Grape crops are a great source of income for farmers.The yield and quality of grapes can be improved by preventing and treating diseases.The farmer’s yield will be dramatically impacted if diseases are found on grape...Grape crops are a great source of income for farmers.The yield and quality of grapes can be improved by preventing and treating diseases.The farmer’s yield will be dramatically impacted if diseases are found on grape leaves.Automatic detection can reduce the chances of leaf diseases affecting other healthy plants.Several studies have been conducted to detect grape leaf diseases,but most fail to engage with end users and integrate the model with real-time mobile applications.This study developed a mobile-based grape leaf disease detection(GLDD)application to identify infected leaves,Grape Guard,based on a TensorFlow Lite(TFLite)model generated from the You Only Look Once(YOLO)v8 model.A public grape leaf disease dataset containing four classes was used to train the model.The results of this study were relied on the YOLO architecture,specifically YOLOv5 and YOLOv8.After extensive experiments with different image sizes,YOLOv8 performed better than YOLOv5.YOLOv8 achieved 99.9%precision,100%recall,99.5%mean average precision(mAP),and 88%mAP50-95 for all classes to detect grape leaf diseases.The Grape Guard android mobile application can accurately detect the grape leaf disease by capturing images from grape vines.展开更多
Polyamines(PAs)and ethylene are involved in the modulation of plant growth and development.However,their roles in fruit-set,especially in exogenous gibberellin(GA_(3))-induced grape parthenocarpic berries,and the rela...Polyamines(PAs)and ethylene are involved in the modulation of plant growth and development.However,their roles in fruit-set,especially in exogenous gibberellin(GA_(3))-induced grape parthenocarpic berries,and the related competitive action mode are poorly understood.For this,we,here performed their content determination,bioinformatics and expression pattern analysis of genes to identify the key ones in the competitive network of polyamines metabolic and ethylene biosynthesis(PMEB)pathways.The content of putrescine(Put)significantly increased;while 1-aminocyclopropanecarboxylic acid(ACC)sharply decreased during the fruit-set process of GA_(3)-induced grape parthenocarpic seedless berries.Totally,twenty-five genes in PMEB pathways,including 20 polyamines metabolic(PM)genes and 5 ethylene biosynthesis(EB)ones were identified in grape,of which 8 PM and 2 EB genes possessed the motifs responsive to phytohormone GA.The expression levels of most PMEB genes kept changing during grape fruit-set generating a competitive action mode of GA_(3)-mediated two metabolic fluxes toward PAs and ethylene synthesis.Exogenous GA_(3)might enhance grape fruit-set of parthenocarpic berries via up-regulation of VvSAMS4,VvSAMDC1/2,VvODC1,VvSPDS1,and VvPAO1 to promote PAs accumulation,whereby repressing the ethylene synthesis by down-regulation of VvACS1 and VvACO_(2).Our findings provide novel insights into GA_(3)-mediated competitive inhibition of ethylene by PAs to promote the fruit-set of parthenocarpic berries in grape,which has important implications for molecular breeding of seedless grape with high fruit-setting rate.展开更多
Anthocyanins are important metabolites that provide a red or blue-purple hue to plants.The biosynthesis of these metabolites is mainly activated by the MYB-bHLH-WD40(MBW)complex and repressed by a wide variety of prot...Anthocyanins are important metabolites that provide a red or blue-purple hue to plants.The biosynthesis of these metabolites is mainly activated by the MYB-bHLH-WD40(MBW)complex and repressed by a wide variety of proteins.Studies have shown that MYB activators activate MYB repressors to balance anthocyanin biosynthesis.However,there is a scarcity of studies investigating this mechanism in grapes.To explore the transcription factors involved in the regulation of anthocyanin biosynthesis,we reanalyzed the RNA-seq database for different developmental stages of‘Muscat Hamburg'berries,and the R2R3-MYB gene,annotated as VvMYB3,was screened.Our study revealed the anthocyanin content of the grape cultivar‘Y73'was higher than that of its parental cultivar MH,and the putative repressor VvMYB3 was found to be highly expressed in‘Y73'by qRT-PCR.The calli transgenic assays demonstrated that the repressive activity of VvMYB3 was conferred by the b HLH-binding motif,as well as by the C1 and C2 motifs.Yeast hybridization and chip-PCR assays revealed that VvMYB3 could repress anthocyanin biosynthesis by competing with VvMYBA1 to bind to VvMYC1 and promoting histone deacetylation of VvUFGT via the C2 motif.However,the expression of VvMYB3 was activated by VvMYBA1,which forms a negative feedback regulatory loop to modulate anthocyanin accumulation.In addition,we found a 408-bp repeat tandem sequence insertion in the VvMYBA1 promoter region of‘Y73'by sequencing.The GUS activity analysis showed that this sequence enhanced the expression of VvMYBA1 and led to an excessive accumulation of anthocyanins.Overall,our results provide insights into the anthocyanin activator-repressor system in grapes that prevents overaccumulation of anthocyanins.展开更多
Evaluating plant stress tolerance and screening key regulatory genes under the combined stresses of high temperature and drought are important for studying plant stress tolerance mechanisms. In this study, the drought...Evaluating plant stress tolerance and screening key regulatory genes under the combined stresses of high temperature and drought are important for studying plant stress tolerance mechanisms. In this study, the drought tolerance of five grape varieties was evaluated under high-temperature conditions to screen key genes for further exploration of resistance mechanisms. By comparing and analysing the morphological characteristics and physiological indicators associated with the response of grapevines to drought stress and integrating them with the membership function to assess the strength of their drought tolerance, the order of drought tolerance was found to be as follows: 420A>110R>Cabernet Sauvignon(CS)>Fercal>188-08. To further analyse the mechanism of differences in drought tolerance, transcriptomic sequencing was performed on the drought-tolerant cultivar 420A, the drought-sensitive cultivar 188-08 and the control cultivar CS. The functional analysis of differential metabolic pathways indicated that the differentially expressed genes were enriched mainly in biological process category, that 420A had higher antioxidant activity. Furthermore, differentially expressed transcription factors were analyzed in five grape varieties. Genes like Vv AGL15, Vv LBD41, and Vv MYB86 showed close associations with drought tolerance,indicating their potential role in regulating drought tolerance and research significance.展开更多
Anthocyanins are important pigments and nutrients in fruits.Red grape is popular because of the high anthocyanin content.Previous studies have identified VvMYBA1 and its homologs as key regulators of fruit color;howev...Anthocyanins are important pigments and nutrients in fruits.Red grape is popular because of the high anthocyanin content.Previous studies have identified VvMYBA1 and its homologs as key regulators of fruit color;however,other transcription factors(TFs)that contribute to fruit color remain poorly understood.The present study identified the R2R3-MYB TF VvMYB24,whose gene expression levels were significantly higher in red berries(L51,Vitis vinifera×Vitis labrusca L.)than in green berries(L20,V.vinifera×V.labrusca L.).Overexpression of VvMYB24 in grape calli increased anthocyanin biosynthesis by upregulating the expression of specific structural genes(VvDFR and VvUFGT).Furthermore,VvMYB24 interacted with VvMYBA1 to form a protein complex that additionally increased the expression of VvDFR and VvUFGT.In addition,light-responsive TF VvHY5 could bind to the VvMYB24 promoters to activate its transcription.Taken together,the results reveal a regulatory module,VvHY5-VvMYB24-VvMYBA1,that influences anthocyanin biosynthesis in grape.展开更多
