Anthocyanin Metabolism Research Articles

Transcriptome Analysis Reveals Roles of Anthocyanin- and Jasmonic Acid-Biosynthetic Pathways in Rapeseed in Response to High Light Stress.

Rapeseed (Brassica napus) is one of the major important oil crops worldwide and is largely cultivated in the Qinghai-Tibetan plateau (QTP), where long and strong solar-radiation is well-known. However, the molecular mechanisms underlying rapeseed’s response to light stress are largely unknown. In the present study, the color of rapeseed seedlings changed from green to purple under high light (HL) stress conditions. Therefore, changes in anthocyanin metabolism and the transcriptome of rapeseed seedlings cultured under normal light (NL) and HL conditions were analyzed to dissect how rapeseed responds to HL at the molecular level. Results indicated that the contents of anthocyanins, especially glucosides of cyanidin, delphinidin, and petunidin, which were determined by liquid chromatography-mass spectrometry (LC-MS), increased by 9.6-, 4.2-, and 59.7-fold in rapeseed seedlings exposed to HL conditions, respectively. Next, RNA-sequencing analysis identified 7390 differentially expressed genes (DEGs), which included 4393 up-regulated and 2997 down-regulated genes. Among the up-regulated genes, many genes related to the anthocyanin-biosynthetic pathway were enriched. For example, genes encoding dihydroflavonol reductase (BnDFR) and anthocyanin synthase (BnANS) were especially induced by HL conditions, which was also confirmed by RT-qPCR analysis. In addition, two PRODUCTION OF ANTHOCYANIN PIGMENTATION 2 (BnPAP2) and GLABRA 3 (BnGL3) genes encoding MYB-type and bHLH-type transcription factors, respectively, whose expression was also up-regulated by HL stress, were found to be associated with the changes in anthocyanin biosynthesis. Many genes involved in the jasmonic acid (JA)-biosynthetic pathway were also up-regulated under HL conditions. This finding, which is in agreement with the well-known positive regulatory role of JA in anthocyanin biosynthesis, suggests that the JA may also play a key role in the responses of rapeseed seedlings to HL. Collectively, these data indicate that anthocyanin biosynthesis-related and JA biosynthesis-related pathways mediate HL responses in rapeseed. These findings collectively provide mechanistic insights into the mechanisms involved in the response of rapeseed to HL stress, and the identified key genes may potentially be used to improve HL tolerance of rapeseed cultivars through genetic engineering or breeding strategies.

A novel transcription factor CmMYB012 inhibits flavone and anthocyanin biosynthesis in response to high temperatures in chrysanthemum

Flavones are among the major colorless pigments synthesized through branches of the flavonoid pathway in plants. However, due to the absence of a gene encoding flavone synthase (FNS) in the model plant Arabidopsis thaliana species, the regulatory mechanism of FNS-catalyzed flavone biosynthesis has rarely been studied in plants. Here, it was found that flavones play a predominant role in the elimination of excess reactive oxygen species (ROS) at high temperatures in colorless plant organs. A novel atypical subgroup 7 (SG7) R2R3-MYB transcription factor, CmMYB012, was found to be induced in response to prolonged high temperatures and to inhibit flavone biosynthesis by directly regulating CmFNS. Moreover, CmMYB012 was also found to inhibit anthocyanin biosynthesis by suppressing the expression of CmCHS, CmDFR, CmANS, and CmUFGT. CmMYB012 overexpression exerted a negative influence on plant fitness and pink flower color formation, while CmMYB012 suppression had the opposite effect in response to high temperatures. Our findings provide new insights into the mechanisms by which high temperatures regulate the metabolism of flavones and anthocyanins to affect plant fitness and flower color formation.

Dietary Antioxidant Anthocyanins Mitigate Type II Diabetes through Improving the Disorder of Glycometabolism and Insulin Resistance.

