Anthocyanin Synthesis Pathway Research Articles

Identification of ANS gene regulating fruit color in jujube (Ziziphus jujuba Mill. cv. Lingwuchangzao) and analysis of its expression under elevated temperature and drought stress

BackgroundThe anthocyanin synthase (ANS) gene is pivotal in plant anthocyanin biosynthesis, significantly influencing fruit coloration. Despite extensive on various studies, there remains a notable absence of research on the regulatory functions of the ANS gene in the ‘Lingwuchangzao’ variety under elevated temperature and drought stress.ResultsIn this study, 107 ZjANS genes were identified. Analyses of their physicochemical properties and gene structures revealed that most are localized in the cytosol and nucleus, with only seven ZjANS genes exhibiting basic characteristics. All genes contained the conserved domains 2OG-FeII_Oxy and DIOX-N. Chromosomal localization and Ka/Ks analysis indicated that these genes are distributed across 12 chromosomes, with 22 tandem gene pairs, suggesting purifying selection. Phylogenetic and synteny analysis demonstrated that ZjANS genes are evolutionarily conserved, forming collinear gene pairs with four species. Cis-acting element analysis identified elements related to anthocyanin synthesis and environmental stress, such as LTR and MBS. Predictions of protein interaction indicated that certain ZjANS proteins interact with MYB, CHI, and GA proteins, potentially involved in the anthocyanin synthesis pathway. Transcriptome and qRT-PCR analysis revealed that thirteen ZjANS genes positively respond to coloring in jujube under elevated temperature and drought stress, with ZjANS17 identified as particularly significant.ConclusionThis study identified 107 ZjANS genes and systematically analyzed their evolutionary conservation. Thirteen ZjANS genes were found to positively influence fruit coloring in ‘Lingwuchangzao’ under elevated temperature and drought stress. These findings provide a foundation for further investigations into ZjANS genes and the breeding new jujube varieties.

PhAN2 regulated Ph3GT silencing changes the flower color and anthocyanin content in petunias.

Anthocyanins are important secondary metabolites in plants. After the formation of anthocyanidins, Flavonoid 3-O-glucosyltransferase (3GT) mediated glycosylation first occurs at the C-3 site, forming a stable anthocyanin 3-O-glucoside. Several studies have investigated the function of 3GT using biochemical methods. However, it is necessary to provide further genetic evidence for the role of Ph3GT in petunia (Petunia hybrida). In addition, there is no information regarding the subcellular localization of Ph3GT and the regulation of transcription factors on Ph3GT. In this study, the full-length Ph3GT gene from petunia (Petunia hybrida) was isolated. We found that Ph3GT is localized in the cytoplasm. Ph3GT exhibited high expression levels in the corollas during the coloring period of petunia flowers. VIGS-mediated Ph3GT silencing resulted in a lighter corolla color and a significant decrease in the anthocyanin content in six petunia cultivars. The silencing of Ph3GT affected the expression levels of eight key genes in the anthocyanin synthesis pathway. Additionally, dual luciferase and yeast one-hybrid assays showed that R2R3-MYB transcription factor PhAN2 directly regulates the transcript of Ph3GT.

UV-B irradiation promotes anthocyanin biosynthesis in the leaves of Lycium ruthenicum Murray.

Anthocyanins are the most valuable pigments in Lycium ruthenicum Murray (L.ruthenicum). Although ultraviolet-B (UV-B) irradiation is a key environmental factor influencing anthocyanin biosynthesis in L. ruthenicum, the deep molecular mechanism remains unclear. Herein, we examined the changes in the total anthocyanin content and transcriptomic characteristics of L. ruthenicum leaves following UV-B irradiation treatment. The results showed a twofold increase in anthocyanin content in the leaves of L. ruthenicum after the treatment. The transcriptome analysis showed that the expression of 24 structural genes identified in the anthocyanin synthesis pathway was up-regulated. In particular, F3'H (Unigene0009145) and C4H (Unigene0046607) exhibit notable up-regulation, suggesting their potential roles in anthocyanin synthesis. Protein interaction network results revealed that MYB1 (Unigene0047706) had the highest connectivity, followed by bHLH (Unigene0014085). Additionally, UVR8 (Unigene0067978) and COP1 (Unigene0008780) were found to be highly involved in UV-B signal transduction. These findings provide new insights into the genetic and biochemical mechanisms that regulate anthocyanin production, and could guide agricultural practices to reduce environmental impacts and improve crop yield and quality.

