showed that MdABI5 specifically interacts with basic helix-loop-helix 3 (MdbHLH3), a positive regulator of anthocyanin biosynthesis. MdABI5 enhanced the binding of MdbHLH3 to its target genes dihydroflavonol 4-reductase (MdDFR) and UDP flavonoid glucosyl transferase (MdUF3GT). In addition, MdABI5 directly bound to the promoter of MdbHLH3 to activate its expression. Moreover, MdABI5 enhanced ABA-promoted interaction between MdMYB1 and MdbHLH3. Finally, antisense suppression of MdbHLH3 significantly reduced anthocyanin biosynthesis promoted by MdABI5, indicating that MdABI5-promoted anthocyanin biosynthesis was dependent on MdbHLH3. Taken together, our data suggest that MdABI5 plays a positive role in ABA-induced anthocyanin biosynthesis by modulating the MdbHLH3-MdMYB1 complex. Our work broadens the regulatory network of ABA-mediated anthocyanin biosynthesis, providing new insights to further study the transcriptional regulatory mechanisms behind this process.

Blue and UV-B light synergistically induce anthocyanin accumulation by co-activating nitrate reductase gene expression in Anthocyanin fruit (Aft) tomato.

The tomato accession LA1996, which carries a dominant allele of anthocyanin fruit (Aft) locus, accumulates anthocyanins in the epidermis of fruits when exposed to sunlight. The involvement of blue, UV-A, UV-B and a combination of these wavelengths on anthocyanin accumulation and the molecular mechanism of their regulation was investigated in LA1996. The most effective treatment for inducing anthocyanin biosynthesis in Aft fruits was co-irradiation with blue and UV-B (blue+UV-B) light. Finding the correlated genes is an important approach towards understanding their molecular mechanisms. In the present study, the nitrate reductase (NR) gene SlNIA was isolated using RNA-seq profiling of Aft fruits given different light treatments. The functions of NR-mediated anthocyanin induction by blue+UV-B were confirmed using a series of chemical treatments, followed by assessment of NR activity and nitric oxide (NO) detection. The expression of NR was highly induced by blue+UV-B, and this specificity was also confirmed with the enzyme activity of NR and the NO concentration. The NR inhibitors, which reduce NO generation, the expression levels of anthocyanin related genes and decreased anthocyanin accumulation in LA1996. Our results suggest that NR plays a key role in blue+UV-B-mediated anthocyanin accumulation in LA1996 fruits.