Insulin resistance (IR) is one of the pathological reasons for type II diabetes mellitus (T2DM). Therefore, it is important to prevent the body from developing T2DM by improving IR and maintaining glucose homeostasis. Anthocyanins (ACNs) are water-soluble pigments and are widely distributed in natural products. This article summarizes research on the bioavailability and metabolism of ACNs. Moreover, we further elaborate on how ACNs reduce IR and hyperglycemia during the development of T2DM based on studies over the past 20 years. Many studies have demonstrated that ACNs are small molecules that target the pancreatic, liver, muscle, and adipose tissues, preventing IR and hyperglycemia. However, the molecular mechanisms are still unclear. Therefore, we envision whether the molecular mechanism of reducing T2DM by ACNs could be more deeply investigated.

Transcriptome and metabolome analysis reveals anthocyanin biosynthesis pathway associated with ramie (Boehmeria nivea (L.) Gaud.) leaf color formation

BackgroundThe bast fiber crop ramie can be used as high-quality forage resources, especially in tropical or subtropical region where there is lack of high-quality protein feed. Hongxuan No.1 (HX_1) is a unique ramie variety with a light reddish brown leaf color, which is obviously different from elite cultivar, Zhongzhu No.1 (ZZ_1, green leaf). While, the regulatory mechanism of color difference or secondary metaboliates synthesis between these two varieties have not been studied.ResultsIn this study, phenotypic, transcriptomic and metabolomic analysis of HX_1 and ZZ_1 were conducted to elucidate the mechanism of leaf color formation. Chromaticity value and pigment content measuring showed that anthocyanin was the main metabolites imparting the different leaf color phenotype between the two varieties. Based on LC/MS, at least 14 anthocyanins were identified in leaves of HX_1 and ZZ_1, and the HX_1 showed the higher relative content of malvidin-, pelargonidin-,and cyanidin-based anthocyanins. Transcriptome and metabolome co-analysis revealed that the up-regulated expression of flavonoids synthesis gene was positively correlated with total anthocyanins accumulation in ramie leaf, and the differentfially expression of “blue gene” (F3’5’H) and the “red gene” (F3’H) in leaves bring out HX_1 metabolic flow more input into the cyanidin branch. Furthermore, the enrichment of glycosylated modification pathway (UGT and AT) and the expression of flavonoid 3-O-glucosyl transferase (UFGT), anthocyanidin reductase (ANR), in leaves were significantly influenced the diversity of anthocyanins between HX_1 and ZZ_1.ConclusionsPhenotypic, transcriptomic and metabolomic analysis of HX_1 and ZZ_1 indicated that the expression levels of genes related to anthocyanin metabolism contribute to the color formation of ramie variety. Anthocyanins are important plant secandary metabilates with many physiological functions, the results of this study will deepened our understanding of ramie leaf color formation, and provided basis for molecular breeding of functional forage ramie.

Identification, through transcriptome analysis, of transcription factors that regulate anthocyanin biosynthesis in different parts of red-fleshed apple ‘May’ fruit

Anthocyanins, a class of flavonoid compounds that provide pigmentation to plant organs, are beneficial components of the human diet and there is an interest in understanding the regulation of their formation in crop plants. The apple cultivar ‘May’ represents an excellent model for the study of anthocyanin metabolism due to the spatial color pattern of its flesh, which transitions sequentially from red to white to red again, from the outside to the inside of the apple fruit. To screen the transcription factors involved in the anthocyanin pathway, transcriptomes from differently colored sections of ‘May’ fruits were analyzed by RNA sequencing. Kyoto Encyclopedia of Genes and Genomes (KEGG) database was used to analyse the gene expression data. The results suggested differentially expressed genes (DEGs) that are related to phytohormones are involved in regulating anthocyanin biosynthesis. K-means clustering analysis revealed 167 common DEGs between different sections of fruit with the same expression pattern as candidates for regulating anthocyanin synthesis. Further analysis showed that nine of the 167 DEGs were annotated as transcription factors and quantitative real time PCR (qRT-PCR) confirmed that their expression was obviously higher in red regions of the fruit, consistent with their roles as hub genes that regulate anthocyanin synthesis. This study provides valuable results for future studies of anthocyanin synthesis in apple fruit.

