Cold tolerance is one of the important traits for grapevine,especially in regions with extremely low temperatures in winter.Vitis amurensis is wild species in the Vitis genus with excellent cold hardiness compared wit...Cold tolerance is one of the important traits for grapevine,especially in regions with extremely low temperatures in winter.Vitis amurensis is wild species in the Vitis genus with excellent cold hardiness compared with Vitis vinifera.However,metabolites that contribute to the cold tolerance of V.amurensis remain unknown.Here,the metabolomics of buds from V.amurensis‘Zuoshan-1'during cold acclimation(CA)were identified,and cold-sensitive cultivar(V.vinifera‘Jingzaojing')was used as the control.The buds were collected in October,November,and December in 2016 and 2018.The cold hardiness of the buds increased during CA in the two grapevines.However,browning was observed only in V.vinifera buds at temperature below-10℃.Among detected metabolites from buds,443 metabolites were overlapped between two years.Forty-four and thirty differentially accumulated metabolites(DAMs)were identified in V.amurensis and V.vinifera,respectively.Ten DAMs including monoacylglycerol(MAG,18:2)isomer 1,trehalose 6-phosphate,and D-glucose showed identical variations in the two grapevines,indicating conserved CA responses within the Vitis genus.Eighteen DAMs exhibited higher accumulation in V.amurensis than in V.vinifera.Maltotetraose,D-glucoronic acid,L-aspartic acid,azelaic acid,and 4-hydroxybenzoic acid were reported to accumulate during CA in other plants.Enhanced cold tolerance was detected in grapevine leaves with exogenous 5 mmol L^(-1)L-aspartic acid and 1%proanthocyanidins.Potential contributions of other DAMs found in V.amurensis such as Cyanidin 3-O-glucoside need to be further elucidated.Thus,eighteen metabolites accumulated in V.amurensis can be used for practical application in improvement of cold resistance in grapevine.Our findings provide new insights into understanding the cold hardiness of V.amurensis.展开更多
The efficacy of the CRISPR/Cas9 system in grapevine(Vitis vinifera L.)has been documented,but the optimization of this system,as well as CRISPR/Cas9-mediated multiplex genome editing,has not been explored in this spec...The efficacy of the CRISPR/Cas9 system in grapevine(Vitis vinifera L.)has been documented,but the optimization of this system,as well as CRISPR/Cas9-mediated multiplex genome editing,has not been explored in this species.Herein,we identified four VvU3 and VvU6 promoters and two ubiquitin(UBQ)promoters in grapevine and demonstrated that the use of the identified VvU3/U6 and UBQ2 promoters could significantly increase the editing efficiency in grape by improving the expression of sgRNA and Cas9,respectively.Furthermore,we conducted multiplex genome editing using the optimized CRISPR/Cas9 vector that contained the conventional multiple sgRNA expression cassettes or the polycistronic tRNA-sgRNA cassette(PTG)by targeting the sugar-related tonoplastic monosaccharide transporter(TMT)family members TMT1 and TMT2,and the overall editing efficiencies were higher than 10%.The simultaneous editing of TMT1 and TMT2 resulted in reduced sugar levels,which indicated the role of these two genes in sugar accumulation in grapes.Moreover,the activities of the VvU3,VvU6,and UBQ2 promoters in tobacco genome editing were demonstrated by editing the phytoene desaturase(PDS)gene in Nicotiana benthamiana leaves.Our study provides materials for the optimization of the CRISPR/Cas9 system.To our knowledge,our simultaneous editing of the grape TMT family genes TMT1 and TMT2 constitutes the first example of multiplex genome editing in grape.The multiplex editing systems described in this manuscript expand the toolbox of grape genome editing,which would facilitate basic research and molecular breeding in grapevine.展开更多