Increasing evidence indicates that a healthy diet plays a protective role against ischemic stroke injury.As an antioxidant food,grape powder(GP)has been shown to have a neuroprotective function after ischemic injury.I...Increasing evidence indicates that a healthy diet plays a protective role against ischemic stroke injury.As an antioxidant food,grape powder(GP)has been shown to have a neuroprotective function after ischemic injury.In the current research,we aimed to determine whether regular GP feeding before brain damage plays a protective role against ischemic damage and its potential mechanism.Mice had GP in drinking water(DW)for4 weeks.Then,fecal samples were collected for 16S rRNA analysis and metabolite profiling.Ischemic stroke was triggered using GP prefeed mice by middle cerebral artery occlusion(MCAo)to compare the stroke-induced infarction size to the DW group.The data showed that 4 weeks of GP pretreatment significantly decreased the ischemic infarct size.Meanwhile,GP treatment altered the gut microbiota by enriching the population of Lachnospiraceae and enhancing a gut metabolite called fiestin,which belongs to the flavonoid group.Mechanistically,we demonstrated that fiestin could inhibit LPS-induced proinflammatory macrophage polarization in vitro.We also showed that regular GP feed could decrease stroke-induced neuroinflammation in vivo.In conclusion,our study demonstrated that 4 weeks of GP feeding attenuates experimental ischemic stroke via the gut-brain-immune axis.展开更多
Grape white rot caused by Coniella vitis is a global concern in the grape industry.pH regulation is essential for cell growth,reproductive processes and pathogenicity in phytopathogenic fungi.In this study,we observed...Grape white rot caused by Coniella vitis is a global concern in the grape industry.pH regulation is essential for cell growth,reproductive processes and pathogenicity in phytopathogenic fungi.In this study,we observed that the growth rate,spore production and virulence of C.vitis significantly declined in alkaline pH,as well as the suppressive effect on secretion of hydrolytic enzymes.Transcriptomic and metabolomic analyses were used to investigate the responses of C.vitis to acidic(pH 5),neutral(pH 7)and alkaline environments(pH 9).We identified 728,1,780 and 3,386 differentially expressed genes(DEGs)at pH 5,pH 7 and pH 9,when compared with the host pH(pH 3),and 2,122 differently expressed metabolites(DEMs)in negative and positive ion mode.Most DEGs were involved in carbohydrate metabolic process,transmembrane transport,tricarboxylic acid cycle,peptide metabolic process,amide biosynthetic process,and organic acid metabolic process.In addition,metabolomic analysis revealed ABC transporters,indole alkaloid biosynthesis,diterpenoid biosynthesis,and carotenoid biosynthesis pathways in response to the pH change.Furthermore,we found that the aspartate synthesis metabolic route associated with the TCA cycle is a key limiting factor for the growth and development of C.vitis in alkaline environments,and aspartate supplementation enables C.vitis to grow in alkaline environments.Plant cell wall-degrading enzymes(PCWDEs)could contribute to the pathogenicity,when C.vitis infected at pH 3.Importantly,aflatrem biosynthesis in acidic environment might contribute to the virulence of C.vitis and has a risk of causing human health problems due to its acute neurotoxic effects.展开更多
In grape breeding programs,the extensive planting of seedlings is a crucial aspect.However,grape seeds display distinct dormancy traits,necessitating a prolonged cold stratification process for dormancy release.In ord...In grape breeding programs,the extensive planting of seedlings is a crucial aspect.However,grape seeds display distinct dormancy traits,necessitating a prolonged cold stratification process for dormancy release.In order to enhance the efficiency of breeding programs,this study presents an innovative in vitro embryo germination technique that eliminates the requirement for cold stratification of seeds.The method involves the disruption of peripheral tissue in grape seed embryos using a straightforward mechanical technique,resulting in the efficient production of a substantial quantity of seed embryos,with a germination rate of up to 88% for these isolated embryos.These embryos are subsequently cultured in vitro to facilitate germination into seedlings,thereby eliminating the need for cold stratification.Consequently,grape seedlings can be obtained within a significantly reduced timeframe of 30-38 d,expediting the overall grape breeding process.This novel approach not only accelerates grape hybridization but also streamlines the selection of new grape varieties,contributing to an efficient and time-sensitive breeding methodology.展开更多
In recent years,fungal diseases affecting grape crops have attracted significant attention.Currently,the assessment of black rot severitymainly depends on the ratio of lesion area to leaf surface area.However,effectiv...In recent years,fungal diseases affecting grape crops have attracted significant attention.Currently,the assessment of black rot severitymainly depends on the ratio of lesion area to leaf surface area.However,effectively and accurately segmenting leaf lesions presents considerable challenges.Existing grape leaf lesion segmentationmodels have several limitations,such as a large number of parameters,long training durations,and limited precision in extracting small lesions and boundary details.To address these issues,we propose an enhanced DeepLabv3+model incorporating Strip Pooling,Content-Guided Fusion,and Convolutional Block Attention Module(SFC_DeepLabv3+),an enhanced lesion segmentation method based on DeepLabv3+.This approach uses the lightweight MobileNetv2 backbone to replace the original Xception,incorporates a lightweight convolutional block attention module,and introduces a content-guided feature fusion module to improve the detection accuracy of small lesions and blurred boundaries.Experimental results showthat the enhancedmodel achieves a mean Intersection overUnion(mIoU)of 90.98%,amean Pixel Accuracy(mPA)of 94.33%,and a precision of 95.84%.This represents relative gains of 2.22%,1.78%,and 0.89%respectively compared to the original model.Additionally,its complexity is significantly reduced without sacrificing performance,the parameter count is reduced to 6.27 M,a decrease of 88.5%compared to the original model,floating point of operations(GFLOPs)drops from 83.62 to 29.00 G,a reduction of 65.1%.Additionally,Frames Per Second(FPS)increases from 63.7 to 74.3 FPS,marking an improvement of 16.7%.Compared to other models,the improved architecture shows faster convergence and superior segmentation accuracy,making it highly suitable for applications in resource-constrained environments.展开更多
