Verticillium dahliae causes significant losses in cotton production.To reveal the mechanism of the defense response to V.dahliae in cotton,transcriptomic analyses were performed using cotton cultivars M138(V.dahliae-r...Verticillium dahliae causes significant losses in cotton production.To reveal the mechanism of the defense response to V.dahliae in cotton,transcriptomic analyses were performed using cotton cultivars M138(V.dahliae-resistant)and P2(V.dahliae-susceptible).The results revealed 11,076 and 6,640 differentially expressed genes(DEGs)in response to V.dahliae,respectively.The weighted gene co-expression network analysis of 4,633 transcription factors(TFs)indicated a“MEblue”module containing 654 TFs that strongly correlate with resistance to V.dahliae.Among these TFs,the ethylene response factor Ghi_A05G10166(GhERF91)was identified as a putative hub gene with a defense response against V.dahliae.A virus-induced gene silencing assay and exogenous application of ethephon showed that GhERF91 is activated by ethylene and positively regulates the response to V.dahliae exposure in cotton.This study provides fundamental transcriptome data and a putative causal gene(GhERF91)associated with resistance to V.dahliae,as well as genetic resources for breeding V.dahliae-resistant cotton.展开更多
Verticillium dahliae is an important fungal pathogen affecting cotton yield and quality.Therefore,the mining of V.dahlia-resistance genes is urgently needed.Proteases and protease inhibitors play crucial roles in plan...Verticillium dahliae is an important fungal pathogen affecting cotton yield and quality.Therefore,the mining of V.dahlia-resistance genes is urgently needed.Proteases and protease inhibitors play crucial roles in plant defense responses.However,the functions and regulatory mechanisms of the protease inhibitor PR6 gene family remain largely unknown.This study provides a comprehensive analysis of the PR6 gene family in the cotton genome.We performed genome-wide identification and functional characterization of the cotton GhPR6 gene family,which belongs to the potato protease inhibitor I family of inhibitors.Thirty-nine PR6s were identified in Gossypium arboreum,G.raimondii,G.barbadense,and G.hirsutum,and they were clustered into four groups.Based on the analysis of pathogen-induced and Ghlmm transcriptome data,Gh PR6-5b was identified as the key gene for V.dahliae resistance.Virus-induced gene silencing experiments revealed that cotton was more sensitive to V.dahliae V991after PR6-5b silencing.The present study established that GhWRKY75 plays an important role in resistance to Verticillium wilt in cotton by positively regulating GhPR6-5b expression by directly binding to the W-box TTGAC(T/C).Our findings established that GhWRKY75 is a potential candidate for improving cotton resistance to V.dahliae,and provide primary information for further investigations and the development of specific strategies to bolster the defense mechanisms of cotton against V.dahliae.展开更多
Verticillium wilt(VW),induced by the soil-borne fungus Verticillium dahliae(Vd),poses a substantial threat to a diverse array of plant species.Employing molecular breeding technology for the development of cotton vari...Verticillium wilt(VW),induced by the soil-borne fungus Verticillium dahliae(Vd),poses a substantial threat to a diverse array of plant species.Employing molecular breeding technology for the development of cotton varieties with heightened resistance to VW stands out as one of the most efficacious protective measures.In this study,we successfully generated two stable transgenic lines of cotton(Gossypium hirsutum L.),VdThitRNAi-1 and VdThit-RNAi-2,using host-induced gene silencing(HIGS)technology to introduce double-stranded RNA(dsRNA)targeting the thiamine transporter protein gene(VdThit).Southern blot analysis confirmed the presence of a single-copy insertion in each line.Microscopic examination showed marked reductions in the colonization and spread of Vd-mCherry in the roots of VdThit-RNAi cotton compared to wild type(WT).The corresponding disease index and fungal biomass of VdThit-RNAi-1/2 also exhibited significant reductions.Real-time quantitative PCR(qRT-PCR)analysis demonstrated a substantial inhibition of VdThit expression following prolonged inoculation of VdThit-RNAi cotton.Small RNA sequencing(sRNA-Seq)analysis revealed the generation of a substantial number of VdThit-specific siRNAs in the VdThit-RNAi transgenic lines.Additionally,the silencing of VdThit by the siVdThit produced by VdThit-RNAi-1/2 resulted in the elevated expression of multiple genes involved in the thiamine biosynthesis pathway in Vd.Under field conditions,VdThit-RNAi transgenic cotton exhibited significantly enhanced disease resistance and yield compared with WT.In summary,our findings underscore the efficacy of HIGS targeting VdThit in restraining the infection and spread of Vd in cotton,thereby potentially enabling the development of cotton breeding as a promising strategy for managing VW.展开更多
