Virus-induced gene silencing(VIGS)and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas)systems are effective technologies for rapid and accurate gene function verification...Virus-induced gene silencing(VIGS)and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas)systems are effective technologies for rapid and accurate gene function verification in modern plant biotechnology.However,the investigation of gene silencing and editing in radish remains limited.In this study,a bleaching phenotype was generated through the knockdown of RsPDS using tobacco rattle virus(TRV)-and turnip yellow mosaic virus(TYMV)-mediated gene silencing vectors.The TYMV-mediated gene silencing efficiency was higher than the TRV-based VIGS system in radish.The expression level of RsPDS was significantly inhibited using VIGS in'NAU-067'radish leaves.The rootless seedlings of‘NAU-067'were infected with Agrobacterium rhizogenes using the 2300GN-Ubi-RsPDS-Cas9 vector with two target sequences.Nine adventitious roots were blue with GUs staining,and four of these adventitious roots were edited at target sequence 1 of the RsPDS gene as indicated by Sanger sequencing.Furthermore,albino lines were generated with A.tumefaciens-mediated transformation of radish cotyledons.Five base substitutions and three base deletions occurred at target sequence 2 in Line 1,and three base insertions and three base substitutions occurred at target sequence 1 in Line 2.This study shows that VIGS and CRISPR/Cas9 techniques can be employed to precisely verify the biological functions of genes in radish,which will facilitate the genetic improvement of vital horticultural traits in radish breeding programs.展开更多
The mechanical characteristics and acoustic behavior of rock masses are greatly influenced by stochastic joints.In this study,numerical models of rock masses incorporating intermittent joints with different numbers an...The mechanical characteristics and acoustic behavior of rock masses are greatly influenced by stochastic joints.In this study,numerical models of rock masses incorporating intermittent joints with different numbers and dip angles were produced using the finite element method(FEM)with the intrinsic cohesive zone model(ICZM).Then,the uniaxial compressive and wave propagation simulations were performed.The results indicate that the joint number and dip angle can affect the mechanical and acoustic properties of the models.The uniaxial compressive strength(UCS)and wave velocity of rock masses decrease monotonically as the joint number increases.However,the wave velocity grows monotonically as the joint dip angle increases.When the joint dip angle is 45°–60°,the UCS of the rock mass is lower than that of other dip angles.The wave velocity parallel to the joints is greater than that perpendicular to the joints.When the dip angle of joints remains unchanged,the UCS and wave velocity are positively related.When the joint dip angle increases,the variation amplitude of the UCS regarding the wave velocity increases.To reveal the effect of the joint distribution on the velocity,a theoretical model was also proposed.According to the theoretical wave velocity,the change in wave velocity of models with various joint numbers and dip angles was consistent with the simulation results.Furthermore,a theoretical indicator(i.e.fabric tensor)was adopted to analyze the variation of the wave velocity and UCS.展开更多
The mechanical characteristics and failure behavior of rocks containing flaws or discontinuities have received wide attention in the field of rock mechanics.When external loads are applied to rock materials,stress-ind...The mechanical characteristics and failure behavior of rocks containing flaws or discontinuities have received wide attention in the field of rock mechanics.When external loads are applied to rock materials,stress-induced cracks would initiate and propagate from the flaws,ultimately leading to the irreversible failure of rocks.To investigate the cracking behavior and the effect of flaw geometries on the mechanical properties of rock materials,a series of samples containing one,two and multiple flaws have been widely investigated in the laboratory.In this paper,the experimental results for pre-cracked rocks under quasistatic compression were systematically reviewed.The progressive failure process of intact rocks is briefly described to reveal the background for experiments on samples with flaws.Then,the nondestructive measurement techniques utilized in experiments,such as acoustic emission(AE),X-ray computed tomography(CT),and digital image correlation(DIC),are summarized.The mechanical characteristics of rocks with different flaw geometries and under different loading conditions,including the geometry of pre-existing flaws,flaw filling condition and confining pressure,are discussed.Furthermore,the cracking process is evaluated from the perspective of crack initiation,coalescence,and failure patterns.展开更多
