Coupling of reduced inorganic fertilizer with plant-based organic fertilizer as a promising fertilizer management strategy for colored rice in tropical regions

Colored rice is a type of high-quality,high-added-value rice that has attracted increasing attention in recent years.The use of large amounts of inorganic nitrogen fertilizer in rice fields results in low fertilizer use efficiency and high environmental pollution.Organic fertilizer is a promising way to improve soil quality and sustain high yields.However,most studies focus on the effect of animal-based organic fertilizers.The effects of different ratios of plantbased organic fertilizer and inorganic fertilizer on the grain yield and quality of colored rice have rarely been reported.Therefore,a two-year field experiment was conducted in 2020 and 2021 to study the effects of replacing inorganic N fertilizers with plant-based organic fertilizers on the yield,nitrogen use efficiency(NUE),and anthocyanin content of two colored rice varieties in a tropical region in China.The experimental treatments included no nitrogen fertilization(T1),100% inorganic nitrogen fertilizer(T2),30%inorganic nitrogen fertilizer substitution with plant-based organic fertilizer(T3),60%inorganic nitrogen fertilizer substitution with plant-based organic fertilizer(T4),and 100% plantbased organic fertilizer(T5).The total nitrogen provided to all the treatments except T1 was the same at 120 kg ha-1.Our results showed that the T3 treatment enhanced the grain yield and anthocyanin content of colored rice by increasing nitrogen use efficiency compared with T2.On average,grain yields were increased by 9 and 8%,while the anthocyanin content increased by 16 and 10% in the two colored rice varieties under T3 across the two years,respectively,as compared with T2.Further study of the residual effect of partial substitution of inorganic fertilizers showed that the substitution of inorganic fertilizer with plant-based organic fertilizer improved the soil physiochemical properties,and thus increased the rice grain yield,in the subsequent seasons.The highest grain yield of the subsequent rice crop was observed under the T5 treatment.Our results suggested that the application of plantbased organic fertilizers can sustain the production of colored rice with high anthocyanin content in tropical regions,which is beneficial in reconciling the relationship between rice production and environmental protection.

Unraveling the regulatory network of flower coloration in soybean:Insights into roles of GmMYBA3 and GmMYBR1

Anthocyanins play crucial roles in pollen protection and pollinator attraction in flowering plants.However,the mechanisms underlying flower color determination and whether floral anthocyanin regulators participate in other processes remain largely unresolved in soybeans(Glycine max).In this study,we investigated the genetic components and mechanisms governing anthocyanin biosynthesis in soybean flowers.Molecular and genetic studies have characterized two antagonistic regulators,the positive activator GmMYBA3 and the negative repressor GmMYBR1,that modulate the gene expression of anthocyanin biosynthesis in soybean flowers.Further findings revealed a regulatory interplay between GmMYBA3 and GmMYBR1 bridged by GmTT8a,highlighting the complexity of anthocyanin regulation in different soybean organs.Exploration of additional soybean cultivars demonstrated the universality of GmMYBA3 and GmMYBR1 in regulating floral anthocyanin biosynthesis-related genes,with GmF3’5’H identified as a crucial determinant of white flower color.This study provides a molecular mechanism underlying soybean flower color determination,paving the way for the molecular modification of soybean flowers to probably enhance their resistance to abiotic stresses and attractiveness to pollinators.

JrATHB-12 mediates JrMYB113 and JrMYB27 to control the anthocyanin levels in different types of red walnut

