Rice false smut(RFS),caused by the ascomycete fungus Ustilaginoidea virens(Cooke)Takahashi,is a notable panicle disease of rice.Mycoviruses represent a category of viruses capable of infecting various fungal groups,ga...Rice false smut(RFS),caused by the ascomycete fungus Ustilaginoidea virens(Cooke)Takahashi,is a notable panicle disease of rice.Mycoviruses represent a category of viruses capable of infecting various fungal groups,garnering interest for their potential application in the biological control of plant fungal diseases.We identified a novel positive-sense single-stranded RNA virus,named Ustilaginoidea virens mitovirus 1(UvMV1),from an abnormal U.virens strain Uv488.The complete genome sequence of UvMV1 is 2474 nt in length and contains a large open reading frame encoding RNA-dependent RNA polymerase.UvMV1 is classified as a new member of the genus Unuamitovirus in the family Mitoviridae based on phylogenetic analysis.It is effectively transmitted vertically through conidia of strain Uv488 and horizontally through hyphal fusion between vegetatively compatible individuals with an 80%transmission efficiency.We further demonstrated that UvMV1 significantly influenced conidiation,colony morphology,growth rate,secondary metabolite production,and the response to environmental stress in U.virens.Furthermore,qRT-PCR results aimed at detecting the expression levels of autophagy-related genes suggested that UvMV1 infection had the potential to induce autophagy in U.virens.Activation or inhibition of autophagy through chemical experiments demonstrated that UvMV1 enhances viral titers by inducing autophagy in U.virens,while inhibition of autophagy results in decreased UvMV1 titers.Transcriptome analyses consistently demonstrated that UvMV1 regulated the expression of genes associated with secondary metabolism,mycelial growth,virulence,and the mitogen-activated protein kinase signaling pathway,thereby influencing both the antiviral response and the virulence of U.virens.The data presented collectively identified a novel mycovirus responsible for inducing growth abnormalities in U.virens,and elucidated the regulatory mechanisms during mycovirus-host fungus interactions,particularly the autophagy pathway.These findings offer new perspectives and potential control strategies for managing RFS in agricultural settings.展开更多
The rice false smut disease, caused by Ustilaginoidea virens, has emerged as a significantglobal threat to rice production. The mechanism of carbon catabolite repression plays a crucial role in theefficient utilizatio...The rice false smut disease, caused by Ustilaginoidea virens, has emerged as a significantglobal threat to rice production. The mechanism of carbon catabolite repression plays a crucial role in theefficient utilization of carbon nutrients and enzyme regulation in the presence of complex nutritionalconditions. Although significant progress has been made in understanding carbon catabolite repression infungi such as Aspergillus nidulans and Magnaporthe oryzae, its role in U. virens remains unclear. Toaddress this knowledge gap, we identified UvCreA, a pivotal component of carbon catabolite repression,in U. virens. Our investigation revealed that UvCreA localized to the nucleus. Deletion of UvCreA resultedin decreased growth and pathogenicity in U. virens. Through RNA-seq analysis, it was found that theknockout of UvCreA led to the up-regulation of 514 genes and down-regulation of 640 genes. Moreover,UvCreA was found to be involved in the transcriptional regulation of pathogenic genes and genesassociated with carbon metabolism in U. virens. In summary, our findings indicated that UvCreA isimportant in fungal development, virulence, and the utilization of carbon sources through transcriptionalregulation, thus making it a critical element of carbon catabolite repression.展开更多
Apple replant disease(ARD) is primarily caused by biotic factors that seriously inhibits the development of apple industry. Therefore, the use of biological control measures to inhibit the main pathogens(such as Fusar...Apple replant disease(ARD) is primarily caused by biotic factors that seriously inhibits the development of apple industry. Therefore, the use of biological control measures to inhibit the main pathogens(such as Fusarium spp.) that cause ARD is of great significance to the sustainable development of the apple industry. Trichoderma virens 6PS-2, which exhibited antagonism toward a variety of pathogens, was screened from the rhizosphere soils of healthy apple trees(Malus robusta) in different replanted orchards in the Yantai and Zibo Cities, Shandong Province, China. Its fermentation extract inhibited the growth of pathogenic Fusarium proliferatum f. sp. Malus domestica MR5, which was proportional to the concentration. These substances also increased the hairy root volume and growth of Arabidopsis thaliana lateral roots. The phenotype of Malus hupehensis seedlings and microbial community structure in rhizosphere soils in greenhouse experiment using Highthroughput sequencing were analyzed, and the field experiment with grafted apple trees were used for further verification. Compared with the application of potato dextrose broth(PDB) medium, application of 6PS-2 spore suspension directly to replanted soils could improve the growth of M. hupehensis seedlings as well as the elongation of grafted apple trees. Concomitant decreases in the gene copy number of Fusarium and increases in the culturable bacteria/fungi were also observed in the greenhouse and field experiments. The abundance of Trichoderma,Bacillus, and Streptomyces increased significantly, but that of Fusarium, Pseudarthrobacter, and Humicola decreased. The content of esters, phenols,furans, and amino acids in root exudates of M. hupehensis seedlings increased, which significantly inhibited the multiplication of Fusarium, but was positively correlated with Bacillus and Trichoderma. In summary, T. virens 6PS-2 not only directly inhibits the activity of pathogenic Fusarium but also secrets secondary metabolites with antifungal and growth-promoting potential. In addition, 6PS-2 spore suspension can also promote the growth of plants to a certain extent, and change the soil microbial community structure of rhizosphere soils. It is believed that T. virens 6PS-2 has the potential for the alleviation of apple replant disease(ARD) in China.展开更多