Radish(Raphanus sativus L.)taproot contains high concentrations of flavonoids,including anthocyanins(ATCs),in redskinned genotypes.However,little information on the genetic regulation of ATC biosynthesis in radish is ...Radish(Raphanus sativus L.)taproot contains high concentrations of flavonoids,including anthocyanins(ATCs),in redskinned genotypes.However,little information on the genetic regulation of ATC biosynthesis in radish is available.A genome-wide association study of radish red skin color was conducted using whole-genome sequencing data derived from 179 radish genotypes.The R2R3-MYB transcription factor production of anthocyanin pigment 2(PAP2)gene was found in the region associated with a leading SNP located on chromosome 2.The amino acid sequence encoded by the RsPAP2 gene was different from those of the other published RsMYB genes responsible for the red skin color of radish.The overexpression of the RsPAP2 gene resulted in ATC accumulation in Arabidopsis and radish,which was accompanied by the upregulation of several ATC-related structural genes.RsPAP2 was found to bind the RsUFGT and RsTT8 promoters,as shown by a dual-luciferase reporter system and a yeast one-hybrid assay.The promoter activities of the RsANS,RsCHI,RsPAL,and RsUFGT genes could be strongly activated by coinfiltration with RsPAP2 and RsTT8.These findings showed the effectiveness of GWAS in identifying candidate genes in radish and demonstrated that RsPAP2 could(either directly or together with its cofactor RsTT8)regulate the transcript levels of ATC-related genes to promote ATC biosynthesis,facilitating the genetic enhancement of ATC contents and other related traits in radish.展开更多
Melatonin(MT)is a tryptophan-derived natural product that plays a vital role in plant response to abiotic stresses,including heavy metals(HMs).However,it remains elusive how exogenous MT mediates lead(Pb)accumulation ...Melatonin(MT)is a tryptophan-derived natural product that plays a vital role in plant response to abiotic stresses,including heavy metals(HMs).However,it remains elusive how exogenous MT mediates lead(Pb)accumulation and detoxification at the methylation and transcriptional levels in radish.In this study,decreased Pb accumulation and increased antioxidant enzyme activity were detected under MT treatment in radish.Single-base resolution maps of DNA methylation under Pb stress(Pb200)and Pb plus MT treatment(Pb_50MT)were first generated.The genome-wide methylation level was increased under Pb stress,while an overall loss of DNA methylation was observed under MT treatment.The differentially methylated region(DMR)-associated genes between Pb_50MT and Pb200 were uniquely enriched in ion binding terms,including cation binding,iron ion binding,and transition metal ion binding.Hyper-DMRs between Pb200 and Control exhibited a decreasing trend of methylation under Pb_50MT treatment.A few critical upregulated antioxidant genes(e.g.,RsAPX2,RsPOD52 and RsGST)exhibited decreased methylation levels under MT treatment,which enabled the radish plants to scavenge lead-induced reactive oxygen species(ROS)and decrease oxidative stress.Notably,several MT-induced HM transporter genes with low methylation(e.g.,RsABCF5,RsYSL7 and RsHMT)and transcription factors(e.g.,RsWRKY41 and RsMYB2)were involved in reducing Pb accumulation in radish roots.These findings could facilitate comprehensive elucidation of the molecular mechanism underlying MT-mediated Pb accumulation and detoxification in radish and other root vegetable crops.展开更多
The fleshy taproot of radish is an important storage organ determining its yield and quality.Taproot thickening is a complex developmental process in radish.However,the molecular mechanisms governing this process rema...The fleshy taproot of radish is an important storage organ determining its yield and quality.Taproot thickening is a complex developmental process in radish.However,the molecular mechanisms governing this process remain unclear at the proteome level.In this study,a comparative proteomic analysis was performed to analyze the proteome changes at three developmental stages of taproot thickening using iTRAQ approach.In total,1862 differentially expressed proteins(DEPs)were identified from 6342 high-confidence proteins,among which 256 up-regulated proteins displayed overlapped accumulation in S1(pre-cortex splitting stage)vs.S2(cortex splitting stage)and S1 vs.S3(expanding stage)pairs,whereas 122 up-regulated proteins displayed overlapped accumulation in S1 vs.S3 and S2 vs.S3 pairs.Gene Ontology(GO)and pathway enrichment analysis showed that these DEPs were mainly involved in several processes such as“starch and sucrose metabolism”,“plant hormone signal transduction”,and“biosynthesis of secondary metabolites”.A high concordance existed between iTRAQ and RT-qPCR at the mRNA expression levels.Furthermore,association analysis showed that 187,181,and 96 DEPs were matched with their corresponding differentially expressed genes(DEGs)in S1 vs.S2,S1 vs.S3,and S2 vs.S3 comparison,respectively.Notably,several functional proteins including cell division cycle 5-like protein(CDC5),expansin B1(EXPB1),and xyloglucan endotransglucosylase/hydrolase protein 24(XTH24)were responsible for cell division and expansion during radish taproot thickening process.These results could facilitate a better understanding of the molecular mechanism underlying taproot thickening,and provide valuable information for the identification of critical genes/proteins responsible for taproot thickening in root vegetable crops.展开更多