Red walnut has broad market prospects because it is richer in anthocyanins than ordinary walnut.However,the mechanism driving anthocyanin biosynthesis in red walnut is still unknown.We studied two types of red walnut,called red walnut 1(R1),with a red pericarp and seed coat,and red walnut 2(R2),with a red seed coat only.R1 mostly contained cyanidin-3-O-galactoside,while R2 contained a various amounts of cyanidin-3-Ogalactoside,cyanidin-3-O-arabinoside,and cyanidin-3-O-glucoside.The LDOX-2(LOC109007163)and LDOX-3(LOC109010746)genes,which encode leucoanthocyanidin dioxygenase/anthocyanidin synthase(LDOX/ANS),were preliminarily indicated as the crucial genes for anthocyanin biosynthesis in R1 and R2,respectively.The MYB differential genes analysis showed that MYB27 and MYB113 are specifically expressed in the red parts of R1 and R2,respectively,and they are regarded as candidate regulatory genes.Ectopic expression in Arabidopsis and transient injection in walnut showed that both MYB27 and MYB113 were located in the nucleus and promoted anthocyanin accumulation,while MYB27 promoted the expression of LDOX-2,and MYB113 promoted the expression of LDOX-3and UAGT-3.Yeast one-hybrid and electrophoretic mobility shift assays showed that MYB27 could only bind to the LDOX-2 promoter,while MYB113 could bind to the promoters of both LDOX-3 and UAGT-3.In addition,we also identified an HD-Zip transcription factor,ATHB-12,which is specifically expressed in the pericarp.After silencing the expression of ATHB-12,the R2 pericarp turned red,and MYB113 expression increased.Further experiments showed that ATHB-12 could specifically interact with MYB113 and bind to its promoter.This suggests that MYB27controls R1 coloration by regulating LDOX-2,while MYB113 controls R2 coloration by regulating LDOX-3 and UAGT-3,but ATHB-12 can specifically bind to and inhibit the MYB113 of the R2 pericarp so that it becomes unpigmented.This study reveals the anthocyanin biosynthetic mechanisms in two different types of red walnut and provides a scientific basis for the selection and breeding of red walnut varieties.

ChMYB1-ChbHLH42-ChTTG1 module regulates abscisic acidinduced anthocyanin biosynthesis in Cerasus humilis

Cerasus humilis is a kind of economic fruit tree peculiar to China,which is widely used in the food,landscape,and pharmaceutical industries.Anthocyanins are a phenolic metabolite that plays an essential role in fruit coloration.However,the regulatory network of C.humilis in anthocyanin biosynthesis is still unclear.In this study,the R2R3-MYB transcription factor ChMYB1 was isolated from the full genome of the species.Yeast one-hybrid,dual-luciferase assays,and GUS staining showed that ChMYB1 significantly increased anthocyanin contents in C.humilis fruit by promoting the expression of ChCHS and ChUFGT by binding MBS(MYB-binding elements).ChMYB1 interacted with ChbHLH42and ChTTG1 to form the MBW complex and further enhanced the expression of ChUFGT.In addition,abscisic acid(ABA)treatment promoted the expression of ChMYB1 and anthocyanin accumulation in C.humilis fruit.Interestingly,ABA treatment enhanced the interaction between ChMYB1 and ChbHLH42.Furthermore,ChABI5 inhibited the interaction between ChMYB1 and ChbHLH42.Our data elucidated the primary molecular mechanism of anthocyanin biosynthesis in C.humilis fruit,deepening the understanding of the regulatory network affecting anthocyanin metabolism in edible fruit crops.

CmMYB3-like negatively regulates anthocyanin biosynthesis and flower color formation during the post-flowering stage in Chrysanthemum morifolium

Color fading caused by a decrease in anthocyanin accumulation during the post-flowering stage significantly affects postharvest quality of chrysanthemum.However,the underlying mechanism by which anthocyanin accumulation decreases during the post-flowering stage still unclear,which greatly restricts design of molecular breeding in chrysanthemum.Here,a chrysanthemum SG7 R2R3 MYB transcription factor(TF),CmMYB3-like,was identified to have a function in regulating anthocyanin biosynthesis during the post-flowering stage.Quantitative real time PCR(qRT-PCR)assays showed that the expression of CmMYB3-like was gradually downregulated when anthocyanin content increased during the flowering stage and was significantly upregulated during the post-flowering stage.Genetic transformation of chrysanthemum and dual-luciferase assays in N.benthamiana leaves showed that CmMYB3-like suppressed anthocyanin accumulation by inhibiting the transcription of CmCHS and CmANS directly and that of CmF3H indirectly.However,overexpression or suppression of CmMYB3-like did not affect the biosynthesis of flavones or flavonols.Genetic transformation of chrysanthemum revealed that the overexpression of CmMYB3-like inhibited anthocyanin accumulation,but its suppression prevented the decrease in anthocyanin accumulation during the post-flowering stage.Our results revealed a crucial role of CmMYB3-like in regulating the color of petals during the post-flowering stage and provided a target gene for molecular design breeding to improve the postharvest quality of chrysanthemum.