Rice false smut,caused by Ustilaginoidea virens,is a devastating disease that greatly reduces rice yield and quality.However,controlling rice false smut disease is challenging due to the unique infection mode of U.vir...Rice false smut,caused by Ustilaginoidea virens,is a devastating disease that greatly reduces rice yield and quality.However,controlling rice false smut disease is challenging due to the unique infection mode of U.virens.Therefore,there is a need for early diagnosis and monitoring techniques to prevent the spread of this disease.Lateral flow strip-based recombinase polymerase amplification(LF-RPA)overcomes the limitations of current U.virens detection technologies,which are time-consuming,require delicate equipment,and have a high false-positive rate.In this study,we used a comparative genomics approach to identify Uv_3611,a specific gene of U.virens,as the target for the LF-RPA assay.The designed primers and probe efffectively detected the genomic DNA(gDNA)of U.virens and demonstrated no cross-reactivity with related pathogens.Under optimal conditions,the LF-RPA assay demonstrated a sensitivity of 10 pg of U.virens gDNA.Additionally,by incorporating a simplified PEG-NaOH method for plant DNA extraction,the LF-RPA assay enabled the detection of U.virens in rice spikelets within 30 min,without the need for specialized equipment.Furthermore,the LF-RPA assay successfully detected U.virens in naturally infected rice and seed samples in the field.Therefore,the LF-RPA assay is sensitive,efficient,and convenient,and could be developed as a kit for monitoring rice false smut disease in the field.展开更多
[Objective] The aim was to study resistance of major and backup rice cul- tivars against Ustilaginoidea virens. [Method] Experiments and surveys were made on resistance of seventeen backup rice cultivars and some majo...[Objective] The aim was to study resistance of major and backup rice cul- tivars against Ustilaginoidea virens. [Method] Experiments and surveys were made on resistance of seventeen backup rice cultivars and some major cultivars in Anhui Province to identify resistance of different rice cultivars. [Result] Yanjing No.9, 80You 226, Tianxie No.l, A01 Xian, Lvjingnuo No.6 were moderate resistant; A03 Xian and Yangjing 636 were susceptible and the rest ten showed moderate susceptibility. Based on surveys on major rice cultivars, most of Liangyou rice series are suscept- able and novel Liangyou Xiang 4 enjoys resistance to certain extent. [Conclusion] The research provided references for resistance of rice against diseases in Anhui Province.展开更多
The sclerotia of Villosiclava virens were found commonly in high altitude and the temperate regions, where the temperatures are relatively low in rice filling stage. To make sure if low temperature induce the scleroti...The sclerotia of Villosiclava virens were found commonly in high altitude and the temperate regions, where the temperatures are relatively low in rice filling stage. To make sure if low temperature induce the sclerotial formation in V. virens, the inoculated rice panicles in laboratory and the diseased rice panicles cut from paddy fields were treated under different temperatures. The results showed that 3 days of night temperature at 15°C were enough to induce the sclerotial formation. The low temperature was much more effective for young balls with intact membranes. After appearance of chlamydospores on the ball surfaces, the sclerotium could not differentiate anymore. The sclerotia began to differentiate below the chlamydospore layer and gradually grew onto the ball surfaces. This suggests that low temperature in the early development stage of false smut balls is an important factor to induce the sclerotial differentiation, and rice cultivars with long growth periods are able to produce more sclerotium-bearing balls, which will produce mass of spores in paddy field in the coming year.展开更多
[Objective] This study aims to screen for the high effective fungicides which could significantly decrease the disease incidence and disease index of rice false smut. [Method] The inhibitory activities of the fungicid...[Objective] This study aims to screen for the high effective fungicides which could significantly decrease the disease incidence and disease index of rice false smut. [Method] The inhibitory activities of the fungicide against mycelial growth of Ustilaginoidea virens were measured to in vitro evaluate the ECho values. And 17 fungicides were sprayed to evaluate the efficacy and effect of the fungicides tested in the field trials on the rice characters, [Result] The results showed that epoxicona- zole, difenoconazole, propiconazole and procloraz exhibited high inhibitory activity against mycelial growth of Ustilaginoidea virens with the ECso values 0.04, 0.07, 0.12 and 0.11 pg/ml, respectively. The results of field trials showed that the efficacy of Wen- quning, and fungicides such as difenoconazole, prochloraz, propiconazole, epoxi- conazole and their mixtures in controlling rice false smut were all 70% or more. [Conclusion] The 17 tested fungicides behaved efficacy in controlling rice false smut and did not cause drug injury on leaves and grains of rice plants, sprayed when flag leaves of rice fully expanded.展开更多
In order to determine population genetic structure and pathogenicity of Ustilaginoidea virens in the major rice-growing areas of Anhui Province, total 92 U. virens strains were collected from 28 rice-planting counties...In order to determine population genetic structure and pathogenicity of Ustilaginoidea virens in the major rice-growing areas of Anhui Province, total 92 U. virens strains were collected from 28 rice-planting counties (cities) of Anhui Province. Their genetic diversity was analyzed by using REP-PCR (repetitive extragenic palindromic sequence PCR), and pathogenicity was determined with artificial inoculation method. The results showed that U. virens in rice-growing regions of Anhui Province had a rich genetic diversity. At the similarity level of 0.76, the 92 U. virens strains could be classified into 7 groups. Significant differences were found in pathogenicity among the 24 U. virens strains belonging to different groups, which showed no association with territorial source of U. virens strain or cluster method adopted by this study. Strain pathogenicity and rice varieties showed significant specificity.展开更多