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.

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.

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.

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 transcriptome analysis identifies RpMYB1 as an activator of anthocyanin biosynthesis in Rehmannia piasezkii

Rehmannia piasezkii is a kind of medicinal plants, of the Orobanchaceae family, and well known for its large pink or purple corolla. However, no research on the molecular mechanism of flower color formation in R. piasezkii has been conducted so far. In this study, we investigated the transcriptome of root, stem, leaf and corollas of R. piasezkii using transcriptome sequencing technology and assembled 144,582 unigenes. A total of 58 anthocyanin biosynthetic genes were identified in the R. piasezkii transcriptome, fourteen of which were highly correlated with anthocyanin content, especially RpF3H2, RpDFR2, RpANS1, RpANS2 and RpUFGT. Totally, 35 MYB genes with FPKM values greater than 5 were identified in the R. piasezkii transcriptome, including an R2R3 MYB transcriptional factor RpMYB1, which belongs to subgroup 6 of the R2R3 MYB family. Agrobacterium-mediated transient expression of Nicotiana benthamiana revealed that overexpression of RpMYB1 could activate the expression of structural genes in anthocyanin synthesis pathway and promote the accumulation of anthocyanins in N. benthamiana leaves, indicating that RpMYB1 is a positive regulator of anthocyanin synthesis. Furthermore, combined transient overexpression of RpMYB1 with RpANS1, RpMYB1+RpANS1 with other structural genes all could further enhance the accumulation of anthocyanins in N. benthamiana leaves. Permanent overexpression of RpMYB1 in R. glutinosa promoted anthocyanin accumulation and expression levels of RgCHS, RgF3H, RgDFR and RgANS. Further evidence from dual-luciferase assay suggested that RpMYB1 could bind to the promoter of RpDFR2 and hence activating its expression. These findings provide insight into the molecular regulation in anthocyanin biosynthesis in R. piasezkii and provide valuable genetic resources for the genetic improvement of flower color.

Integrated Transcriptome and Metabolome to Elucidate the Mechanism of Aluminum-Induced Blue-Turning of Hydrangea Sepals

Hydrangea macrophylla is an ornamental plant with varied calyx colors. Interestingly, from red, to purple, to blue, the colors of all Hydrangea macrophylla are formed by unique delphinidin-3-O-glucoside and aluminum ions (Al3+) and 5-O-p-coumaroylquinic acid. The sepals of ‘Blue Mama’ changed from pink to blue, and the contents of delphinidin-3-O-glucoside and aluminum ions increased under 3 g/L aluminum sulfate treatment. However, the mechanism of the effect of aluminum ions on the synthesis and metabolism of anthocyanins in Hydrangea macrophylla is still unclear. In this project, transcriptome sequencing and anthocyanin metabolome analysis were performed on the sepals of ‘Blue Mama’ during flower development at the bud stage (S1), discoloration stage (S2) and full-bloom stage (S3) under aluminum treatment. It was found that delphinidin, delphinidin-3-O-glucoside and delphinidin-3-O-galactoside were the main differential metabolites. The structural genes CHS, F3H, ANS, DFR and BZI in the anthocyanin synthesis pathway were up-regulated with the deepening in sepal color. There was no significant difference between the aluminum treatment and the non-aluminum treatment groups. However, seven transcription factors were up-regulated and expressed to regulate anthocyanin synthesis genes CHS, F3H, BZI and 4CL, promoting the sepals to turn blue. The KEGG enrichment pathway analysis of differentially expressed genes showed that the glutathione metabolism and the ABC transporter pathway were closely related to anthocyanin synthesis and aluminum-ion transport. GST (Hma1.2p1_0158F.1_g069560.gene) may be involved in the vacuolar transport of anthocyanins. The expression of anthocyanin transporter genes ABCC1 (Hma1.2p1_0021F.1_g014400.gene), ABCC2 (Hma1.2p1_0491F.1_g164450.gene) and aluminum transporter gene ALS3 (Hma1.2p1_0111F.1_g053440.gene) were significantly up-regulated in the aluminum treatment group, which may be an important reason for promoting the transport of anthocyanin and aluminum ions to vacuoles and making the sepals blue. These results preliminarily clarified the mechanism of aluminum ion in the synthesis and transport of anthocyanin in Hydrangea macrophylla, laying a foundation for the further study of the formation mechanism of ‘blue complex’ in Hydrangea macrophylla.