Lignin metabolism plays a pivotal role in plant defense against pathogens and is always positively correlated as a response to pathogen infection. Thus, understanding resistance genes against plant pathogens depends o...Lignin metabolism plays a pivotal role in plant defense against pathogens and is always positively correlated as a response to pathogen infection. Thus, understanding resistance genes against plant pathogens depends on a genetic analysis of the lignin response. This study used eight Upland cotton lines to construct a multi-parent advanced generation intercross(MAGIC) population(n=280), which exhibited peculiar characteristics from the convergence of various alleles coding for advantageous traits. In order to measure the lignin response to Verticillium wilt(LRVW), the artificial disease nursery(ADN) and rotation nursery(RN) were prepared for MAGIC population planting in four environments. The stem lignin contents were collected, and the LRVW was measured with the lignin value of ADN/RN in each environment, which showed significant variations. We employed 9 323 high-quality single-nucleotide polymorphism(SNP) markers obtained from the Cotton-SNP63K array for genotyping the MAGIC population. The SNPs were distributed through the whole genome with 4.78 SNP/Mb density, ranging from 1.14(ChrA06) to 10.08(ChrD08). In addition, a genome-wide association study was performed using a Mixed Linear Model(MLM) for LRVW. Three stable quantitative trait loci(QTLs), qLRVW-A04, qLRVW-A10, and qLRVW-D05, were identified in more than two environments. Two key candidate genes, Ghi_D05G01046 and Ghi_D05G01221, were selected within the QTLs through the combination of variations in the coding sequence, induced expression patterns, and function annotations. Both genes presented nonsynonymous mutations in coding regions and were strongly induced by Verticillium dahliae. Ghi_D05G01046 encodes a leucine-rich extensin(LRx) protein involved in Arabidopsis cell wall biosynthesis and organization. Ghi_D05G01221 encodes a transcriptional co-repressor novel interactor of novel interactor of jasmonic acid ZIM-domain(JAZ–NINJA), which functions in the jasmonic acid(JA) signaling pathway. In summary, the study creates valuable genetic resources for breeding and QTL mapping and opens up a new perspective to uncover the genetic basis of VW resistance in Upland cotton.展开更多
The severity of Verticillium wilt on cotton caused by defoliating strains of Verticillium dahliae has gradually increased and threatens production worldwide. Identification of the molecular components of leaf defoliat...The severity of Verticillium wilt on cotton caused by defoliating strains of Verticillium dahliae has gradually increased and threatens production worldwide. Identification of the molecular components of leaf defoliation may increase cotton tolerance to V. dahliae. Ethylene, a major player in plant physiological processes, is often associated with senescence and defoliation of plants. We investigated the cotton–V.dahliae interaction with a focus on the role of ethylene in defoliation and defense against V. dahliae.Cotton plants inoculated with V. dahliae isolate V991, a defoliating strain, accumulated more ethylene and showed increased disease symptoms than those inoculated with a non-defoliating strain. In cotton with a transiently silenced ethylene synthesis gene(GhACOs) and signaling gene(GhEINs) during cotton–V. dahliae interaction, ethylene produced was derived from cotton and more ethylene increased cotton susceptibility and defoliation rate. Overexpression of AtCTR1, a negative regulator in ethylene signaling, in cotton reduced sensitivity to ethylene and increased plant resistance to V. dahliae.Collectively, the results indicated precise regulation of ethylene synthesis or signaling pathways improve cotton resistant to Verticillium wilt.展开更多
