Regulation Of Anthocyanin Synthesis Research Articles

Genome-Wide Identification and Analysis of Anthocyanidin Reductase Gene Family in Lychee (Litchi chinensis Sonn.).

Anthocyanidin reductase (ANR) is a key enzyme regulating anthocyanin synthesis and accumulation in plants. Here, lychee ANR genes were globally identified, their sequence and phylogenetic characteristics were analyzed, and their spatiotemporal expression patterns were characterized. A total of 51 ANR family members were identified in the lychee genome. The length of the encoded amino acid residues ranged from 87 aa to 289 aa, the molecular weight ranged from 9.49 KD to 32.40 KD, and the isoelectric point (pI) ranged from 4.83 to 9.33. Most of the members were acidic proteins. Most members of the LcANR family were located in the cytoplasm. The 51 LcANR family members were unevenly distributed in 11 chromosomes, and their exons and motif conserved structures were significantly different from each other. Promoters in over 90% of LcANR members contained anaerobically induced response elements, and 88% contained photoresponsive elements. Most LcANR family members had low expression in nine lychee tissues and organs (root, young leaf, bud, female flower, male flower, pericarp, pulp, seed, and calli), and some members showed tissue-specific expression patterns. The expression of one gene, LITCHI029356.m1, decreased with the increase of anthocyanin accumulation in 'Feizixiao' and 'Ziniangxi' pericarp, which was negatively correlated with pericarp coloring. The identified LcANR gene was heterologously expressed in tobacco K326, and the function of the LcANR gene was verified. This study provides a basis for the further study of LcANR function, particularly the role in lychee pericarp coloration.

Sequence and epigenetic variations of R2R3-MYB transcription factors determine the diversity of taproot skin and flesh colors in different cultivated types of radish (Raphanus sativus L.).

This study found that three paralogous R2R3-MYB transcription factors exhibit functional divergence among different subspecies and cultivated types in radish. Cultivated radish taproots exhibit a wide range of color variations due to unique anthocyanin accumulation patterns in various tissues. This study investigated the universal principles of taproot color regulation that developed during domestication of different subspecies and cultivated types. The key candidate genes RsMYB1 and RsMYB2, which control anthocyanin accumulation in radish taproots, were identified using bulked segregant analysis in two genetic populations. We introduced the RsMYB1-RsF3'H-RsMYB1Met genetic model to elucidate the complex and unstable genetic regulation of taproot flesh color in Xinlimei radish. Furthermore, we analyzed the expression patterns of three R2R3-MYB transcription factors in lines with different taproot colors and investigated the relationship between RsMYB haplotypes and anthocyanin accumulation in a natural population of 56 germplasms. The results revealed that three paralogous RsMYBs underwent functional divergence during radish domestication, with RsMYB1 regulating the red flesh of Xinlimei radish, and RsMYB2 and RsMYB3 regulating the red skin of East Asian big long radish (R. sativus var. hortensis) and European small radish (R. sativus var. sativus), respectively. Moreover, RsMYB1-H1, RsMYB2-H10, and RsMYB3-H6 were identified as the primary haplotypes exerting regulatory functions on anthocyanin synthesis. These findings provide an understanding of the genetic mechanisms regulating anthocyanin synthesis in radish and offer a potential strategy for early prediction of color variations in breeding programs.

AcMYB266, a key regulator of the red coloration in pineapple peel: a case of subfunctionalization in tandem duplicated genes.

Red fruit peel is an attractive target for pineapple breeding. Various pineapple accessions with distinct red coloration patterns exist; however, the precise molecular mechanism accounting for these differences remains unknown, which hinders the pineapple breeding process from combining high fruit quality with red peel. In this study, we characterized a transcription factor, AcMYB266, which is preferentially expressed in pineapple peel and positively regulates anthocyanin accumulation. Transgenic pineapple, Arabidopsis, and tobacco plants overexpressing AcMYB266 exhibited significant anthocyanin accumulation. Conversely, transient silencing of this gene led to decreased anthocyanin accumulation in pineapple red bracts. In-depth analysis indicated that variations of AcMYB266 sequences in the promoter instead of the protein-coding region seem to contribute to different red coloration patterns in peels of three representative pineapple varieties. In addition, we found that AcMYB266 was located in a cluster of four MYB genes exclusive to and conserved in Ananas species. Of this cluster, each was proved to regulate anthocyanin synthesis in different pineapple tissues, illustrating an interesting case of gene subfunctionalization after tandem duplication. In summary, we have characterized AcMYB266 as a key regulator of pineapple red fruit peel and identified an MYB cluster whose members were subfunctionalized to specifically regulate the red coloration of different pineapple tissues. The present study will assist in establishing a theoretical mechanism for pineapple breeding for red fruit peel and provide an interesting case for the investigation of gene subfunctionalization in plants.