Previous studies have already shown that Raman spectroscopy can be used in the encoding of suspension array technology.However,almost all existing convolutional neural network-based decoding approaches rely on supervi...Previous studies have already shown that Raman spectroscopy can be used in the encoding of suspension array technology.However,almost all existing convolutional neural network-based decoding approaches rely on supervision with ground truth,and may not be well generalized to unseen datasets,which were collected under different experimental conditions,applying with the same coded material.In this study,we propose an improved model based on CyCADA,named as Detail constraint Cycle Domain Adaptive Model(DCDA).DCDA implements the clasification of unseen datasets through domain adaptation,adapts representations at the encode level with decoder-share,and enforces coding features while leveraging a feat loss.To improve detailed structural constraints,DCDA takes downsample connection and skips connection.Our model improves the poor generalization of existing models and saves the cost of the labeling process for unseen target datasets.Compared with other models,extensive experiments and ablation studies show the superiority of DCDA in terms of classification stability and generalization.The model proposed by the research achieves a classification with an accuracy of 100%when applied in datasets,in which the spectrum in the source domain is far less than the target domain.展开更多
The growth and development of taproots are inhibited by cold stress in radish(Raphanus sativus L.).Ethylene-responsive element binding factors(ERF)are key participators in the cold stress response and growth regulatio...The growth and development of taproots are inhibited by cold stress in radish(Raphanus sativus L.).Ethylene-responsive element binding factors(ERF)are key participators in the cold stress response and growth regulation of plants.However,the function of ERF genes in cold tolerance and root development in radish remains elusive.Here,we showed that the secondary growth of radish taproots was inhibited by cold stress.Comparative transcriptome analysis demonstrated that the RsERF40 gene is an important regulator of the cold stress response and root growth regulation.The cold tolerance of transgenic Arabidopsis plants overexpressing the RsERF40 gene was significantly improved.Overexpressing RsERF40 in the cold-sensitive radish genotype and silencing RsERF40 in the cold-tolerant radish genotype indicated that RsERF40 was beneficial for alleviating oxidative damage under cold stress in radish.Transgenic Arabidopsis seedlings showed an increase in the elongation and radial growth of dark-grown roots.RT-qPCR analysis showed that the expression of the cold-related genes(CORs)RsCOR78 and RsCOR413PM1 and the cell wall strengthening-related genes RsCESA6 and RsEXPB3 was upregulated in transgenic Arabidopsis seedlings.Yeast one-hybrid(Y1H)and dual-luciferase reporter assays(DLA)revealed that RsERF40 directly regulates RsCOR78,RsCOR413PM1,RsCESA6 and RsEXPB3 expression,illustrating that RsERF40 enhances cold tolerance and taproot growth by modulating osmotic adjustment and cell wall mechanical strength in radish.In this study,the RsERF40-regulon was firstly found to be a new cold response pathway independent of the CBF-COR pathway conferring cold stress tolerance with increasing radish taproot growth.These results provided novel insight into the molecular mechanism underlying cold stress response and would facilitate the genetic improvement of cold tolerance in radish and other root vegetable crops.展开更多
Heterosis,a crucial biological phenomenon,plays a vital role in determining the yield and quality of plants.Radish,an important root vegetable crop,exhibits notable heterosis in terms of root yield and quality.Neverth...Heterosis,a crucial biological phenomenon,plays a vital role in determining the yield and quality of plants.Radish,an important root vegetable crop,exhibits notable heterosis in terms of root yield and quality.Nevertheless,the molecular mechanism underlying the formation of heterosis in radish remains unclear.Herein,both the transcriptome and DNA methylome analyses were performed on the F_(1)hybrids and parental lines.Expression level dominance(ELD)genes and allele-specific expression(ASE)genes together significantly contribute to