Grape white rot is a fungal disease caused by Coniella diplodiella(Speg.)Sacc.that seriously affects fruit quality and yield;however,the underlying mechanism governing the plant response to C.diplodiella pathogens is ...Grape white rot is a fungal disease caused by Coniella diplodiella(Speg.)Sacc.that seriously affects fruit quality and yield;however,the underlying mechanism governing the plant response to C.diplodiella pathogens is still poorly understood.Here,we characterized a homeodomain(HD)transcription factor from grape(Vitis vinifera),VvOCP3,and demonstrated its signifcance in C.diplodiella resistance.Expression analysis showed that VvOCP3 expression was signifcantly down-regulated upon inoculation with C.diplodiella.Functional analysis with transient injection in grape berries and stable overexpression in grape calli demonstrated that VvOCP3 negatively regulates grape resistance to C.diplodiella.Further studies showed that VvOCP3 directly binds to the promoter of VvPR1(pathogenesis-related protein 1)and inhibits its expression,resulting in reduced resistance to C.diplodiella.In addition,VvOCP3 can interact with the type 2C protein phosphatase VvABI1,which is a negative modulator of the ABA signaling pathway.In summary,our findings suggest that VvOCP3 plays a crucial role in regulating white rot resistance in grape,and offer theoretical guidance for developing grape cultivars with enhanced C.diplodiella resistance by regulating the expression of VvOCP3.展开更多
Seedlessness has always been a valuable quality characteristic of edible grape varieties.Although the production of seedless grapes has been ongoing for decades,the genetic complexity of seedless grapes is not yet ful...Seedlessness has always been a valuable quality characteristic of edible grape varieties.Although the production of seedless grapes has been ongoing for decades,the genetic complexity of seedless grapes is not yet fully understood.Therefore,determining the genetic mechanisms and key regulatory genes of seedless grapes is of great significance for seedless grape breeding and meeting market demands.The emergence of high-throughput analysis software offers greater possibilities for mining genes related to plant organ development.Specifically,to mine a greater number of candidate genes related to grape seed traits,this study used the seed trait parameters analyzed by Tomato Analyzer as the target trait and then used a genome-wide association study(GWAS)to mine candidate genes.In the two-year analysis using principal component analysis(PCA),we extracted five principal components with a cumulative contribution rate of 96.586%.The cumulative contribution rate for component 1 reached 87.352%.Correlation analysis revealed correlation coefficients ranging from 0.54 to 0.98 among the seven basic traits.The GWAS results indicated that 370 SNP loci were significantly correlated with seed traits.These SNP loci were distributed on 18 chromosomes,except for chromosome 4,with most SNP loci distributed on chromosome 18.Based on the physical location of single nucleotide polymorphism(SNP)markers significantly associated with seed-related traits in the grape reference genome,candidate genes are screened within the range of linkage disequilibrium(LD)attenuation distance,both upstream and downstream of the significant SNP loci.These candidate genes were mainly transcription factor-related genes(VvMADS4 and VvMADS5),ubiquitin ligase-related genes(E3 ubiquitin ligase BIG BROTHER),serine/threonine protein kinase-related genes,and carbohydrate metabolism-related genes(Sucrose Synthase 2)and simultaneously controlled multiple(at least two or more)seed traits.These results indicate that seed traits are jointly regulated by some genes involved in seed morphology regulation.In this work,we identified new gene loci related to grape seed traits.Identifying molecular markers closely related to these seed traits is of great significance for breeding seedless grape varieties.展开更多
Climate change is significantly altering viticultural practices worldwide,with profound implications for the accumulation of polyphenolic compounds that determine wine’s sensory and health properties.This review summ...Climate change is significantly altering viticultural practices worldwide,with profound implications for the accumulation of polyphenolic compounds that determine wine’s sensory and health properties.This review summarizes the effects of climate change,particularly rising temperatures,shifting precipitation patterns,and altered light conditions-on polyphenol synthesis in Vitis amurensis(V.amurensis)grapes from Northeast China,the country’s highest-latitude wine region.Key findings reveal that:(1)Temperature increases accelerate phenological stages but differentially impact polyphenols,suppressing anthocyanins and flavonols while promoting tannins;(2)Precipitation variability induces water stress that can enhance anthocyanin content under moderate drought but reduce quality during extreme events;(3)Declining sunshine duration may limit polyphenol production,though certain cultivars(e.g.,Beibinghong)exhibit adaptability to low light conditions.The region’s unique climatic trends-stronger winter warming and reduced summer precipitation-paradoxically offer potential benefits by extending the growing season while minimizing heat stress during critical ripening periods.It is highlighted how V.amurensis,with its cold hardiness and naturally high polyphenol content(notably anthocyanins and resveratrol),could become increasingly valuable under climate change.However,strategic adaptation through cultivar selection,vineyard management,and stress-responsive breeding will be critical to maintain wine quality.This synthesis provides a framework for understanding climate-polyphenol dynamics in cool-climate viticulture and outlines research priorities to safeguard the future of Northeast China’s distinctive wine industry.展开更多
Southern China has high temperatures and receives concentrated rainfall;therefore,the two-crop-a-year cultivation system has been applied to grape production so as to resolve the problem of relative seasonal surplus o...Southern China has high temperatures and receives concentrated rainfall;therefore,the two-crop-a-year cultivation system has been applied to grape production so as to resolve the problem of relative seasonal surplus of grape yield.However,a common issue associated with this technique is the tendency of the second season fruits to be smaller than the first season fruits.We here used the first and second season fruits of‘Summer Black'at different ripening stages as research materials.Phenotypic and histological analyses revealed fewer cell number occurring between 7 and 14 days post anthesis(DPA)in the second season fruits,which ultimately resulted in a smaller fruit size compared with the first season fruits.To unravel the mechanism underlying this phenomenon,first and second season fruits of four time periods(7,14,21,and 28 DPA)were selected for RNA-seq analysis.This analysis identified 10431 differentially expressed genes(DEGs).These DEGs were classified into 9 clusters through GO and KEGG enrichment analyses.Then the time-ordered gene co-expression network(TO-GCN)analysis with the breadth-first search algorithm showed that DEGs in the GCN were divided into 8 levels.The DEGs of early berry development(L1-L3)were enriched in heat stress-and cell division-related pathways.The field investigation of effective accumulated temperature confirmed that the growth and development of the second season fruits were subjected to high temperature stress during 7-14 DPA.Moreover,based on the results of interactive analysis of TO-GCN and transcriptional regulation prediction of L1-L3 genes,we constructed a unique hierarchical regulatory network for the heat stress regulation of berry size.The expression level of 5 candidate genes was verified through qRT-PCR.Vitvi10g00469(HSFB2A),Vitvi16g00982(HSFB2A),Vitvi02g00387(HSFB2B),Vitvi15g01542(NTL9),and Vitvi06g00592(DIVARICATA)were upregulated in 7-14 DPA,whereas Vitvi18g00777(HSFB4)was downregulated in 7 DPA.These results suggest that during intense cell division,heat stress might act as a major factor causing a reduction in cell number,thereby ultimately resulting in the smaller size of the second season fruits.展开更多