Smart Colorimetric Corn Starch Films Combined with Anthocyanin-Loaded Glutenin-Carboxymethyl Chitosan Nanocomplexes for Freshness Monitoring of Chilled Pork

In this study,intelligent,pH-responsive colorimetric films were prepared by encapsulating anthocyanins in nanocomplexes prepared from glutenin and carboxymethyl chitosan.These nanocomplexes were added to a corn starch matrix and used in the freshness monitoring of chilled pork.The effects of anthocyanin-loaded nanocomplexes on the physical,structural,and functional characteristics of the films were investigated.The addition of anthocyanin-loaded nanocomplexes increased the tensile strength,elongation at break,hydrophobicity,and light transmittance of the films while decreasing their water vapor permeability.This is because new hydrogen bonds are formed between the film components,resulting in a more homogeneous and dense structure.The colorimetric film has a significant color response to pH changes.These films were used in experiments on the freshness of chilled pork,in which the pH changes with changing freshness states.The results show that the colorimetric film can monitor changes in the freshness of chilled pork in real time,where orange,pink,and green represent the fresh,secondary fresh,and putrefied states of pork,respectively.Therefore,the intelligent colorimetric film developed in this study has good application potential in the food industry.

Distribution of antioxidants and phenolic compounds in flour milling fractions from hard red winter wheat

Mature wheat kernels contain three main parts:endosperm,bran,and germ.Flour milling results in multiple streams that are chemically different;however,the distribution of antioxidants and phenolic compounds has not been well documented in terms of conventional milling by-product streams.In this study,multiple analytical methods were used to investigate antioxidant activity and phenolic compound compositions of hard red winter wheat(whole ground wheat),the parts of a wheat kernel(bran,flour,germ),and wheat by-product streams(mill feed,red dog,shorts)for the first time.For each mill stream,phenolic compounds(total,flavonoid,and anthocyanin contents)were determined and antioxidant activities were evaluated with 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical-scavenging activity,ferric reducing/antioxidant power(FRAP),and total antioxidant capacity assays.Significant differences(P<0.05)were observed in phenolic concentrations among fractions of bran,flour,and germ milled from the same kernels and noted that germ accounts for the majority of antioxidant properties,whereas bran contains a substantial portion of phenolic compounds and anthocyanins.Mill feed was high in phenolic content(5.29 mg FAE/g),total antioxidant capacity(866 mg/g),and antioxidant activity(up to 75% DPPH inhibition and 20.26μmol FeSO_(4)/g).The comprehensive information on distribution of antioxidants and phenolic compounds provides insights for future human consumption of commonly produced co-products from flour milling,and for selecting and using different milling fractions to make foods with improved nutritional properties.

Blueberry anthocyanins extract attenuates oxidative stress and angiogenesis on an in vitro high glucose-induced retinopathy model through the miR-33/GLCCI1 axis

Background:Diabetes retinopathy(DR)is a complication of diabetes that affects patients’vision.Previous studies have found blueberry anthocyanins extract(BAE)can inhibit the progression of DR,but its mechanism is not completely clear.Methods:To study the role of BAE in diabetes retinopathy,we treated human retinal endothelial cells(HRCECs)with 30 mM high glucose to simulate the microenvironment of diabetes retinopathy and used BAE to intervene the in vitro high glucose-induced retinopathy model.HRCEC cell viability and apoptosis rates were examined by Cell Counting Kit 8(CCK-8)assay and flow cytometry assay.The binding sites between miR-33 and glucocorticoid-induced transcript 1(GLCCI1)were assessed by luciferase reporter assay.Retinal neovascularization and oxidative stress contribute to diabetic retinopathy.The tubule formation assay was applied to detect the retinal neovascularization.The oxidative stress in the HRCECs was manifested by the reactive oxygen species(ROS)level,the malondialdehyde(MDA)level,and the superoxide dismutase(SOD)activity.Results:Compared with HRCECs cells cultured under normal conditions,high glucose(HG)can induce oxidative stress in HRCRCs,specifically manifested in the increase of ROS and MDA levels,and the decrease of SOD activity.BAE relieved the tubule formation in n the HRCEC.BAE also relieved the ROS and MDA levels and increased the SOD activity.Luciferase reporter assay revealed that GLCCI1 is a target molecule downstream of miR-33.In HRCEC,BAE significantly inhibited the expression of miR-33 induced by HG.miR-33 mimic inhibited the BAE’s effects on oxidative stress and angiogenesis in an in vitro high glucose-induced retinopathy model.Conclusion:BAE alleviated the oxidative stress and microangiogenesis of HRCEC by regulating the miR-33/GLCCI1 axis.

