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BnaWRKY75 positively regulates the resistance against sclerotinia sclerotiorum in ornamental Brassica napus

Horticultural Plant Journal 2024年第3期10卷 784-796页

作者： Kexin Yu Yijie Zhang Xiaoyuan Fei Luyue Ma Rehman Sarwar Xiaoli Tan Zheng Wang School of Life Sciences Jiangsu UniversityZhenjiangJiangsu 212013China School of Food and Biological Engineering Jiangsu University ZhenjiangJiangsu 212013China Key Laboratory for Theory and Technology of Intelligent Agricultural Machinery and Equipment Jiangsu UniversityZhenjiangJiangsu 212013China

With the development of tourism at home and abroad,Rapeseed(Brassica napus)has become an important ornamental ***,its ornamental value at the inflorescence stage is greatly reduced by sclerotinia *** of important genes in the defense responses is critical for molecular breeding,which is an important strategy for controlling the *** this study,we isolated a *** WRKY transcription factor gene,***75 was found to encode a nucleus-localized protein and exhibited relatively high expression in the *** thaliana transgenic plants expressing BnaWRKY75 showed enhanced resistance to ***,and both ProBnaWRKY75:GUS and gene expression analyses showed that BnaWRKY75 was highly responsive to *** infection,indicating the involvement of BnaWRKY75 in response to this ***,overexpression(OE)of BnaWRKY75 in *** significantly enhanced the resistance to ***,whereas the resistance was reduced in RNAi transgenic *** ***,the BnaWRKY75-OE *** plants exhibited constitutive activation of salicylic acid-,jasmonic acid-,and ethylene-mediated defense responses and the inhibition of both H_(2)O_(2)and O_(2)·^(-)accumulation in response to this *** contrast,BnaWRKY75-RNAi plants showed a reverse pattern,suggesting that BnaWRKY75 is involved in hormonal signaling pathways and in the control of reactive oxygen species *** conclusion,these data indicate that BnaWRKY75,a regulator of multiple defense responses,positively regulates resistance against ***,which may guide the improvement of resistance in rapeseed.

关键词： Brassica napus BnaWRKY75 Defense response sclerotinia sclerotiorum

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SsdchA is a novel secretory cellobiohydrolase driving pathogenicity in sclerotinia sclerotiorum

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The Crop Journal 2024年第2期12卷 493-502页

作者： Yangui Chen Yijuan Ding Siqi Zhao Nan Yang Zhaohui Wu Ping Zhang Hongmei Liao Mengquan Dong Yang Yu Huafang Wan Wei Qian Integrative Science Center of Germplasm Creation in Western China(Chongqing)Science City College of Agronomy and BiotechnologySouthwest UniversityChongqing 400715China Academy of Agricultural Sciences Southwest UniversityChongqing 400715China Engineering Research Center of South Upland Agriculture Ministry of EducationChongqing 400715China College of Plant Protection Southwest UniversityChongqing 400715China

The necrotrophic fungus, sclerotinia sclerotiorum, employs an array of cell wall-degrading enzymes(CWDEs), including cellulase, to dismantle host cell walls. However, the molecular mechanisms through which S. sclerotiorum degrades cellulose remain elusive. Here, we unveil a novel secretory cellobiohydrolase, SsdchA, characterized by a signal peptide and a Glyco_hydro_7(GH7) domain. SsdchA exhibits a robust expression of during early infection stages. Interestingly, colony morphology and growth rates remain unaffected across the wild-type, SsdchA deletion strains and SsdchA overexpression strains on potato dextrose agar(PDA) medium. Nevertheless, the pathogenicity and cellobiohydrolase activity decreased in the SsdchA deletion strains, but enhanced in the SsdchA overexpression strains. Moreover,the heterologous expression of SsdchA in Arabidopsis thaliana leads to reduced cellulose content and heightened susceptibility to S. sclerotiorum. Collectively, our data underscore the pivotal role of the novel cellobiohydrolase SsdchA in the pathogenicity of S. sclerotiorum.