‘Concord’,the most well-known juice grape with a parentage of the North American grape species Vitis labrusca L.,possesses a special‘foxy’aroma predominantly resulted from the accumulation of methyl anthranilate(M...‘Concord’,the most well-known juice grape with a parentage of the North American grape species Vitis labrusca L.,possesses a special‘foxy’aroma predominantly resulted from the accumulation of methyl anthranilate(MA)in berries.This aroma,however,is often perceived as an undesirable attribute by wine consumers and rarely noticeable in the common table and wine grape species V.vinifera.Here we discovered homology-induced promoter indels as a major genetic mechanism for species-specific regulation of a key‘foxy’aroma gene,anthraniloyl-CoA:methanol acyltransferase(AMAT),that is responsible for MA biosynthesis.We found the absence of a 426-bp and/or a 42-bp sequence in AMAT promoters highly associated with high levels of AMAT expression and MA accumulation in‘Concord’and other V.labrusca-derived grapes.These promoter variants,all with direct and inverted repeats,were further confirmed in more than 1,300 Vitis germplasm.Moreover,functional impact of these indels was validated in transgenic Arabidopsis.Superimposed on the promoter regulation,large structural changes including exonic insertion of a retrotransposon were present at the AMAT locus in some V.vinifera grapes.Elucidation of the AMAT genetic regulation advances our understanding of the‘foxy’aroma trait and makes it genetically trackable and amenable in grapevine breeding.展开更多
Grape is a widely cultivated crop with high economic value.Most cultivars derived from mild or cooler climates may not withstand increasing heat stress.Therefore,dissecting the mechanisms of heat tolerance in grapes i...Grape is a widely cultivated crop with high economic value.Most cultivars derived from mild or cooler climates may not withstand increasing heat stress.Therefore,dissecting the mechanisms of heat tolerance in grapes is of particular significance.Here,we performed comparative transcriptome analysis of Vitis davidii‘Tangwei’(heat tolerant)and Vitis vinifera‘Jingxiu’(heat sensitive)grapevines after exposure to 25°C,40°C,or 45°C for 2 h.More differentially expressed genes(DEGs)were detected in‘Tangwei’than in‘Jingxiu’in response to heat stress,and the number of DEGs increased with increasing treatment temperatures.We identified a class B Heat Shock Factor,HSFB1,which was significantly upregulated in‘Tangwei’,but not in‘Jingxiu’,at high temperature.VdHSFB1 from‘Tangwei’and VvHSFB1 from‘Jingxiu’differ in only one amino acid,and both showed similar transcriptional repression activities.Overexpression and RNA interference of HSFB1 in grape indicated that HSFB1 positively regulates the heat tolerance.Moreover,the heat tolerance of HSFB1-overexpressing plants was positively correlated to HSFB1 expression level.The activity of the VdHSFB1 promoter is higher than that of VvHSFB1 under both normal and high temperatures.Promoter analysis showed that more TATA-box and AT∼TATA-box cis-elements are present in the VdHSFB1 promoter than the VvHSFB1 promoter.The promoter sequence variations between VdHSFB1 and VvHSFB1 likely determine the HSFB1 expression levels that inf luence heat tolerance of the two grape germplasms with contrasting thermotolerance.Collectively,we validated the role of HSFB1 in heat tolerance,and the knowledge gained will advance our ability to breed heat-tolerant grape cultivars.展开更多
Histone H3 lysine 27 trimethylation(H3K27me3) is a histone modification associated with transcriptional repression. However, insights into the genome-wide pattern of H3K27me3 in grapevines are limited. Here, anti-H3K2...Histone H3 lysine 27 trimethylation(H3K27me3) is a histone modification associated with transcriptional repression. However, insights into the genome-wide pattern of H3K27me3 in grapevines are limited. Here, anti-H3K27 chromatin immunoprecipitation(ChIP), high-throughput sequencing, and transcriptome analysis were performed using leaves of Vitis amurensis. The leaves were treated at 4°C for 2 h and 24 h and used to investigate changes in H3K27me3 under chilling treatment. The results show that H3K27me3 is well-distributed both in gene regions(-50%) and in the intergenic region(-50%) in the grapevine genome(Vitis vinifera ‘Pinot Noir PN40024'). H3K27me3 was found to be localized in8 368 annotated gene regions in all