heterosis,primarily through energy metabolism and plant hormone signal transduction pathway.An increase in average methylation level in the F_(1)hybrids was observed compared to the parental lines.Interestingly,a negative correlation was found between the methylation level of differentially expressed genes(DEGs)in gene body regions and their expression levels in NAU-LB and the F_(1)hybrids.Moreover,the hybrids were more sensitive to the 5-azacytidine than their parents,and the root weight and total sugar content in the F_(1)hybrids were dramatically decreased compared to the control.Immunolocalization results indicated that the auxin content of the F_(1)hybrids were reduced under 5-azacytidine treatment.Proliferating cell nuclear antigen immunolocalization revealed significant inhibition of vascular cambium activity in both the hybrids and parental lines.Notably,the expression profiles of a few differentially methylated DEGs including RsSUS1,RsSUC2a,RsIAA7,and RsIAA18,were significantly increased in the root of hybrids compared to their parents,suggesting a potential role of DNA methylation in yield heterosis.Collectively,these findings would provide valuable insight into the molecular mechanism underlying taproot yield heterosis and facilitate the genetic improvement of taproot yield and quality in radish breeding programs.展开更多
基金This work was supported by Jiangsu Seed Industry Revitalization Project,China[JBGS(2021)071]Fundamental Research Funds for the Central Universities,China(YDZX2023019)+3 种基金the National Natural Science Foundation of China(32172579)the earmarked fund for Jiangsu Agricultural Industry Technology System,China[JATS(2023)421]the Jiangsu Postgraduate Scientific Research Innovation Plan,China(KYCX21_0610-2021)the Project Founded by the Priority Academic Program Development of Jiangsu Higher Education Institutions,China(PAPD).
文摘Virus-induced gene silencing(VIGS)and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas)systems are effective technologies for rapid and accurate gene function verification in modern plant biotechnology.However,the investigation of gene silencing and editing in radish remains limited.In this study,a bleaching phenotype was generated through the knockdown of RsPDS using tobacco rattle virus(TRV)-and turnip yellow mosaic virus(TYMV)-mediated gene silencing vectors.The TYMV-mediated gene silencing efficiency was higher than the TRV-based VIGS system in radish.The expression level of RsPDS was significantly inhibited using VIGS in'NAU-067'radish leaves.The rootless seedlings of‘NAU-067'were infected with Agrobacterium rhizogenes using the 2300GN-Ubi-RsPDS-Cas9 vector with two target sequences.Nine adventitious roots were blue with GUs staining,and four of these adventitious roots were edited at target sequence 1 of the RsPDS gene as indicated by Sanger sequencing.Furthermore,albino lines were generated with A.tumefaciens-mediated transformation of radish cotyledons.Five base substitutions and three base deletions occurred at target sequence 2 in Line 1,and three base insertions and three base substitutions occurred at target sequence 1 in Line 2.This study shows that VIGS and CRISPR/Cas9 techniques can be employed to precisely verify the biological functions of genes in radish,which will facilitate the genetic improvement of vital horticultural traits in radish breeding programs.
基金financial support from the National Key R&D Program of China(Grant No.2020YFA0711802).
文摘The mechanical characteristics and acoustic behavior of rock masses are greatly influenced by stochastic joints.In this study,numerical models of rock masses incorporating intermittent joints with different numbers and dip angles were produced using the finite element method(FEM)with the intrinsic cohesive zone model(ICZM).Then,the uniaxial compressive and wave propagation simulations were performed.The results indicate that the joint number and dip angle can affect the mechanical and acoustic properties of the models.The uniaxial compressive strength(UCS)and wave velocity of rock masses decrease monotonically as the joint number increases.However,the wave velocity grows monotonically as the joint dip angle increases.When the joint dip angle is 45°–60°,the UCS of the rock mass is lower than that of other dip angles.The wave velocity parallel to the joints is greater than that perpendicular to the joints.When the dip angle of joints remains unchanged,the UCS and wave velocity are positively related.When the joint dip angle increases,the variation amplitude of the UCS regarding the wave velocity increases.To reveal the effect of the joint distribution on the velocity,a theoretical model was also proposed.According to the theoretical wave velocity,the change in wave velocity of models with various joint numbers and dip angles was consistent with the simulation results.Furthermore,a theoretical indicator(i.e.fabric tensor)was adopted to analyze the variation of the wave velocity and UCS.
