Anthocyanin Synthesis Research Articles

Multi-omics analysis reveals anthocyanin synthesis is associated with drought stress tolerance in Chaenomeles speciosa flowers

Drought is one of the most severe abiotic stresses that limit plant growth. Chaenomeles speciosa, a Rosaceae plant of high ornamental value, exhibits noticeable changes in petal color under drought conditions. A multi-omics joint analysis was employed to examine the mechanism of flower color changes under drought conditions. We detected that the main reason for the change in flower color is the synthesis of anthocyanins, while ethylene (ET) and abscisic acid (ABA) also significantly increase. While ET and ABA are known to promote anthocyanin synthesis, their coordinated regulation under drought conditions remains unclear. This study aims to validate the hypothesis that ET/ABA mediated responses to drought regulate anthocyanin synthesis. The study inferred that the CsERF2, CsbZIP24–CsMYB6 signaling module plays a crucial role in the precise and dynamic regulation of anthocyanin synthesis. Yeast heterologous expression is used to demonstrate the drought tolerance function of candidate genes. Also, we performed transient expression analysis in petals suggests that these genes significantly contribute to anthocyanin accumulation under drought conditions. Moreover, Yeast one-hybrid and dual luciferase assays indicated that CsERF2 and CsbZIP24 can directly activate the expression of CsMYB6. Yeast two-hybrid, bimolecular fluorescence complementation, and Split-luciferase assays revealed interactions between CsERF2 and CsbZIP24 proteins, promoting anthocyanin biosynthesis. The study proposes a regulatory model for the mechanism of flower color change and aimed at exploring drought resistant ornamental plants and achieving their sustainable development.

Integrated Phenotypic Physiology and Transcriptome Analysis Revealed the Molecular Genetic Basis of Anthocyanin Accumulation in Purple Pak-Choi

Purple Pak-choi is rich in anthocyanins, which have both ornamental and edible health functions, and has been used more and more widely in facility cultivation. In order to further clarify the molecular mechanism of purple Pak-choi, two Pak-choi inbred lines (‘PQC’ and ‘HYYTC’) were selected for the determination of pigment content and transcriptome analysis, and the key genes controlling the formation of purple character in leaves of Pak-choi were discovered. The results of pigment determination showed that the anthocyanin content of ‘PQC’ was 0.29 mg/g, which was about 100 times than ‘HYYTC’; The chlorophyll content of ‘HYYTC’ was 2.25 mg/g, while ‘PQC’ only contained 1.05 mg/g. A total of 20 structural genes related to anthocyanin biosynthesis and 28 transcriptional regulatory genes were identified by transcriptome analysis. Weighted gene co-expression network analysis (WGCNA) was used to construct the weight network analysis map of 14 genes. The results showed that the cinnamate hydroxylase gene (BraA04002213, BrC4H3), flavanone-3- hydroxylase (BraA09004531, BrF3H1), and chalcone synthetase (BraA10002265, BrCHS1) were the core genes involved in the anthocyanin synthesis pathway of purple Pak-choi. The results identified the key genes controlling the formation of purple leaf traits, which laid a foundation for further analysis of the molecular mechanism of anthocyanin accumulation in purple Pak-choi and provided a theoretical basis for leaf color regulation.

Biosynthesis and Extraction of Chlorophyll, Carotenoids, Anthocyanins, and Betalaine In Vivo and In Vitro.

