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Phytopathology
American Phytopathological Society
Phytopathology

American Phytopathological Society

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    Identification of a Novel ToxA Haplotype of Pyrenophora tritici-repentis from Japan

    Hafez, MohamedDespins, ThereseNakajima, KaoriAboukhaddour, Reem...
    6页
    查看更多>>摘要：Pyrenophora tritici-repentis was described first as a pathogen of wheat (tan spot) in Japan in the 1920s, but, since then, no reports on P. tritici-repentis race structure or its effectors in Japan have been published. In this study, 10 single-spore isolates of P. tritici-repentis were collected from bread wheat in Japan. These isolates were evaluated for virulence on four differential wheat genotypes and tested for the presence/absence of the effector-encoding genes, ToxA and ToxB, in multiplex PCR assays. These isolates were identified as ToxA producers, of which eight were designated as race 2 (ToxA producers) and two were classified as race 1 (ToxA and ToxC producers) based on their virulence patterns. Sequence analysis of the ToxA amplicons from these 10 isolates indicated the presence of a novel ToxA haplotype (denoted PtrA2). A comparative sequence analysis and resequencing of ToxA from reference P. tritici-repentis isolates showed that all previously published ToxA haplotypes in P. tritici-repentis were identical, and are hence denoted PtrA1 in this study. A total of 163 PtrToxA sequences from global origins were already deposited in GenBank and were confirmed identical to PtrA1. Sequence variation in PtrA1 and PtrA2 open reading frames were found at three positions: one synonymous mutation at position 412 (C/G) and two nonsynonymous mutations at positions 342 and 362 that alter amino acid sequence. These mutations did not seem to affect the necrosis development on a ToxA-sensitive wheat genotype when rated for symptoms 5 to 7 days after inoculation. This is the first report correctly confirming the presence of an additional novel ToxA haplotype in P. tritici-repentis for which we have predicted its isoform and updated the ToxA haplotype evolutionary network.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    Development of Agrobacterium tumefaciens Infiltration of Infectious Clones of Grapevine Geminivirus A Directly into Greenhouse-Grown Grapevine and Nicotiana benthamiana Plants

    Kuo, Yen-WenBednarska, AlicjaAl Rwahnih, MaherFalk, Bryce W....
    7页
    查看更多>>摘要：Grapevine virus infectious clones are important tools for fundamental studies, but also because of their potential for translational applications for grapevine improvement. Although several grapevine virus infectious clones have been developed, there has been difficulty in directly infecting mature grapevine plants, and many of the viruses used still cause disease symptoms in grapevine plants, making them less likely candidates for biotechnological applications in grapes. Here, we developed an improved Agrobacterium tumefaciens infiltration method that can be used to deliver DNA plasmids and viral infectious clones directly into approximately 20- to 40-cm-high (above soil) greenhouse-grown grapevine plants. We also developed infectious clones for two isolates of grapevine geminivirus A (GGVA): Longyan (China; GenBank accession KX570611; GGVA-76) and Super Hamburg (Japan; GenBank accession KX570610; GGVA-93). Neither virus caused any obvious symptoms when inoculated to plants of grapevine varieties Colombard, Salt Creek, Cabernet Sauvignon, and Vaccarese. However, the two GGVA isolates induced different symptom severity and viral titer in Nicotiana benthamiana plants. The two GGVA isolates used here were found to accumulate to different titers in different parts/branches of the infected grapevine plants. The GGVA infectious clones and the improved grapevine infiltration technique developed here provide new, valuable tools that can be applied to grapevine plants, possibly even for translational applications such as disease management and desired trait improvements.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    Identification of Genetic and Chemical Factors Affecting Type III Secretion System Expression in Pseudomonas syringae pv. actinidiae Biovar 3 Using a Luciferase Reporter Construct