Application of ABA and GA3 alleviated browning of litchi (Litchi chinensis Sonn.) via different strategies

Browning of harvested litchi causes fast quality losses that are closely associated with anthocyanin degradation. Abscisic acid (ABA) and gibberellins (GA) are antagonistic in regulating plant growth and development, but it is unclear whether and how they affect anthocyanin metabolism in harvested litchi fruit. Here, application of ABA and GA3 to harvested litchi reduced browning and maintained higher anthocyanin content relative to the untreated control during storage at 20 °C. Transcriptome profiling showed 2362 and 6304 differentially expressed genes (DEGs) were induced in response to ABA and GA3 treatment, respectively, implying the metabolism pathways regulated by ABA and GA are quite different. The flavonoid and phenylpropanoid biosynthesis are two of the 30 most enriched KEGG pathways for ABA-treated fruit compared with the control, but not for GA3-treated fruit. ABA upregulated key DEGs involved in phenylpropanoid biosynthesis and anthocyanin synthesis pathways, phenylalanine ammonia-lyase (PAL), cinnamic acid 4-hydroxylase (C4H), chalcone synthase (CHS) and UDP-flavonoid glucosyl transferase (UFGT), but GA3 downregulated them. ABA upregulated DEGs related to anthocyanin degradation and transport, laccase (LAC), peroxidase (POD), and glutathione S-transferase (GST), whereas GA3 downregulated them. Of the 29 different anthocyanin-related metabolites identified by LC–MS, ABA and GA3 caused increase of four and two, respectively. Taken together, ABA alleviated browning mainly by promoting anthocyanin synthesis, whereas GA3 inhibited anthocyanin degradation. These findings add to understanding of the roles of ABA and GA in regulating anthocyanin metabolism of plants in senescent stages and provides new approaches to pericarp browning prevention in litchi.

Transcriptomic, proteomic and LC-MS analyses reveal anthocyanin biosynthesis during litchi pericarp browning

Anthocyanin is a widely distributed flavonoid pigment that confers plant red color. The biosynthesis of anthocyanin associated with onset of color in developing plants has been well documented. However, whether anthocyanin biosynthesis pathway still functions in fruit at senescent stage remains unclear. In this study we investigated the metabolism of anthocyanin in litchi, a red fruit whose color fades fast after harvest. We showed as the pericarp browning index increased, the total anthocyanin contents decreased in litchi during storage at 20 °C. RNA-seq and proteomics analysis identified 4769 differentially expressed genes (DEG) and 84 differentially expressed proteins (DEP), respectively, in stored fruit compared with that before storage. Among them, 4 DEGs and 1 DEP involved in anthocyanin degradation, and 10 DEGs and 2 DEPs in anthocyanin biosynthesis pathway were upregulated. In particular, chalcone synthase (CHS), the first enzyme involved anthocyanin biosynthesis, and peroxidase (POD), a major enzyme for anthocyanin degradation, showed increased accumulation at both transcript and protein levels. LC-MS analysis showed that the accumulation of five anthocyanins and derivatives were decreased, while one anthocyanin increased in litchi stored for 6 d. These results suggest that anthocyanin degradation is accompanied by biosynthesis during pericarp browning, and that the browning of litchi resulted from more degradation than biosynthesis. The work provides new insight into mechanism underlying litchi browning during storage and possibilities for developing new browning prevention techniques. Additionally, from the evolutional perspective, this finding might imply that plants at senescent stage still try to preserve and disseminate the seeds against stresses.

Bioavailability, Absorption, and Metabolism of Pelargonidin-Based Anthocyanins Using Sprague-Dawley Rats and Caco-2 Cell Monolayers.