关键词： Cellobiohydrolase Cellulose Pathogenicity sclerotinia sclerotiorum Secretory protein SsdchA

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Physiological and biochemical characteristics of boscalid resistant isolates of sclerotinia sclerotiorum from asparagus lettuce

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Journal of Integrative Agriculture 2023年第12期22卷 3694-3708页

作者： SHI Dong-ya LI Feng-jie ZHANG Zhi-hui XU Qiao-nan CAO Ying-ying Jane Ifunanya MBADIANYA LI Xin WANG Jin CHEN Chang-jun Department of Pesticide Science College of Plant ProtectionNanjing Agricultural UniversityNanjing 210095P.R.China Jiangsu Pesticide Research Institute Co. Ltd.Nanjing 210046P.R.China

Laboratory mutants of sclerotinia sclerotiorum(Lib)de Bary,resistant to boscalid,have been extensively ***,the resistance situation in the lettuce field remains largely *** this study,among the 172 *** isolates collected from asparagus lettuce field in Jiangsu Province,China,132 isolates(76.74%)exhibited low-level resistance to boscalid(Bos^(LR)),with a discriminatory dose of 5μg *** comparison to the boscalid-sensitive(BosS)isolates,most Bos^(LR)isolates demonstrated a slightly superior biological fitness,as evidenced by data on mycelial growth,sclerotium production and ***,most Bos^(LR)isolates showed comparable levels of oxalic acid(OA)accumulation,increased exopolysaccharide(EPS)content and reduced membrane permeability when compared to the BosS ***,their responses to distinct stress factors diverged ***,the effectiveness of boscalid in controlling Bos^(LR)isolates on radish was diminished compared to its efficacy on BosS *** mutations were identified in the SDH genes of Bos^(LR)isolates,revealing the existence of three resistant genotypes:I(^(A11V)at SDHB,SDHB^(A11V)),II(^(Q38R)at SDHC,SDHC^(Q38R))and III(SDHB^(A11V)+SDHC^(Q38R)).Importantly,no cross-resistance was observed between boscalid and other fungicides such as thifluzamide,pydiflumetofen,fluazinam,or *** molecular docking analysis indicated that the docking total score(DTS)of the type I resistant isolates(1.3993)was lower than that of the sensitive isolates(1.7499),implying a reduced affinity between SDHB and boscalid as a potential mechanism underlying the boscalid resistance in *** findings contribute to an enhanced comprehension of boscalid’s mode of action and furnish valuable insights into the management of boscalid resistance.

关键词： sclerotinia sclerotiorum boscalid asparagus lettuce SDHB^(A11V) SDHC^(Q38R)

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Host-induced gene silencing of multiple pathogenic factors of sclerotinia sclerotiorum confers resistance to sclerotinia rot in Brassica napus

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The Crop Journal 2022年第3期10卷 661-671页

作者： Jian Wu Shengliang Yin Li Lin Dongxiao Liu Sichao Ren Wenjing Zhang Wencheng Meng Peipei Chen Qinfu Sun Yujie Fang Cunxu Wei Youping Wang Key Laboratory of Plant Functional Genomics of the Ministry of Education Yangzhou UniversityYangzhouChina Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding Yangzhou UniversityYangzhouChina