Transcriptomic Analysis Reveals the Formation Mechanism of Anthocyanins Light-Independent Synthesis in Chrysanthemum

Chrysanthemum×morifolium is a horticultural crop which plays a vital role in theflower industry with signifi-cant economic value and has a cultivation history of over three thousand years in China.The accumulation of anthocyanins is always affected by light.Here,we revealed that anthocyanin accumulation is highly dependent on light in‘2021135’genotype chrysanthemum,while it is light-independent in‘2001402’genotype chrysanthe-mum.However,no literature has been reported regarding the non-photosensitive chrysanthemum in anthocya-nins light-independent synthesis pathways.Through the phenotype analysis of 44 F1 generations,we found that light-independence is a dominant trait which can be stable inherited by progeny.The transcriptome of the rayflorets of‘2021135’and‘2001402’under light and bagging treatment were sequenced and analyzed.Based on weighted gene co-expression network analysis(WGCNA),K-means analysis,and Real-Time Quantitative Poly-merase Chain Reaction(RT-qPCR)analysis,16 genes were highly correlated with the anthocyanin content.The anthocyanin content of rayflorets treated with different light-quality conditions indicated that blue light signifi-cantly affected anthocyanin accumulations.Through Yeast one-hybrid analysis,CmBIC1.1 and CmBIC1.2 can directly regulate the anthocyanin structural gene CmCHS2.In our study,we revealed the important characteristics of light-independent anthocyanin synthesis in chrysanthemums and screened regulatory factors in light-depen-dent and light-independent anthocyanin synthesis pathways.The results laid the groundwork for subsequent ana-lysis of the molecular mechanism involved in the light-independent synthesis of anthocyanins in chrysanthemums.

Analysis and Verification of the Conserved MYB Binding Element in the DFR Promoter in Compositae

Anthocyanins,ubiquitous in the Compositae family,are regulated by MYB(v-myb avian myeloblastosis viral oncogene homolog),playing an important role in anthocyanin synthesis.In this study,we analyzed the regulation pathway in which the MYB protein of subgroup 6 promotes dihydroflavonol reductase(DFR)expression in Compositae,and validated this law in Saussurea medusa through yeast one-hybrid experiments.Our results showed that MYB and DFR underwent purification selection,DFR promoter analysis revealed the presence of MYB binding site(GAGTTGAATGG)and bHLH binding site(CANNTG)at the sense strand of 84–116 nucleotide residues from the start codon.These two motifs were separated by 9–10 nucleotide residues,as existed in the DFR promoters of many Compositae plants.Furthermore,the yeast one-hybrid experiment demonstrated that SmMYB1 can activate the promoter of SmDFR.Our results provide a reference for further functional study of DFR in Compositae.

Metabolomic Analysis of the Anthocyanins Associated with Different Colors of Cymbidium goeringii in Guizhou, China

Cymbidium goeringii is an economically important ornamental plant,and flower color is one of the main features of C.goeringii that contributes to its high economic value.To clarify the molecular mechanisms underlying the role of anthocyanins in mediating differences in color among varieties,liquid chromatography–tandem mass spectrometry was used to perform anthocyanin-targeted metabolomics of seven C.goeringii varieties,including‘Jin Qian Yuan’(JQY),‘Jin Xiu Qian Yuan’(JXQY),‘Miao Jiang Su Die’(MJSD),‘Qian Ming Su’(QMS),‘Shi Chan’(SC),and‘Yang Ming Su’(YMS),as well as the C.goeringii.We detected 64 anthocyanins,including cyanidins,delphinidins,malvidins,pelargonidins,peonidins,petunidins,procyanidins,and flavonoids.We identified six shared differentially accumulated metabolites(DAMs),including cyanidin-3-O-rutinoside,delphinidin-3-Osophoroside,pelargonidin-3-O-rutinoside,peonidin-3-O-(6-O-malonyl-beta-D-glucoside),peonidin-3-Osophoroside,and chalcone.Most DAMs were enriched in the anthocyanin biosynthesis pathway.Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the differentially expressed metabolites were significantly enriched in the anthocyanin biosynthesis pathway.Analysis of the content of differentially expressed metabolites indicated that peonidin-3-O-(6-O-malonyl-beta-D-glucoside)was the key metabolite underlying color differences among C.goeringii varieties.Procyanidin B2,pelargonidin-3-O-galactoside,and naringenin might also affect the color formation of JQY and QMS,SC,and MJSD,respectively.The results of this study shed light on the metabolic mechanism underlying flower color differences in C.goeringii at the molecular level.Our findings will aid future studies of the mechanism of flower color regulation in C.goeringii and have implications for the breeding of new varieties.