The present study is aimed to clarify the absorption and metabolism properties of pelargonidin-based anthocyanins. Results showed that pelargonidin-3-O-rutinoside (Pg3R) was absorbed in its intact form after oral administration and reached a maximum plasma concentration of 175.38 ± 55.95 nM at 60 min. Three main metabolites were identified in plasma, including Pg3R-monoglucuronide (m/z 755.2046), Pg3R-hydroxylated (m/z 595.1644), and Pg3R-demethylated (m/z 565.1569) metabolites. The plasma concentration of the Pg3R-demethylated metabolite (57.04 ± 23.15 nM) was much higher than that of other two metabolites, indicating that demethylation was the main metabolic pathway for Pg3R, while the glucuronide conjugate was detected as the dominant metabolic form of pelargonidin-3-O-glucoside (Pg3G). The bioavailability of Pg3R (1.13%) was fourfold higher than that of Pg3G (0.28%), demonstrating that anthocyanins linked to the rutinoside may exhibit higher bioavailability than that of glucoside. In vitro transport study unveiled that passive diffusion and active efflux were involved in the absorption of Pg3R and Pg3G.

Elevated CO2 differently suppresses the arsenic oxide nanoparticles-induced stress in C3 (Hordeum vulgare) and C4 (Zea maize) plants via altered homeostasis in metabolites specifically proline and anthocyanin metabolism

Nano-sized arsenic oxide nanoparticles (As2O3-NP) limit crop growth and productivity. As2O3-NP represent a strong environmental hazard. The predicted rise in future atmospheric CO2 could boost plant growth both under optimal and heavy metal stress conditions. So far, the phytotoxicity of As2O3-NP and their interaction with eCO2 were not investigated at physiological and metabolic levels in crop species groups such as C3 and C4. We investigated how eCO2 level (620 ppm) alleviated soil As2O3-NP toxicity induced growth and mitigated oxidative damages through analysing photosynthetic parameters, primary (sugars and amino acids) and secondary (phenolics, flavonoids and anthocyanins) metabolism in C3 (barley) and C4 (maize) plants. Compared to maize, barley accumulated higher As2O3-NP level, which inhibited growth and induced oxidative damage particularly in barley (increased H2O2 and lipid peroxidation). Interestingly, eCO2 differently mitigated As2O3-NP toxicity on photosynthesis, which consequently improved sugar metabolism. Moreover, high carbon availability in eCO2 treated plants directed to produce osmo-protectant (soluble sugars and proline) and antioxidants (anthocyanins and tocopherols). In the line with increased proline and anthocyanins, their metabolism was also improved. Notable differences occurred between the two plant species. The ornithine pathway was preferred in maize while in barley proline accumulation was mainly through glutamate pathway. Moreover, under As2O3-NP stress, barley preferentially accumulated anthocyanins while maize accumulated total phenolics and flavonoids. This work contributes to improving our understanding of the differences in growth, physiological and biochemical responses of major crops of two functional photosynthetic groups (C3 and C4 plants) under ambient and elevated CO2 grown under As2O3-NP stress.

Calcium Positively Mediates Blue Light-Induced Anthocyanin Accumulation in Hypocotyl of Soybean Sprouts.