sclerotinia sclerotiorum is generally considered one of the most economically damaging pathogens in oilseed rape(Brassica napus).Breeding for sclerotinia resistance is challenging,as no immune germplasm available in *** is desirable to develop new breeding *** the present study,hostinduced gene silencing(HIGS),developed based on RNA interference(RNAi),was applied to protect *** from *** *** pathogenicity genes,the endo-polygalacturonase gene(Ss PG1),cellobiohydrolase gene(Ss CBH),and oxaloacetate acetylhydrolase gene(Ss OAH1),were chosen as HIGS ***-incubation of synthesized double-stranded RNAs(ds RNAs)with *** in liquid medium significantly reduced the transcript levels of the target *** to plant surfaces of ds RNA targeting the three genes conferred effective protection against *** transgenic *** plants expressing small interfering RNAs with sequence identity to Ss PG1,Ss CBH,and Ss OAH1 were *** transgenic *** prevented the expression of *** target genes,slowed pathogenicity-factor accumulation,impeded fungal growth,and suppressed appressorium formation,thereby conferring resistance to *** silencing of Ss PG1,Ss CBH,and Ss OAH1 by stable expression of a chimeric hairpin RNAi construct in *** led to enhanced protection phenotypes(with disease lesion size reduced by 36.8%–43.7%).We conclude that HIGS of pathogenic-factor genes of *** is a promising strategy for controlling sclerotinia rot in oilseed rape.

关键词： sclerotinia sclerotiorum Brassica napus SIGS HIGS RNAi

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sclerotinia sclerotiorum virulence is affected by mycelial age via reduction in oxalate biosynthesis

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Journal of Integrative Agriculture 2016年第5期15卷 1034-1045页

作者： WANG Ji-peng XU You-ping ZANG Xian-peng LI Shuang-sheng CAI Xin-zhong Institute of Biotechnology College of Agriculture and Biotechnology Zhejiang University Center of Analysis and Measurement Zhejiang University

sclerotinia sclerotiorum is one of the most devastating necrotrophic phytopathogens. Virulence of the hyphae of this fungus at different ages varies significantly. Molecular mechanisms underlying this functional distinction are largely unknown. In this study, we confirmed the effect of mycelial culture time/age on virulence in two host plants and elucidated its molecular and morphological basis. The virulence of the S. sclerotiorum mycelia in plants dramatically decreases along with the increase of the mycelial age. Three-day-old mycelia lost the virulence in plants. Comparative proteomics analyses revealed that metabolism pathways were comprehensively reprogrammed to suppress the oxalic acid（OA） accumulation in old mycelia. The oxaloacetate acetylhydrolase（OAH）, which catalyzes OA biosynthesis, was identified in the S. sclerotiorum genome. Both gene expression and protein accumulation of OAH in old mycelia were strongly repressed. Moreover, in planta OA accumulation was strikingly reduced in old mycelia-inoculated plants compared with young vegetative mycelia-inoculated plants. Furthermore, supply with 10 mmol L^（-1） OA enabled the old mycelia infect the host plants, demonstrating that loss of virulence of old mycelia is mainly caused by being unable to accumulate OA. Additionally, aerial mycelia started to develop from 0.5-day-old vegetative mycelia and dominated over 1-day-old mycelia grown on potato dextrose agar plates. They were much smaller in hypha diameter and grew significantly slower than young vegetative mycelia when subcultured, which did not maintain to progenies. Collectively, our results reveal that S. sclerotiorum aerial hyphae-dominant old mycelia fail to accumulate OA and thereby lose the virulence in host plants.

关键词： sclerotinia sclerotiorum mycelial age oxalic acid virulence proteomics

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Proteomics analysis of the phytopathogenic fungus sclerotinia sclerotiorum:a narrative review

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Journal of Bio-X Research 2022年第3期5卷 112-117页