detected samples(leaves at normal temperature and under chilling treatments) and mainly enriched in gene bodies with the adjacent promoter and downstream areas. The short-term chilling treatments(4°C for 2 h) induced 2 793 gains and 305losses in H3K27me3 modification. Subsequently, 97.3% of the alterations were restored to original levels after 24 h treatment. The ChIP-qPCR for five differential peaks showed similar results to the data for ChIP-seq, indicating that the chilling-induced H3K27me3 modification is reliable.Integrative analysis of transcriptome and ChIP-seq results showed that the expression of H3K27me3 target genes was significantly lower than those of non-target genes, indicating transcriptional repression of H3K27me3 in grapevine leaves. Furthermore, histone methylation alterations were detected in 82 genes and were related to either repression or activation of their expression during chilling stress. The findings provide the genome-wide H3K27me3 patterns in grapevines and shed light on uncovering its regulation in chilling stress responses.展开更多
A colchicine-induced autotetraploid grapevine exhibiting potentially valuable agronomic traits for grape production and breeding,including self-pruning,was identified.This study investigated DNA methylation variation ...A colchicine-induced autotetraploid grapevine exhibiting potentially valuable agronomic traits for grape production and breeding,including self-pruning,was identified.This study investigated DNA methylation variation and its role in gene expression during self-pruning in the autotetraploid grapevine.We used RNA-Seq to estimate differentially expressed genes between diploid and autotetraploid grapevine shoot tips.The genes showing increases in the autotetraploid were mainly related to stress response pathways,whereas those showing decreases in the autotetraploid were related to biological metabolism and biosynthesis.Whole-genome bisulfite sequencing was performed to produce single-base methylomes for the diploid and autotetraploid grapevines.Comparison between the methylomes revealed that they were conserved in CG and CHG contexts.In the autotetraploid grapevine,hypodifferentially methylated regions(DMRs)and hyper-DMRs in the gene body increased or decreased gene expression,respectively.Our results indicated that a hypo-DMR in the ACO1 gene body increased its expression and might promote self-pruning.This study reports that hypo-DMRs in the gene body increase gene expression in plants and reveals the mechanism underlying the changes in the modifications affecting gene expression during genome duplication.Overall,our results provide valuable information for understanding the relationships between DNA methylation,gene expression,and autotetraploid breeding in grape.展开更多
Anthocyanins are essential for the quality of perennial horticultural crops,such as grapes.In grapes,ELONGATED HYPOCOTYL 5(HY5)and MYBA1 are two critical transcription factors that regulate anthocyanin biosynthesis.Ou...Anthocyanins are essential for the quality of perennial horticultural crops,such as grapes.In grapes,ELONGATED HYPOCOTYL 5(HY5)and MYBA1 are two critical transcription factors that regulate anthocyanin biosynthesis.Our previous work has shown that Vitis vinifera B-box protein 44(VvBBX44)inhibits anthocyanin synthesis and represses VvHY5 expression in grape calli.However,the regulatory mechanism underlying this regulation was unclear.In this study,we found that loss of VvBBX44 function resulted in increased anthocyanin accumulation in grapevine callus.VvBBX44 directly represses VvMYBA1,which activates VvBBX44.VvMYBA1,but not VvBBX44,directly modulates the expression of grape UDP flavonoid 3-O-glucosyltransferase(VvUFGT).We demonstrated that VvBBX44 represses the transcriptional activation of VvUFGT and VvBBX44 induced by VvMYBA1.However,VvBBX44 and VvMYBA1 did not physically interact in yeast.The application of exogenous anthocyanin stimulated VvBBX44 expression in grapevine suspension cells and tobacco leaves.These findings suggest that VvBBX44 and VvMYBA1 form a transcriptional feedback loop to prevent overaccumulation of anthocyanin and reduce metabolic costs.Our work sheds light on the complex regulatory network that controls anthocyanin biosynthesis in grapevine.展开更多