基金financial support from the National Key R&D Program of China(Grant No.2020YFA0711802)。
文摘The mechanical characteristics and failure behavior of rocks containing flaws or discontinuities have received wide attention in the field of rock mechanics.When external loads are applied to rock materials,stress-induced cracks would initiate and propagate from the flaws,ultimately leading to the irreversible failure of rocks.To investigate the cracking behavior and the effect of flaw geometries on the mechanical properties of rock materials,a series of samples containing one,two and multiple flaws have been widely investigated in the laboratory.In this paper,the experimental results for pre-cracked rocks under quasistatic compression were systematically reviewed.The progressive failure process of intact rocks is briefly described to reveal the background for experiments on samples with flaws.Then,the nondestructive measurement techniques utilized in experiments,such as acoustic emission(AE),X-ray computed tomography(CT),and digital image correlation(DIC),are summarized.The mechanical characteristics of rocks with different flaw geometries and under different loading conditions,including the geometry of pre-existing flaws,flaw filling condition and confining pressure,are discussed.Furthermore,the cracking process is evaluated from the perspective of crack initiation,coalescence,and failure patterns.
基金supported by grants from the Jiangsu Agricultural S&T Innovation Fund[CX(20)3144],the National Key Technology R&D Program of China(2017YFD0101806/03)the Fundamental Research Funds for the Central Universities(KYZZ201910)+1 种基金the Jiangsu Agricultural Industry Technology System(JATS2020)the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Radish(Raphanus sativus L.)taproot contains high concentrations of flavonoids,including anthocyanins(ATCs),in redskinned genotypes.However,little information on the genetic regulation of ATC biosynthesis in radish is available.A genome-wide association study of radish red skin color was conducted using whole-genome sequencing data derived from 179 radish genotypes.The R2R3-MYB transcription factor production of anthocyanin pigment 2(PAP2)gene was found in the region associated with a leading SNP located on chromosome 2.The amino acid sequence encoded by the RsPAP2 gene was different from those of the other published RsMYB genes responsible for the red skin color of radish.The overexpression of the RsPAP2 gene resulted in ATC accumulation in Arabidopsis and radish,which was accompanied by the upregulation of several ATC-related structural genes.RsPAP2 was found to bind the RsUFGT and RsTT8 promoters,as shown by a dual-luciferase reporter system and a yeast one-hybrid assay.The promoter activities of the RsANS,RsCHI,RsPAL,and RsUFGT genes could be strongly activated by coinfiltration with RsPAP2 and RsTT8.These findings showed the effectiveness of GWAS in identifying candidate genes in radish and demonstrated that RsPAP2 could(either directly or together with its cofactor RsTT8)regulate the transcript levels of ATC-related genes to promote ATC biosynthesis,facilitating the genetic enhancement of ATC contents and other related traits in radish.