As natural bioactive compounds, plant pigments play crucial roles not only in plant phenotype, growth, development, and adaptation to stress but also hold unique value in biotechnology, healthcare, and industrial applications. There is growing interest in the biosynthesis and acquisition of plant pigments. Thus, this paper explores emerging extraction methods of natural pigments and elucidates the biosynthesis pathways of four key plant pigments, chlorophylls, carotenoids, anthocyanins, and betalaine in vivo and in vitro. We comprehensively discuss the application of solvent, supercritical fluid [extraction], ultrasonic, and microwave-assisted extraction techniques, as well as introducing key enzymes, precursors, and synthetic pathways involved in pigment synthesis. δ-Aminolevulinic acid represents a pivotal initiating enzyme for chlorophyll synthesis, whereas isopentenylpyrophosphate, (IPP) and dimethylallyl pyrophosphate, (DMAPP) are closely associated with carotenoid biosynthesis. Phenylalanine and tyrosine are critical substances for anthocyanin and betalaine synthesis, respectively. Hence, crucial genes such as chlI, crtB, PGT8, CYP76AD1, and BvDODA can be employed for heterologous biosynthesis in vitro to meet the demand for increased plant pigment amount. As a pivotal determinant of plant coloration, an in-depth exploration into the high-quality acquisition of plant pigments can provide a basis for developing superior pigments and offer new insights into increasing pigment yield.

Loss-of-function mutation in anthocyanidin reductase activates the anthocyanin synthesis pathway in strawberry

Fruit color substantially affects consumer preferences, with darker red strawberries being economically more valuable due to their higher anthocyanin content. However, the molecular basis for the dark red coloration remains unclear. Through screening of an ethyl methanesulfonate mutant library, we identified a rg418 mutant, that demonstrated anthocyanin accumulation during early fruit development stages. Furthermore, the ripening fruits of this mutant had higher anthocyanin content than wild-type (WT) fruits. An analysis of flavonoid content in WT and rg418 mutant fruits revealed substantial changes in metabolic fluxes, with the mutant exhibiting increased levels of anthocyanins and flavonols and decreased levels of proanthocyanidins. Bulked sergeant analysis sequencing indicated that the mutant gene was anthocyanidin reductase (ANR), a key gene in the proanthocyanidin synthesis pathway. Furthermore, transcriptome sequencing revealed the increased expression of MYB105 during the early development stage of mutant fruits, which promoted the expression of UFGT (UDP-glucose flavonoid 3-O-glucosyltransferase), a key gene involved in anthocyanin synthesis, thus substantially enhancing the anthocyanin content in the mutant fruits. Additionally, mutating ANR in a white-fruited strawberry variant (myb10 mutant) resulted in appealing pink-colored fruits, suggesting the diverse roles of ANR in fruit color regulation. Our study provides valuable theoretical insights for improving strawberry fruit color.

Inhibitors of abscisic acid synthesis or signaling affect anthocyanin synthesis and photoreceptors in grape berries

Effects of endogenous abscisic acid (ABA) and ABA signal transduction on anthocyanin and photoreceptors in grape berries (Vitis labruscana) were examined using NDGA, the inhibitor of the rate-limiting ABA biosynthesis enzyme in grapes, and PANMe, an ABA receptor interaction antagonist. Both NDGA and PANMe treatments decreased anthocyanin concentrations. The expressions of VvSnRK2.6, VvABF2, and VvPP2C9 in NDGA treatment were not significantly different from the untreated control; by contrast, PANMe treatment decreased the expressions of VvSnRK2.6, VvABF2, and VvPP2C9. Both NDGA and PANMe treatments decreased the expressions of VvPhot2 and VvCRYa. These results suggest that both endogenous ABA concentrations and ABA signaling affect anthocyanin synthesis and photoreceptor genes such as VvPhot2 and VvCRYa in grape skins.

The Genome-Wide Identification of the Dihydroflavonol 4-Reductase (DFR) Gene Family and Its Expression Analysis in Different Fruit Coloring Stages of Strawberry.