Molecular mechanism of abscisic acid signaling response factor VcbZIP55 to promote anthocyanin biosynthesis in blueberry (Vaccinium corymbosum)

A high content of anthocyanin in blueberry (Vaccinium corymbosum) is an important indicator to evaluate fruit quality. Abscisic acid (ABA) can promote anthocyanin biosynthesis, but since the molecular mechanism is unclear, clarifying the mechanism will improve for blueberry breeding and cultivation regulation. VcbZIP55 regulating anthocyanin synthesis in blueberry were screened and mined using the published Isoform-sequencing, RNA-Seq and qRT-PCR at different fruit developmental stages. Blueberry genetic transformation and transgenic experiments confirmed that VcbZIP55 could promote anthocyanin biosynthesis in blueberry adventitious buds, tobacco leaves, blueberry leaves and blueberry fruit. VcbZIP55 responded to ABA signals and its expression was upregulated in blueberry fruit. In addition, using VcbZIP55 for Yeast one hybrid assay (Y1H) and transient expression in tobacco leaves demonstrated an interaction between VcbZIP55 and a G-Box motif on the VcMYB1 promoter to activate the expression of VcMYB1. This study will lay the theoretical foundation for the molecular mechanisms of phytohormone regulation responsible for anthocyanin synthesis and provide theoretical support for blueberry quality improvement.

Transcription factors FtMYB22 negatively regulates anthocyanin synthesis by forming an MBW ternary complex in Tartary buckwheat

AbstractAnthocyanins are important flavonoid compounds that are abundant in Tartary buckwheat, providing a variety of bright colors and enhancing abiotic stress resistance. Our study found that FtMYB22 specifically affected anthocyanin anabolism. In this study, overexpression of the FtMYB22 transcription factor in tobacco significantly reduced the accumulation of flavonol in the petals of transgenic tobacco plants. Moreover, it significantly downregulated the expression of the genes involved in anthocyanin biosynthesis in tobacco. Overexpression of the FtMYB22 transcription factor in Tartary buckwheat hairy roots significantly inhibited anthocyanin and total flavonoids biosynthesis, and downregulated the expression of the anthocyanin biosynthesis genes. Comparing wild‐type and transgenic hairy roots, FtMYB22 overexpression significantly inhibited anthocyanin accumulation and downregulated most flavonoid synthesis pathway gene expression. Yeast two‐hybrid assays demonstrated that FtMYB22 could interact with the FtbHLH family member FtTT8 and the FtWD40 family member FtTTG1 to form a ternary MBW complex to inhibit anthocyanin synthesis. Therefore, FtMYB22 can be exploited as a target site for further gene editing and molecular breeding to improve the resilience and quality of Tartary buckwheat.

Comparative Transcriptome Analysis Reveals Changes in Gene Expression Associated with Anthocyanin Metabolism in Begonia semperflorens under Light Conditions