Association analysis of BSA-seq, BSR-seq, and RNA-seq reveals key genes involved in purple leaf formation in a tea population (Camellia sinensis).

Purple tea, rich in anthocyanins, has a variety of health benefits and is attracting global interest. However, the regulation mechanism of anthocyanin in purple tea populations has not been extensively studied. In this experiment, RNA-seq, BSA-seq, and BSR-seq were performed using 30 individuals with extreme colors (dark-purple and green) in an F 1 population of 'Zijuan' and 'Jinxuan'. The results show that 459 genes were differentially expressed in purple and green leaves, among which genes involved in the anthocyanin synthesis and transport pathway, such as CHS, F3H, ANS, MYB75, GST, MATE, and ABCC, were highly expressed in purple leaves. Moreover, there were multiple SNP/InDel variation sites on chromosomes 2 and 14 of the tea plant, as identified by BSA-seq. The integrated analysis identified two highly expressed genes (CsANS and CsMYB75) with SNP/InDel site variations in the purple tea plants. By silencing leaves, we proved that CsMYB75 could positively regulate anthocyanin accumulation and expression of related structural genes in tea plants. A 181-bp InDel in the CsMYB75 promoter was also found to be co-segregating with leaf color. The results of this study provide a theoretical reference for the molecular mechanism of anthocyanin accumulation in purple tea plants and contribute to the creation of new tea cultivars with high anthocyanin content.

Investigating the regulatory role of HvANT2 in anthocyanin biosynthesis through protein-motif interaction in Qingke.

Currently, there are no reports on the HvbHLH gene family in the recent barley genome (Morex_V3). Furthermore, the structural genes related to anthocyanin synthesis that interact with HvANT2 have yet to be fully identified. In this study, a bioinformatics approach was used to systematically analyze the HvbHLH gene family. The expression of this gene family was analyzed through RNA sequencing (RNA-seq), and the gene with the most significant expression level, HvANT2, was analyzed using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in different tissues of two differently colored varieties. Finally, structural genes related to anthocyanin synthesis and their interactions with HvANT2 were verified using a yeast one-hybrid (Y1H) assay. The study identified 161 bHLH genes, designated as HvbHLH1 to HvbHLH161, from the most recent barley genome available. Evolutionary tree analysis categorized barley bHLH TFs into 21 subfamilies, demonstrating a pronounced similarity to rice and maize. Through RNA-Seq analysis of purple and white grain Qingke, we discovered a significant transcription factor (TF), HvANT2 (HvbHLH78), associated with anthocyanin biosynthesis. Subsequently, HvANT2 protein-motifs interaction assays revealed 41 interacting motifs, three of which were validated through Y1H experiments. These validated motifs were found in the promoter regions of key structural genes (CHI, F3'H, and GT) integral to the anthocyanin synthesis pathway. These findings provide substantial evidence for the pivotal role of HvANT2 TF in anthocyanin biosynthesis.