Rice false smut disease, which is caused by the fungus Ustilaginoidea virens, is currently one of the most devastating rice fungal diseases in the world. Rice false smut disease not only causes severe yield loss and g...Rice false smut disease, which is caused by the fungus Ustilaginoidea virens, is currently one of the most devastating rice fungal diseases in the world. Rice false smut disease not only causes severe yield loss and grain quality reduction, but also threatens food safety due to its production of mycotoxins. In this review, the most recent progresses regarding the life cycle, infection processes, genome and genetic diversity, pathogenic gene and disease resistance in rice were summarized in order to provide theoretical basis for the control of U. virens. We also proposed some future directions and key questions that need to be addressed for a better understanding of the molecular mechanism that leads to rice false smut disease and the prospects for sustainable control of rice false smut.展开更多
Korean spruce (Picea koraiensis Sieb. E1 Zucc.) is one of the main afforestation species in northern China. Seedling quality is a critical factor at planting time. To test whether the synergistic growth enhancement ...Korean spruce (Picea koraiensis Sieb. E1 Zucc.) is one of the main afforestation species in northern China. Seedling quality is a critical factor at planting time. To test whether the synergistic growth enhancement of Scotch pine (P. sylvestris var. mongolica) seedlings brought by the plant beneficial fungus Trichoderma virens (J.H. Mill., Giddens and A.A. Foster) Arx and ectomycorrhizal fungus (Suillus luteus (L.) Roussel.) can also benefit Korean spruce seed- lings, we examined the effects of S. luteus and T. virens on the growth of P. koraiensis seedlings and drought resistance of P. sylvestris var. mongolica in peat soils. The two fungi were added to sterilized peat soil in pots, and the plants were grown for 4 months. Seedling growth and physiological variables, including mycorrhizal colonization rate of roots, biomass, and chlorophyll content, were examined. The colonization rate of the mycorrhizal fungus on P. koraiensis exceeded 65 %, and the synergism between S. luteus and T. virens enhanced most of the variables for P. koraiensis seedlings after inoculation with S. luteus then 30 days later with T. virens as in our published results for seedlings of P. sylvestris var. mongolica. When seedlings of P. sylvestris var. mongolica were inoculated with this sequence, they became more drought tolerant. T. virens also induced S. luteus to produce -l,3-glucanase and chitinase. This inocu- lation sequence at planting can thus improve the quality of P. sylvestris var. mongolica and P. koraiensis seedlings and substantiates our previous results.展开更多
Transcription factors(TFs)orchestrate the regulation of cellular gene expression and thereby determine cell functionality.In this study,we analyzed the distribution of TFs containing domains,which named as ZnFTFs,both...Transcription factors(TFs)orchestrate the regulation of cellular gene expression and thereby determine cell functionality.In this study,we analyzed the distribution of TFs containing domains,which named as ZnFTFs,both in ascomycete and basidiomycete fungi.We found that ZnFTFs were widely distributed in these fungal species,but there was more expansion of the ZnFTF class in Ascomycota than Basidiomycota.We identified 40 ZnFTFs in Ustilaginoidea virens,and demonstrated the involvement of UvZnFTF1 in vegetative growth,conidiation,pigment biosynthesis and pathogenicity.RNA-Seq analysis suggested that UvZnFTF1 may regulate different nutrient metabolism pathways,the production of secondary metabolites,and the expression of pathogen-host interaction genes and secreted protein-encodi ng genes.Analysis of the distributi on of differe nt fungal TFs in U.virens further dem on strated that UvZnFTFs make up a large TF family and may play essential biological roles in U.virens.展开更多
In this research,the conditions for extraction of phenolics from leaves of Ficus virens were optimized using response surface methodology(RSM).The extraction abilities of phenolics(EAP) and flavonoids(EAF),the 2,2-dip...In this research,the conditions for extraction of phenolics from leaves of Ficus virens were optimized using response surface methodology(RSM).The extraction abilities of phenolics(EAP) and flavonoids(EAF),the 2,2-diphenyl-1-pierylhydrazyl(DPPH) free-radical scavenging potential,and the ferric reducing/antioxidant power(FRAP) were used as quality indicators.The results of single-factor experiments showed that temperature,ethanol concentration,extraction time,and the number of extraction cycles were the main influencing variables,and these provided key information for the central composite design.The results of RSM fitted well to a second degree polynomial model and more than 98% of the variability was explained.The ideal extraction conditions for EAP,EAF,DPPH free-radical scavenging potential,and FRAP were obtained.Considering the four quality indicators overall,the ideal extraction conditions were 58% ethanol at 57 °C for 37 min with three extraction cycles.At the ideal extraction conditions,the values of EAP,EAF,DPPH free-radical scavenging potential,and FRAP were 5.72%,3.09%,58.88 mg ascorbic acid equivalent(AAE)/g dry weight(DW),and 15.86 mg AAE/g DW,respectively.In addition,linear correlations were observed between EAP,EAF,and antioxidant potential.展开更多