The quality of planting materials is the foundation for productivity,longevity,and berry quality of perennial grapevines with a long lifespan.Manipulating the nursery light spectrum may speed up the production of heal...The quality of planting materials is the foundation for productivity,longevity,and berry quality of perennial grapevines with a long lifespan.Manipulating the nursery light spectrum may speed up the production of healthy and high-quality planting vines but the underlying mechanisms remain elusive.Herein,the effects of different monochromatic lights(green,blue,and red)on grapevine growth,leaf photosynthesis,whole-plant carbon allocation,and transcriptome reprograming were investigated with white light as control.Results showed that blue and red lights were favorable for plantlet growth in comparison with white light.Blue light repressed excessive growth,significantly increased the maximum net photosynthetic rate(Pn)of leaves by 39.58%and leaf specific weight by 38.29%.Red light increased the dry weight of the stem by 53.60%,the starch content of the leaf by 53.63%,and the sucrose content of the stem by 230%.Green light reduced all photosynthetic indexes of the grape plantlet.Photosynthetic photon flux density(PPFD)/Ci–Pn curves indicated that blue light affected photosynthetic rate depending on the light intensity and CO2 concentration.RNA-seq analysis of different organs(leaf,stem,and root)revealed a systematic transcriptome remodeling and VvCOP1(CONSTITUTIVELY PHOTOMORPHOGENIC 1),VvHY5(ELONGATED HYPOCOTYL5),VvHYH(HY5 HOMOLOG),VvELIP(early light-induced protein)and VvPIF3(PHYTOCHROME INTERACTING FACTOR 3)may play important roles in this shoot-to-root signaling.Furthermore,the correlation network between differential expression genes and physiological traits indicated that VvpsbS(photosystem II subunit S),Vvpsb28(photosystem II subunit 28),VvHYH,VvSUS4(sucrose synthase 4),and VvALDA(fructose-bisphosphate aldolase)were pertinent candidate genes in responses to different light qualities.Our results provide a foundation for optimizing the light recipe of grape plantlets and strengthen the understanding of light signaling and carbon metabolism under different monochromatic lights.展开更多
The widely appreciated muscat flavor of grapes and wine is mainly attributable to the monoterpenes that accumulate in ripe grape berries.To identify quantitative trait loci(QTL)for grape berry monoterpene content,an F...The widely appreciated muscat flavor of grapes and wine is mainly attributable to the monoterpenes that accumulate in ripe grape berries.To identify quantitative trait loci(QTL)for grape berry monoterpene content,an F1 mapping population was constructed by a cross between two grapevine genotypes,one with neutral aroma berries(cv.‘Beifeng’)and the other with a pronounced muscat aroma(elite Vitis vinifera line‘3–34’).A high-density genetic linkage map spanning 1563.7 cM was constructed using 3332 SNP markers that were assigned to 19 linkage groups.Monoterpenes were extracted from the berry of the F1 progeny,then identified and quantified by gas chromatography–mass spectrometry.Twelve stable QTLs associated with the amounts of 11 monoterpenes in berries were thus identified.In parallel,the levels of RNA in berries from 34 diverse cultivars were estimated by RNA sequencing and compared to the monoterpene content of the berries.The expression of five genes mapping to stable QTLs correlated well with the monoterpene content of berries.These genes,including the basic leucine zipper VvbZIP61 gene on chromosome 12,are therefore considered as potentially being involved in monoterpene metabolism.Overexpression of VvbZIP61 in Vitis amurensis callus through Agrobacterium-mediated transformation significantly increased the accumulation of several monoterpenes in the callus,including nerol,linalool,geranial,geraniol,β-myrcene,and D-limonene.It is hypothesized that VvbZIP61 expression acts to increase muscat flavor in grapes.These results advance our understanding of the genetic control of monoterpene biosynthesis in grapes and provide important information for the marker-assisted selection of aroma compounds in grape breeding.展开更多