作者： Oluwatobi Sarah Otun Ikechukwu Achilonu Khayalethu Ntushelo Department of Molecular and Cell Biology University of Witwatersrand Department of Agriculture and Animal Health Science CampusUniversity of South AfricaJohannesburgSouth Africa

sclerotinia sclerotiorum(Lib.)de Bary is a necrotrophic plant pathogen that causes cottony rot,watery soft rot,stem rot,white mold,and other disease symptoms in over 700 plant hosts around the *** of economically important crops,the lack of resistant cultivars,and the general challenge of controlling diseases caused by this broad-based pathogen call for continued ***,in recent years,mass spectrometry-based proteomics analyses have been used to acquire a fundamental and in-depth molecular understanding of this fungal *** this review,we describe the characteristics of the sclerotinia sclerotiorum pathogen and examine its virulence factors,secreted proteins,and host suppression ***,we review recent proteomics studies and extrapolate their primary findings for the identification and functional characterization of sclerotinia sclerotiorum ***,we discuss key findings that shape the understanding of the virulent factors and pathogenesis of sclerotinia sclerotiorum and outline directions for future proteomic investigations of plant pathogens.

关键词： fungal pathogenesis mass spectrometry-based proteomics plant pathogen sclerotinia sclerotiorum virulent proteins

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Maleic acid and malonic acid reduced the pathogenicity of sclerotinia sclerotiorum by inhibiting mycelial growth,sclerotia formation and virulence factors

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Stress Biology 2023年第1期3卷 495-506页

作者： Yu-chen Fei Qin Cheng Huan Zhang Chuang Han Xu Wang Yan-feng Li Shi-qian Li Xiao-hu Zhao State Key Laboratory of Agricultural Microbiology/College of Resources and Environment Huazhong Agricultural UniversityWuhan 430070China Fujian Universities and Colleges Engineering Research Center of Modern Facility Agriculture Fuqing 350300China Institute of Quality Standard and Monitoring Technology for Agro-Products of Guangdong Academy of Agricultural Sciences Guangzhou 510640China

sclerotinia sclerotiorum is a necrotrophic plant pathogenic fungus with broad distribution and host *** compounds derived from plant extracts have been proven to be effective in controlling *** this study,the mycelial growth of *** was effectively inhibited by maleic acid,malonic acid,and their combination at a concentration of 2 mg/mL,with respective inhibition rates of 32.5%,9.98%,and 67.6%.The treatment of detached leaves with the two acids resulted in a decrease in lesion ***,maleic acid and malonic acid decreased the number of sclerotia while simultaneously increasing their *** two acids also disrupted the cell structure of sclerotia,leading to sheet-like electron-thin *** a molecular level,maleic acid reduced oxalic acid secretion,upregulated the expression of Ss-Odc2 and downregulated CWDE10,Ss-Bi1 and ***,malonic acid downregulated CWDE2 and *** findings verified that maleic acid and malonic acid could effectively inhibit ***,providing promising evidence for the development of an environmentally friendly biocontrol agent.

关键词： sclerotinia sclerotiorum Maleic acid Malonic acid Inhibition

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Inhibitory effect of chitosan on growth of the fungal phytopathogen,sclerotinia sclerotiorum,and sclerotinia rot of carrot

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Journal of Integrative Agriculture 2015年第4期14卷 691-697页

作者： WANG Qing ZUO Jin-hua WANG Qian NA Yang GAO Li-pu Beijing Vegetable Research Center Beijing Academy of Agriculture and Forestry Sciences/Beijing Key Laboratory of Fruits and Vegetable Storage and Processing/Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China)Ministry of Agriculture/Key Laboratory of Urban Agriculture (North)Ministry of Agriculture

The antifungal activity of chitosan on a common fungal phytopathogen, sclerotinia sclerotiorum, and the control effect on sclerotinia rot of carrot were investigated. Mycelial growth and fungal biomass were strongly inhibited by chitosan. Using propidium iodide stain combined with fluorescent microscopy, the plasma membrane of chitosan-treated S. sclerotiorum mycelia was observed to be markedly damaged. Concomitantly, protein leakage and lipid peroxidation was also found to be significantly higher in chitosan-treated mycelia compared to the control. Chitosan provided an effective control of sclerotinia rot of carrot, with induction of activity of defense-related enzymes including polyphenoloxidase and peroxidase. These data suggest that the effects of chitosan on sclerotinia rot of carrot may be associated with the direct damage to the plasma membrane and lipid peroxidation of S. sclerotiorum, and the elicitation of defense response in carrot.