Soybean sprouts are a flavorful microgreen that can be eaten all year round and are widely favored in Southeast Asia. In this study, the regulatory mechanism of calcium on anthocyanin biosynthesis in soybean sprouts under blue light was investigated. The results showed that blue light, with a short wavelength, effectively induced anthocyanin accumulation in the hypocotyl of soybean sprout cultivar “Dongnong 690.” Calcium supplementation further enhanced anthocyanin content, which was obviously inhibited by LaCl3 and neomycin treatment. Moreover, exogenous calcium changed the metabolism of anthocyanins, and seven anthocyanin compounds were detected. The trend of calcium fluorescence intensity in hypocotyl cells, as well as that of the inositol 1,4,5-trisphosphate and calmodulin content, was consistent with that of anthocyanins content. Specific spatial distribution patterns of calcium antimonate precipitation were observed in the ultrastructure of hypocotyl cells under different conditions. Furthermore, calcium application upregulated the expression of genes related to anthocyanin biosynthesis, and calcium inhibitors suppressed these genes. Finally, transcriptomics was performed to gain global insights into the molecular regulation mechanism of calcium-associated anthocyanin production. Genes from the flavonoid biosynthesis pathway were distinctly enriched among the differentially expressed genes, and weighted gene co-expression network analysis showed that two MYBs were related to the accumulation of anthocyanins. These results indicated that calcium released from apoplast and intracellular stores in specific spatial-temporal features promote blue light-induced anthocyanin accumulation by upregulation of the expression of genes related to anthocyanin synthesis of “Dongnong 690” hypocotyl. The findings deepen the understanding of the calcium regulation mechanism of blue light-induced anthocyanin accumulation in soybean sprouts, which will help growers produce high-quality foods beneficial for human health.

Chemical Hemisynthesis of Sulfated Cyanidin-3-O-Glucoside and Cyanidin Metabolites.

The metabolism of anthocyanins in humans is still not fully understood, which is partly due to the lack of reference compounds. It is known that sulfation is one way of the complex phase II biotransformation mechanism. Therefore, cyanidin-3-O-glucoside and the cyanidin aglycone were chemically converted to their sulfates by reaction with sulfur trioxide-N-triethylamine complex in dimethylformamide. The reaction products were characterized by UHPLC coupled to linear ion trap and IMS-QTOF mass spectrometry. Based on MS data, retention times, and UV-Vis spectra, the compounds could tentatively be assigned to A-, C-, or B-ring sulfates. Analysis of urine samples from two volunteers after ingestion of commercial blackberry nectar demonstrated the presence of two sulfated derivatives of the cyanidin aglycone and one sulfated derivative of the cyanidin-3-O-glucoside. It was found that both the A ring and the B ring are sulfated by human enzymes. This study marks an important step toward a better understanding of anthocyanin metabolism.

Subcellular Localization and Vesicular Structures of Anthocyanin Pigmentation by Fluorescence Imaging of Black Rice (Oryza sativa L.) Stigma Protoplast.

Anthocyanins belong to the group of flavonoid compounds broadly distributed in plant species responsible for attractive colors. In black rice (Oryza sativa L.), they are present in the stems, leaves, stigmas, and caryopsis. However, there is still no scientific evidence supporting the existence of compartmentalization and trafficking of anthocyanin inside the cells. In the current study, we took advantage of autofluorescence with anthocyanin’s unique excitation/emission properties to elucidate the subcellular localization of anthocyanin and report on the in planta characterization of anthocyanin prevacuolar vesicles (APV) and anthocyanic vacuolar inclusion (AVI) structure. Protoplasts were isolated from the stigma of black and brown rice and imaging using a confocal microscope. Our result showed the fluorescence displaying magenta color in purple stigma and no fluorescence in white stigma when excitation was provided by a helium–neon 552 nm and emission long pass 610–670 nm laser. The fluorescence was distributed throughout the cell, mainly in the central vacuole. Fluorescent images revealed two pools of anthocyanin inside the cells. The diffuse pools were largely found inside the vacuole lumen, while the body structures could be observed mostly inside the cytoplasm (APV) and slightly inside the vacuole (AVI) with different shapes, sizes, and color intensity. Based on their sizes, AVI could be grouped into small (Ф < 0.5 um), middle (Ф between 0.5 and 1 um), and large size (Ф > 1 um). Together, these results provided evidence about the sequestration and trafficking of anthocyanin from the cytoplasm to the central vacuole and the existence of different transport mechanisms of anthocyanin. Our results suggest that stigma cells are an excellent system for in vivo studying of anthocyanin in rice and provide a good foundation for understanding anthocyanin metabolism in plants, sequestration, and trafficking in black rice.