Anthocyanins, recognized as stress indicators, particularly under high-light conditions, play a pivotal role in plant stress responses. The advent of transcriptomics has opened avenues to elucidate the mechanisms underlying high light-induced anthocyanin biosynthesis. This study delved into transcriptomic changes in Begonia semperflorens leaves under varying light intensities: 950–9600 lx (TL_100), 6800–7000 lx (HS_75), and 4300–4500 lx (LS_25). To confirm the expression profiles of the key genes, we chose 12 critical genes associated with anthocyanin production for quantitative reverse transcription PCR (qRT-qPCR) analysis. Following this, we measured the levels of anthocyanins to substantiate the findings from the gene expression analysis. The transcriptome assembly in this study was extensive, yielding 43,038 unigenes that collectively spanned about 49.83 million base pairs, with an average unigene length of 1157 bp and an N50 value of 1685 bp. This assembly facilitated a thorough functional annotation across seven distinct protein databases, leading to the classification of 16,363 unigenes into 58 different families of transcription factors. Our comparative analysis of the transcriptomes highlighted a substantial number of differentially expressed genes (DEGs): 5411 DEGs between HS_75 and TL_100 conditions, with 3078 showing increased expression and 2333 showing decreased expression; 4701 DEGs between LS_25 and TL_100, consisting of 2648 up-regulated and 2053 down-regulated genes; and 6558 DEGs between LS_25 and HS_75, with 3032 genes up-regulated and 3526 down-regulated. These DEGs were significantly involved in critical pathways, such as anthocyanin synthesis, plant hormone signaling, and other regulatory mechanisms. This study suggests that genes, including F3′H, MYB102, and SWEET1, could play vital roles in regulating anthocyanin synthesis in response to various light conditions, potentially impacting the expression levels of other genes, like WRKYs, ATHB12, and those similar to HSP.

Genome-Wide Analysis of R2R3-MYB Genes and Functional Characterization of SmMYB75 in Eggplant Fruit Implications for Crop Improvement and Nutritional Enhancement.

R2R3-MYB represents a substantial gene family that plays diverse roles in plant development. In this study, 102 SmR2R3-MYB genes were identified from eggplant fruit and classified into 31 subfamilies. Analysis indicated that segmental duplication events played a pivotal role in the expansion of the SmR2R3-MYB gene family. Furthermore, the prediction of miRNAs targeting SmR2R3-MYB genes revealed that 60 SmR2R3-MYBs are targeted by 57 miRNAs, with specific miRNAs displaying varying numbers of target genes, providing valuable insights into the regulatory functions of miRNAs in plant growth, development, and responses to stress conditions. Through expression profile analysis under various treatment conditions, including low temperature (4 °C), plant hormone (ABA, Abscisic acid), and drought stress (PEG, Polyethylene glycol), diverse and complex regulatory mechanisms governing SmR2R3-MYB gene expression were elucidated. Notably, EGP21875.1 and EGP21874.1 exhibited upregulation in expression under all treatment conditions. Transcriptome and metabolome analyses demonstrated that, apart from anthocyanins (delphinidin-3-O-glucoside, cyanidin-3-O-(6-O-p-coumaroyl)-glucoside, and malvidin-3-O-(6-O-p-coumaroyl)-glucoside), overexpression of SmMYB75 could also elevate the content of various beneficial compounds, such as flavonoids, phenolic acids, and terpenes, in eggplant pulp. This comprehensive study enhances our understanding of SmR2R3-MYB gene functions and provides a strong basis for further research on their roles in regulating anthocyanin synthesis and improving eggplant fruit quality.

The R2R3-MYB Transcriptional Repressor TgMYB4 Negatively Regulates Anthocyanin Biosynthesis in Tulips (Tulipa gesneriana L.).

Anthocyanins play a paramount role in color variation and significantly contribute to the economic value of ornamental plants. The conserved activation complex MYB-bHLH-WD40 (MBW; MYB: v-myb avian myeloblastosis viral oncogene homolog; bHLH: basic helix-loop-helix protein; WD40:WD-repeat protein) involved in anthocyanin biosynthesis has been thoroughly researched, but there have been limited investigations into the function of repressor factors. In this study, we characterized TgMYB4, an R2R3-MYB transcriptional repressor which is highly expressed during petal coloration in red petal cultivars. TgMYB4-overexpressing tobaccos exhibited white or light pink petals with less anthocyanin accumulation compared to control plants. TgMYB4 was found to inhibit the transcription of ANTHOCYANIDIN SYNTHASE (TfANS1) and DIHYDRO-FLAVONOL-4-REDUCTASE (AtDFR), although it did not bind to their promoters. Moreover, the TgMYB4 protein was able to compete with the MYB activator to bind to the :bHLHprotein, thereby suppressing the function of the activator MBW complex. These findings demonstrate that TgMYB4 plays a suppressive role in the regulation of anthocyanin synthesis during flower pigmentation.