Verticillium wilt,caused by Verticillium dahliae,seriously restricts the yield and quality improvement of cotton.Previous studies have revealed the involvement of WRKY members in plant defense against V.dahliae,but th...Verticillium wilt,caused by Verticillium dahliae,seriously restricts the yield and quality improvement of cotton.Previous studies have revealed the involvement of WRKY members in plant defense against V.dahliae,but the underlying mechanisms involved need to be further elucidated.Here,we demonstrated that Gossypium hirsutum WRKY DNA-binding protein 33(GhWRKY33) functions as a negative regulator in plant defense against V.dahliae.GhWRKY33 expression is induced rapidly by V.dahliae and methyl jasmonate,and overexpression of GhWRKY33 reduces plant tolerance to V.dahliae in Arabidopsis.Quantitative RT-PCR analysis revealed that expression of several JA-associated genes was significantly repressed in GhWRKY33 overexpressing transgenic plants.Yeast one-hybrid analysis revealed that GhWRKY33 may repress the transcription of both AtERF1 and GhERF2 through its binding to their promoters.Protein-protein interaction analysis suggested that GhWRKY33 interacts with G.hirsutum JASMONATE ZIM-domain protein 3(GhJAZ3).Similarly,overexpression of GhJAZ3 also decreases plant tolerance to V.dahliae.Furthermore,GhJAZ3 acts synergistically with GhWRKY33 to suppress both AtERF1 and GhERF2 expression.Our results imply that GhWRKY33 may negatively regulate plant tolerance to V.dahliae via the JA-mediated signaling pathway.展开更多
基金supported by the fund for National Key Research and Development Program of China(2023YFD2301203-05)the BTNYGG,China(NYHXGG,2023AA102)the Key Programs for Science and Technology Development of Shihezi City,Xinjiang Production and Construction Corps,China(2022NY01)。
文摘Verticillium dahliae causes significant losses in cotton production.To reveal the mechanism of the defense response to V.dahliae in cotton,transcriptomic analyses were performed using cotton cultivars M138(V.dahliae-resistant)and P2(V.dahliae-susceptible).The results revealed 11,076 and 6,640 differentially expressed genes(DEGs)in response to V.dahliae,respectively.The weighted gene co-expression network analysis of 4,633 transcription factors(TFs)indicated a“MEblue”module containing 654 TFs that strongly correlate with resistance to V.dahliae.Among these TFs,the ethylene response factor Ghi_A05G10166(GhERF91)was identified as a putative hub gene with a defense response against V.dahliae.A virus-induced gene silencing assay and exogenous application of ethephon showed that GhERF91 is activated by ethylene and positively regulates the response to V.dahliae exposure in cotton.This study provides fundamental transcriptome data and a putative causal gene(GhERF91)associated with resistance to V.dahliae,as well as genetic resources for breeding V.dahliae-resistant cotton.
基金supported by the National Key R&D Program of China(2022YFD1200300)the National Nature Science Youth Science Fund Project,China(31801412)+2 种基金the Key R&D Program of Shandong Province,China(2021LZGC026)the Agricultural Science and Technology Innovation Project of Shandong Academy of Agricultural Sciences,China(CXGC2023G02)the Shandong Provincial Program,China(WST2020011)。
文摘Verticillium dahliae is an important fungal pathogen affecting cotton yield and quality.Therefore,the mining of V.dahlia-resistance genes is urgently needed.Proteases and protease inhibitors play crucial roles in plant defense responses.However,the functions and regulatory mechanisms of the protease inhibitor PR6 gene family remain largely unknown.This study provides a comprehensive analysis of the PR6 gene family in the cotton genome.We performed genome-wide identification and functional characterization of the cotton GhPR6 gene family,which belongs to the potato protease inhibitor I family of inhibitors.Thirty-nine PR6s were identified in Gossypium arboreum,G.raimondii,G.barbadense,and G.hirsutum,and they were clustered into four groups.Based on the analysis of pathogen-induced and Ghlmm transcriptome data,Gh PR6-5b was identified as the key gene for V.dahliae resistance.Virus-induced gene silencing experiments revealed that cotton was more sensitive to V.dahliae V991after PR6-5b silencing.The present study established that GhWRKY75 plays an important role in resistance to Verticillium wilt in cotton by positively regulating GhPR6-5b expression by directly binding to the W-box TTGAC(T/C).Our findings established that GhWRKY75 is a potential candidate for improving cotton resistance to V.dahliae,and provide primary information for further investigations and the development of specific strategies to bolster the defense mechanisms of cotton against V.dahliae.