Discussion of the Composition of Jaboticaba in Different Processes

This study discusses the composition analysis and comparison of Jaboticaba under different processing conditions for fermented liquid, jam, fruit powder, and fruit vinegar. The differences and comparison of the contents of total polyphenols, anthocyanins, resveratrol, superoxide dismutase, small molecular peptides and ellagic acid in their products were analyzed. After analysis and comparison of results, the content of total polyphenols was found to be the highest 44.82 ± 0.89 (mg/g) after being fermented directly from fresh fruit. The fresh fruit was dried at low temperature and ground into powder, with a maximum ellagic acid content of 12.799 ± 0.12 (mg/g). Fresh fruit was then made into jam, with the highest anthocyanin content of 57.39 ± 1.20 (mg/g). The fruit vinegar was produced by fermentation. Except for the content of small molecule peptides, which was similar to the content of fermentation broth and fruit powder, they were 20.63 ± 1.61, 23.84 ± 2.51 and 20.52 ± 1.21 (mg/g), and the rest of the composition was not as good as other samples. In the fresh fruit fermentation broth samples, resveratrol was produced and detected, and its content was 2.14 (mg/L), while it was not detected in other samples, and a superoxide dismutase (SOD)-like enzyme was detected in the fresh fruit fermentation broth. The highest activity was 49002.5 units/mL. Jaboticaba is prepared using a fermentation broth processing method with fresh fruit, and its total polyphenol content is higher than that of fruit powder, jam, and fruit vinegar. The commercial value of Jaboticaba and the establishment of composition content data are relatively improved, which is more promising for the future.

Mapping of Purple Gene in Spears of Asparagus(Asparagus officinalis L.)

[Objectives]This study was conducted to find out regulatory genes related to purple in spears of asparagus(Asparagus officinalis L.).[Methods]The stable asparagus inbred line JX1513-5(the base of the spear is purple)and JLV1718-7(the base of the spear is green)were used as parents to study the genetic law of purple/green traits in their offspring.[Results]The results showed that the purple in the basal part of asparagus spear was controlled by a pair of alleles,and purple was dominant over green.The F 2 segregation population was resequenced by the bulk segregation analysis(BSA)method,and the purple trait in the basal part of asparagus spear was located in the interval of 24.51-25.08 Mb on Chr07 chromosome,which included 47 genes.According to the annotation information,three candidate genes were screened out:LOC109849403,LOC109849430 and LOC109849442.The candidate genes were verified by real-time fluorescence quantitative PCR(qRT-PCR),and finally LOC109849442 was obtained as the candidate gene for controlling the purple/green trait in the basal part of asparagus spear.[Conclusions]This study lays a foundation for the breeding of new asparagus varieties and molecular marker-assisted breeding.

Quantification of Total Phenols, Total Flavonoids, Total Anthocyanins and Evaluation of Antioxidant and Antiradical Activities of Detarium Senegalense Extracts from Chad

The aim of the present work is to assess the value of Detarium Senegalense by determining the content of total phenols, total flavonoids and total anthocyanins, and by evaluating the free radical scavenging activity of Detarium Senegalense extracts. For this purpose, sequential extraction using solvents of increasing polarity was essential. The various extracts obtained underwent phytochemical and biochemical analyses. Phytochemical screening revealed the presence of flavonoids, alkaloids, tannins, polyphenols, anthocyanins and steroids/terpenes. Quantitative analysis of total polyphenols, total flavonoids and total anthocyanins yielded the following results: total flavonoids (0.803 ± 0029 mg EQ/100g P for acetone extract of roots and 0.871 ± 0.401 mg EQ/100g P for methanol extract of leaves);total polyphenols (23.298 ± 12.68 mg EAG/100g P for acetone extract of roots and 24.69 ± 0.49 401 mg EAG/100g P for methanol extract of leaves);total monomeric anthocyanins (44.697 ± 0.939 mg EC3G/100g P and 16.699 ± 0.193 mg EC3G/100g P respectively for acetone and methanol extracts of stem bark). DPPH free radical scavenging activity was 1.674 ± 0.023 mg/mL for the acetone extract and 0.934 ± 0.24 mg/mL for the methanol extract of roots. .