关键词： antifungal activity carrot chitosan plasma membrane sclerotinia sclerotiorum

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A novel Arabidopsis miRNA, ath-miR38-3P, is involved in response to sclerotinia sclerotiorum infection

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Journal of Integrative Agriculture 2016年第11期15卷 2556-2562页

作者： ZHAO Xu SHAN Ya-nan ZHAO Yan WANG Ai-rong WANG Zong-hua Fujian University Key Laboratory for Functional Genomics of Plant Fungal Pathogens Fujian Agriculture and Forestry University Key Laboratory of Bio-Pesticide and Chemistry Biology Ministry of Education

Plant defense responses against penetration or colonization of pathogens are mediated by activation and repression of a large array of genes. Host endogenous small RNAs are essential in gene expression reprogramming process. We identified a new Arabidopsis micro RNA（mi RNA） ath-mi R38-3P by high-throughput sequencing and further confirmed it by Northern blot assay. Interestingly, ath-miR38-3P was highly induced after infection of the pathogen sclerotinia sclerotiorum. Further analysis based on the mi RNA target database demonstrated that ath-mi R38-3P might target to five putative genes： AT2G03140, AT5G59430, AT5G66320, AT1G36620 and AT3G03820. To confirm the target, we conducted the quantitative real-time PCR to observe the expression pattern of each candidate gene. The results showed that only AT3G03820 was down-regulated after inoculation of S. sclerotiorum. In addition, overexpression of ath-miR38-3P down-regulates AT3G03820, suggesting AT3G03820 might represent the target for ath-mi R38-3P. Our results may provide the useful information for further studying the biological function of a novel ath-mi R38-3P and its targets in Arabidopsis-sclerotinia interaction.

关键词： miRNA Arabidopsis thaliana sclerotinia sclerotiorum miRNA-target gene

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Changes in Anatomical Features and Protein Pattern of Sunflower Partially Resistant and Susceptible Lines During Infection By Virulence Factors of sclerotinia sclerotiorum

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Phyton-International Journal of Experimental Botany 2019年第2期88卷 149-159页

作者： Maryam Monazzah Sattar Tahmasebi Enferadi Zohre Rabiei Alessandro Mattiello Department of Plant Molecular Biotechnology Institute of Agricultural BiotechnologyNational Institute of Genetic Engineering and BiotechnologyTehranPO Box 14925/16Iran Department of Agri-Food Animal and Environmental SciencesUniversity of UdineUdineItaly.

Helianthus annuus *** an oil seed crop is widely grown throughout the *** of the most destructive diseases of sunflower is stem rot caused by sclerotinia *** acid is the major virulence factor of this necrotrophic *** is important to further investigate plant responses to this non-specific ***,in the present study,we compared the patterns of total soluble proteins and xylem morphology of partially resistant and susceptible sunflower lines after treatment with sclerotinia culture *** basal stems of both lines were treated with 40 mM oxalic acid(pH 3.7)of fungus culture filtrate and samples were collected at 24,48 and 72 hours post *** SDS-PAGE protein pattern new protein bands appeared in both lines after *** observations suggest induction of stress-related proteins upon culture filtrate *** identities of these new proteins need to be more clarify in future *** changes in xylem morphology and degree of lignification of both lines was studied by light microscopy and microtome sectioning techniques after treatment with *** culture *** investigations revealed changes in xylem diameter and xylem lignification of treated lines at various time *** lignin deposition in xylem vessels of partially resistant line has been observed after *** addition,the size of xylem vessels in partially resistant line has been sharply decreased upon pathogen filtrate *** results of this study will help us gain a more complete understanding of resistance mechanisms to this cosmopolitan and devastating pathogen.

关键词： Microtome Xylem Helianthus annuus sclerotinia sclerotiorum Stem rot

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