    Zhi, TaihuiLiu, QuanhongXie, TingDing, Yue...
    10页
    查看更多>>摘要：The type III secretion system (T3SS) is a key factor in the pathogenesis of Pseudomonas syringae pv. actinidiae biovar 3 (Psa3), the causal agent of a global kiwifruit bacterial canker pandemic. To monitor the T3SS expression levels in Psa3, we constructed a luciferase reporter plasmid-expressing HrpA(Psa)(3)-NLuc fusion protein. The expression of HrpA-NLuc was induced in hrp-inducing conditions whereas the level of luciferase activity correlated with the expression of hrp/hrc genes in Psa3 confirmed the reliability of the reporter construct. Based on the readout of the NLuc reporter construct, three small molecule compounds 4-methoxy-cinnamic acid, sulforaphane, and ferulic acid were determined as T3SS inhibitors in Psa3, whereas sodium acetate was determined to be a T3SS inducer. Moreover, the aqueous extract of fruit inhibited the accumulation of HrpA-NLuc in Psa3 in medium and in planta. Additionally, the T3SS inhibitors suppress Psa3 virulence, whereas the T3SS inducer promotes Psa3 virulence on kiwifruit. Thus, our findings may provide clues to why the fruit is not infected by Psa3, and the Psa3 T3SS inhibitors have potential as alternatives to current nonspecific antimicrobials for disease management.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    The Copper-Binding Protein CutC Is Involved in Copper Homeostasis and Affects Virulence in the Xylem-Limited Pathogen Xylella fastidiosa

    Cobine, Paul A.de la Fuente, LeonardoZhu, XinyuGe, Qing...
    10页
    查看更多>>摘要：Copper (Cu) is an essential element that can be toxic if homeostasis is disrupted. Xylella fastidiosa, a xylem-limited plant pathogenic bacterium that causes disease in many economically important crops worldwide, has been exposed to Cu stress caused by wide application of Cu-containing antimicrobials used to control other diseases. However, X. fastidiosa Cu homeostasis mechanisms are still poorly understood. The potentially Cu-related protein CutC, which is involved in Cu tolerance in Escherichia coli and humans, has not been analyzed functionally in plant pathogenic bacteria. We demonstrate that recombinantly expressed X. fastidiosa CutC binds Cu and deletion of cutC gene (PD0586) in X. fastidiosa showed increased sensitivity to Cu-shock compared with wild type (WT) strain TemeculaL. When infecting plants in the greenhouse, cutC mutant showed decreased disease incidence and severity compared with WT but adding Cu exaggerated severity. Interestingly, the inoculation of cutC mutant caused reduced symptoms in the acropetal regions of plants. We hypothesize that X. fastidiosa cutC is involved in Cu homeostasis by binding Cu in cells, leading to Cu detoxification, which is crucial to withstand Cu-shock stress. Unveiling the role of cutC gene in X. fastidiosa facilitates further understanding of Cu homeostasis in bacterial pathogens.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    Taxonomic Refinement of Xanthomonas arboricola

    Zarei, SadeghTaghavi, S. MohsenRahimi, TourajMafakheri, Hamzeh...
    10页
    查看更多>>摘要：Xanthomonas arboricola comprises a number of economically important fruit tree pathogens classified within different pathovars. Dozens of nonpathogenic and taxonomically unvalidated strains are also designated as X. arboricola, leading to a complicated taxonomic status in the species. In this study, we have evaluated the whole-genome resources of all available Xanthomonas spp. strains designated as X. arboricola in the public databases to refine the members of the species based on DNA similarity indexes and core genome-based phylogeny. Our results show that, of the nine validly described pathovars within X. arboricola, pathotype strains of seven pathovars are taxonomically genuine, belonging to the core clade of the species regardless of their pathogenicity on the host of isolation (thus the validity of pathovar status). However, strains of X. arboricola pv. guizotiae and X. arboricola pv. populi do not belong to X. arboricola because of the low DNA similarities between the type strain of the species and the pathotype strains of these two pathovars. Thus, we propose to elevate the two pathovars to the rank of a species as X. guizotiae sp. nov. with the type strain CFBP 7408(T) and X. populina sp. nov. with the type strain CFBP 3123(T). In addition, other mislabeled strains of X. arboricola were scattered within Xanthomonas spp. that belong to previously described species or represent novel species that await formal description.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    Identification and Mapping of bs8, a Novel Locus Conferring Resistance to Bacterial Spot Caused by Xanthomonas gardneri