基金supported by the National Key Research and Development Program of China(Grant No.2018YFD1000300)the National Natural Science Foundation of China(Grant Nos.32272691 and 32025032)+1 种基金the Grape Breeding Project of Ningxia(Grant No.NXNYYZ202101-04)the Major Program of Technological Innovation in Hubei Province(Grant No.2022BBA0022)。
文摘Cold tolerance is one of the important traits for grapevine,especially in regions with extremely low temperatures in winter.Vitis amurensis is wild species in the Vitis genus with excellent cold hardiness compared with Vitis vinifera.However,metabolites that contribute to the cold tolerance of V.amurensis remain unknown.Here,the metabolomics of buds from V.amurensis‘Zuoshan-1'during cold acclimation(CA)were identified,and cold-sensitive cultivar(V.vinifera‘Jingzaojing')was used as the control.The buds were collected in October,November,and December in 2016 and 2018.The cold hardiness of the buds increased during CA in the two grapevines.However,browning was observed only in V.vinifera buds at temperature below-10℃.Among detected metabolites from buds,443 metabolites were overlapped between two years.Forty-four and thirty differentially accumulated metabolites(DAMs)were identified in V.amurensis and V.vinifera,respectively.Ten DAMs including monoacylglycerol(MAG,18:2)isomer 1,trehalose 6-phosphate,and D-glucose showed identical variations in the two grapevines,indicating conserved CA responses within the Vitis genus.Eighteen DAMs exhibited higher accumulation in V.amurensis than in V.vinifera.Maltotetraose,D-glucoronic acid,L-aspartic acid,azelaic acid,and 4-hydroxybenzoic acid were reported to accumulate during CA in other plants.Enhanced cold tolerance was detected in grapevine leaves with exogenous 5 mmol L^(-1)L-aspartic acid and 1%proanthocyanidins.Potential contributions of other DAMs found in V.amurensis such as Cyanidin 3-O-glucoside need to be further elucidated.Thus,eighteen metabolites accumulated in V.amurensis can be used for practical application in improvement of cold resistance in grapevine.Our findings provide new insights into understanding the cold hardiness of V.amurensis.
基金the National Natural Science Foundation of China(31772266 and 32001994)the Bureau of International Cooperation of the Chinese Academy of Sciences(151111KYSB20170032).
文摘The efficacy of the CRISPR/Cas9 system in grapevine(Vitis vinifera L.)has been documented,but the optimization of this system,as well as CRISPR/Cas9-mediated multiplex genome editing,has not been explored in this species.Herein,we identified four VvU3 and VvU6 promoters and two ubiquitin(UBQ)promoters in grapevine and demonstrated that the use of the identified VvU3/U6 and UBQ2 promoters could significantly increase the editing efficiency in grape by improving the expression of sgRNA and Cas9,respectively.Furthermore,we conducted multiplex genome editing using the optimized CRISPR/Cas9 vector that contained the conventional multiple sgRNA expression cassettes or the polycistronic tRNA-sgRNA cassette(PTG)by targeting the sugar-related tonoplastic monosaccharide transporter(TMT)family members TMT1 and TMT2,and the overall editing efficiencies were higher than 10%.The simultaneous editing of TMT1 and TMT2 resulted in reduced sugar levels,which indicated the role of these two genes in sugar accumulation in grapes.Moreover,the activities of the VvU3,VvU6,and UBQ2 promoters in tobacco genome editing were demonstrated by editing the phytoene desaturase(PDS)gene in Nicotiana benthamiana leaves.Our study provides materials for the optimization of the CRISPR/Cas9 system.To our knowledge,our simultaneous editing of the grape TMT family genes TMT1 and TMT2 constitutes the first example of multiplex genome editing in grape.The multiplex editing systems described in this manuscript expand the toolbox of grape genome editing,which would facilitate basic research and molecular breeding in grapevine.
基金supported by the USDA Agricultural Research Service.