基金supported by the National Key Research and Development Program of China(2022YFD1200300)the National Natural Science Foundation of China(32072376 and 32372515)+3 种基金Winall Hi-tech Seed Co.,Ltd.,China(GMLM2023)the Nanfan Special Project of Chinese Academy of Agricultural Sciences(CAAS)(ZDXM2303 and YBXM2415)the Natural Science Foundation of Hebei Province,China(C2022204205)the Agricultural Science and Technology Innovation Program of CAAS。
文摘Verticillium wilt(VW),induced by the soil-borne fungus Verticillium dahliae(Vd),poses a substantial threat to a diverse array of plant species.Employing molecular breeding technology for the development of cotton varieties with heightened resistance to VW stands out as one of the most efficacious protective measures.In this study,we successfully generated two stable transgenic lines of cotton(Gossypium hirsutum L.),VdThitRNAi-1 and VdThit-RNAi-2,using host-induced gene silencing(HIGS)technology to introduce double-stranded RNA(dsRNA)targeting the thiamine transporter protein gene(VdThit).Southern blot analysis confirmed the presence of a single-copy insertion in each line.Microscopic examination showed marked reductions in the colonization and spread of Vd-mCherry in the roots of VdThit-RNAi cotton compared to wild type(WT).The corresponding disease index and fungal biomass of VdThit-RNAi-1/2 also exhibited significant reductions.Real-time quantitative PCR(qRT-PCR)analysis demonstrated a substantial inhibition of VdThit expression following prolonged inoculation of VdThit-RNAi cotton.Small RNA sequencing(sRNA-Seq)analysis revealed the generation of a substantial number of VdThit-specific siRNAs in the VdThit-RNAi transgenic lines.Additionally,the silencing of VdThit by the siVdThit produced by VdThit-RNAi-1/2 resulted in the elevated expression of multiple genes involved in the thiamine biosynthesis pathway in Vd.Under field conditions,VdThit-RNAi transgenic cotton exhibited significantly enhanced disease resistance and yield compared with WT.In summary,our findings underscore the efficacy of HIGS targeting VdThit in restraining the infection and spread of Vd in cotton,thereby potentially enabling the development of cotton breeding as a promising strategy for managing VW.