A light responsive transcription factor CsbHLH89 positively regulates anthocyanidin synthesis in tea (Camellia sinensis)

Anthocyanins are a class of secondary metabolites with rich physiological functions and many benefits for the human body. Although there have been numerous reports on anthocyanin synthesis in tea plants, the mechanism by which bHLH transcription factors regulate anthocyanin synthesis in tea plants is still unclear. In this study, we identified CsbHLH89, a basic helix–loop–helix (bHLH) transcription factor that is located in the nucleus and cytoplasm and has transcriptional activation activity. CsbHLH89 directly binds to G-box elements and activates the CsCHS, CsFLS, and CsDFR promoters in vitro and in vivo. Further experiments demonstrated that CsbHLH89 could be involved in the synthesis of tea plant anthocyanins as homodimers or heterodimers. Our research results show that CsbHLH89 can positively regulate anthocyanin synthesis in tea. In addition, it was found that CsHY5, a light signal transduction factor, could bind to the promoter of CsbHLH89 to indirectly promote anthocyanin accumulation. Silencing of CsHY5 and CsbHLH89 by antisense oligonucleotide, respectively, revealed that the expression of structural genes and transcription factors were down-regulated. Shading treatment of tea plants revealed a significant reduction in anthocyanin content and down-regulation of the expression of genes involved in anthocyanin synthesis. Our research provides new insight into the regulatory network of anthocyanin synthesis in tea and a new strategy for breeding rich anthocyanin.

RHA2b-mediated MYB30 degradation facilitates MYB75-regulated, sucrose-induced anthocyanin biosynthesis in Arabidopsis seedlings

Anthocyanins play diverse roles in plant physiology and stress adaptation. In Arabidopsis, the MYB–bHLH–WD40 (MBW) complex has a crucial role in the regulation of anthocyanin synthesis. Here, we report that the R2R3-MYB transcription factor MYB30 and the ubiquitin E3 ligase RHA2b participate in anthocyanin biosynthesis through regulation of the MBW complex. MYB30 was found to negatively regulate sucrose-induced anthocyanin biosynthesis in Arabidopsis seedlings. Expression of multiple genes involved in flavonoid or anthocyanin biosynthesis was affected in the myb30 mutant, and MYB30 directly repressed the expression of MYB75, which encodes a core component of the MBW complex, by binding to its promoter. Moreover, MYB30 physically interacted with MYB75 to inhibit its activity by repressing MBW complex assembly. In addition, sucrose treatment significantly promoted MYB30 degradation via the action of RHA2b. The ubiquitination and degradation of MYB30 were significantly attenuated in the rha2b mutant under high-sucrose treatment, and further analysis showed that MYB75 directly promoted RHA2b expression in response to high sucrose. Our work thus reveals an anthocyanin biosynthetic regulatory module, RHA2b–MYB30, that controls the function of the MBW complex via MYB75. The repression of MYB75 by MYB30 is released by MYB75-induced RHA2b expression, thus ensuring the self-activation of MYB75 when anthocyanin synthesis is needed.

Eggplant transcription factor SmMYB5 integrates jasmonate and light signaling during anthocyanin biosynthesis.