Dihydroflavonol 4-reductase (DFR) significantly influences the modification of flower color. To explore the role of DFR in the synthesis of strawberry anthocyanins, in this study, we downloaded the CDS sequences of the DFR gene family from the Arabidopsis genome database TAIR; the DFR family of forest strawberry was compared; then, a functional domain screen was performed using NCBI; the selected strawberry DFR genes were analyzed; and the expression characteristics of the family members were studied by qRT-PCR. The results showed that there are 57 members of the DFR gene family in strawberry, which are mainly expressed in the cytoplasm and chloroplast; most of them are hydrophilic proteins; and the secondary structure of the protein is mainly composed of α-helices and random coils. The analysis revealed that FvDFR genes mostly contain light, hormone, abiotic stress, and meristem response elements. From the results of the qRT-PCR analysis, the relative expression of each member of the FvDFR gene was significantly different, which was expressed throughout the process of fruit coloring. Most genes had the highest expression levels in the full coloring stage (S4). The expression of FvDFR30, FvDFR54, and FvDFR56 during the S4 period was 8, 2.4, and 2.4 times higher than during the S1 period, indicating that the DFR gene plays a key role in regulating the fruit coloration of strawberry. In the strawberry genome, 57 members of the strawberry DFR gene family were identified. The higher the DFR gene expression, the higher the anthocyanin content, and the DFR gene may be the key gene in anthocyanin synthesis. Collectively, the DFR gene is closely related to fruit coloring, which lays a foundation for further exploring the function of the DFR gene family.

The ERF transcription factor ZbERF3 promotes ethylene-induced anthocyanin biosynthesis in Zanthoxylum bungeanum

Ethylene regulates fruit ripening, and in Zanthoxylum bungeanum, fruit color deepened with increasing of ethylene during fruit ripening. However, the molecular mechanism of this physiological process was still unclear. In this study, through the combined analysis of transcriptome and metabolome, it was found that ethylene release was consistent with anthocyanin synthesis, and ethylene response factors (ERFs) were significantly related to anthocyanin biosynthesis during the fruit ripening of Z. bungeanum. Ethylene treatment significantly induced fruit coloration and promoted anthocyanin synthesis and the expression of ZbERF3. Furthermore, Yeast one-hybrid assays and Luciferase reporter assays demonstrated that ZbERF3 directly bound to the promoter of ZbMYB17 and transcriptionally activated its expression. What's more, it was demonstrated that ZbMYB17 directly bound to the promoter of ZbANS, promoting anthocyanin biosynthesis. Overall, this study revealed the mechanism of ERF and MYB synergistically regulating the coloration of Z. bungeanum fruit.

NAC Transcription Factor PpNAP4 Promotes Chlorophyll Degradation and Anthocyanin Synthesis in the Skin of Peach Fruit.

Chlorophyll (Chl) catabolism and anthocyanin synthesis play pivotal roles in determining the final skin color of fruits during maturation. However, in peach (Prunus persica) fruit, the regulatory mechanism governing skin color, especially the Chl catabolism, remains largely elusive. In this study, we identified ten Chl catabolic genes (CCGs), with PpSGR emerging as a key regulator in Chl degradation in peaches. Furthermore, a NAC-like, activated by AP3/P1 (NAP) transcription factor (TF), PpNAP4, was identified as a positive modulator of Chl breakdown. PpNAP4 induced the expression of PpSGR and other CCGs, including PpPPH, PpPAO, and PpTIC55-2, by directly binding to their promoters. Overexpression of PpNAP4 resulted in a heightened expression of these genes and accelerated Chl degradation. Notably, PpNAP4 also positively regulated the expression of PpANS and PpMYB10.1, one key structural gene and a core transcriptional regulator of anthocyanin synthesis, thereby contributing to fruit coloration. In summary, our findings elucidate that PpNAP4 serves as a pivotal regulator in determining the final skin color of peach by orchestrating Chl degradation and anthocyanin accumulation through direct activation of multiple CCGs and anthocyanin related genes.