    Sharma, AnujMinsavage, Gerald, VGill, Upinder S.Hutton, Samuel F....
    11页
    查看更多>>摘要：Although cultivars possessing recessive resistance alleles provide effective control of bacterial spot of pepper (Capsicum annuum), the deployed resistance gene, bs5, is ineffective against Xanthomonas gardneri, one of the pathogenic species. Resistance against X. gardneri was identified in C. annuum accession PI 163192, and this study sought to characterize this novel resistance and to map the resistance gene(s) to the pepper genome. We crossed PI 163192 with the susceptible cultivar Early Calwonder (ECW) to develop resistant near-isogenic lines (NILs) of ECW, designated ECW80R. The novel resistance in ECW80R was determined to be quantitative, recessively inherited, and non-hypersensitive-response causing, and inhibits lesion expansion and chlorosis. Presence of the resistance in NILs decreased the in planta bacterial population by ninefold compared with ECW. Bulked segregant analysis of resistant and susceptible individuals from an F-2 population using whole genome single nucleotide polymorphisms identified a major resistance locus within an approximate 6-Mbp interval on the subtelomeric region of chromosome 11. We developed markers spanning this region and used these to genotype backcross F-2 populations, which further delimited the resistance locus within a 2.3-Mbp interval. The novel resistance locus has been designated bs8. ECW80R and the linked markers developed in this study should prove useful for breeders seeking to advance this resistance into commercially relevant germplasm and for pyramiding bs8 with other resistance alleles such as bs5 and bs6. The allele bs8 will help prolong the durability of bacterial spot resistance in pepper and improve resistance to multiple species of Xanthomonas.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    Efficacy of Fungicides Used to Manage Downy Mildew in Cucumber Assessed with Multiple Meta-Analysis Techniques

    Keinath, Anthony P.Toporek, Sean M.
    8页
    查看更多>>摘要：A nationwide, quantitative synthesis of fungicide efficacy data on management of cucurbit downy mildew (CDM) caused by Pseudoperonospora cubensis is needed to broadly evaluate fungicide performance. Three-level meta-analysis, three-level meta-regression, and network meta-analyses were conducted on data from 46 cucumber (Cucumis sativus) CDM fungicide efficacy studies conducted in the eastern United States retrieved from Plant Disease Management Reports published between 2009 and 2018. Three response variables were examined in each analysis: disease severity, marketable yield, and total yield, from which percent disease control and percent yield return compared with nontreated controls was calculated. Moderator variables used in the three-level meta-analysis or three-level meta-regression included year, disease pressure, number of fungicide applications, and slicing or pickling cucumbers. In the network meta-analysis, fungicides were grouped by common combinations of Fungicide Resistance Action Committee Codes and modes of action. Overall, fungicides significantly (P < 0.001) reduced disease severity and increased marketable and total yields, resulting in a mean 54.0% disease control and 61.9% marketable and 73.3% total yield return. Subgroup differences were observed for several fungicide applications, control plot disease severity, and cucumber type for marketable yield. Based on the meta-regression analysis for disease severity by year, fungicide efficacy has been decreasing from 2009 to 2018, potentially indicating broad development of fungicide resistance over time. Treatments containing quinone inside inhibitors, pyridinylmethyl-benzamides, and protectants and treatments containing oxysterol binding protein inhibitors and protectants most effectively reduced disease severity. The most effective fungicide combinations for disease control did not always result in the highest yield return.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    Rapidly Increasing Boscalid Resistance in Corynespora cassiicola in China

    Sun, BingxueZhu, GuangxueXie, XuewenChai, Ali...
    8页
    查看更多>>摘要：Corynespora leaf spot caused by Corynespora cassiicola is an important foliar disease in cucumber. Succinate dehydrogenase inhibitors are the main fungicides used to control this disease. With the application of succinate dehydrogenase inhibitors (SDHIs) in the field, boscalid-resistant isolates have been continuously detected in the field. Resistance monitoring programs were performed to investigate the frequency and genotypes of resistant isolates. In our resistance monitoring, the frequency of resistant isolates rapidly increased from 9.68 to 85.88% in 2005 to 2020. Nine genotypes conferring SDHI resistance were found in resistant isolates, with different levels of resistance to SDHIs: B-H278R, B-H278L, B-H278Y, B-I280V, C-N75S, C-S73P, D-D95E, D-H105R, and D-G109V. The first sdh mutation was detected in Hebei Province in China, conferring an amino acid substitution at codon 278 in the sdhB subunit from histidine to tyrosine (B-H278Y), and it was the dominant resistance genotype in 2014 to 2015. Subsequently, other genotypes were gradually detected in the field, and the dominant mutations varied across years and across regions. The newest genotype (B-H278L) conferring SDHI resistance was found in 2020. To the best of our knowledge, this is the first report of C. cassiicola in cucumber. To date, multiple resistance to SDHIs, quinone outside inhibitors, benzimidazole fungicides, and dicarboximide fungicides have been detected, accounting for 75.64% of SDHI-resistant isolates. Therefore, the above four fungicides must be strictly restricted, and further monitoring work in other provinces with more isolates should be performed in the future.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    Development, Validation, and Utility of Species-Specific Diagnostic Markers for Detection of Peronospora belbahrii