Low light conditions severely suppress anthocyanin synthesis in fruit skins, leading to compromised fruit quality in eggplant (Solanum melongena L.) production. In this study, we found that exogenous methyl-jasmonate (MeJA) application can effectively rescue the poor coloration of the eggplant pericarp under low light conditions. However, the regulatory relationship between jasmonate and light signaling for regulating anthocyanin synthesis remains unclear. Here, we identified a JA response factor, SmMYB5, as an anthocyanin positive regulator by applying RNA-sequencing and characterization of transgenic plants. Firstly, we resolved that SmMYB5 can interact with TRANSPARENT TESTA8 (SmTT8), an anthocyanin-promoted BASIC HELIX-LOOP-HELIX (bHLH) transcription factor, to form the SmMYB5-SmTT8 complex and activate CHALCONE SYNTHASE (SmCHS), FLAVANONE-3-HYDROXYLASE (SmF3H), and ANTHOCYANIN SYNTHASE (SmANS) promoters by direct binding. Secondly, we revealed that JA signaling repressors JASMONATE ZIM DOMAIN5 (SmJAZ5) and SmJAZ10 can interfere with the stability and transcriptional activity of SmMYB5-SmTT8 by interacting with SmMYB5. JA can partially rescue the transcriptional activation of SmF3H and SmANS promoters by inducing SmJAZ5/10 degradation. Thirdly, we demonstrated that the protein abundance of SmMYB5 is regulated by light. CONSTITUTIVELY PHOTOMORPHOGENIC1 (SmCOP1) interacts with SmMYB5 to trigger SmMYB5 degradation via the 26S proteasome pathway. Finally, we delineated a light-dependent JA-SmMYB5 signaling pathway that promotes anthocyanin synthesis in eggplant fruit skins. These results provide insights into the mechanism of the integration of JA and light signals in regulating secondary metabolite synthesis in plants.

Genome-Wide Analysis of the bHLH Gene Family in Loropetalum chinense var. rubrum: Identification, Classification, Evolution, and Diversity of Expression Patterns under Cultivation.

The basic helix-loop-helix (bHLH) transcription factor family is the second-largest transcription factor family in plants. Members of this family are involved in the processes of growth and development, secondary metabolic biosynthesis, signal transduction, and plant resistance. Loropetalum chinense var. rubrum is a critical woody plant with higher ornamental and economic values, which has been used as ornamental architecture and traditional Chinese herbal medicine plants. However, the bHLH transcription factors in Loropetalum chinense var. rubrum (L. chinense var. rubrum) have not yet been systematically demonstrated, and their role in the biosynthesis of anthocyanin is still unclear. Here, we identified 165 potential LcbHLHs genes by using two methods, and they were unequally distributed on chromosomes 1 to 12 of the genome of L. chinense var. rubrum. Based on an evolutionary comparison with proteins from Arabidopsis and Oryza sativa, these bHLH proteins were categorized into 21 subfamilies. Most LcbHLHs in a particular subfamily had similar gene structures and conserved motifs. The Gene Ontology annotation and Cis-elements predicted that LcbHLHs had many molecular functions and were involved in processes of plant growth, including the biosynthesis of flavonoids and anthocyanins. Transcriptomic analysis revealed different expression patterns among different tissues and cultivars of L. chinense var. rubrum. Many LcbHLHs were expressed in the leaves, and only a few genes were highly expressed in the flowers. Six LcbHLHs candidate genes were identified by bioinformatics analysis and expression analysis. Further Real-time quantitative PCR analysis and protein interaction network analysis showed that LcbHLH156, which is one of the candidate proteins belonging to the IIIf subfamily, could interact with proteins related to anthocyanin synthesis. Therefore, LcbHLH156 was transiently expressed in L. chinense var. rubrum to verify its function in regulating anthocyanin synthesis. Compared with the control group, red pigment accumulation appeared at the wound after injection, and the total anthocyanin content increased at the wound of leaves. These results lay a foundation for the research of the regulation mechanism of leaf colors in L. chinense var. rubrum and also provide a basis for the function of the LcbHLH family.

Analysis of bZIP gene family in lotus (Nelumbo) and functional study of NnbZIP36 in regulating anthocyanin synthesis