Genome-wide identification of the phenylalanine ammonia-lyase gene from Epimedium Pubescens Maxim. (Berberidaceae): novel insight into the evolution of the PAL gene family

BackgroundPhenylalanine ammonia-lyase (PAL) serves as a key gateway enzyme, bridging primary metabolism and the phenylpropanoid pathway, and thus playing an indispensable role in flavonoid, anthocyanin and lignin biosynthesis. PAL gene families have been extensively studied across species using public genomes. However, a comprehensive exploration of PAL genes in Epimedium species, especially those involved in prenylated flavonol glycoside, anthocyanin, or lignin biosynthesis, is still lacking. Moreover, an in-depth investigation into PAL gene family evolution is warranted.ResultsSeven PAL genes (EpPAL1-EpPAL7) were identified. EpPAL2 and EpPAL3 exhibit low sequence identity to other EpPALs (ranging from 61.09 to 64.38%) and contain two unique introns, indicating distinct evolutionary origins. They evolve at a rate ~ 10 to ~ 54 times slower compared to EpPAL1 and EpPAL4-7, suggesting strong purifying selection. EpPAL1 evolved independently and is another ancestral gene. EpPAL1 formed EpPAL4 through segmental duplication, which lead to EpPAL5 and EpPAL6 through tandem duplications, and EpPAL7 through transposed duplication, shaping modern EpPALs. Correlation analysis suggests EpPAL1, EpPAL2 and EpPAL3 play important roles in prenylated flavonol glycosides biosynthesis, with EpPAL2 and EpPAL3 strongly correlated with both Epimedin C and total prenylated flavonol glycosides. EpPAL1, EpPAL2 and EpPAL3 may play a role in anthocyanin biosynthesis in leaves. EpPAL2, EpPAL3, EpPAL6, and EpPAL7 might be engaged in anthocyanin production in petals, and EpPAL2 and EpPAL3 might also contribute to anthocyanin synthesis in sepals. Further experiments are needed to confirm these hypotheses. Novel insights into the evolution of PAL gene family suggest that it might have evolved from a monophyletic group in bryophytes to large-scale sequence differentiation in gymnosperms, basal angiosperms, and Magnoliidae. Ancestral gene duplications and vertical inheritance from gymnosperms to angiosperms likely occurred during PAL evolution. Most early-diverging eudicotyledons and monocotyledons have distinct histories, while modern angiosperm PAL gene families share similar patterns and lack distant gene types.ConclusionsEpPAL2 and EpPAL3 may play crucial roles in biosynthesis of prenylated flavonol glycosides and anthocyanins in leaves and flowers. This study provides novel insights into PAL gene family evolution. The findings on PAL genes in E. pubescens will aid in synthetic biology research on prenylated flavonol glycosides production.

Blue maize with pigmented germ: Phytochemical compounds and tortilla color

Blue maize is used in the production of various traditional foods, and its phytochemical composition has been claimed to possess health benefits. In this study, two blue maize hybrids with pigmented germ grown in five environments were studied under the hypothesis that the germ could have a different anthocyanin profile from that of anthocyanins synthesized in the aleurone layer, and that those in the germ could increase the total anthocyanin content in the whole grain. The percentage of pigmented germ, total anthocyanin content (TA) and total soluble phenols in the germ, whole grain and tortilla were evaluated to determine how tortilla color is modified. For the first time, the anthocyanin and fatty acid profiles of pigmented germ were determined. In the anthocyanin profile, anthocyanins derived from peonidin stood out, making 50.7 %. The most abundant fatty acid was linoleic acid (40.6 %). Whole kernel TA content increased when the maize had a higher percentage of pigmented germ, with minimal changes when grain was transformed to tortilla, resulting in darker tortillas. The large variation in TA among environments highlights the importance of identifying the environments that most favor anthocyanin synthesis.

Understanding the molecular regulation of flavonoid 3’-hydroxylase in anthocyanin synthesis: insights from purple qingke