Preharvest long-term exposure to UV-B radiation promotes fruit ripening and modifies stage-specific anthocyanin metabolism in highbush blueberry

Ultraviolet-B (UV-B) light (280–315 nm) is an important environmental signal that regulates plant development and photomorphogenesis, while also affecting the flavonoid pathway, including anthocyanin biosynthesis. Regarding the effects of UV-B radiation on fruits, the effects of a short-term or postharvest irradiation on fruit quality have been well-documented, but the effects of a long-term preharvest UV-B irradiation on fruit growth and coloration remain unclear. Thus, in this study, we investigated the effects of a long-term treatment involving an environmentally relevant UV-B dose on highbush blueberry (Vaccinium corymbosum) fruit. The preharvest UV-B treatment quickly promoted fruit growth and sugar accumulation, which is not commonly observed in other fruit tree species. The UV-B exposure also accelerated fruit ripening and coloration. The dual-luciferase assay proved that in blueberries, expression of VcUFGT encoding anthocyanin biosynthesis key enzyme, is positively and negatively regulated by VcMYBA1 and VcMYBC2, respectively. Throughout the fruit development stage, the UV-B treatment up-regulated VcMYBPA1 expression, which increased VcUFGT expression via VcMYBA1. In the green fruit stage, the UV-B treatment increased HY5 encoding UV receptor, which up-regulates VcMYBPA1 and down-regulates VcMYBC2, thereby promotes the accumulation of anthocyanins. On the other hand, excessive anthocyanin synthesis was inhibited by increased VcMYBC2 levels in mature fruits when exposed to UV-B light through HY5-independent pathway. In conclusion, anthocyanin-related MYB activators and repressor may coordinately balance the accumulation of anthocyanins in blueberry fruits, with UV-B treatments possibly influencing their effects in a stage-specific manner. The potential utility of preharvest UV-B treatments for improving blueberry fruit quality is discussed herein.

Phenolic Metabolites in the Urine and Plasma of Healthy Men After Acute Intake of Purple Potato Extract Rich in Methoxysubstituted Monoacylated Anthocyanins

Structurally stable acylated anthocyanins have potential in various food applications but the effects of acylation and methoxysubstitution on anthocyanin metabolism are poorly understood. This is the first study thoroughly investigating phenolic metabolites, their time-wise changes, and pharmacokinetics following an acute intake of methoxysubstituted monoacylated anthocyanins. Healthy male volunteers (n = 17) consumed a yellow potato meal with and without purple potato extract rich in acylated anthocyanins (152mg) and hydroxycinnamic acid conjugates (140mg). Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) is used for identification and quantification of metabolites from serially collected urine and plasma. While the parent anthocyanins are not detected, 28 phenolic metabolites from urine and 14 from plasma are quantified, including hydroxybenzoic and hydroxycinnamic acids and protocatechuic acid sulfates and glucuronides; three (catechol, gallic acid-4-O-glucuronide, and 2-methoxybenzoic acid) are detected for the first time after anthocyanin-rich food. Urinary hippuric acid is the most abundant with an increase of 139µM mM-1 creatinine after the treatment. A large additional set of tentatively identified phenolic metabolites are detected. Late urinary peak time values suggest colonic degradation. Acylated anthocyanins are more bioavailable than earlier reported after extensive degradation in human and/or colonial metabolism to phenolic metabolites, which may be further conjugated and demethylated.

Genetic analysis of seed coat colour in adzuki bean (Vigna angularis L.)