The AnUFGT1 Is Involved in the Anthurium ‘Alabama’ Anthocyanidin Deficiency

Anthurium is the second largest tropical flower crop in the world. The international market has urgent demand for anthurium varieties with different spathe colors, which mainly arises from the types and contents of anthocyanin. The flavonoid 3-O-glycosyltransferase (UF3GT) gene is the key enzyme involved in promoting anthocyanin accumulation through glycosylation downstream of the anthocyanin synthesis pathway (ASP). Abnormal functioning of UFGT usually results in a reduction in or loss of anthocyanins. The aim of this study was to reveal the role of one anthurium UFGT gene (AnUFGT1) in ‘Xueyu’ (X), an anthocyanin-deficient mutant of ‘Alabama’. Metabolome analysis was used to analyze the metabolic products in the ASP to determine the possible key link of the anthocyanin deletion mutation. Agrobacterium-mediated transformation of Arabidopsis UFGT functionally deficient mutant (ufgt) and ‘X’ validated the function of AnUFGT1. The results of comparative metabolome analysis of ‘X’ and ‘Alabama’ showed that there was no significant difference in product levels upstream of ASP. The expression levels of AnUFGT1 were significantly greater in ‘Alabama’ than in ‘X’. The overexpression of AnUFGT1 in ufgt significantly increased its anthocyanin contents. The overexpression of AnUFGT1 in ‘X’, mediated by a new injection method, can only promote the synthesis of trace anthocyanins. These results showed that AnUFGT1 could fully compensate the phenotype of ufgt, but only partially compensate the anthocyanidin-deficient phenotype of anthurium mutant X. This difference suggested that anthocyanin-deletion mutations in anthurium ‘X’ are associated with AnUFGT1, but AnUFGT1 is not the only factor. There should be other factors interacting with AnUFGT1 that cause anthocyanin deficiency.

Integrated transcriptome and metabolome analysis reveals anthocyanin biosynthesis mechanisms in pepper (Capsicum annuum L.) leaves under continuous blue light irradiation

BackgroundDifferent metabolic compounds give pepper leaves and fruits their diverse colors. Anthocyanin accumulation is the main cause of the purple color of pepper leaves. The light environment is a critical factor affecting anthocyanin biosynthesis. It is essential that we understand how to use light to regulate anthocyanin biosynthesis in plants.ResultPepper leaves were significantly blue–purple only in continuous blue light or white light (with a blue light component) irradiation treatments, and the anthocyanin content of pepper leaves increased significantly after continuous blue light irradiation. This green-to-purple phenotype change in pepper leaves was due to the expression of different genes. We found that the anthocyanin synthesis precursor-related genes PAL and 4CL, as well as the structural genes F3H, DFR, ANS, BZ1, and F3’5’H in the anthocyanin synthesis pathway, had high expression under continuous blue light irradiation. Similarly, the expression of transcription factors MYB1R1-like, MYB48, MYB4-like isoform X1, bHLH143-like, and bHLH92-like isoform X3, and circadian rhythm-related genes LHY and COP1, were significantly increased after continuous blue light irradiation. A correlation network analysis revealed that these transcription factors and circadian rhythm-related genes were positively correlated with structural genes in the anthocyanin synthesis pathway. Metabolomic analysis showed that delphinidin-3-O-glucoside and delphinidin-3-O-rutinoside were significantly higher under continuous blue light irradiation relative to other light treatments. We selected 12 genes involved in anthocyanin synthesis in pepper leaves for qRT-PCR analysis, and the accuracy of the RNA-seq results was confirmed.ConclusionsIn this study, we found that blue light and 24-hour irradiation together induced the expression of key genes and the accumulation of metabolites in the anthocyanin synthesis pathway, thus promoting anthocyanin biosynthesis in pepper leaves. These results provide a basis for future study of the mechanisms of light quality and photoperiod in anthocyanin synthesis and metabolism, and our study may serve as a valuable reference for screening light ratios that regulate anthocyanin biosynthesis in plants.

IbMYC2 Contributes to Salt and Drought Stress Tolerance via Modulating Anthocyanin Accumulation and ROS-Scavenging System in Sweet Potato.