    Childs, K. L.Tian, M.Quesada-Ocampo, L. M.Standish, J. R....
    9页
    查看更多>>摘要：Peronospora belbahrii is an oomycete and the cause of basil downy mildew, one of the most destructive diseases affecting basil production worldwide. Disease management is challenging due to wind-dispersed sporangia and contaminated seed; therefore, identifying P. belbahrii in seed lots before sale or planting or in the field before symptoms develop could allow for timely deployment of disease management strategies. In this study, a draft genome assembly and next-generation sequencing reads for P. belbahrii, as well as publicly available DNA-seq and RNA-seq reads of several other downy mildew pathogens, were incorporated into a bioinformatics pipeline to predict P. belbahrii-specific diagnostic markers. The specificity of each candidate marker was validated against a diverse DNA collection of P. belbahrii, host tissue, and related oomycetes using PCR. Two species-specific markers were identified and used as templates to develop a highly sensitive probe-based real-time quantitative PCR (qPCR) assay that could detect P. belbahrii in leaf tissue and seed samples. Both markers were capable of reliably detecting as low as 500 fg/mu l of P. belbahrii genomic DNA and as few as 10 sporangia. The qPCR assay was then validated with seed samples collected from a basil cultivar experiment. In total, 48 seed samples were collected and tested; P. belbahrii was detected in samples of all cultivars at estimated concentrations of 600 fg/mu l up to 250 pg/mu l and at as few as 10 sporangia up to >1,000 sporangia. The markers and assays are valuable for diagnostics and identifying P. belbahrii-contaminated seed lots to mitigate the effects of future basil downy mildew epidemics.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc

    The Diversity and Abundance of Soft Rot Pectobacteriaceae Along the Durance River Stream in the Southeast of France Revealed by Multiple Seasonal Surveys

    Ben Moussa, HajarBertrand, ClaireRochelle-Newall, EmmaFiorini, Sarah...
    10页
    查看更多>>摘要：Although irrigation water is frequently assessed for the presence of plant pathogens, large spatial and temporal surveys that provide clues on the diversity and circulation of pathogens are missing. We evaluate the diversity of soft rot Pectobacteriaceae (SRP) of the genera Dickeya and Pectobacterium over 2 years in a temperate, mixed-use watershed. The abundance of isolated strains correlates with the agricultural gradient along the watershed with a positive correlation found with temperature, nitrate, and dissolved organic carbon water concentration. We characterized 582 strains by amplification and sequencing of the gapA gene. Multilocus sequence analysis, performed with three housekeeping genes for 99 strains, and core genome analysis of 38 sequenced strains, confirmed for all the strains but one, the taxonomic assignation obtained with the sole gapA sequence. Pectobacterium spp. (549 isolates) were far more abundant than Dickeya spp. (33 isolates). Dickeya spp. were only observed in the lower part of the river when water temperature was >19 degrees C, and we experimentally confirmed a decreased fitness of several Dickeya spp. at 8 degrees C in river water. D. oryzae dominates the Dickeya spp. and P. versatile and P. aquaticum dominate the Pectobacterium spp., but their repartition along the watershed was different, with P. versatile being the only species regularly recovered all along the watershed. Excepting P. versatile, the Dickeya and Pectobacterium spp. responsible for disease outbreak on crops were less abundant or rarely detected. This work sheds light on the various ecological behaviors of different SRP types in stream water and indicates that SRP occupation is geographically structured.
      原文链接:
    • NSTL
    • Amer Phytopathological Soc