AbstractSeed coat colour is an important quality trait, domestication trait and morphological marker, and is closely associated with flavonoid and anthocyanin metabolism pathways. The seed coat colour of adzuki bean, an important legume crop, influences the processing quality of its paste, the commodity and its nutritional quality. In this study, the genetic relationships of seed coat colour were analysed using 12 hybridized combinations of F2 individuals and four F3 families derived from hybridized combinations between the accessions of eight seed coat colours. The loci of the colour traits were analysed based on phenotypes and using the chi-square test. Ivory colour is recessive to red and is controlled by a single R locus. Black, black mottle on grey, black mottle on red, light brown, golden and brown are all dominant to red. The phenotypes of black mottle on red, light brown, golden and brown are all controlled by a single genetic locus. Black mottle on grey is controlled by two loci. Black is controlled with two loci, and the black locus shows dominant epistasis to another locus. A genetic model of these seed coat colours was predicted. Our results will be important for gene mapping and cloning of seed coat colour characters and for providing further insight into the regulatory network of seed coat colour.

OsRR6, a type-A response regulator in rice, mediates cytokinin, light and stress responses when over-expressed in Arabidopsis

Plants have evolved a complex network of components that sense and respond to diverse signals. In the present study, we have characterized OsRR6, a type-A response regulator, which is part of the two-component sensor-regulator machinery in rice. The expression of OsRR6 is induced by exogenous cytokinin and various abiotic stress treatments, including drought, cold and salinity stress. Organ-specific expression analysis revealed that its expression is high in anther and low in shoot apical meristem. The Arabidopsis plants constitutively expressing OsRR6 (OsRR6OX) exhibited reduced cytokinin sensitivity, adventitious root formation and enhanced anthocyanin accumulation in seeds. OsRR6OX plants were more tolerant to drought and salinity conditions when compared to wild-type. The hypocotyl growth in OsRR6OX seedlings was significantly inhibited under red, far-red and blue-light conditions and also a decline in transcript levels of OsRR6 was observed in rice under the above monochromatic as well as white light treatments. Transcriptome profiling revealed that the genes associated with defense responses and anthocyanin metabolism are up-regulated in OsRR6OX seedlings. Comparative transcriptome analysis showed that the genes associated with phenylpropanoid and triterpenoid biosynthesis are enriched among differentially expressed genes in OsRR6OX seedlings of Arabidopsis, which is in conformity with reanalysis of the transcriptome data performed in rice transgenics for OsRR6. Further, genes like DREB1A/CBF3, COR15A, KIN1, ERD10 and RD29A are significantly upregulated in OsRR6OX seedlings when subjected to ABA and abiotic stress treatments. Thus, a negative regulator of cytokinin signaling, OsRR6, plays a positive role in imparting abiotic stress tolerance.

Anti-aging effects of the fermented anthocyanin extracts of purple sweet potato on Caenorhabditis elegans.

Anthocyanins have anti-inflammatory, anticarcinogenic and antioxidant properties and anti-aging effects as well as potential application as pigments. The metabolism of anthocyanins in fermented food has attracted increasing attention. However, the effect of lactic acid bacteria (LAB) fermentation on its anti-aging activity remains mostly unknown. The current study aimed to investigate the compositions, antioxidant activities and anti-aging effect of fermented purple sweet potato anthocyanins (FSPA) on aging Caenorhabditis elegans compared to raw purple sweet potato anthocyanins (PSPA). Results showed that anthocyanins were degraded into more bioavailable phenolic acids by Weissella confusa fermentation. PSPA and FSPA can extend the lifespan of C. elegans by 26.7% and 37.5%, respectively, through improving the activity of antioxidant enzymes as well as decreasing MDA content, ROS levels and lipofuscin accumulation. Pretreatment of the worms with PSPA and FSPA induced their potential to resist to thermal tolerance and oxidative stress, and FSPA exerted a higher anti-stress effect than PSPA. Moreover, FSPA supplementation upregulated the mRNA expressions of genes daf-16, hsp-16.2, sir-2.1, skn-1 and sod-3 and downregulated the expression of daf-2 in the nematodes, whereas PSPA only induced the increase in the expressions of sir-2.1, skn-1 and sod-3. Overall, FSPA can improve stress resistance and extend the lifespan of C. elegans by both insulin/IGF-1 signaling pathway and dietary restriction pathway, providing a theoretical basis for the application of PSPA in fermented food as functional pigments.