Basic helix-loop-helix (bHLH) transcription factors extensively affect various physiological processes in plant metabolism, growth, and abiotic stress. However, the regulation mechanism of bHLH transcription factors in balancing anthocyanin biosynthesis and abiotic stress in sweet potato (Ipomoea batata (L.) Lam.) remains unclear. Previously, transcriptome analysis revealed the genes that were differentially expressed among the purple-fleshed sweet potato cultivar 'Jingshu 6' and its anthocyanin-rich mutant 'JS6-5'. Here, we selected one of these potential genes, IbMYC2, which belongs to the bHLH transcription factor family, for subsequent analyses. The expression of IbMYC2 in the JS6-5 storage roots is almost four-fold higher than Jingshu 6 and significantly induced by hydrogen peroxide (H2O2), methyl jasmonate (MeJA), NaCl, and polyethylene glycol (PEG)6000. Overexpression of IbMYC2 significantly enhances anthocyanin production and exhibits a certain antioxidant capacity, thereby improving salt and drought tolerance. In contrast, reducing IbMYC2 expression increases its susceptibility. Our data showed that IbMYC2 could elevate the expression of anthocyanin synthesis pathway genes by binding to IbCHI and IbDFR promoters. Additionally, overexpressing IbMYC2 activates genes encoding reactive oxygen species (ROS)-scavenging and proline synthesis enzymes under salt and drought conditions. Taken together, these results demonstrate that the IbMYC2 gene exercises a significant impact on crop quality and stress resistance.

Integrated Transcriptome and Metabolome Analyses Reveal Bamboo Culm Color Formation Mechanisms Involved in Anthocyanin Biosynthetic in Phyllostachys nigra.

Phyllostachys nigra has green young culms (S1) and purple black mature culms (S4). Anthocyanins are the principal pigment responsible for color presentation in ornamental plants. We employ a multi-omics approach to investigate the regulatory mechanisms of anthocyanins in Ph. nigra. Firstly, we found that the pigments of the culm of Ph. nigra accumulated only in one to four layers of cells below the epidermis. The levels of total anthocyanins and total flavonoids gradually increased during the process of bamboo culm color formation. Metabolomics analysis indicated that the predominant pigment metabolites observed were petunidin 3-O-glucoside and malvidin O-hexoside, exhibiting a significant increase of up to 9.36-fold and 13.23-fold, respectively, during pigmentation of Ph. nigra culm. Transcriptomics sequencing has revealed that genes involved in flavonoid biosynthesis, phenylpropanoid biosynthesis, and starch and sucrose metabolism pathways were significantly enriched, leading to color formation. A total of 62 differentially expressed structural genes associated with anthocyanin synthesis were identified. Notably, PnANS2, PnUFGT2, PnCHI2, and PnCHS1 showed significant correlations with anthocyanin metabolites. Additionally, certain transcription factors such as PnMYB6 and PnMYB1 showed significant positive or negative correlations with anthocyanins. With the accumulation of sucrose, the expression of PnMYB6 is enhanced, which in turn triggers the expression of anthocyanin biosynthesis genes. Based on these findings, we propose that these key genes primarily regulate the anthocyanin synthesis pathway in the culm and contribute to the accumulation of anthocyanin, ultimately resulting in the purple-black coloration of Ph. nigra.

Exploration of cross-cultivar group characteristics of a new cultivar of Prunus mume 'Zhizhang Guhong Chongcui'