文摘‘Concord’,the most well-known juice grape with a parentage of the North American grape species Vitis labrusca L.,possesses a special‘foxy’aroma predominantly resulted from the accumulation of methyl anthranilate(MA)in berries.This aroma,however,is often perceived as an undesirable attribute by wine consumers and rarely noticeable in the common table and wine grape species V.vinifera.Here we discovered homology-induced promoter indels as a major genetic mechanism for species-specific regulation of a key‘foxy’aroma gene,anthraniloyl-CoA:methanol acyltransferase(AMAT),that is responsible for MA biosynthesis.We found the absence of a 426-bp and/or a 42-bp sequence in AMAT promoters highly associated with high levels of AMAT expression and MA accumulation in‘Concord’and other V.labrusca-derived grapes.These promoter variants,all with direct and inverted repeats,were further confirmed in more than 1,300 Vitis germplasm.Moreover,functional impact of these indels was validated in transgenic Arabidopsis.Superimposed on the promoter regulation,large structural changes including exonic insertion of a retrotransposon were present at the AMAT locus in some V.vinifera grapes.Elucidation of the AMAT genetic regulation advances our understanding of the‘foxy’aroma trait and makes it genetically trackable and amenable in grapevine breeding.
基金supported by the National Natural Science Foundation of China(Grant no.U21A20227)the National Key Research and Development Program of China(Grant no.2018YFD1000300)the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant no.XDA23080602).
文摘Grape is a widely cultivated crop with high economic value.Most cultivars derived from mild or cooler climates may not withstand increasing heat stress.Therefore,dissecting the mechanisms of heat tolerance in grapes is of particular significance.Here,we performed comparative transcriptome analysis of Vitis davidii‘Tangwei’(heat tolerant)and Vitis vinifera‘Jingxiu’(heat sensitive)grapevines after exposure to 25°C,40°C,or 45°C for 2 h.More differentially expressed genes(DEGs)were detected in‘Tangwei’than in‘Jingxiu’in response to heat stress,and the number of DEGs increased with increasing treatment temperatures.We identified a class B Heat Shock Factor,HSFB1,which was significantly upregulated in‘Tangwei’,but not in‘Jingxiu’,at high temperature.VdHSFB1 from‘Tangwei’and VvHSFB1 from‘Jingxiu’differ in only one amino acid,and both showed similar transcriptional repression activities.Overexpression and RNA interference of HSFB1 in grape indicated that HSFB1 positively regulates the heat tolerance.Moreover,the heat tolerance of HSFB1-overexpressing plants was positively correlated to HSFB1 expression level.The activity of the VdHSFB1 promoter is higher than that of VvHSFB1 under both normal and high temperatures.Promoter analysis showed that more TATA-box and AT∼TATA-box cis-elements are present in the VdHSFB1 promoter than the VvHSFB1 promoter.The promoter sequence variations between VdHSFB1 and VvHSFB1 likely determine the HSFB1 expression levels that inf luence heat tolerance of the two grape germplasms with contrasting thermotolerance.Collectively,we validated the role of HSFB1 in heat tolerance,and the knowledge gained will advance our ability to breed heat-tolerant grape cultivars.
基金supported by the National Key Research and Development Program of China (Grant No. 2018YFD1000300)the National Natural Science Foundation of China (Grant No. 32025032)+1 种基金the Grape Breeding Project of Ningxia (Grant No. NXNYYZ202101-04)Major Program of Technological Innovation in Hubei Province (Grant No. 2019ABA093).
文摘Histone H3 lysine 27 trimethylation(H3K27me3) is a histone modification associated with transcriptional repression. However, insights into the genome-wide pattern of H3K27me3 in grapevines are limited. Here, anti-H3K27 chromatin immunoprecipitation(ChIP), high-throughput sequencing, and transcriptome analysis were performed using leaves of Vitis amurensis. The leaves were treated at 4°C for 2 h and 24 h and used to investigate changes in H3K27me3 under chilling treatment. The results show that H3K27me3 is well-distributed both in gene regions(-50%) and in the intergenic region(-50%) in the grapevine genome(Vitis vinifera ‘Pinot Noir PN40024'). H3K27me3 was found to be localized in8 368 annotated gene regions in all detected samples(leaves at normal temperature and under chilling treatments) and mainly enriched in gene bodies with the adjacent promoter and downstream areas. The short-term chilling treatments(4°C for 2 h) induced 2 793 gains and 305losses in H3K27me3 modification. Subsequently, 97.3% of the alterations were restored to original levels after 24 h treatment. The ChIP-qPCR for five differential peaks showed similar results to the data for ChIP-seq, indicating that the chilling-induced H3K27me3 modification is reliable.Integrative analysis of transcriptome and ChIP-seq results showed that the expression of H3K27me3 target genes was significantly lower than those of non-target genes, indicating transcriptional repression of H3K27me3 in grapevine leaves. Furthermore, histone methylation alterations were detected in 82 genes and were related to either repression or activation of their expression during chilling stress. The findings provide the genome-wide H3K27me3 patterns in grapevines and shed light on uncovering its regulation in chilling stress responses.