BackgroundThe Flavonoid 3’-hydroxylase gene(F3’H) is an important structural gene in the anthocyanin synthesis pathway of plants, which has been proven to be involved in the color formation of organs such as leaves, flowers, and fruits in many plants. However, the mechanism and function in barley are still unclear.ResultsIn order to explore the molecular mechanism of the grain color formation of purple qingke, we used the cultivated qingke variety Nierumzha (purple grain) and the selected qingke variety Kunlun 10 (white grain) to conduct transcriptomic sequencing at the early milk, late milk and soft dough stage. Weighted Gene Co-expression Network Analysis (WGCNA) was used to construct weighted gene co-expression network related to grain color formation, and three key modules (brown, yellow, and turquoise modules) related to purple grain of qingke were selected. F3’H (HORVU1Hr1G094880) was selected from the hub gene of the module for the yeast library, yeast two-hybrid (Y2H), subcellular localization and other studies. It was found that in purple qingke, HvnF3’H mainly distributed in the cytoplasm and cell membrane and interacted with several stress proteins such as methyltransferase protein and zinc finger protein.ConclusionsThe results of this study provide reference for the regulation mechanism of anthocyanin-related genes in purple grain qingke.

Telomere-to-telomere, gap-free assembly of the Rosa rugosa reference genome

Rosa, a genus esteemed worldwide for its ornamental plants, has encountered barriers in functional genomic studies and further genetic enhancement due to incomplete sequences and floating regions in previously sequenced genomes. Our groundbreaking study introduced a meticulously assembled, continuous, and fully bridged reference genome for Rosa rugosa, constructed through a sophisticated combination of PacBio High-Fidelity, ONT ultra-long reads, and Hi-C data. This robust assembly spanned 444.55 Mb and encompassed 34 109 protein-coding genes. We have uniquely assembled each chromosome into single, gap-free structures, successfully identifying all 14 telomeres and seven centromeres, a feat not achieved previously. The centromeric regions were distinguished by tandem repeats, primarily composed of centromere-specific 159-bp monomers, and a significant enrichment of ATHILA/Gypsy long terminal repeat retrotransposons in proximal regions. Our research highlighted recent tandem duplications as instrumental in bolstering R. rugosa's stress tolerance, environmental adaptability, and enhanced anthocyanin synthesis. Furthermore, our study ventured into uncharted territory by predicting transcription factors potentially regulating anthocyanin biosynthesis through the employment of gene co-expression networks, providing new avenues for research. This comprehensive reference genome not only serves as a cornerstone for in-depth exploration of genomic architecture and functionalities in R. rugosa but also acts as a catalyst for innovative breeding strategies and genetic refinement within the genus.

Metabolomics Uncovers the Mechanisms of Nitrogen Response to Anthocyanins Synthesis and Grain Quality of Colored Grain Wheat (Triticum aestivum L.).

Nitrogen (N) is a key factor for plant growth and affects anthocyanin synthesis. This study aimed to clarify the potential mechanisms of N levels (LN, 0 kg·ha-1; MN, 150 kg·ha-1; HN, 225 kg·ha-1) in anthocyanin synthesis and grain quality of colored grain wheat. HN increased the yield component traits and grain morphology traits in colored grain wheat while decreasing the processing and nutrient quality traits. Most quality traits were significantly negatively correlated with the yield composition and morphological traits. Anthocyanin was more accumulated under LN conditions, but other related yield and morphological traits of colored grain wheat declined. The anthocyanin content was the highest in blue wheat, followed by that in purple wheat. Cyanidin-3-O-(6-O-malonyl-β-d-glucoside) and cyanidin-3-O-rutinoside were the predominant anthocyanins in blue and purple wheat. The identified anthocyanin-related metabolites were associated with flavonoid biosynthesis, anthocyanin biosynthesis, and secondary metabolite biosynthesis. Therefore, the study provided information for optimizing nitrogen fertilizer management in producing high quality colored wheat and verified the close relationship between anthocyanin and low N condition.

Combined Metabolome and Transcriptome Analysis Revealed the Accumulation of Anthocyanins in Grape Berry (Vitis vinifera L.) under High-Temperature Stress.