'Zhizhang Guhong Chongcui' is a new cultivar of Prunus mume with cross-cultivar group characteristics. It has typical characteristics of cinnabar purple cultivar group and green calyx cultivar group. It has green calyx, white flower, and light purple xylem, but the mechanism remains unclear. In order to clarify the causes of its cross-cultivar group traits, the color phenotype, anthocyanin content and the expression levels of genes related to anthocyanin synthesis pathway of 'Zhizhang Guhong Chongcui', 'Yuxi Zhusha' and 'Yuxi Bian Lü'e' were determined. It was found that the red degree of petals, sepals and fresh xylem in branches was positively correlated with the total anthocyanin content. MYBɑ1, MYB1, and bHLH3 were the key transcription factor genes that affected the redness of the three cultivars of flowers and xylem. The transcription factors further promoted the high expression of structural genes F3'H, DFR, ANS and UFGT, thereby promoting the production of red traits. Combined with phenotype, anthocyanin content and qRT-PCR results, it was speculated that the white color of petals of 'Zhizhang Guhong Chongcui' were derived from the high expression of FLS, F3'5'H, LAR and ANR genes in other branches of cyanidin synthesis pathway, and the low expression of GST gene. The green color of sepals might be originated from the relatively low expression of F3'H, DFR and ANS genes. The red color of xylem might be derived from the high expression of ANS and UFGT genes. This study made a preliminary explanation for the characteristics of the cross-cultivar group of 'Zhizhang Guhong Chongcui', and provided a reference for molecular breeding of flower color and xylem color of Prunus mume.

Metabolomics and transcriptomics reveal molecular mechanisms of anthocyanin accumulation in 'Nanhong' pear (Pyrus ussuriensis) peel at different temperatures

The bright red hue of the 'Nanhong' pear's pericarp is a key sensory attribute, yet its susceptibility to discoloration at harvest poses challenges for maintaining its commercial appeal. This study, employs quality metrics, metabolomics, and transcriptomics to examine fruit quality and anthocyanin metabolism, along with gene expression involved in anthocyanin biosynthesis, under dark conditions at 0 °C, 10 °C, 20 °C, and 30 °C for up to 21 d. Findings indicate that 0 °C is optimal for long-term storage, while 10 °C is preferable for retail display. LC-MS/MS identified 42 anthocyanin metabolites cyanidin-3-O-galactoside (cya-3-O-gal) being the primary pigment in the pericarp. Discoloration was primarily associated with cyanidin-type and peonidin-type anthocyanins. Transcriptome analysis identified a large number of differentially expressed genes (DEGs) in different comparison groups, KEGG pathway analysis highlighted the significant enrichment of DEGs in phenylpropane and flavonoid metabolic pathways. Key genes in the anthocyanin synthesis pathway, including PuPAL, PuCHS, PuCHI, PuF3`H, PuANS, and PuDFR were markedly down-regulated at 20 °C and 30 °C. Transcription factors such as AP2/ERF, WRKY, MYB, NAC, C2H2, bHLH,and bZIP played an influential part in this process. The study also established that fruit senescence and high-temperature stress were the primary causes of pericarp discoloration at these temperatures. Consequently, identifying optimal storage conditions that preserve fruit quality, delay senescence, and sustain anthocyanin accumulation is crucial. In this study, storage at 10 °C demonstrated the most favorable outcomes for maintaining the pericarp's red color during the marketing period.

The Impact of Different Nitrogen Levels on the Tuber Yield and Anthocyanin Synthesis of Purple Potatoes

The biosynthesis of anthocyanins is influenced by external environmental conditions such as light, temperature, and nitrogen level, with nitrogen level being a key factor in anthocyanin synthesis and accumulation. Nitrogen level regulates the transcription factors involved in the anthocyanin synthesis pathway, with low nitrogen levels promoting anthocyanin accumulation, while high nitrogen levels have the opposite effect. Purple potatoes are a type of cultivated crop that is rich in anthocyanins and has unique economic value. Nitrogen fertilizer is crucial to improve the agronomic traits, yield, quality, and anthocyanin content of purple potatoes. In this study, the impact of four different nitrogen concentrations—0 kg/hm2 (N0), 90 kg/hm2 (N1), 225 kg/hm2 (N2) and 360 kg/hm2 (N3)—on the agronomic traits, yield, quality, and anthocyanin content of purple potatoes, ‘Huasong 66’, at different stages were investigated by using physiological index measurement and RNA-seq technology. It was found that the purple potato ‘Huasong 66’ was more sensitive to low nitrogen (N1). Under N1 level of nitrogen fertilization, ‘Huasong 66’ possessed the finest agronomic traits, yield, and quality, and the total anthocyanins in the tubers were significantly increased. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that nitrogen levels in purple potato tubers primarily affect genes related to nutrient transport and metabolism by regulating carbon and nitrogen metabolism, enzyme catalysis and binding, and signal transduction. In addition, nine candidate genes related to the anthocyanin synthesis pathway had been preliminarily screened. These results provide a basis to understand the impact of different nitrogen levels on the tuber yield and anthocyanin synthesis of purple potatoes.