基金supported by grants from the National Natural Science Foundation of China(31601766)National Key Technology R&D Program of China(2018YFD1000800)+2 种基金the earmarked fund for Jiangsu Agricultural Industry Technology System(JATS[2020]410)the Jiangsu Agricultural Science and Technology Innovation Fund(CX(20)3144)the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Melatonin(MT)is a tryptophan-derived natural product that plays a vital role in plant response to abiotic stresses,including heavy metals(HMs).However,it remains elusive how exogenous MT mediates lead(Pb)accumulation and detoxification at the methylation and transcriptional levels in radish.In this study,decreased Pb accumulation and increased antioxidant enzyme activity were detected under MT treatment in radish.Single-base resolution maps of DNA methylation under Pb stress(Pb200)and Pb plus MT treatment(Pb_50MT)were first generated.The genome-wide methylation level was increased under Pb stress,while an overall loss of DNA methylation was observed under MT treatment.The differentially methylated region(DMR)-associated genes between Pb_50MT and Pb200 were uniquely enriched in ion binding terms,including cation binding,iron ion binding,and transition metal ion binding.Hyper-DMRs between Pb200 and Control exhibited a decreasing trend of methylation under Pb_50MT treatment.A few critical upregulated antioxidant genes(e.g.,RsAPX2,RsPOD52 and RsGST)exhibited decreased methylation levels under MT treatment,which enabled the radish plants to scavenge lead-induced reactive oxygen species(ROS)and decrease oxidative stress.Notably,several MT-induced HM transporter genes with low methylation(e.g.,RsABCF5,RsYSL7 and RsHMT)and transcription factors(e.g.,RsWRKY41 and RsMYB2)were involved in reducing Pb accumulation in radish roots.These findings could facilitate comprehensive elucidation of the molecular mechanism underlying MT-mediated Pb accumulation and detoxification in radish and other root vegetable crops.
基金This work was in part supported by grants from National Key Technology R&D Program of China(2017YFD0101803,2017YFD0101806)Key Technology R&D Program of Jiangsu Province(BE2016379)+2 种基金the Jiangsu Agricultural Science and Technology Innovation Fund[CX(16)1012]the Fundamental Research Funds for the Central Universities(KJQN201656,KJQN201734)Project of Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,MOA,China(IVF201706).
文摘The fleshy taproot of radish is an important storage organ determining its yield and quality.Taproot thickening is a complex developmental process in radish.However,the molecular mechanisms governing this process remain unclear at the proteome level.In this study,a comparative proteomic analysis was performed to analyze the proteome changes at three developmental stages of taproot thickening using iTRAQ approach.In total,1862 differentially expressed proteins(DEPs)were identified from 6342 high-confidence proteins,among which 256 up-regulated proteins displayed overlapped accumulation in S1(pre-cortex splitting stage)vs.S2(cortex splitting stage)and S1 vs.S3(expanding stage)pairs,whereas 122 up-regulated proteins displayed overlapped accumulation in S1 vs.S3 and S2 vs.S3 pairs.Gene Ontology(GO)and pathway enrichment analysis showed that these DEPs were mainly involved in several processes such as“starch and sucrose metabolism”,“plant hormone signal transduction”,and“biosynthesis of secondary metabolites”.A high concordance existed between iTRAQ and RT-qPCR at the mRNA expression levels.Furthermore,association analysis showed that 187,181,and 96 DEPs were matched with their corresponding differentially expressed genes(DEGs)in S1 vs.S2,S1 vs.S3,and S2 vs.S3 comparison,respectively.Notably,several functional proteins including cell division cycle 5-like protein(CDC5),expansin B1(EXPB1),and xyloglucan endotransglucosylase/hydrolase protein 24(XTH24)were responsible for cell division and expansion during radish taproot thickening process.These results could facilitate a better understanding of the molecular mechanism underlying taproot thickening,and provide valuable information for the identification of critical genes/proteins responsible for taproot thickening in root vegetable crops.
基金The authors gratefully acknowledge the financial support from the National Natural Science Foundation of China under Grant 81871395.
文摘Previous studies have already shown that Raman spectroscopy can be used in the encoding of suspension array technology.However,almost all existing convolutional neural network-based decoding approaches rely on supervision with ground truth,and may not be well generalized to unseen datasets,which were collected under different experimental conditions,applying with the same coded material.In this study,we propose an improved model based on CyCADA,named as Detail constraint Cycle Domain Adaptive Model(DCDA).DCDA implements the clasification of unseen datasets through domain adaptation,adapts representations at the encode level with decoder-share,and enforces coding features while leveraging a feat loss.To improve detailed structural constraints,DCDA takes downsample connection and skips connection.Our model improves the poor generalization of existing models and saves the cost of the labeling process for unseen target datasets.Compared with other models,extensive experiments and ablation studies show the superiority of DCDA in terms of classification stability and generalization.The model proposed by the research achieves a classification with an accuracy of 100%when applied in datasets,in which the spectrum in the source domain is far less than the target domain.