Ustilaginoidea virens is the causal agent of rice false smut,which can be a highly destructive disease of rice.The plasma membrane phosphatase Psr1 proteins,which act as a regulator of the salinity stress response in ...Ustilaginoidea virens is the causal agent of rice false smut,which can be a highly destructive disease of rice.The plasma membrane phosphatase Psr1 proteins,which act as a regulator of the salinity stress response in yeast,are widely distributed across fungi,but their functional characterization is sketchy.In this study,we characterized the functions of Psr1 protein,UvPsr1,in U.virens.Analyses of the AUvpsr1 and its complementation strain showed that UvPsr1 is required for normal mycelial growth,conidiation and tolerance to oxidative,osmotic and cell wall stresses.When rice panicles were inoculated with the AUvpsr1 strains,no symptoms of false smut disease developed,showing that UvPSR1 also contributes to the pathogenicity of the fungus.The deletion mutant of UvPSR1 also appeared to produce a smaller titer of toxic compounds able to inhibit elongation of the germinated seeds.In conclusion,our results indicated that UvPsr1 is a new pathogenic factor of U.virens.展开更多
Rice false smut,which is caused by Ustilaginoidea virens,is an emerging disease of rice spikelets in rice-growing areas worldwide.However,the infection mechanism of U.virens on rice spikelets is still unclear.Here,we ...Rice false smut,which is caused by Ustilaginoidea virens,is an emerging disease of rice spikelets in rice-growing areas worldwide.However,the infection mechanism of U.virens on rice spikelets is still unclear.Here,we characterized a suppressor of mitogen-activated protein kinase kinase or ERK kinase(MEK)null(UvSMEKI)in U.virens that is conserved among filamentous fungi.Compared with wild type U.virens strain P-1,UvSMEKI deletion mutants were defective in pathogenicity and conidial germination.In addition,conidiation of UvSMEKI deletion mutants was significantly reduced on yeast extract tryptone(YT)plates,but inc「eased in YT broth compared with the wild type.Compared with UvSMEKI expression level during the vegetative mycelia and conidiation stages,UvSMEKI dramatically increased during infection of rice florets.Surprisingly,the UvSMEKI deletion mutants exhibited higher tolerance to H_(2)O_(2) and NaCl.In summary,presented evidence suggested that UvSMEKI positively regulated pathogenicity,conidial germination and conidiation in YT broth,and negatively regulated conidiation on YT medium and tolerance to oxidative and osmotic stresses.The results enhanee our understanding of the regulatory mechanism of pathogenicity of U.virens,and present a potential molecular target for blocking rice infection by U.virens.展开更多
Some stress response-related genes have been identified in Ustilaginoidea virens,but it is not clear whether and how defects of stress responses affect the pathogenesis processes of U.virens.In this study,we identifie...Some stress response-related genes have been identified in Ustilaginoidea virens,but it is not clear whether and how defects of stress responses affect the pathogenesis processes of U.virens.In this study,we identified a general stress response factor UvWHI2 as a homolog of Saccharomyces cerevisiae Whi2 in U.virens.The relative expression level of Uv Whi2 was significantly up-regulated during infection,suggesting that UvWHI2 may be involved in pathogenesis.Furthermore,knockout of Uv Whi2 showed decreased mycelial growth,increased conidiation in the potato sucrose medium and a defect in pathogenicity.In addition,the RNA-Seq and phenotypic analysis showed that UvWHI2 was involved in response to oxidative,hyperosmotic,cell wall stress and nutrient limitation.Further studies revealed that the defects of stress responses of the?Uvwhi2 mutant affected the formation of secondary spores on the nutrient limited surface and the rice surface,resulting in a significant reduction of pathogenicity of U.virens.Our results suggest that UvWHI2 is necessary for fungal growth,stress responses and the formation of secondary spores in U.virens.In addition,the defects of stress responses can affect the formation of secondary spores on the rice surface,and then compromise the pathogenicity of U.virens.展开更多
The fungus U. virens is the causal agent of rice false smut disease. The green fluorescent protein (GFP) was used to mark this fungus in order to visualize and analyze the colonization and infection processes in vivo....The fungus U. virens is the causal agent of rice false smut disease. The green fluorescent protein (GFP) was used to mark this fungus in order to visualize and analyze the colonization and infection processes in vivo. Using epifluorescence microscopy colonization and infection on rice roots were visualized in vivo. After inoculation for 2 to 15 d, it was observed that the conidia and their germ-tubes had penetrated into epidermis of young roots. The hyphae were found inside the root xylem 18 d after inoculation. Generally, the transformed fungus colonized the rhizosphere, the cortex as well as the vascular tissues with symptoms of root necrosis observed. The results of this work show that U. virens colonize not only rice panicles but also the roots.展开更多
bZIP proteins are widely distributed in eukaryotic organisms and regulate a diverse range of physiological processes.Several bZIP proteins have previously been identified in Ustilaginoidea virens.However,the biologica...bZIP proteins are widely distributed in eukaryotic organisms and regulate a diverse range of physiological processes.Several bZIP proteins have previously been identified in Ustilaginoidea virens.However,the biological roles of these bZIP proteins in this pathogen are still unknown.Here,one of these bZIP protein coding genes,UvATF21,was functionally characterized.Targeted deletion of UvATF21resulted in reduced conidiation and pathogenicity despite of the increased vegetative growth.The deletion mutants also significantly decreased the sensitivity to osmotic and oxidative stresses.Interestingly,deletion of UvATF21 exhibited different performances to cell wall integrity stress.These results indicated that UvATF21 played crucial roles in vegetative growth,conidiation,stress response,and full virulence in U.virens.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.32072363 and 32360648).