基金supported by the National Key Research and Development Program of China(2018YFD1000105)the grants from the National Key Research and Development Program of China,the National Science Foundation of China(31772266)+1 种基金Agricultural Breeding Project of Ningxia Hui Autonomous Region(NXNYYZ20150203)STS Project of the Chinese Academy of Sciences(KFJ-STS-ZDTP-025).
文摘A colchicine-induced autotetraploid grapevine exhibiting potentially valuable agronomic traits for grape production and breeding,including self-pruning,was identified.This study investigated DNA methylation variation and its role in gene expression during self-pruning in the autotetraploid grapevine.We used RNA-Seq to estimate differentially expressed genes between diploid and autotetraploid grapevine shoot tips.The genes showing increases in the autotetraploid were mainly related to stress response pathways,whereas those showing decreases in the autotetraploid were related to biological metabolism and biosynthesis.Whole-genome bisulfite sequencing was performed to produce single-base methylomes for the diploid and autotetraploid grapevines.Comparison between the methylomes revealed that they were conserved in CG and CHG contexts.In the autotetraploid grapevine,hypodifferentially methylated regions(DMRs)and hyper-DMRs in the gene body increased or decreased gene expression,respectively.Our results indicated that a hypo-DMR in the ACO1 gene body increased its expression and might promote self-pruning.This study reports that hypo-DMRs in the gene body increase gene expression in plants and reveals the mechanism underlying the changes in the modifications affecting gene expression during genome duplication.Overall,our results provide valuable information for understanding the relationships between DNA methylation,gene expression,and autotetraploid breeding in grape.
基金We thank professor Yu-Jin Hao(College of Horticulture Science and Engineering,Shandong Agricultural University)for providing the plasmid for the EMSA experiment.All data generated and analyzed in this study are shown in the article or attached as supplementary data.All the materials used in the study are available upon reasonable request fromthe corresponding author.This work was supported by the National Natural Science Foundation of China(Grant no.U21A20227)the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant no.XDA23080602).Research conducted as part of the LIA INNOGRAPE International Associated Laboratory.
文摘Anthocyanins are essential for the quality of perennial horticultural crops,such as grapes.In grapes,ELONGATED HYPOCOTYL 5(HY5)and MYBA1 are two critical transcription factors that regulate anthocyanin biosynthesis.Our previous work has shown that Vitis vinifera B-box protein 44(VvBBX44)inhibits anthocyanin synthesis and represses VvHY5 expression in grape calli.However,the regulatory mechanism underlying this regulation was unclear.In this study,we found that loss of VvBBX44 function resulted in increased anthocyanin accumulation in grapevine callus.VvBBX44 directly represses VvMYBA1,which activates VvBBX44.VvMYBA1,but not VvBBX44,directly modulates the expression of grape UDP flavonoid 3-O-glucosyltransferase(VvUFGT).We demonstrated that VvBBX44 represses the transcriptional activation of VvUFGT and VvBBX44 induced by VvMYBA1.However,VvBBX44 and VvMYBA1 did not physically interact in yeast.The application of exogenous anthocyanin stimulated VvBBX44 expression in grapevine suspension cells and tobacco leaves.These findings suggest that VvBBX44 and VvMYBA1 form a transcriptional feedback loop to prevent overaccumulation of anthocyanin and reduce metabolic costs.Our work sheds light on the complex regulatory network that controls anthocyanin biosynthesis in grapevine.