基金financed by the National Natural Science Foundation of China (31760402 and 31771844)the Innovation Leadership Program in Sciences and Technologies for Young and Middle-aged Scientists of Xinjiang Production and Construction Corps, China (2019CB027)。
文摘Lignin metabolism plays a pivotal role in plant defense against pathogens and is always positively correlated as a response to pathogen infection. Thus, understanding resistance genes against plant pathogens depends on a genetic analysis of the lignin response. This study used eight Upland cotton lines to construct a multi-parent advanced generation intercross(MAGIC) population(n=280), which exhibited peculiar characteristics from the convergence of various alleles coding for advantageous traits. In order to measure the lignin response to Verticillium wilt(LRVW), the artificial disease nursery(ADN) and rotation nursery(RN) were prepared for MAGIC population planting in four environments. The stem lignin contents were collected, and the LRVW was measured with the lignin value of ADN/RN in each environment, which showed significant variations. We employed 9 323 high-quality single-nucleotide polymorphism(SNP) markers obtained from the Cotton-SNP63K array for genotyping the MAGIC population. The SNPs were distributed through the whole genome with 4.78 SNP/Mb density, ranging from 1.14(ChrA06) to 10.08(ChrD08). In addition, a genome-wide association study was performed using a Mixed Linear Model(MLM) for LRVW. Three stable quantitative trait loci(QTLs), qLRVW-A04, qLRVW-A10, and qLRVW-D05, were identified in more than two environments. Two key candidate genes, Ghi_D05G01046 and Ghi_D05G01221, were selected within the QTLs through the combination of variations in the coding sequence, induced expression patterns, and function annotations. Both genes presented nonsynonymous mutations in coding regions and were strongly induced by Verticillium dahliae. Ghi_D05G01046 encodes a leucine-rich extensin(LRx) protein involved in Arabidopsis cell wall biosynthesis and organization. Ghi_D05G01221 encodes a transcriptional co-repressor novel interactor of novel interactor of jasmonic acid ZIM-domain(JAZ–NINJA), which functions in the jasmonic acid(JA) signaling pathway. In summary, the study creates valuable genetic resources for breeding and QTL mapping and opens up a new perspective to uncover the genetic basis of VW resistance in Upland cotton.
基金supported by the National Key Research and Development Project of China (2018YFD0100403)the National Natural Science Foundation of China (U1703231)。
文摘The severity of Verticillium wilt on cotton caused by defoliating strains of Verticillium dahliae has gradually increased and threatens production worldwide. Identification of the molecular components of leaf defoliation may increase cotton tolerance to V. dahliae. Ethylene, a major player in plant physiological processes, is often associated with senescence and defoliation of plants. We investigated the cotton–V.dahliae interaction with a focus on the role of ethylene in defoliation and defense against V. dahliae.Cotton plants inoculated with V. dahliae isolate V991, a defoliating strain, accumulated more ethylene and showed increased disease symptoms than those inoculated with a non-defoliating strain. In cotton with a transiently silenced ethylene synthesis gene(GhACOs) and signaling gene(GhEINs) during cotton–V. dahliae interaction, ethylene produced was derived from cotton and more ethylene increased cotton susceptibility and defoliation rate. Overexpression of AtCTR1, a negative regulator in ethylene signaling, in cotton reduced sensitivity to ethylene and increased plant resistance to V. dahliae.Collectively, the results indicated precise regulation of ethylene synthesis or signaling pathways improve cotton resistant to Verticillium wilt.
基金This work was supported by the National key R&D plan(2016YFD0101006)Yunnan Fundamental Research Projects(2019FA010).
文摘Verticillium wilt,caused by Verticillium dahliae,seriously restricts the yield and quality improvement of cotton.Previous studies have revealed the involvement of WRKY members in plant defense against V.dahliae,but the underlying mechanisms involved need to be further elucidated.Here,we demonstrated that Gossypium hirsutum WRKY DNA-binding protein 33(GhWRKY33) functions as a negative regulator in plant defense against V.dahliae.GhWRKY33 expression is induced rapidly by V.dahliae and methyl jasmonate,and overexpression of GhWRKY33 reduces plant tolerance to V.dahliae in Arabidopsis.Quantitative RT-PCR analysis revealed that expression of several JA-associated genes was significantly repressed in GhWRKY33 overexpressing transgenic plants.Yeast one-hybrid analysis revealed that GhWRKY33 may repress the transcription of both AtERF1 and GhERF2 through its binding to their promoters.Protein-protein interaction analysis suggested that GhWRKY33 interacts with G.hirsutum JASMONATE ZIM-domain protein 3(GhJAZ3).Similarly,overexpression of GhJAZ3 also decreases plant tolerance to V.dahliae.Furthermore,GhJAZ3 acts synergistically with GhWRKY33 to suppress both AtERF1 and GhERF2 expression.Our results imply that GhWRKY33 may negatively regulate plant tolerance to V.dahliae via the JA-mediated signaling pathway.