文摘Rice false smut(RFS),caused by the ascomycete fungus Ustilaginoidea virens(Cooke)Takahashi,is a notable panicle disease of rice.Mycoviruses represent a category of viruses capable of infecting various fungal groups,garnering interest for their potential application in the biological control of plant fungal diseases.We identified a novel positive-sense single-stranded RNA virus,named Ustilaginoidea virens mitovirus 1(UvMV1),from an abnormal U.virens strain Uv488.The complete genome sequence of UvMV1 is 2474 nt in length and contains a large open reading frame encoding RNA-dependent RNA polymerase.UvMV1 is classified as a new member of the genus Unuamitovirus in the family Mitoviridae based on phylogenetic analysis.It is effectively transmitted vertically through conidia of strain Uv488 and horizontally through hyphal fusion between vegetatively compatible individuals with an 80%transmission efficiency.We further demonstrated that UvMV1 significantly influenced conidiation,colony morphology,growth rate,secondary metabolite production,and the response to environmental stress in U.virens.Furthermore,qRT-PCR results aimed at detecting the expression levels of autophagy-related genes suggested that UvMV1 infection had the potential to induce autophagy in U.virens.Activation or inhibition of autophagy through chemical experiments demonstrated that UvMV1 enhances viral titers by inducing autophagy in U.virens,while inhibition of autophagy results in decreased UvMV1 titers.Transcriptome analyses consistently demonstrated that UvMV1 regulated the expression of genes associated with secondary metabolism,mycelial growth,virulence,and the mitogen-activated protein kinase signaling pathway,thereby influencing both the antiviral response and the virulence of U.virens.The data presented collectively identified a novel mycovirus responsible for inducing growth abnormalities in U.virens,and elucidated the regulatory mechanisms during mycovirus-host fungus interactions,particularly the autophagy pathway.These findings offer new perspectives and potential control strategies for managing RFS in agricultural settings.
基金the Key Projects of Zhejiang Provincial Natural Science Foundation,China(Grant No.LZ23C130002)the National Natural Science Foundation of China(Grant No.32100161)+3 种基金the Zhejiang Science and Technology Major Program on Rice New Variety Breeding,China(Grant No.2021C02063)the Key R&D Project of China National Rice Research Institute(Grant No.CNRRI-2020-04)the Chinese Academy of Agricultural Sciences under the Agricultural Sciences and Technologies Innovation Program,the Youth innovation Program of Chinese Academy of Agricultural Sciences(Grant No.Y2023QC22)the Joint Open Competitive Project of the Yazhou Bay Seed Laboratory and China National Seed Company Limited(Grant Nos.B23YQ1514 and B23CQ15EP).
文摘The rice false smut disease, caused by Ustilaginoidea virens, has emerged as a significantglobal threat to rice production. The mechanism of carbon catabolite repression plays a crucial role in theefficient utilization of carbon nutrients and enzyme regulation in the presence of complex nutritionalconditions. Although significant progress has been made in understanding carbon catabolite repression infungi such as Aspergillus nidulans and Magnaporthe oryzae, its role in U. virens remains unclear. Toaddress this knowledge gap, we identified UvCreA, a pivotal component of carbon catabolite repression,in U. virens. Our investigation revealed that UvCreA localized to the nucleus. Deletion of UvCreA resultedin decreased growth and pathogenicity in U. virens. Through RNA-seq analysis, it was found that theknockout of UvCreA led to the up-regulation of 514 genes and down-regulation of 640 genes. Moreover,UvCreA was found to be involved in the transcriptional regulation of pathogenic genes and genesassociated with carbon metabolism in U. virens. In summary, our findings indicated that UvCreA isimportant in fungal development, virulence, and the utilization of carbon sources through transcriptionalregulation, thus making it a critical element of carbon catabolite repression.
基金supported by China Agriculture Research System of MOF and MARA (Grant No.CARS-27)Shandong Agricultural Major Applied Technology Innovation Project (Grant No.SD2019ZZ008)+5 种基金Qingchuang Science and Technology Support Project of Shandong Colleges and Universities (Grant No.2019KJF020)Natural Science Foundation of Shandong Province (Grant No.ZR2020MC131)the National Key Research and Development Program of China (Grant No.2020YFD1000201)Taishan Scholars Funded Project (Grant No.ts20190923)the National Natural Science Foundation of China (Grant No.32072510)the Fruit Innovation Team in Shandong Province,China (Grant No.SDAIT-06-07)。
文摘Apple replant disease(ARD) is primarily caused by biotic factors that seriously inhibits the development of apple industry. Therefore, the use of biological control measures to inhibit the main pathogens(such as Fusarium spp.) that cause ARD is of great significance to the sustainable development of the apple industry. Trichoderma virens 6PS-2, which exhibited antagonism toward a variety of pathogens, was screened from the rhizosphere soils of healthy apple trees(Malus robusta) in different replanted orchards in the Yantai and Zibo Cities, Shandong Province, China. Its fermentation extract inhibited the growth of pathogenic Fusarium proliferatum f. sp. Malus domestica MR5, which was proportional to the concentration. These substances also increased the hairy root volume and growth of Arabidopsis thaliana lateral roots. The phenotype of Malus hupehensis seedlings and microbial community structure in rhizosphere soils in greenhouse experiment using Highthroughput sequencing were analyzed, and the field experiment with grafted apple trees were used for further verification. Compared with the application of potato dextrose broth(PDB) medium, application of 6PS-2 spore suspension directly to replanted soils could improve the growth of M. hupehensis seedlings as well as the elongation of grafted apple trees. Concomitant decreases in the gene copy number of Fusarium and increases in the culturable bacteria/fungi were also observed in the greenhouse and field experiments. The abundance of Trichoderma,Bacillus, and Streptomyces increased significantly, but that of Fusarium, Pseudarthrobacter, and Humicola decreased. The content of esters, phenols,furans, and amino acids in root exudates of M. hupehensis seedlings increased, which significantly inhibited the multiplication of Fusarium, but was positively correlated with Bacillus and Trichoderma. In summary, T. virens 6PS-2 not only directly inhibits the activity of pathogenic Fusarium but also secrets secondary metabolites with antifungal and growth-promoting potential. In addition, 6PS-2 spore suspension can also promote the growth of plants to a certain extent, and change the soil microbial community structure of rhizosphere soils. It is believed that T. virens 6PS-2 has the potential for the alleviation of apple replant disease(ARD) in China.