CRISPR-Cas9 has emerged as a powerful tool for gene editing,and it has been widely used in plant functional genomics research and crop genetic breeding(Chen et al.2019).The target specificity of CRISPR-Cas9 relies on ...CRISPR-Cas9 has emerged as a powerful tool for gene editing,and it has been widely used in plant functional genomics research and crop genetic breeding(Chen et al.2019).The target specificity of CRISPR-Cas9 relies on the 20-base-pair single guide RNA(sgRNA),which makes creating plant-specific mutant libraries through large-scale synthesis of sgRNAs targeting multiple genes or even the whole genome relatively quick and straightforward.展开更多
The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the p...The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the present study was to elucidate the evolutionary characteristics of the miR399 gene family in grape and to verify the cleavage effect on the target genes.Grape miR399s were identified by miRNA sequencing and retrieval from the miRBase database.The mature sequences and precursor sequences were subjected to phylogenetic analysis to reconstruct evolutionary trees,as well as secondary structure analysis of the precursor sequence,and prediction of target genes.The cisacting elements in the miR399 promoter were predicted and the cleavage effect of grape miR399b on its target genes was verified.The grape miR399 family comprised nine precursor sequences and nine mature sequences.The precursor sequences formed a typical and stable stem—loop structure.The minimum folding free energy ranged from-55.70 kcal·mol^(-1)to-37.40 kcal·mol^(-1).Multiple sequence alignment revealed that the miR399 family was highly conserved.The grape miR399 family was phylogenetically closely related to peach,apple,and citrus miR399s.Grape miR399s were predicted to target inorganic phosphate transporter 1—3,phospholipase D delta-like,and beta-glucuronosyltransferase.The cleavage effect of grape miR399b on the target genes was verified by means of a dual-luciferase assay and 5’RLM-RACE.Histochemical GUS staining showed that the promoter activity of miR399b was promoted by GA3treatment.展开更多
We determined whether the inclusion of 100 g/kg dry matter of grape pomace silage (GPS) and grape pomace bran (GPB) as substitutes for other traditional fiber sources in the diet of steers (Charolais x Nellore) would ...We determined whether the inclusion of 100 g/kg dry matter of grape pomace silage (GPS) and grape pomace bran (GPB) as substitutes for other traditional fiber sources in the diet of steers (Charolais x Nellore) would improve carcass characteristics, meat quality and composition, and shelf life. Twenty-four animals (248 ± 19.32 kg of initial body weight) were fed a high concentrate diet for 121 days. Carcass characteristics were measured, and the longissimus dorsi muscle was analyzed for fatty acid (FA) profile and composition. The meat was sliced and stored in air-permeable packages for 10 days. On each sampling day (d 1, 3, 7, and 10), oxidative stability, bacterial load, lipid and protein oxidation, and staining were analyzed. The experimental diets influenced the pH of cold carcasses only. The GPS group had a higher pH than the control. The GPS and GPB groups showed improved oxidant status (i.e., lower lipid peroxidation and concentrations of reactive oxygen species were in the meat of both groups than in control). On the first day of storage, the antioxidant enzyme glutathione S-transferase activity was more significant in the meat of the GPS and GPB groups than in the control. The bacterial loads in the meat were attenuated by GPS inclusion;there were lower total coliform counts and a trend toward lower counts for enterobacteria in the control group. The diets altered the FA profile of the meat;i.e., the GPB diet allowed for a more significant amount of the n-6 omegas in the meat, while the GPS diet showed a tendency for a more significant amount of n-6 and 9 omegas. Both diets (GPS and GPB) increased the amounts of long-chain FAs. The GPS diet decreased saturated FA levels. We conclude that the dietary treatments GPS and GPB are a promising alternative to maintain meat quality standards throughout in real-world retail conditions. These treatments gave rise to an improvement in the nutritional value of the meat due to the more significant amounts of FAs that improve human health.展开更多
Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR...Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR)family,comprising highly conserved ligand-gated ion channels,regulates plant growth and development in response to stress.In this study,11 members of the VvGLR gene family in grapes were identified using whole-genome sequence analysis.Bioinformatic methods were employed to analyze the basic physical and chemical properties,phylogenetic trees,conserved domains,motifs,expression patterns,and evolutionary relationships.Phylogenetic and collinear analyses revealed that the VvGLRs were divided into three subgroups,showing the high conservation of the grape GLR family.These members exhibited 2 glutamate receptor binding regions(GABAb and GluR)and 3-4 transmembrane regions(M1,M2,M3,and M4).Real-time quantitative PCR analysis demonstrated the sensitivity of all VvGLRs to low temperature and salt stress.Subsequent localization studies in Nicotiana tabacum verified that VvGLR3.1 and VvGLR3.2 proteins were located on the cell membrane and cell nucleus.Additionally,yeast transformation experiments confirmed the functionality of VvGLR3.1 and VvGLR3.2 in response to low temperature and salt stress.Thesefindings highlight the significant role of the GLR family,a highly conserved group of ion channels,in enhancing grape stress resistance.This study offers new insights into the grape GLR gene family,providing fundamental knowledge for further functional analysis and breeding of stress-resistant grapevines.展开更多
The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the under...The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes(DEGs) by comparing the transcriptomes of folic acid(FA)-treated and water-treated(CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced(AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology(GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes,particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2(VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries.Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.展开更多
文摘Grape crops are a great source of income for farmers.The yield and quality of grapes can be improved by preventing and treating diseases.The farmer’s yield will be dramatically impacted if diseases are found on grape leaves.Automatic detection can reduce the chances of leaf diseases affecting other healthy plants.Several studies have been conducted to detect grape leaf diseases,but most fail to engage with end users and integrate the model with real-time mobile applications.This study developed a mobile-based grape leaf disease detection(GLDD)application to identify infected leaves,Grape Guard,based on a TensorFlow Lite(TFLite)model generated from the You Only Look Once(YOLO)v8 model.A public grape leaf disease dataset containing four classes was used to train the model.The results of this study were relied on the YOLO architecture,specifically YOLOv5 and YOLOv8.After extensive experiments with different image sizes,YOLOv8 performed better than YOLOv5.YOLOv8 achieved 99.9%precision,100%recall,99.5%mean average precision(mAP),and 88%mAP50-95 for all classes to detect grape leaf diseases.The Grape Guard android mobile application can accurately detect the grape leaf disease by capturing images from grape vines.