Protective effect of BlingLife®-berry extract on blue lightinduced eye damage and its mechanism

This study aims to investigate the protective effects of BlingLife®-berry extract on the eyes.BlingLife®-berry extract is a mixture of high-quality natural berries,including blackcurrant,aromia,bilberry and maquiberry.The main active ingredient responsible for theeye-protective effects is anthocyanins.Anthocyanins are natural water-soluble pigments belonging to the flavonoid class,and theyhave multiple benefits,including improving vision,anti-inflammatory and antibacterial properties,and antioxidant effects.Dueto the dual benefits of anthocyanins in protecting vision and eliminating free radicals,this study explores the comprehensive eye protective effects of BlingLife®-berry extract in terms of combating blue light-induced eye damage and oxidative stress-inducedvisual fatigue.The results provide robust evidence for the in vivo effects and further technological applications of BlingLife®-berry extract.

The roles of R2R3-MYBs in regulating complex pigmentation patterns in flowers

Pigmentation patterns are ubiquitous in nature.Visually striking pigmentation patterns are not only aesthetically appealing,but also crucial to pollinator interaction and plant fitness.The formation of complex floral pigmentation patterns mainly relies on the spatiotemporal expression of R2R3-MYB transcription factors and is often associated with certain floral development programs,such as floral organ identity,symmetry,which likely provide key information to initiate the patterning.For a complex pigmentation pattern to form,at least a pair of activator and inhibitor is required,despite their interaction might vary depending on the system being investigated.The regulation of pigmentation pattern involves multiple molecular mechanisms,such as transcriptional regulation,small RNA,transposon-mediated gene silencing,and methylation of gene body.Identifying these regulators can be facilitated by using single-cell and spatial transcriptomics as well as innovative plant transformation technologies.Moreover,plant organ development and pigmentation patterns are often interdependent,but current methods of describing patterns are static.Therefore,more precise and quantitative measurements are needed to elucidate the developmental mechanisms underlying complex pigmentation patterns in flowers.

The MYB transcription factor RcMYB1 plays a central role in rose anthocyanin biosynthesis

Rose(Rosa hybrida)is one of most famous ornamental plants in the world,and its commodity value largely depends on its flower color.However,the regulatory mechanism underlying rose flower color is still unclear.In this study,we found that a key R2R3-MYB transcription factor,RcMYB1,plays a central role in rose anthocyanin biosynthesis.Overexpression of RcMYB1 significantly promoted anthocyanin accumulation in both white rose petals and tobacco leaves.In 35S:RcMYB1 transgenic lines,a significant accumulation of anthocyanins occurred in leaves and petioles.We further identified two MBW complexes(RcMYB1-RcBHLH42-RcTTG1;RcMYB1-RcEGL1-RcTTG1)associated with anthocyanin accumulation.Yeast one-hybrid and luciferase assays showed that RcMYB1 could active its own gene promoter and those of other EBGs(early anthocyanin biosynthesis genes)and LBGs(late anthocyanin biosynthesis genes).In addition,both of the MBW complexes enhanced the transcriptional activity of RcMYB1 and LBGs.Interestingly,our results also indicate that RcMYB1 is involved in the metabolic regulation of carotenoids and volatile aroma.In summary,we found that RcMYB1 widely participates in the transcriptional regulation of ABGs(anthocyanin biosynthesis genes),indicative of its central role in the regulation of anthocyanin accumulation in rose.Our results provide a theoretical basis for the further improvement of the flower color trait in rose by breeding or genetic modification.