基金supported by grants from the Jiangsu Agricultural Science and Technology Innovation Fund,China(JASTIF)(CX(21)3012)to Haifeng Zhang。
文摘Rice false smut,caused by Ustilaginoidea virens,is a devastating disease that greatly reduces rice yield and quality.However,controlling rice false smut disease is challenging due to the unique infection mode of U.virens.Therefore,there is a need for early diagnosis and monitoring techniques to prevent the spread of this disease.Lateral flow strip-based recombinase polymerase amplification(LF-RPA)overcomes the limitations of current U.virens detection technologies,which are time-consuming,require delicate equipment,and have a high false-positive rate.In this study,we used a comparative genomics approach to identify Uv_3611,a specific gene of U.virens,as the target for the LF-RPA assay.The designed primers and probe efffectively detected the genomic DNA(gDNA)of U.virens and demonstrated no cross-reactivity with related pathogens.Under optimal conditions,the LF-RPA assay demonstrated a sensitivity of 10 pg of U.virens gDNA.Additionally,by incorporating a simplified PEG-NaOH method for plant DNA extraction,the LF-RPA assay enabled the detection of U.virens in rice spikelets within 30 min,without the need for specialized equipment.Furthermore,the LF-RPA assay successfully detected U.virens in naturally infected rice and seed samples in the field.Therefore,the LF-RPA assay is sensitive,efficient,and convenient,and could be developed as a kit for monitoring rice false smut disease in the field.
基金Supported by Demonstration and Extension Project of Technology Controlling Ustilaginoidea virens(11E1110)~~
文摘[Objective] The aim was to study resistance of major and backup rice cul- tivars against Ustilaginoidea virens. [Method] Experiments and surveys were made on resistance of seventeen backup rice cultivars and some major cultivars in Anhui Province to identify resistance of different rice cultivars. [Result] Yanjing No.9, 80You 226, Tianxie No.l, A01 Xian, Lvjingnuo No.6 were moderate resistant; A03 Xian and Yangjing 636 were susceptible and the rest ten showed moderate susceptibility. Based on surveys on major rice cultivars, most of Liangyou rice series are suscept- able and novel Liangyou Xiang 4 enjoys resistance to certain extent. [Conclusion] The research provided references for resistance of rice against diseases in Anhui Province.
基金financially supported by the National Natural Science Foundation of China(31271999)the Special Fund for Agro-scientific Research in the Public Interest,China(200903039-5)
文摘The sclerotia of Villosiclava virens were found commonly in high altitude and the temperate regions, where the temperatures are relatively low in rice filling stage. To make sure if low temperature induce the sclerotial formation in V. virens, the inoculated rice panicles in laboratory and the diseased rice panicles cut from paddy fields were treated under different temperatures. The results showed that 3 days of night temperature at 15°C were enough to induce the sclerotial formation. The low temperature was much more effective for young balls with intact membranes. After appearance of chlamydospores on the ball surfaces, the sclerotium could not differentiate anymore. The sclerotia began to differentiate below the chlamydospore layer and gradually grew onto the ball surfaces. This suggests that low temperature in the early development stage of false smut balls is an important factor to induce the sclerotial differentiation, and rice cultivars with long growth periods are able to produce more sclerotium-bearing balls, which will produce mass of spores in paddy field in the coming year.
基金Supported by Agricultural S&T Innovation:Genetic Structure Analysis and Parasitism Fitness of U.virens in Anhui Province(14B1148)Special Fund for Talent Development:Pesticide Innovation and High-efficient Application Technology(13C1109)~~
文摘[Objective] This study aims to screen for the high effective fungicides which could significantly decrease the disease incidence and disease index of rice false smut. [Method] The inhibitory activities of the fungicide against mycelial growth of Ustilaginoidea virens were measured to in vitro evaluate the ECho values. And 17 fungicides were sprayed to evaluate the efficacy and effect of the fungicides tested in the field trials on the rice characters, [Result] The results showed that epoxicona- zole, difenoconazole, propiconazole and procloraz exhibited high inhibitory activity against mycelial growth of Ustilaginoidea virens with the ECso values 0.04, 0.07, 0.12 and 0.11 pg/ml, respectively. The results of field trials showed that the efficacy of Wen- quning, and fungicides such as difenoconazole, prochloraz, propiconazole, epoxi- conazole and their mixtures in controlling rice false smut were all 70% or more. [Conclusion] The 17 tested fungicides behaved efficacy in controlling rice false smut and did not cause drug injury on leaves and grains of rice plants, sprayed when flag leaves of rice fully expanded.
基金Supported by Agricultural Science and Technology Innovation Fund of Anhui Province(14B1148)Special Fund for Talent Development in Anhui Province(13C1109)Science and Technology Major Project of Anhui Province(15CZZ03132)~~
文摘In order to determine population genetic structure and pathogenicity of Ustilaginoidea virens in the major rice-growing areas of Anhui Province, total 92 U. virens strains were collected from 28 rice-planting counties (cities) of Anhui Province. Their genetic diversity was analyzed by using REP-PCR (repetitive extragenic palindromic sequence PCR), and pathogenicity was determined with artificial inoculation method. The results showed that U. virens in rice-growing regions of Anhui Province had a rich genetic diversity. At the similarity level of 0.76, the 92 U. virens strains could be classified into 7 groups. Significant differences were found in pathogenicity among the 24 U. virens strains belonging to different groups, which showed no association with territorial source of U. virens strain or cluster method adopted by this study. Strain pathogenicity and rice varieties showed significant specificity.