BackgroundThe basic leucine zipper (bZIP) family is a predominant group of transcription factors in plants, involved in regulating plant growth, development, and response to stressors. Additionally, the bZIP gene family has a key role in anthocyanin production. Despite the significant role of bZIP genes in plants, their potential contribution in lotus remains understudied.ResultsA total of 124 bZIP genes (59 NnbZIPs and 65 NlbZIPs) were identified from genomes of two lotus species. These genes were classified into 13 groups according to the grouping principle of the Arabidopsis bZIP gene family. Analysis of promoter cis-acting elements indicated that most bZIP gene family members in lotus are associated with response to abiotic stresses. The promoters of some bZIP genes contain MYB binding sites that regulate anthocyanin synthesis. We examined the anthocyanin content of the petals from three different colored lotus, combined with transcriptome data analysis and qRT-PCR results, showing that the expression trends of NnbZIP36 and the homologous gene NlbZIP38 were significantly correlated with the anthocyanin content in lotus petals. Furthermore, we found that overexpression of NnbZIP36 in Arabidopsis promoted anthocyanin accumulation by upregulating the expression of genes (4CL, CHI, CHS, F3H, F3'H, DFR, ANS and UF3GT) related to anthocyanin synthesis.ConclusionsOur study enhances the understanding of the bZIP gene family in lotus and provides evidence for the role of NnbZIP36 in regulating anthocyanin synthesis. This study also sets the stage for future investigations into the mechanism by which the bZIP gene family regulates anthocyanin biosynthesis in lotus.

Transcriptomic and Targeted Metabolomics Analysis of Detached Lycium ruthenicum Leaves Reveals Mechanisms of Anthocyanin Biosynthesis Induction through Light Quality and Sucrose Treatments.

Light quality and sucrose-induced osmotic stress are known to cause anthocyanin synthesis in detached Lycium ruthenicum leaves. To identify the mechanisms by which the kind of light quality and sucrose concentration are induced, here, we conducted transcriptome sequencing in detached L. ruthenicum leaves treated with different qualities of light and sucrose concentrations. Leaves treated with blue light or sucrose showed a significantly increased total anthocyanins content compared to those treated with white light. Delphinidin-3-O-rutinoside and delphinidin-3-O-glucoside production were differentially regulated by the BL(-S), BL(+S), and WL(+S) treatments. The structural genes CHS, CHI, F3'H, F3'5'H, ANS, and UFGT were significantly up-regulated in leaves treated with blue light or sucrose. Leaves treated with blue light additionally showed up-regulation of the light photoreceptors CRY1, PIF3, COP1, and HY5. The anthocyanin-related genes NCED1, PYR/PYL, PP2C, SnRK2, and ABI5 were significantly up-regulated in leaves treated with sucrose, promoting adaptability to sucrose osmotic stress. Co-expression and cis-regulatory analyses suggested that HY5 and ABI5 could regulate LrMYB44 and LrMYB48 through binding to the G-box element and ABRE element, respectively, inducing anthocyanin synthesis in response to blue light or sucrose treatment. Candidate genes responsive to blue light or sucrose osmotic stress in the anthocyanin biosynthesis pathway were validated through quantitative reverse transcription PCR. These findings deepen our understanding of the mechanisms by which blue light and sucrose-induced osmotic stress regulate anthocyanin synthesis, providing valuable target genes for the future improvement in anthocyanin production in L. ruthenicum.

Molecular Regulatory Network of Anthocyanin Accumulation in Black Radish Skin as Revealed by Transcriptome and Metabonome Analysis.

To understand the coloring mechanism in black radish, the integrated metabolome and transcriptome analyses of root skin from a black recombinant inbred line (RIL 1901) and a white RIL (RIL 1911) were carried out. A total of 172 flavonoids were detected, and the analysis results revealed that there were 12 flavonoid metabolites in radish root skin, including flavonols, flavones, and anthocyanins. The relative concentrations of most flavonoids in RIL 1901 were higher than those in RIL 1911. Meanwhile, the radish root skin also contained 16 types of anthocyanins, 12 of which were cyanidin and its derivatives, and the concentration of cyanidin 3-o-glucoside was very high at different development stages of black radish. Therefore, the accumulation of cyanidin and its derivatives resulted in the black root skin of radish. In addition, a module positively related to anthocyanin accumulation and candidate genes that regulate anthocyanin synthesis was identified by the weighted gene co-expression network analysis (WGCNA). Among them, structural genes (RsCHS, RsCHI, RsDFR, and RsUGT75C1) and transcription factors (TFs) (RsTT8, RsWRKY44L, RsMYB114, and RsMYB308L) may be crucial for the anthocyanin synthesis in the root skin of black radish. The anthocyanin biosynthesis pathway in the root skin of black radish was constructed based on the expression of genes related to flavonoid and anthocyanin biosynthesis pathways (Ko00941 and Ko00942) and the relative expressions of metabolites. In conclusion, this study not only casts new light on the synthesis and accumulation of anthocyanins in the root skin of black radish but also provides a molecular basis for accelerating the cultivation of new black radish varieties.