Dietary supplementation of bilberry anthocyanin on growth performance,intestinal mucosal barrier and cecal microbes of chickens challenged with Salmonella Typhimurium

Background Anthocyanins(AC)showed positive effects on improving the intestinal health and alleviating intestinal pathogen infections,therefore,an experiment was conducted to explore the protective effects of supplemented AC on Salmonella-infected chickens.Methods A total of 240 hatchling chickens were randomly allocated to 4 treatments,each with 6 replicates.Birds were fed a basal diet supplemented with 0(CON,and ST),100(ACL)and 400(ACH)mg/kg of AC for d 60,and orally challenged with PBS(CON)or 10^(9) CFU/bird(ST,ACL,ACH)Salmonella Typhimurium at d 14 and 16.Results(1)Compared with birds in ST,AC supplementation increased the body weight(BW)at d 18 and the average daily gain(ADG)from d 1 to 18 of the Salmonella-infected chickens(P<0.05);(2)AC decreased the number of Salmonella cells in the liver and spleen,the contents of NO in plasma and inflammatory cytokines in ileal mucosa of Salmonella-infected chickens(P<0.05);(3)Salmonella infection decreased the ileal villi height,villi height to crypt depth(V/C),and the expression of zonulaoccludins-1(ZO-1),claudin-1,occludin,and mucin 2(MUC2)in ileal mucosa.AC supplementation relieved these adverse effects,and decreased ileal crypt depth(P<0.05);(4)In cecal microbiota of Salmonella-infected chickens,AC increased(P<0.05)the alpha-diversity(Chao1,Pd,Shannon and Sobs indexes)and the relative abundance of Firmicutes,and decreased(P<0.05)the relative abundance of Proteobacteria and Bacteroidota and the enrichment of drug antimicrobial resistance,infectious bacterial disease,and immune disease pathways.Conclusions Dietary AC protected chicken against Salmonella infection via inhibiting the Salmonella colonization in liver and spleen,suppressing secretion of inflammatory cytokines,up-regulating the expression of ileal barrier-related genes,and ameliorating the composition and function of cecal microbes.Under conditions here used,100 mg/kg bilberry anthocyanin was recommended.

Systematic analysis of MYB transcription factors and the role of LuMYB216 in regulating anthocyanin biosynthesis in the flowers of flax(Linum usitatissimum L.)

Anthocyanin is an important pigment that affects plant color and nutritional quality.MYBs play an important role in plant anthocyanin synthesis and accumulation.However,the regulatory function of MYB transcription factors in anthocyanin synthesis in flax flowers is still unclear.In this study,402 MYB transcription factors were identified in the flax genome.These MYB members are unevenly distributed on 15 chromosomes.The R2R3-LuMYB members were divided into 32phylogenetic subfamilies.qRT-PCR analysis showed that seven R2R3-LuMYB genes in the adjacent subfamily of the evolutionary tree had similar expression patterns,among which Lu MYB216 was highly expressed in the petals of different colors.Moreover,gene editing of LuMYB216 in flax showed that the petal color,anther color and seed coat color of mutant plants were significantly lighter than those of wild-type plants,and the anthocyanin content of lumyb216 mutant plants was significantly reduced.Correlation analysis indicated that LuMYB216 was significantly positively correlated with the upstream regulator bHLH30.This study systematically analyzed the MYB gene family in flax,laying a foundation for studying the regulation of LuMYB216 in flax flower anthocyanin synthesis.

Interaction of AcMADS68 with transcription factors regulates anthocyanin biosynthesis in red-fleshed kiwifruit

In red-fleshed kiwifruit,anthocyanin pigmentation is a crucial commercial trait.The MYB-bHLH-WD40(MBW)complex and other transcription factors regulate its accumulation.Herein,a new SEP gene,AcMADS68,was identified as a regulatory candidate for anthocyanin biosynthesis in the kiwifruit by transcriptome data and bioinformatic analyses.AcMADS68 alone could not induce the accumulation of anthocyanin both in Actinidia arguta fruit and tobacco leaves.However,in combination with AcMYBF110,AcMYB123,and AcbHLH1,AcMADS68 co-overexpression increased anthocyanin biosynthesis,whereas its silencing reduced anthocyanin accumulation.The results of the dual-luciferase reporter,firefly luciferase complementation,yeast two-hybrid and co-immunoprecipitation assays showed that AcMADS68 could interact with both AcMYBF110 and AcMYB123 but not with AcbHLH1,thereby co-regulating anthocyanin biosynthesis by promoting the activation of the target genes,including AcANS,AcF3GT1,and AcGST1.Moreover,AcMADS68 also could activate the promoter of AcbHLH1 surported by dual-luciferase reporter and yeast one-hybrid assays,thereby further amplifying the regulation signals from the MBW complex,thus resulting in enhanced anthocyanin accumulation in the kiwifruit.These findings may facilitate better elucidation of various regulatory mechanisms underlying anthocyanin accumulation and contribute to the quality enhancement of red-fleshed kiwifruit.