基金funded by the Zhejiang Provincial Natural Science Foundation of China (Grant Nos. LQ19C140004 and LQ19C130007)the Chinese Academy of Agricultural Sciences under the ’Elite Youth’ Programthe Agricultural Sciences and Technologies Innovation Program of China (CAAS-ASTIP-2016-CNRRI)
文摘Rice false smut disease, which is caused by the fungus Ustilaginoidea virens, is currently one of the most devastating rice fungal diseases in the world. Rice false smut disease not only causes severe yield loss and grain quality reduction, but also threatens food safety due to its production of mycotoxins. In this review, the most recent progresses regarding the life cycle, infection processes, genome and genetic diversity, pathogenic gene and disease resistance in rice were summarized in order to provide theoretical basis for the control of U. virens. We also proposed some future directions and key questions that need to be addressed for a better understanding of the molecular mechanism that leads to rice false smut disease and the prospects for sustainable control of rice false smut.
基金the National Natural Science Foundation of China (31170597, 31200484)the National Fiveyear Science and Technology Research Project (2012BAD19B0801)+1 种基金the Fundamental Research Funds for the Central Universities (2572014AA30)the National Forestry Bureau ‘‘948’’ Project (2009-4-39) for financial support
文摘Korean spruce (Picea koraiensis Sieb. E1 Zucc.) is one of the main afforestation species in northern China. Seedling quality is a critical factor at planting time. To test whether the synergistic growth enhancement of Scotch pine (P. sylvestris var. mongolica) seedlings brought by the plant beneficial fungus Trichoderma virens (J.H. Mill., Giddens and A.A. Foster) Arx and ectomycorrhizal fungus (Suillus luteus (L.) Roussel.) can also benefit Korean spruce seed- lings, we examined the effects of S. luteus and T. virens on the growth of P. koraiensis seedlings and drought resistance of P. sylvestris var. mongolica in peat soils. The two fungi were added to sterilized peat soil in pots, and the plants were grown for 4 months. Seedling growth and physiological variables, including mycorrhizal colonization rate of roots, biomass, and chlorophyll content, were examined. The colonization rate of the mycorrhizal fungus on P. koraiensis exceeded 65 %, and the synergism between S. luteus and T. virens enhanced most of the variables for P. koraiensis seedlings after inoculation with S. luteus then 30 days later with T. virens as in our published results for seedlings of P. sylvestris var. mongolica. When seedlings of P. sylvestris var. mongolica were inoculated with this sequence, they became more drought tolerant. T. virens also induced S. luteus to produce -l,3-glucanase and chitinase. This inocu- lation sequence at planting can thus improve the quality of P. sylvestris var. mongolica and P. koraiensis seedlings and substantiates our previous results.
基金supported by the National Natural Science Foundation of China(Grant No.31601593)the Young Elite Scientist Sponsorship of China Association for Science and Technology(Grant No.YESS20170108)the Natural Science Foundation of Jiangsu Province,China(Grant No.BK20160588).
文摘Transcription factors(TFs)orchestrate the regulation of cellular gene expression and thereby determine cell functionality.In this study,we analyzed the distribution of TFs containing domains,which named as ZnFTFs,both in ascomycete and basidiomycete fungi.We found that ZnFTFs were widely distributed in these fungal species,but there was more expansion of the ZnFTF class in Ascomycota than Basidiomycota.We identified 40 ZnFTFs in Ustilaginoidea virens,and demonstrated the involvement of UvZnFTF1 in vegetative growth,conidiation,pigment biosynthesis and pathogenicity.RNA-Seq analysis suggested that UvZnFTF1 may regulate different nutrient metabolism pathways,the production of secondary metabolites,and the expression of pathogen-host interaction genes and secreted protein-encodi ng genes.Analysis of the distributi on of differe nt fungal TFs in U.virens further dem on strated that UvZnFTFs make up a large TF family and may play essential biological roles in U.virens.
基金Project supported by the National Natural Science Foundation of China(No.31070522)the Science and Technology Foundation of Fujian Province(No.2010N5013),China
文摘In this research,the conditions for extraction of phenolics from leaves of Ficus virens were optimized using response surface methodology(RSM).The extraction abilities of phenolics(EAP) and flavonoids(EAF),the 2,2-diphenyl-1-pierylhydrazyl(DPPH) free-radical scavenging potential,and the ferric reducing/antioxidant power(FRAP) were used as quality indicators.The results of single-factor experiments showed that temperature,ethanol concentration,extraction time,and the number of extraction cycles were the main influencing variables,and these provided key information for the central composite design.The results of RSM fitted well to a second degree polynomial model and more than 98% of the variability was explained.The ideal extraction conditions for EAP,EAF,DPPH free-radical scavenging potential,and FRAP were obtained.Considering the four quality indicators overall,the ideal extraction conditions were 58% ethanol at 57 °C for 37 min with three extraction cycles.At the ideal extraction conditions,the values of EAP,EAF,DPPH free-radical scavenging potential,and FRAP were 5.72%,3.09%,58.88 mg ascorbic acid equivalent(AAE)/g dry weight(DW),and 15.86 mg AAE/g DW,respectively.In addition,linear correlations were observed between EAP,EAF,and antioxidant potential.