基金supported by grants from Jiangsu province seed industry revitalization of the leading project(JBGS[2021]086)the National Natural Science Funds(31972373,32272647,32202433)+1 种基金the Provincial Natural Science Foundation of Jiangsu(BK20200541)the Priority Academic Program Development of Jiangsu Higher Education Institutions,China(PAPD),and Postgraduate Research&Practice Innovation Program of Jiangsu Province,China(KYCX22_0754,SJCX23-0209).
文摘Polyamines(PAs)and ethylene are involved in the modulation of plant growth and development.However,their roles in fruit-set,especially in exogenous gibberellin(GA_(3))-induced grape parthenocarpic berries,and the related competitive action mode are poorly understood.For this,we,here performed their content determination,bioinformatics and expression pattern analysis of genes to identify the key ones in the competitive network of polyamines metabolic and ethylene biosynthesis(PMEB)pathways.The content of putrescine(Put)significantly increased;while 1-aminocyclopropanecarboxylic acid(ACC)sharply decreased during the fruit-set process of GA_(3)-induced grape parthenocarpic seedless berries.Totally,twenty-five genes in PMEB pathways,including 20 polyamines metabolic(PM)genes and 5 ethylene biosynthesis(EB)ones were identified in grape,of which 8 PM and 2 EB genes possessed the motifs responsive to phytohormone GA.The expression levels of most PMEB genes kept changing during grape fruit-set generating a competitive action mode of GA_(3)-mediated two metabolic fluxes toward PAs and ethylene synthesis.Exogenous GA_(3)might enhance grape fruit-set of parthenocarpic berries via up-regulation of VvSAMS4,VvSAMDC1/2,VvODC1,VvSPDS1,and VvPAO1 to promote PAs accumulation,whereby repressing the ethylene synthesis by down-regulation of VvACS1 and VvACO_(2).Our findings provide novel insights into GA_(3)-mediated competitive inhibition of ethylene by PAs to promote the fruit-set of parthenocarpic berries in grape,which has important implications for molecular breeding of seedless grape with high fruit-setting rate.
基金supported by the National Natural Science Foundation of China(Grant No.32202438)the China Agriculture Research System(Grant No.CARS-29)the Agricultural Science and Technology Innovation Program(Grant No.CAASASTIP-ZFRI)。
文摘Anthocyanins are important metabolites that provide a red or blue-purple hue to plants.The biosynthesis of these metabolites is mainly activated by the MYB-bHLH-WD40(MBW)complex and repressed by a wide variety of proteins.Studies have shown that MYB activators activate MYB repressors to balance anthocyanin biosynthesis.However,there is a scarcity of studies investigating this mechanism in grapes.To explore the transcription factors involved in the regulation of anthocyanin biosynthesis,we reanalyzed the RNA-seq database for different developmental stages of‘Muscat Hamburg'berries,and the R2R3-MYB gene,annotated as VvMYB3,was screened.Our study revealed the anthocyanin content of the grape cultivar‘Y73'was higher than that of its parental cultivar MH,and the putative repressor VvMYB3 was found to be highly expressed in‘Y73'by qRT-PCR.The calli transgenic assays demonstrated that the repressive activity of VvMYB3 was conferred by the b HLH-binding motif,as well as by the C1 and C2 motifs.Yeast hybridization and chip-PCR assays revealed that VvMYB3 could repress anthocyanin biosynthesis by competing with VvMYBA1 to bind to VvMYC1 and promoting histone deacetylation of VvUFGT via the C2 motif.However,the expression of VvMYB3 was activated by VvMYBA1,which forms a negative feedback regulatory loop to modulate anthocyanin accumulation.In addition,we found a 408-bp repeat tandem sequence insertion in the VvMYBA1 promoter region of‘Y73'by sequencing.The GUS activity analysis showed that this sequence enhanced the expression of VvMYBA1 and led to an excessive accumulation of anthocyanins.Overall,our results provide insights into the anthocyanin activator-repressor system in grapes that prevents overaccumulation of anthocyanins.
基金supported by the Major Innovation Project of Shandong Province, China (2022CXGC010605)the National Natural Science Foundation of China (32172518 and 32002023)+1 种基金the National Key R&D Program of China (2023YFD2301103)the Key R&D Projects in Ningxia Hui Autonomous Region, China (2022BBF02014)。
文摘Evaluating plant stress tolerance and screening key regulatory genes under the combined stresses of high temperature and drought are important for studying plant stress tolerance mechanisms. In this study, the drought tolerance of five grape varieties was evaluated under high-temperature conditions to screen key genes for further exploration of resistance mechanisms. By comparing and analysing the morphological characteristics and physiological indicators associated with the response of grapevines to drought stress and integrating them with the membership function to assess the strength of their drought tolerance, the order of drought tolerance was found to be as follows: 420A>110R>Cabernet Sauvignon(CS)>Fercal>188-08. To further analyse the mechanism of differences in drought tolerance, transcriptomic sequencing was performed on the drought-tolerant cultivar 420A, the drought-sensitive cultivar 188-08 and the control cultivar CS. The functional analysis of differential metabolic pathways indicated that the differentially expressed genes were enriched mainly in biological process category, that 420A had higher antioxidant activity. Furthermore, differentially expressed transcription factors were analyzed in five grape varieties. Genes like Vv AGL15, Vv LBD41, and Vv MYB86 showed close associations with drought tolerance,indicating their potential role in regulating drought tolerance and research significance.
基金supported by the National Natural Science Foundation of China(Grant No.31972368)the China Agriculture Research System(Grant No.CARS-29-yc-6)+1 种基金the Major Agricultural Science Projects of Liaoning Province(Grant No.2023JH1/10200004)the Science and Technology Program of Shenyang(Grant No.23-410-2-03).
文摘Anthocyanins are important pigments and nutrients in fruits.Red grape is popular because of the high anthocyanin content.Previous studies have identified VvMYBA1 and its homologs as key regulators of fruit color;however,other transcription factors(TFs)that contribute to fruit color remain poorly understood.The present study identified the R2R3-MYB TF VvMYB24,whose gene expression levels were significantly higher in red berries(L51,Vitis vinifera×Vitis labrusca L.)than in green berries(L20,V.vinifera×V.labrusca L.).Overexpression of VvMYB24 in grape calli increased anthocyanin biosynthesis by upregulating the expression of specific structural genes(VvDFR and VvUFGT).Furthermore,VvMYB24 interacted with VvMYBA1 to form a protein complex that additionally increased the expression of VvDFR and VvUFGT.In addition,light-responsive TF VvHY5 could bind to the VvMYB24 promoters to activate its transcription.Taken together,the results reveal a regulatory module,VvHY5-VvMYB24-VvMYBA1,that influences anthocyanin biosynthesis in grape.