基金supported by National Natural Science Foundation of China(U20A2041)the National Key R&D Program of China(2019YFD1000100,2021YFE0109500)+1 种基金the Agricultural Breeding Project of Ningxia Hui Autonomous Region(NXNYYZ202101)the CAS Youth Interdisciplinary Team(JCTD-2022-06).
文摘The quality of planting materials is the foundation for productivity,longevity,and berry quality of perennial grapevines with a long lifespan.Manipulating the nursery light spectrum may speed up the production of healthy and high-quality planting vines but the underlying mechanisms remain elusive.Herein,the effects of different monochromatic lights(green,blue,and red)on grapevine growth,leaf photosynthesis,whole-plant carbon allocation,and transcriptome reprograming were investigated with white light as control.Results showed that blue and red lights were favorable for plantlet growth in comparison with white light.Blue light repressed excessive growth,significantly increased the maximum net photosynthetic rate(Pn)of leaves by 39.58%and leaf specific weight by 38.29%.Red light increased the dry weight of the stem by 53.60%,the starch content of the leaf by 53.63%,and the sucrose content of the stem by 230%.Green light reduced all photosynthetic indexes of the grape plantlet.Photosynthetic photon flux density(PPFD)/Ci–Pn curves indicated that blue light affected photosynthetic rate depending on the light intensity and CO2 concentration.RNA-seq analysis of different organs(leaf,stem,and root)revealed a systematic transcriptome remodeling and VvCOP1(CONSTITUTIVELY PHOTOMORPHOGENIC 1),VvHY5(ELONGATED HYPOCOTYL5),VvHYH(HY5 HOMOLOG),VvELIP(early light-induced protein)and VvPIF3(PHYTOCHROME INTERACTING FACTOR 3)may play important roles in this shoot-to-root signaling.Furthermore,the correlation network between differential expression genes and physiological traits indicated that VvpsbS(photosystem II subunit S),Vvpsb28(photosystem II subunit 28),VvHYH,VvSUS4(sucrose synthase 4),and VvALDA(fructose-bisphosphate aldolase)were pertinent candidate genes in responses to different light qualities.Our results provide a foundation for optimizing the light recipe of grape plantlets and strengthen the understanding of light signaling and carbon metabolism under different monochromatic lights.
基金supported by the National Key Research and Development Program(2018YFD1000205)the National Science Foundation of China(31372028)+1 种基金the Agricultural Breeding Project of Ningxia Hui Autonomous Region NXNYYZ202101the Alliance of International Science Organizations(ANSO-CR-PP-2020-04).
文摘The widely appreciated muscat flavor of grapes and wine is mainly attributable to the monoterpenes that accumulate in ripe grape berries.To identify quantitative trait loci(QTL)for grape berry monoterpene content,an F1 mapping population was constructed by a cross between two grapevine genotypes,one with neutral aroma berries(cv.‘Beifeng’)and the other with a pronounced muscat aroma(elite Vitis vinifera line‘3–34’).A high-density genetic linkage map spanning 1563.7 cM was constructed using 3332 SNP markers that were assigned to 19 linkage groups.Monoterpenes were extracted from the berry of the F1 progeny,then identified and quantified by gas chromatography–mass spectrometry.Twelve stable QTLs associated with the amounts of 11 monoterpenes in berries were thus identified.In parallel,the levels of RNA in berries from 34 diverse cultivars were estimated by RNA sequencing and compared to the monoterpene content of the berries.The expression of five genes mapping to stable QTLs correlated well with the monoterpene content of berries.These genes,including the basic leucine zipper VvbZIP61 gene on chromosome 12,are therefore considered as potentially being involved in monoterpene metabolism.Overexpression of VvbZIP61 in Vitis amurensis callus through Agrobacterium-mediated transformation significantly increased the accumulation of several monoterpenes in the callus,including nerol,linalool,geranial,geraniol,β-myrcene,and D-limonene.It is hypothesized that VvbZIP61 expression acts to increase muscat flavor in grapes.These results advance our understanding of the genetic control of monoterpene biosynthesis in grapes and provide important information for the marker-assisted selection of aroma compounds in grape breeding.