基金funded by grants from the National Key Technology R&D Program of China(2018YFD1000800)National Natural Science Foundation of China(32172579)+3 种基金Jiangsu Seed Industry Revitalization Project[JBGS(2021)071]the earmarked fund for Jiangsu Agricultural Industry Technology System[JATS(2022)463]Jiangsu Agricultural Science and Technology Innovation Fund(CX(21)2020)the Project Founded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘The growth and development of taproots are inhibited by cold stress in radish(Raphanus sativus L.).Ethylene-responsive element binding factors(ERF)are key participators in the cold stress response and growth regulation of plants.However,the function of ERF genes in cold tolerance and root development in radish remains elusive.Here,we showed that the secondary growth of radish taproots was inhibited by cold stress.Comparative transcriptome analysis demonstrated that the RsERF40 gene is an important regulator of the cold stress response and root growth regulation.The cold tolerance of transgenic Arabidopsis plants overexpressing the RsERF40 gene was significantly improved.Overexpressing RsERF40 in the cold-sensitive radish genotype and silencing RsERF40 in the cold-tolerant radish genotype indicated that RsERF40 was beneficial for alleviating oxidative damage under cold stress in radish.Transgenic Arabidopsis seedlings showed an increase in the elongation and radial growth of dark-grown roots.RT-qPCR analysis showed that the expression of the cold-related genes(CORs)RsCOR78 and RsCOR413PM1 and the cell wall strengthening-related genes RsCESA6 and RsEXPB3 was upregulated in transgenic Arabidopsis seedlings.Yeast one-hybrid(Y1H)and dual-luciferase reporter assays(DLA)revealed that RsERF40 directly regulates RsCOR78,RsCOR413PM1,RsCESA6 and RsEXPB3 expression,illustrating that RsERF40 enhances cold tolerance and taproot growth by modulating osmotic adjustment and cell wall mechanical strength in radish.In this study,the RsERF40-regulon was firstly found to be a new cold response pathway independent of the CBF-COR pathway conferring cold stress tolerance with increasing radish taproot growth.These results provided novel insight into the molecular mechanism underlying cold stress response and would facilitate the genetic improvement of cold tolerance in radish and other root vegetable crops.
基金supported by grants from the Jiangsu Seed Industry Revitalization Project[Grant No.JBGS(2021)071]Key Technology R&D Program of Jiangsu Province(Grant No.BE2023366)+3 种基金National Natural Science Foundation of China(Grant No.32172579)the earmarked fund for Jiangsu Agricultural Industry Technology System[Grant No.JATS(2023)421]the Fundamental Research Funds for the Central Universities(Grant No.YDZX2023019)the Project Founded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘Heterosis,a crucial biological phenomenon,plays a vital role in determining the yield and quality of plants.Radish,an important root vegetable crop,exhibits notable heterosis in terms of root yield and quality.Nevertheless,the molecular mechanism underlying the formation of heterosis in radish remains unclear.Herein,both the transcriptome and DNA methylome analyses were performed on the F_(1)hybrids and parental lines.Expression level dominance(ELD)genes and allele-specific expression(ASE)genes together significantly contribute to heterosis,primarily through energy metabolism and plant hormone signal transduction pathway.An increase in average methylation level in the F_(1)hybrids was observed compared to the parental lines.Interestingly,a negative correlation was found between the methylation level of differentially expressed genes(DEGs)in gene body regions and their expression levels in NAU-LB and the F_(1)hybrids.Moreover,the hybrids were more sensitive to the 5-azacytidine than their parents,and the root weight and total sugar content in the F_(1)hybrids were dramatically decreased compared to the control.Immunolocalization results indicated that the auxin content of the F_(1)hybrids were reduced under 5-azacytidine treatment.Proliferating cell nuclear antigen immunolocalization revealed significant inhibition of vascular cambium activity in both the hybrids and parental lines.Notably,the expression profiles of a few differentially methylated DEGs including RsSUS1,RsSUC2a,RsIAA7,and RsIAA18,were significantly increased in the root of hybrids compared to their parents,suggesting a potential role of DNA methylation in yield heterosis.Collectively,these findings would provide valuable insight into the molecular mechanism underlying taproot yield heterosis and facilitate the genetic improvement of taproot yield and quality in radish breeding programs.