基金supported by the National Natural Science Foundation of China(82001248,82471329)2023 Beijing Demonstration Project of Collaborative Research of Traditional Chinese and Western Medicine for Major and Difficult Diseases-Migraine(2023BJSZDYNJBXTGG-015)。
文摘Increasing evidence indicates that a healthy diet plays a protective role against ischemic stroke injury.As an antioxidant food,grape powder(GP)has been shown to have a neuroprotective function after ischemic injury.In the current research,we aimed to determine whether regular GP feeding before brain damage plays a protective role against ischemic damage and its potential mechanism.Mice had GP in drinking water(DW)for4 weeks.Then,fecal samples were collected for 16S rRNA analysis and metabolite profiling.Ischemic stroke was triggered using GP prefeed mice by middle cerebral artery occlusion(MCAo)to compare the stroke-induced infarction size to the DW group.The data showed that 4 weeks of GP pretreatment significantly decreased the ischemic infarct size.Meanwhile,GP treatment altered the gut microbiota by enriching the population of Lachnospiraceae and enhancing a gut metabolite called fiestin,which belongs to the flavonoid group.Mechanistically,we demonstrated that fiestin could inhibit LPS-induced proinflammatory macrophage polarization in vitro.We also showed that regular GP feed could decrease stroke-induced neuroinflammation in vivo.In conclusion,our study demonstrated that 4 weeks of GP feeding attenuates experimental ischemic stroke via the gut-brain-immune axis.
基金supported by the Shandong Provincial Natural Science Foundation,China(ZR2021QC131)the Shandong Province Key Research and Development Plan,China(2022TZXD001102)+1 种基金the Shandong Province Demonstration Project for Model Construction in Rural Revitalization Service,China(2022DXAL0226)the Innovation Project of Shandong Academy of Agricultural Sciences,China(CXGC2023F15,CXGC2023A41,and CXGC2023A47)。
文摘Grape white rot caused by Coniella vitis is a global concern in the grape industry.pH regulation is essential for cell growth,reproductive processes and pathogenicity in phytopathogenic fungi.In this study,we observed that the growth rate,spore production and virulence of C.vitis significantly declined in alkaline pH,as well as the suppressive effect on secretion of hydrolytic enzymes.Transcriptomic and metabolomic analyses were used to investigate the responses of C.vitis to acidic(pH 5),neutral(pH 7)and alkaline environments(pH 9).We identified 728,1,780 and 3,386 differentially expressed genes(DEGs)at pH 5,pH 7 and pH 9,when compared with the host pH(pH 3),and 2,122 differently expressed metabolites(DEMs)in negative and positive ion mode.Most DEGs were involved in carbohydrate metabolic process,transmembrane transport,tricarboxylic acid cycle,peptide metabolic process,amide biosynthetic process,and organic acid metabolic process.In addition,metabolomic analysis revealed ABC transporters,indole alkaloid biosynthesis,diterpenoid biosynthesis,and carotenoid biosynthesis pathways in response to the pH change.Furthermore,we found that the aspartate synthesis metabolic route associated with the TCA cycle is a key limiting factor for the growth and development of C.vitis in alkaline environments,and aspartate supplementation enables C.vitis to grow in alkaline environments.Plant cell wall-degrading enzymes(PCWDEs)could contribute to the pathogenicity,when C.vitis infected at pH 3.Importantly,aflatrem biosynthesis in acidic environment might contribute to the virulence of C.vitis and has a risk of causing human health problems due to its acute neurotoxic effects.
基金supported by Natural Science Foundation of Beijing Academy of Agriculture and Forestry Sciences(Grant No.QNJJ202301)the Youth Research Foundation of Institute of Forestry and Pomology+2 种基金Beijing Academy of Agriculture and Forestry Science(Grant No.LGJJ202202)the BAAFS Funding for the Development of Distinguished Scientist(Grant No.JKZX202402)the Beijing Natural Science Foundation(Grant No.6242019)。
文摘In grape breeding programs,the extensive planting of seedlings is a crucial aspect.However,grape seeds display distinct dormancy traits,necessitating a prolonged cold stratification process for dormancy release.In order to enhance the efficiency of breeding programs,this study presents an innovative in vitro embryo germination technique that eliminates the requirement for cold stratification of seeds.The method involves the disruption of peripheral tissue in grape seed embryos using a straightforward mechanical technique,resulting in the efficient production of a substantial quantity of seed embryos,with a germination rate of up to 88% for these isolated embryos.These embryos are subsequently cultured in vitro to facilitate germination into seedlings,thereby eliminating the need for cold stratification.Consequently,grape seedlings can be obtained within a significantly reduced timeframe of 30-38 d,expediting the overall grape breeding process.This novel approach not only accelerates grape hybridization but also streamlines the selection of new grape varieties,contributing to an efficient and time-sensitive breeding methodology.
基金supported by the following grants:Zhejiang A&F University Research Development Fund(Talent Initiation Project No.2021LFR048)and 2023 University-Enterprise Joint Research Program(Grant No.LHYFZ2302)from the Modern Agricultural and Forestry Artificial Intelligence Industry Academy.
文摘In recent years,fungal diseases affecting grape crops have attracted significant attention.Currently,the assessment of black rot severitymainly depends on the ratio of lesion area to leaf surface area.However,effectively and accurately segmenting leaf lesions presents considerable challenges.Existing grape leaf lesion segmentationmodels have several limitations,such as a large number of parameters,long training durations,and limited precision in extracting small lesions and boundary details.To address these issues,we propose an enhanced DeepLabv3+model incorporating Strip Pooling,Content-Guided Fusion,and Convolutional Block Attention Module(SFC_DeepLabv3+),an enhanced lesion segmentation method based on DeepLabv3+.This approach uses the lightweight MobileNetv2 backbone to replace the original Xception,incorporates a lightweight convolutional block attention module,and introduces a content-guided feature fusion module to improve the detection accuracy of small lesions and blurred boundaries.Experimental results showthat the enhancedmodel achieves a mean Intersection overUnion(mIoU)of 90.98%,amean Pixel Accuracy(mPA)of 94.33%,and a precision of 95.84%.This represents relative gains of 2.22%,1.78%,and 0.89%respectively compared to the original model.Additionally,its complexity is significantly reduced without sacrificing performance,the parameter count is reduced to 6.27 M,a decrease of 88.5%compared to the original model,floating point of operations(GFLOPs)drops from 83.62 to 29.00 G,a reduction of 65.1%.Additionally,Frames Per Second(FPS)increases from 63.7 to 74.3 FPS,marking an improvement of 16.7%.Compared to other models,the improved architecture shows faster convergence and superior segmentation accuracy,making it highly suitable for applications in resource-constrained environments.