In grape (Vitis vinifera L.) cultivation, high temperatures (HTs) usually reduce the accumulation of anthocyanins. In order to elucidate the regulatory mechanism of anthocyanin biosynthesis under high-temperature environments, we investigated the effects of HT stress at veraison (5% coloring of grape ears) on fruit coloration and anthocyanin biosynthesis in 'Summer Black' (XH) and 'Flame seedless' (FL) grapevines. Compared to the control group (35 °C), the total anthocyanin content of XH and FL grapes subjected to a high-temperature (HT) treatment group (40 °C) decreased significantly as the HT treatment continued, but showed an upward trend with fruit development. However, the concentration of procyanidins increased significantly following HT treatment but decreased with fruit development. Nonetheless, FL grapes showed some resistance to the HT condition, producing anthocyanin content at ripeness comparable to the control group, demonstrating a greater adaptability to HT conditions than XH grapes. Based on the CIRG index, at stage S4, the fruit of FL was classified as dark red, while XH was classified as blue-black in the control group. Anthocyanin-targeted metabonomics identified eight different types of anthocyanins accumulating in the peels of XH and FL grapes during ripening, including cyanidins, delphinidins, malvidins, pelargonidins, peonidins, petunidins, procyanidins, and flavonoids. Malvidins were the most abundant in the two grape varieties, with malvidin-3-O-glucoside being more sensitive to high temperatures. HT treatment also down-regulated the expression of structural genes and regulators involved in the anthocyanin synthesis pathways. We used the WGCNA method to identify two modules that were significantly correlated with total anthocyanin and procyanidin contents. Among them, MYBCS1, bHLH137, WRKY65, WRKY75, MYB113-like, bZIP44, and GST3 were predicted to be involved in grape anthocyanin biosynthesis. In conclusion, this study conducted in-depth research on the HT inhibition of the biosynthesis of anthocyanins in XH and FL grapes, for reference.

Identification of a recessive gene RgM4G52 conferring red glume, stem, and rachis in a Triticum boeoticum mutant.

Anthocyanins are plant secondary metabolites belonging to the polyphenol class of natural water-soluble phytopigments. The accumulation of anthocyanins in different plant tissues can improve plant survival under adverse conditions. In addition, plants with the resulting colorful morphology can be utilized as landscape plants. Triticum boeoticum (syn. Triticum monococcum ssp. aegilopoides, 2n=2x=14, AbAb) serves as a valuable genetic resource for the improvement of its close relative common wheat in terms of enhancing resilience to various biotic and abiotic stresses. In our previous study, the EMS-mutagenized mutant Z2921 with a red glume, stem, and rachis was generated from T. boeoticum G52, which has a green glume, stem, and rachis. In this study, the F1, F2, and F2:3 generations of a cross between mutant-type Z2921 and wild-type G52 were developed. A single recessive gene, tentatively designated RgM4G52, was identified in Z2921 via genetic analysis. Using bulked segregant exome capture sequencing (BSE-Seq) analysis, RgM4G52 was mapped to chromosome 6AL and was flanked by the markers KASP-58 and KASP-26 within a 3.40-cM genetic interval corresponding to 1.71-Mb and 1.61-Mb physical regions in the Chinese Spring (IWGSC RefSeq v1.1) and Triticum boeoticum (TA299) reference genomes, respectively, in which seven and four genes related to anthocyanin synthesis development were annotated. Unlike previously reported color morphology-related genes, RgM4G52 is a recessive gene that can simultaneously control the color of glumes, stems, and rachis in wild einkorn. In addition, a synthetic Triticum dicoccum-T. boeoticum amphiploid Syn-ABAb-34, derived from the colchicine treatment of F1 hybrids between tetraploid wheat PI 352367 (T. dicoccum, AABB) and Z2921, expressed the red stems of Z2921. The flanking markers of RgM4G52 developed in this study could be useful for developing additional common wheat lines with red stems, laying the foundation for marker-assisted breeding and the fine mapping of RgM4G52.