基金This research was funded by Zhejiang Provincial Natural Science Foundation of China(Grant No.LQ19C140004)Key Research and Development Project of Zhejiang Province(Grant No.2019C02018)the Chinese Academy of Agricultural Sciences under the‘Elite Youth’program,and the Agricultural Sciences and Technologies Innovation Program.We thank Prof.HUANG Junbin of Huazhong Agriculture University(Wuhan,China)for providing inoculum of the U.virens strain HWD-2.
文摘Ustilaginoidea virens is the causal agent of rice false smut,which can be a highly destructive disease of rice.The plasma membrane phosphatase Psr1 proteins,which act as a regulator of the salinity stress response in yeast,are widely distributed across fungi,but their functional characterization is sketchy.In this study,we characterized the functions of Psr1 protein,UvPsr1,in U.virens.Analyses of the AUvpsr1 and its complementation strain showed that UvPsr1 is required for normal mycelial growth,conidiation and tolerance to oxidative,osmotic and cell wall stresses.When rice panicles were inoculated with the AUvpsr1 strains,no symptoms of false smut disease developed,showing that UvPSR1 also contributes to the pathogenicity of the fungus.The deletion mutant of UvPSR1 also appeared to produce a smaller titer of toxic compounds able to inhibit elongation of the germinated seeds.In conclusion,our results indicated that UvPsr1 is a new pathogenic factor of U.virens.
基金supported by the National Key Research and Development Project in China(Grant No.2016YFD200805)National Natural Science Foundation of China(Grant Nos.31301624 and 31571961).
文摘Rice false smut,which is caused by Ustilaginoidea virens,is an emerging disease of rice spikelets in rice-growing areas worldwide.However,the infection mechanism of U.virens on rice spikelets is still unclear.Here,we characterized a suppressor of mitogen-activated protein kinase kinase or ERK kinase(MEK)null(UvSMEKI)in U.virens that is conserved among filamentous fungi.Compared with wild type U.virens strain P-1,UvSMEKI deletion mutants were defective in pathogenicity and conidial germination.In addition,conidiation of UvSMEKI deletion mutants was significantly reduced on yeast extract tryptone(YT)plates,but inc「eased in YT broth compared with the wild type.Compared with UvSMEKI expression level during the vegetative mycelia and conidiation stages,UvSMEKI dramatically increased during infection of rice florets.Surprisingly,the UvSMEKI deletion mutants exhibited higher tolerance to H_(2)O_(2) and NaCl.In summary,presented evidence suggested that UvSMEKI positively regulated pathogenicity,conidial germination and conidiation in YT broth,and negatively regulated conidiation on YT medium and tolerance to oxidative and osmotic stresses.The results enhanee our understanding of the regulatory mechanism of pathogenicity of U.virens,and present a potential molecular target for blocking rice infection by U.virens.
基金funded by the Zhejiang Provincial Natural Science Foundation of China(Grant No.LQ19C140004)the Key Research and Development Project of Zhejiang Province,China(Grant No.2019C02018)Key Research and Development Project of China National Rice Research Institute(Grant No.CNRRI-2020-04)。
文摘Some stress response-related genes have been identified in Ustilaginoidea virens,but it is not clear whether and how defects of stress responses affect the pathogenesis processes of U.virens.In this study,we identified a general stress response factor UvWHI2 as a homolog of Saccharomyces cerevisiae Whi2 in U.virens.The relative expression level of Uv Whi2 was significantly up-regulated during infection,suggesting that UvWHI2 may be involved in pathogenesis.Furthermore,knockout of Uv Whi2 showed decreased mycelial growth,increased conidiation in the potato sucrose medium and a defect in pathogenicity.In addition,the RNA-Seq and phenotypic analysis showed that UvWHI2 was involved in response to oxidative,hyperosmotic,cell wall stress and nutrient limitation.Further studies revealed that the defects of stress responses of the?Uvwhi2 mutant affected the formation of secondary spores on the nutrient limited surface and the rice surface,resulting in a significant reduction of pathogenicity of U.virens.Our results suggest that UvWHI2 is necessary for fungal growth,stress responses and the formation of secondary spores in U.virens.In addition,the defects of stress responses can affect the formation of secondary spores on the rice surface,and then compromise the pathogenicity of U.virens.
文摘The fungus U. virens is the causal agent of rice false smut disease. The green fluorescent protein (GFP) was used to mark this fungus in order to visualize and analyze the colonization and infection processes in vivo. Using epifluorescence microscopy colonization and infection on rice roots were visualized in vivo. After inoculation for 2 to 15 d, it was observed that the conidia and their germ-tubes had penetrated into epidermis of young roots. The hyphae were found inside the root xylem 18 d after inoculation. Generally, the transformed fungus colonized the rhizosphere, the cortex as well as the vascular tissues with symptoms of root necrosis observed. The results of this work show that U. virens colonize not only rice panicles but also the roots.
基金supported by the National Natural Science Foundation of China(Grant No.31701736)Key Research and Development Program of Hubei Province,China(Grant No.2021BBA236)National Key Research and Development Program,China(Grant No.2016YFD0300700)。
文摘bZIP proteins are widely distributed in eukaryotic organisms and regulate a diverse range of physiological processes.Several bZIP proteins have previously been identified in Ustilaginoidea virens.However,the biological roles of these bZIP proteins in this pathogen are still unknown.Here,one of these bZIP protein coding genes,UvATF21,was functionally characterized.Targeted deletion of UvATF21resulted in reduced conidiation and pathogenicity despite of the increased vegetative growth.The deletion mutants also significantly decreased the sensitivity to osmotic and oxidative stresses.Interestingly,deletion of UvATF21 exhibited different performances to cell wall integrity stress.These results indicated that UvATF21 played crucial roles in vegetative growth,conidiation,stress response,and full virulence in U.virens.