基金funded by the National Natural Science Foundation of China(31972368)the China Agriculture Research System(CARS-29-yc-6)+1 种基金the Major Agricultural Science Projects of Liaoning Province,China(2023JH1/10200004)the Science and Technology Program of Shenyang,China(23-410-2-03)。
文摘Grape white rot is a fungal disease caused by Coniella diplodiella(Speg.)Sacc.that seriously affects fruit quality and yield;however,the underlying mechanism governing the plant response to C.diplodiella pathogens is still poorly understood.Here,we characterized a homeodomain(HD)transcription factor from grape(Vitis vinifera),VvOCP3,and demonstrated its signifcance in C.diplodiella resistance.Expression analysis showed that VvOCP3 expression was signifcantly down-regulated upon inoculation with C.diplodiella.Functional analysis with transient injection in grape berries and stable overexpression in grape calli demonstrated that VvOCP3 negatively regulates grape resistance to C.diplodiella.Further studies showed that VvOCP3 directly binds to the promoter of VvPR1(pathogenesis-related protein 1)and inhibits its expression,resulting in reduced resistance to C.diplodiella.In addition,VvOCP3 can interact with the type 2C protein phosphatase VvABI1,which is a negative modulator of the ABA signaling pathway.In summary,our findings suggest that VvOCP3 plays a crucial role in regulating white rot resistance in grape,and offer theoretical guidance for developing grape cultivars with enhanced C.diplodiella resistance by regulating the expression of VvOCP3.
基金supported by grants from the Basic Research Business Fees for Public Welfare in Xinjiang Autonomous Region(Mining candidate genes related to grape seed traits based on GWAS,KY2023028)the Xinjiang Uygur Autonomous Region Tianchi Talent-Young Doctor for Chuan Zhang(Revealing the domestication history of Xinjiang native grape varieties and genetic analysis of important agronomic traits)the Xinjiang Academy of Agricultural Sciences Youth Science and Technology Backbone Innovation Ability Training Project(xjnkq-2023006).
文摘Seedlessness has always been a valuable quality characteristic of edible grape varieties.Although the production of seedless grapes has been ongoing for decades,the genetic complexity of seedless grapes is not yet fully understood.Therefore,determining the genetic mechanisms and key regulatory genes of seedless grapes is of great significance for seedless grape breeding and meeting market demands.The emergence of high-throughput analysis software offers greater possibilities for mining genes related to plant organ development.Specifically,to mine a greater number of candidate genes related to grape seed traits,this study used the seed trait parameters analyzed by Tomato Analyzer as the target trait and then used a genome-wide association study(GWAS)to mine candidate genes.In the two-year analysis using principal component analysis(PCA),we extracted five principal components with a cumulative contribution rate of 96.586%.The cumulative contribution rate for component 1 reached 87.352%.Correlation analysis revealed correlation coefficients ranging from 0.54 to 0.98 among the seven basic traits.The GWAS results indicated that 370 SNP loci were significantly correlated with seed traits.These SNP loci were distributed on 18 chromosomes,except for chromosome 4,with most SNP loci distributed on chromosome 18.Based on the physical location of single nucleotide polymorphism(SNP)markers significantly associated with seed-related traits in the grape reference genome,candidate genes are screened within the range of linkage disequilibrium(LD)attenuation distance,both upstream and downstream of the significant SNP loci.These candidate genes were mainly transcription factor-related genes(VvMADS4 and VvMADS5),ubiquitin ligase-related genes(E3 ubiquitin ligase BIG BROTHER),serine/threonine protein kinase-related genes,and carbohydrate metabolism-related genes(Sucrose Synthase 2)and simultaneously controlled multiple(at least two or more)seed traits.These results indicate that seed traits are jointly regulated by some genes involved in seed morphology regulation.In this work,we identified new gene loci related to grape seed traits.Identifying molecular markers closely related to these seed traits is of great significance for breeding seedless grape varieties.
文摘Climate change is significantly altering viticultural practices worldwide,with profound implications for the accumulation of polyphenolic compounds that determine wine’s sensory and health properties.This review summarizes the effects of climate change,particularly rising temperatures,shifting precipitation patterns,and altered light conditions-on polyphenol synthesis in Vitis amurensis(V.amurensis)grapes from Northeast China,the country’s highest-latitude wine region.Key findings reveal that:(1)Temperature increases accelerate phenological stages but differentially impact polyphenols,suppressing anthocyanins and flavonols while promoting tannins;(2)Precipitation variability induces water stress that can enhance anthocyanin content under moderate drought but reduce quality during extreme events;(3)Declining sunshine duration may limit polyphenol production,though certain cultivars(e.g.,Beibinghong)exhibit adaptability to low light conditions.The region’s unique climatic trends-stronger winter warming and reduced summer precipitation-paradoxically offer potential benefits by extending the growing season while minimizing heat stress during critical ripening periods.It is highlighted how V.amurensis,with its cold hardiness and naturally high polyphenol content(notably anthocyanins and resveratrol),could become increasingly valuable under climate change.However,strategic adaptation through cultivar selection,vineyard management,and stress-responsive breeding will be critical to maintain wine quality.This synthesis provides a framework for understanding climate-polyphenol dynamics in cool-climate viticulture and outlines research priorities to safeguard the future of Northeast China’s distinctive wine industry.
基金supported by National Technology System for Grape Industry(Grant No.CARS-29-zp-9)National Key Research and Development Program of China(Grant No.2021YFD1200200)Postgraduate Scientific Research Innovation Project of Hunan Province(Grant No.CX20230701)。
文摘Southern China has high temperatures and receives concentrated rainfall;therefore,the two-crop-a-year cultivation system has been applied to grape production so as to resolve the problem of relative seasonal surplus of grape yield.However,a common issue associated with this technique is the tendency of the second season fruits to be smaller than the first season fruits.We here used the first and second season fruits of‘Summer Black'at different ripening stages as research materials.Phenotypic and histological analyses revealed fewer cell number occurring between 7 and 14 days post anthesis(DPA)in the second season fruits,which ultimately resulted in a smaller fruit size compared with the first season fruits.To unravel the mechanism underlying this phenomenon,first and second season fruits of four time periods(7,14,21,and 28 DPA)were selected for RNA-seq analysis.This analysis identified 10431 differentially expressed genes(DEGs).These DEGs were classified into 9 clusters through GO and KEGG enrichment analyses.Then the time-ordered gene co-expression network(TO-GCN)analysis with the breadth-first search algorithm showed that DEGs in the GCN were divided into 8 levels.The DEGs of early berry development(L1-L3)were enriched in heat stress-and cell division-related pathways.The field investigation of effective accumulated temperature confirmed that the growth and development of the second season fruits were subjected to high temperature stress during 7-14 DPA.Moreover,based on the results of interactive analysis of TO-GCN and transcriptional regulation prediction of L1-L3 genes,we constructed a unique hierarchical regulatory network for the heat stress regulation of berry size.The expression level of 5 candidate genes was verified through qRT-PCR.Vitvi10g00469(HSFB2A),Vitvi16g00982(HSFB2A),Vitvi02g00387(HSFB2B),Vitvi15g01542(NTL9),and Vitvi06g00592(DIVARICATA)were upregulated in 7-14 DPA,whereas Vitvi18g00777(HSFB4)was downregulated in 7 DPA.These results suggest that during intense cell division,heat stress might act as a major factor causing a reduction in cell number,thereby ultimately resulting in the smaller size of the second season fruits.