Effect of Co-Application of Chinese Milk Vetch and Iron-Modified Biochar on Rice in Antimony-Polluted Soil

Antimony (Sb) toxicity is a serious concern across the globe due to its hazardous impacts on plants and living organisms. The co-application of Chinese milk vetch (CMV) and biochar (BC) is a common agricultural practice, however, the effects of combined CMV and BC in mitigating Sb toxicity and bio-availability remain unclear. Therefore, this study investigated the impacts of CMV, rape straw biochar (RBC), and iron-modified biochar (FMB) and their combinations on rice productivity, physiological, and biochemical functioning of rice and Sb availability. Antimony toxicity caused a marked reduction in rice growth and productivity by decreasing chlorophyll, and anthocyanin synthesis, leaf water contents, osmolyte synthesis and antioxidant activities while, increasing hydrogen peroxide (H2O2), electrolyte leakage (EL), and malondialdehyde (MDA) production and Sb accumulation. Co-application of CMV and FMB increased biomass (29.50%) and grain yield (51.07%) of rice by increasing chlorophyll, and anthocyanin synthesis, leaf water contents, osmolyte synthesis, antioxidant activities, and decreasing production of H2O2, EL, and MDA and Sb accumulation in roots (90.41%) and shoots (96.38%). Furthermore, the combined addition of CMV and FMB also reduced the soil available Sb by 75.57% which resulted in less accumulation of Sb in plant parts and improved growth and yield. Given these facts, these findings indicate that co-application of CMV and FMB is a promising approach to remediate Sb-polluted soils and improve sustainable and safer rice productivity.

Combining transcriptomics and HPLC to uncover variations in quality formation between ‘Benihoppe’ and ‘Fenyu No.1’ strawberries

‘Benihoppe’ and ‘Fenyu No.1’ are representative varieties of red and pink strawberries in China, possess distinct hue and flavor profiles. This study analyzed the underlying biochemical and molecular differences of two varieties utilizing transcriptomics and high-performance liquid chromatography (HPLC). Ripening ‘Benihoppe’ fruits accumulated more sucrose and pelargonin-3-glucoside (P3G) with a little cyanidin and higher firmness. Whereas ripening ‘Fenyu No.1’ fruits contained more fructose, glucose, malic acid and ascorbic acid (AsA), but less P3G and citric acid. Moreover, genotype significantly influenced phenolic compounds contents in strawberries. Transcriptome analysis revealed that pectin degradation (PL, PG, PE), sucrose synthesis (CWINV, SUS, TPS) and citric acid metabolism (α-OGDH, ICDH, GAD, GS, GDH, PEPCK, AST) were weakened in ‘Benihoppe’ fruit. In contrast, the synthesis of sucrose (CWINH, SPS), citric acid (CS, PEPC), anthocyanin (F3H, F3′H, F3′5′H, DFR, UFGT and ANS), and citric acid transport (V-ATPase) was enhanced. In ‘Fenyu No.1’ fruit, the degradation of sucrose, citric acid, and pectin was enhanced, along with the synthesis of malic acid (ME) and ascorbic acid (PMM, MDHAR and GaLUR). However, anthocyanins synthesis, glucose metabolism (HK, G6PI, PFK, G6PDH, PGK, PGM, ENO, PK), fructose metabolism (FK), citric acid synthesis and transport, and AsA degradation (AO, APX) were relatively weak. RT-qPCR results corroborated the transcriptome data. In conclusion, this study revealed the distinctions and characteristics of strawberries with different fruit colors regarding texture, flavor and color formation processes. These findings offer valuable insights for regulating metabolic pathways and identifying key candidate genes to improve strawberry quality.

Exogenous inoculation of Pediococcus pentosaceus SSC12 positively regulates anthocyanin biosynthesis to alleviate browning of postharvest litchi (Litchi chinensis Sonn.)