A novel R2R3-MYB transcription factor PbMYB1L of Pyrus bretschneideri regulates cold tolerance and anthocyanin accumulation.

PbMYB1L enhances the cold tolerance and anthocyanin accumulation of transgenic Arabidopsis by regulating the expression of genes related to the cold-responsive genes pathway and anthocyanin synthesis pathway. MYB transcription factors (TFs) have been demonstrated to play diverse roles in plant growth and development. In the present study, we identified a novel R2R3-MYB transcription factor, PbMYB1L, from the peel of 'Red Zaosu' pear (Pyrus bretschneideri), which was induced by cold stress and acted as a positive regulator in anthocyanin biosynthesis. Notably, the transgenic Arabidopsis lines exhibited enhanced tolerance to cold stress. Compared to the Arabidopsis wild-type plants, the transgenic lines displayed longer primary roots and reduced reactive oxygen species (ROS) levels including O2-, hydrogen peroxide (H2O2), and malondialdehyde (MDA). Furthermore, significant upregulation of key cold-responsive genes AtCBF1, AtCBF2, AtCBF3, AtCBF4, and AtKIN1 was observed in the transgenic plants under cold stress conditions compared to wild type. Arabidopsis plants overexpressing PbMYB1L had significant anthocyanin accumulation in leaves after cold treatment with quantitative results indicating higher expression of anthocyanin structural genes compared to wild type. These findings suggest that PbMYB1L not only plays a vital role in conferring cold tolerance but also acts as a crucial regulator of anthocyanin biosynthesis.

Prevention of drought damage through zeolite treatments onVitis vinifera: A promising sustainable solution for soil management

AbstractViticultural activity is intrinsically interlocked with climatic fluctuations and awful subjected to soil management. Increasing temperatures and reduced precipitation originate several alterations in the metabolic plant pathway and a depletion in grapes quality. The main aspiration of soil management is to keep and retain a microenvironment that considers all vine requirements to upgrade the balance between vegetative growth and cluster quality intensifying its resilience towards abiotic stress. Aiming to establish match and dissimilar drought stress responses among Sangiovese grapevines, we scouted ecophysiological and biochemical alterations after zeolite and irrigation soil application (combined and/or alone), using as control plants naturally exposed to abiotic stress in a Chianti Classico vineyard. When grapevines were exposed to water stress alone, the main reactions encompassed dwindled leaf midday water potential (Ψleaf) and increased proline content. These were accompanied by augments in zeaxanthin and lutein leaf contents, symptomatic of oxidative stress. Positive metabolic alterations in grapes triggered by zeolite and/or water showed some influence on the ripening of the berry differently depending on the year. They created a situation of resilience in berry avoiding arrest in the anthocyanin synthesis pathway during the warmest season and produced a more balanced maturation, closer to the objectives of the modern producer of classic Chianti (i.e. fresher and less alcoholic wines) during the fresher one. Under severe drought, it was activated the shikimic acid pathway and in stressed vines treatment, (WS) a reduction in malic acid content was found. Sugar and phenolic compounds were enhanced by zeolitic and undrought treatments. A negative effect enkindled by the drought and high temperatures combined with stress was the accumulation of quercetin‐3‐O‐rutinoside (Qrut), quercetin‐3‐O‐galactoside (Qgala), quercetin‐3‐O‐glucoside (Qgluc) and quercetin‐3‐O‐glucoronide (Qgluco) in WS treatment. These results summarize that clinoptilolite applied to the soil, both combined with water as well in purity, noteworthy protectorship from more detrimental repercussions of drought‐stressed grapevines, suggesting that new sustainable approaches in soil management are useful for the wellness of vines.