Physiological response of anthocyanin synthesis to different light intensities in blueberry.

Fruit color is an important economic character of blueberry, determined by the amount of anthocyanin content. Anthocyanin synthesis within the blueberry fruits is significantly affected by light. To reveal the physiological response mechanism of anthocyanin synthesis in blueberry fruits in different light intensities, four light intensities (100% (CK), 75%, 50% and 25%) were set for the 'O'Neal' southern highbush blueberry as the experimental material in our study. The relationship between endogenous hormones content, associated enzyme activities, and variations with the anthocyanin content in blueberry fruits under various light intensities during the white fruit stage (S1), purple fruit stage (S2), and blue fruit stage (S3) were studied. The results showed that adequate light could significantly promote anthocyanin synthesis in blueberry fruits (P < 0.05). Blueberry fruits had an anthocyanin content that was 1.76~24.13 times higher under 100% light intensity than it was under non-full light intensity. Different light intensities significantly affected the content of endogenous hormones and the activity of associated enzymes in anthocyanin synthesis pathway (P < 0.05). Among them, the JA (jasmonic acid) content and PAL (phenylalanine ammonia lyase) activity of fruits under 100% light intensity were 2.49%~41.83% and 2.47%~48.48% higher than those under other light intensity, respectively. And a significant correlation was found between the variations in anthocyanin content in fruits and the content or activities of JA, ABA (abscisic acid), ETH (ethylene), GA3 (gibberellin 3), IAA (indoleacetic acid), PAL, CHI (chalcone isomerase), DFR (dihydroflavonol reductase) and UFGT (UDP-glucose: flavonoid 3-glucosyltransferase) (P < 0.05). It indicated that 100% light intensity significantly promoted anthocyanin synthesis in blueberry fruits by affecting endogenous hormones content and associated enzyme activities in the anthocyanin synthesis pathway. This study will lay a foundation for further research on the molecular mechanism of light intensity regulating anthocyanin synthesis in blueberry.

Effects of Light Intensity on Endogenous Hormones and Key Enzyme Activities of Anthocyanin Synthesis in Blueberry Leaves

Plant anthocyanin is a secondary metabolite widely distributed in the roots, stems, leaves, flowers and fruits of plants, and its synthesis is significantly affected by light intensity. To reveal the physiological response mechanism of anthocyanin synthesis in blueberry leaves at different light intensities, four light intensities (100% (CK), 75%, 50% and 25%) were set for the ‘O’Neal’ southern highbush blueberry as the experimental material in our study. The relationship between endogenous hormone contents, key enzyme activities, and variations in the anthocyanin content in blueberry leaves under various light intensities during the white fruit stage (S1), purple fruit stage (S2) and blue fruit stage (S3) of fruit development were studied. The results showed that the anthocyanin content of blueberry leaves increased first and then decreased, and decreased first and then increased with the increase in light intensity and development stage, respectively. The appropriate light intensity could significantly promote the synthesis of anthocyanin, and the anthocyanin content in leaves treated with 75% light intensity was 1.09~4.08 times that of other light intensity treatments. The content or activities of gibberellin (GA3), indoleacetic acid (IAA), jasmonic acid (JA), abscisic acid (ABA), ethylene (ETH), phenylalanine ammonia lyase (PAL), chalcone isomerase (CHI), dihydroflavonol reductase (DFR) and UDP-glucose: flavonoid 3-glucosyltransferase (UFGT) were significantly or extremely significantly correlated with the content of anthocyanin in leaves. This indicated that light intensity significantly promoted anthocyanin synthesis in blueberry leaves by affecting endogenous hormone contents and key enzyme activities in the anthocyanin synthesis pathway. This study lays a foundation for further research on the molecular mechanism of light intensity regulating anthocyanin synthesis in blueberry leaves.