Litchi is prone to pericarp browning after-harvest, which seriously affects the quality of litchi and causes huge loss. Pericarp browning is closely related to anthocyanin metabolism. Lactic acid bacteria (LAB), a kind of probiotics, is used for food preservation. The aim of this study was to investigate whether and how inoculation of LAB strain Pediococcus pentosaceus SSC12 of litchi fruit affect the pericarp browning and anthocyanin metabolism in postharvest through transcriptome and metabolome analyses. This is the first work showing that strain SSC12 suppressed development of browning and maintained the coloration of litchi pericarp at 10 ± 1 °C. Strain SSC12 upregulated 8 key differentially expressed genes (DEGs) associated with anthocyanin biosynthesis and transport, while downregulated 3 key DEGs involved in anthocyanin degradation in anthocyanin synthesis pathway. Strain SSC12 caused increase in 9 different anthocyanin-related metabolites levels and phenylalanine ammonia lyase activity, disease in peroxidase, laccase and polyphenol oxidase activities. Taken together, strain SSC12 suppressed browning of litchi via elevateing anthocyanin synthesis and transport and inhibiting anthocyanin degradation. These findings suggested that strain SSC12 might serve as a potential probiotic strains to alternative for chemical reagents in postharvest fruit preservation and contribute to the exploration of newgreen approachs for postharvest losses.

De novo biosynthesis of anthocyanins in Saccharomyces cerevisiae using metabolic pathway synthases from blueberry

BackgroundAnthocyanins are water-soluble flavonoids in plants, which give plants bright colors and are widely used as food coloring agents, nutrients, and cosmetic additives. There are several limitations for traditional techniques of collecting anthocyanins from plant tissues, including species, origin, season, and technology. The benefits of using engineering microbial production of natural products include ease of use, controllability, and high efficiency.ResultsIn this study, ten genes encoding enzymes involved in the anthocyanin biosynthetic pathway were successfully cloned from anthocyanin-rich plant materials blueberry fruit and purple round eggplant rind. The Yeast Fab Assembly technology was utilized to construct the transcriptional units of these genes under different promoters. The transcriptional units of PAL and C4H, 4CL and CHS were fused and inserted into Chr. XVI and IV of yeast strain JDY52 respectively using homologous recombination to gain Strain A. The fragments containing the transcriptional units of CHI and F3H, F3’H and DFR were inserted into Chr. III and XVI to gain Strain B1. Strain B2 has the transcriptional units of ANS and 3GT in Chr. IV. Several anthocyanidins, including cyanidin, peonidin, pelargonidin, petunidin, and malvidin, were detected by LC-MS/MS following the predicted outcomes of the de novo biosynthesis of anthocyanins in S. cerevisiae using a multi-strain co-culture technique.ConclusionsWe propose a novel concept for advancing the heterologous de novo anthocyanin biosynthetic pathway, as well as fundamental information and a theoretical framework for the ensuing optimization of the microbial synthesis of anthocyanins.

Identification and characterization of HCT genes in blackberry: Overexpression of RuHCT1 enhances anthocyanin contents

Emerging as a new generation of economic fruit, blackberries are rich in flavonoids and anthocyanins. Acylation can change the pharmacological activity and structural stability of anthocyanins and is of great significance to plant growth and secondary metabolism. In this study, based on the transcriptome data of the blackberry fruit, five blackberry hydroxycinnamoyl transferase (HCT) genes were screened and identified, all of which belong to the BAHD superfamily. Among them, RuHCT1, RuHCT2 and RuHCT3 have similar tertiary structures and functions. RuHCT1, the gene with the largest difference in expression between immature and mature blackberry fruits, was selected for overexpression research. The results showed that the content of anthocyanin in the leaves of transgenic plants was significantly increased, while the content of secondary metabolites such as lignan and chlorogenic acid was decreased. Moreover, the expression levels of the NtCCOMT, NtCHS and NtCYP98A genes in transgenic tobacco were lower than those in nontransgenic plants. In conclusion, RuHCT1 may be an important structural gene regulating anthocyanin synthesis in blackberry. RuHCT1 may also inhibit lignan synthesis and affect the expression of other key structural genes. This study provides a theoretical reference regarding the biosynthesis and regulatory mechanisms of anthocyanins in blackberry, contributing to the breeding of new blackberry cultivars with high anthocyanin content.