基金supported by the National Natural Science Foundation of China(32272670 and 31972986)the Key Research and Development Program of Shaanxi Province,China(2023-YBNY-059)。
文摘CRISPR-Cas9 has emerged as a powerful tool for gene editing,and it has been widely used in plant functional genomics research and crop genetic breeding(Chen et al.2019).The target specificity of CRISPR-Cas9 relies on the 20-base-pair single guide RNA(sgRNA),which makes creating plant-specific mutant libraries through large-scale synthesis of sgRNAs targeting multiple genes or even the whole genome relatively quick and straightforward.
基金supported by Natural Science Foundation of China(Grant No.U1904113)National Key Research and Development Program of China(Grant No.2018YFD1000105)+3 种基金Program for Innovative Research Team(in Science and Technology)in University of Henan Province(Grant No.21IRTSTHN021)Program for Science&Technology Innovation Talents in Universities of Henan Province(Grant No.21HASTIT035)Scientific and technological breakthroughs in Henan Province(Grant No.222102110083)PhD Research Startup Foundation of Henan University of Science and Technology(Grant Nos.13480067,13480068)。
文摘The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the present study was to elucidate the evolutionary characteristics of the miR399 gene family in grape and to verify the cleavage effect on the target genes.Grape miR399s were identified by miRNA sequencing and retrieval from the miRBase database.The mature sequences and precursor sequences were subjected to phylogenetic analysis to reconstruct evolutionary trees,as well as secondary structure analysis of the precursor sequence,and prediction of target genes.The cisacting elements in the miR399 promoter were predicted and the cleavage effect of grape miR399b on its target genes was verified.The grape miR399 family comprised nine precursor sequences and nine mature sequences.The precursor sequences formed a typical and stable stem—loop structure.The minimum folding free energy ranged from-55.70 kcal·mol^(-1)to-37.40 kcal·mol^(-1).Multiple sequence alignment revealed that the miR399 family was highly conserved.The grape miR399 family was phylogenetically closely related to peach,apple,and citrus miR399s.Grape miR399s were predicted to target inorganic phosphate transporter 1—3,phospholipase D delta-like,and beta-glucuronosyltransferase.The cleavage effect of grape miR399b on the target genes was verified by means of a dual-luciferase assay and 5’RLM-RACE.Histochemical GUS staining showed that the promoter activity of miR399b was promoted by GA3treatment.
文摘We determined whether the inclusion of 100 g/kg dry matter of grape pomace silage (GPS) and grape pomace bran (GPB) as substitutes for other traditional fiber sources in the diet of steers (Charolais x Nellore) would improve carcass characteristics, meat quality and composition, and shelf life. Twenty-four animals (248 ± 19.32 kg of initial body weight) were fed a high concentrate diet for 121 days. Carcass characteristics were measured, and the longissimus dorsi muscle was analyzed for fatty acid (FA) profile and composition. The meat was sliced and stored in air-permeable packages for 10 days. On each sampling day (d 1, 3, 7, and 10), oxidative stability, bacterial load, lipid and protein oxidation, and staining were analyzed. The experimental diets influenced the pH of cold carcasses only. The GPS group had a higher pH than the control. The GPS and GPB groups showed improved oxidant status (i.e., lower lipid peroxidation and concentrations of reactive oxygen species were in the meat of both groups than in control). On the first day of storage, the antioxidant enzyme glutathione S-transferase activity was more significant in the meat of the GPS and GPB groups than in the control. The bacterial loads in the meat were attenuated by GPS inclusion;there were lower total coliform counts and a trend toward lower counts for enterobacteria in the control group. The diets altered the FA profile of the meat;i.e., the GPB diet allowed for a more significant amount of the n-6 omegas in the meat, while the GPS diet showed a tendency for a more significant amount of n-6 and 9 omegas. Both diets (GPS and GPB) increased the amounts of long-chain FAs. The GPS diet decreased saturated FA levels. We conclude that the dietary treatments GPS and GPB are a promising alternative to maintain meat quality standards throughout in real-world retail conditions. These treatments gave rise to an improvement in the nutritional value of the meat due to the more significant amounts of FAs that improve human health.
基金This research was funded by the Natural Science Foundation of Shandong Province of China(ZR2022MC144).
文摘Grapes,one of the oldest tree species globally,are rich in vitamins.However,environmental conditions such as low temperature and soil salinization significantly affect grape yield and quality.The glutamate receptor(GLR)family,comprising highly conserved ligand-gated ion channels,regulates plant growth and development in response to stress.In this study,11 members of the VvGLR gene family in grapes were identified using whole-genome sequence analysis.Bioinformatic methods were employed to analyze the basic physical and chemical properties,phylogenetic trees,conserved domains,motifs,expression patterns,and evolutionary relationships.Phylogenetic and collinear analyses revealed that the VvGLRs were divided into three subgroups,showing the high conservation of the grape GLR family.These members exhibited 2 glutamate receptor binding regions(GABAb and GluR)and 3-4 transmembrane regions(M1,M2,M3,and M4).Real-time quantitative PCR analysis demonstrated the sensitivity of all VvGLRs to low temperature and salt stress.Subsequent localization studies in Nicotiana tabacum verified that VvGLR3.1 and VvGLR3.2 proteins were located on the cell membrane and cell nucleus.Additionally,yeast transformation experiments confirmed the functionality of VvGLR3.1 and VvGLR3.2 in response to low temperature and salt stress.Thesefindings highlight the significant role of the GLR family,a highly conserved group of ion channels,in enhancing grape stress resistance.This study offers new insights into the grape GLR gene family,providing fundamental knowledge for further functional analysis and breeding of stress-resistant grapevines.
基金financially supported by the National Natural Science Foundation of China(32202560 and 32302470)the Program for Innovative Research Team(in Science and Technology)in University of Henan Province+6 种基金China(21IRTSTHN021)the Natural Science Foundation of HenanChina(232300421112)the Program for Science&Technology Innovation Talents in Universities of Henan ProvinceChina(21HASTIT035)the PhD Research Startup Foundation of Henan University of Science and TechnologyChina(13480068 and 13480067)。
文摘The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes(DEGs) by comparing the transcriptomes of folic acid(FA)-treated and water-treated(CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced(AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology(GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes,particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2(VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries.Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.