The coloring mechanism of a novel golden variety in Populus deltoides based on the RGB color mode.

Compared to flower color and red leaf mutants, the mechanism of yellow leaf formation in woody plants is more complicated and less understood. Poplars are common and widely distributed perennial trees. Here, we obtained a golden leaf mutant poplar variety (JHY) and compared it with its original green leaf poplar (Populus sp. Linn. '2025') in terms of phenotype, pigment content, the transcriptomes and metabolomes. Through transcriptome sequencing, we found that the chlorophyll degradation genes, and the genes in anthocyanin metabolism in JHY were up-regulated compared to L2025 and the carotenoid synthesis genes were down-regulated. Further based on HPLC-ESI-MS/MS technology, 16 differentially expressed anthocyanins were identified, 14 reddish anthocyanins of which were significantly up-regulated. According to these results, we proposed a coloring mechanism of JHY based on the RGB color mode. The yellow color of carotenoids and the red color of anthocyanins overlay each other, combined with a reduction in chlorophyll, turning the leaves golden. This study systematic analyzes the causes of golden leaf formation through the omics integration approach, which will provide reference for the breeding of golden leaf trees.

Genome-wide identification and expression profiling of MYB transcription factor genes in radish (Raphanus sativus L.)

Radish (Raphanus sativus L.), an important root vegetable crop of the Brassicaceae family, has a high level of anthocyanin accumulation in its pigment root tissues. It was reported that MYB transcription factors (TFs) play vital roles in plant development and anthocyanin metabolism, and the PAP1/2 could promote expression of anthocyanin biosynthesis genes. In this study, a total of 187 radish MYB genes (RsMYBs) were identified in the radish genome and clustered into 32 subfamilies. Among them, 159 RsMYBs were localized on nine radish chromosomes. Interestingly, 14 RsMYBs exhibited differential expression profiles in different taproot developmental stages among four differently colored radish lines. A number of RsMYBs were highly expressed in the pigmented root tissues at the maturity stage, several genes including RsMYB41, RsMYB117, and RsMYB132 being homologous to PAP1/2, showed high expression levels in the red skin of NAU-YH (red skin-white flesh) taproot, while RsMYB65 and RsMYB159 were highly expressed in the purple root skin of NAU-YZH (purple skin-red flesh), indicating that these RsMYBs might positively regulate the process of anthocyanin accumulation in radish taproot. These results would provide valuable information for further functional characterization of RsMYBs, and facilitate clarifying the molecular mechanism underlying anthocyanin biosynthesis in radish.

Integrated Physiological and Transcriptomic Analyses Reveal a Regulatory Network of Anthocyanin Metabolism Contributing to the Ornamental Value in a Novel Hybrid Cultivar of Camellia Japonica.

Camellia japonica is a plant species with great ornamental and gardening values. A novel hybrid cultivar Chunjiang Hongxia (Camellia japonica cv. Chunjiang Hongxia, CH) possesses vivid red leaves from an early growth stage to a prolonged period and is, therefore, commercially valuable. The molecular mechanism underlying this red-leaf phenotype in C. japonica cv. CH is largely unknown. Here, we investigated the leaf coloration process, photosynthetic pigments contents, and different types of anthocyanin compounds in three growth stages of the hybrid cultivar CH and its parental cultivars. The gene co-expression network and differential expression analysis from the transcriptome data indicated that the changes of leaf color were strongly correlated to the anthocyanin metabolic processes in different leaf growth stages. Genes with expression patterns associated with leaf color changes were also discussed. Together, physiological and transcriptomic analyses uncovered the regulatory network of metabolism processes involved in the modulation of the ornamentally valuable red-leaf phenotype and provided the potential candidate genes for future molecular breeding of ornamental plants such as Camellia japonica.