Exogeneous selenium enhances anthocyanin synthesis during grain development of colored-grain wheat

Anthocyanins and selenium (Se) play critical roles in antioxidant, anticancer, antibacterial, and antiviral treatments. Previous studies indicate that colored-grain wheat accumulates more Se than regular wheat, and Se synergistically promotes anthocyanin synthesis. However, the mechanism through which Se regulates anthocyanin synthesis remains unclear. We studied anthocyanin accumulation during the grain-filling stage of colored-grain wheat development by employing transcriptomics and metabolomics. We show that Se biofortification increased the concentrations of Se, anthocyanin, chlorophyll a and b, and carotenoids in colored-grain wheat. Genes related to biosynthesis of anthocyanins, phenylpropanoids biosynthesis, and flavonoids biosynthesis were significantly upregulated after Se treatment, which led to the accumulation of anthocyanin metabolites in colored-grain wheat. Genetic alterations in the expression profiles of several genes and transcription factors were observed, which slowed down lignin and proanthocyanidin biosynthesis and accelerated anthocyanin synthesis. Our results deepen the understanding of anthocyanin metabolism in Se-treated colored-grain wheat, which will likely promote harvest of these varieties.

Metabolome and transcriptome analyses of anthocyanin biosynthesis reveal key metabolites and candidate genes in purple wheat (Triticum aestivum L.).

Wheat (Triticum aestivum L.) is continuously subjected to genetic improvement to optimize grain quality. Purple wheat has recently gained attention because of its high anthocyanin and nutrient content. In this study, we performed an integrated transcriptome and metabolome analysis of the inbred wheat lines ZM152 (white wheat line) and ZM163 (purple wheat line) to elucidate molecular networks and identify potential genes regulating anthocyanin synthesis. A total of 564 metabolites were detected, of which 47 metabolite contents differed significantly between the two lines. Twenty-five flavonoids, including four anthocyanins, were significantly higher in purple wheat. High contents of cyanidin 3-rutinoside and malvidin 3-glucoside might contribute to the purple coloration of the wheat grains. Consistently, gene ontology and pathway enrichment analyses revealed that flavonoid and anthocyanin biosynthesis were mostly enriched, and the expression of anthocyanin structural genes was specifically upregulated in purple wheat lines, while transcription factors (TFs) were mostly downregulated in purple wheat lines. Especially, the correlation analysis showed the anthocyanin synthesis-related genes CHS (TraesCS2B02G048400) and UFGT (TraesCS7A02G155400) were likely regulated negatively by the TFs MYB4 (TraesCS1A02G268800, TraesCS1B02G279400), TT8 (TraesCS1D02G094200, TraesCS1B02G113100, and TraesCS1A02G102400), which thus could be considered important regulatory genes in the anthocyanin biosynthesis pathway of purple wheat lines. In summary, these results offer new insights into anthocyanin biosynthesis and accumulation of purple wheat, and provide very useful candidate genes for future colored wheat breeding.

Identification of PAL Gene in Purple Cabbage and Functional Analysis Related to Anthocyanin Synthesis

Anthocyanin is a characteristic nutrient of purple cabbage, and phenylalanine ammonia-lyase (PAL) is the rate-limiting enzyme for the synthesis of anthocyanin by the phenylpropane pathway, which is an important part of plant secondary metabolism. In this research, 7 BrPAL, 8 BoPAL, and 15 BnPAL genes from genomes of Brassica rapa, Brassica oleracea, and Brassica napus, divided into four subgroups, evolved from 4 PAL genes in Arabidopsis. The amplification and evolution of the BrPAL gene are due to segmental duplication and purifying selection. BrPAL genes clustered in the same clade have similar intron/exon structures and motifs. The cis-regulatory elements are divided into four categories: light, growth and development, stress and hormones. The qRT-PCR assays showed that most BrPAL genes were upregulated by UVA, low temperature and MeJA and downregulated by FR, high temperature, salt, PEG, IAA, ABA and GA, and there was a positive correlation between anthocyanin content and gene expression. This study can be used as a source for the function of the cabbage PAL gene and its molecular mechanism of regulating anthocyanin synthesis and provides a theoretical basis for the molecular breeding of cabbage.