Fire blight, caused by Erwinia amylovora, is an economically important disease in apples and pears worldwide. This pathogen relies on the type III secretion system (T3SS) to cause disease. Compounds that inhibit the function of the T3SS (T3SS inhibitors) have emerged as alternative strategies for bacterial plant disease management, as they block bacterial virulence without affecting growth, unlike traditional antibiotics. In this study, we investigated the mode of action of a T3SS inhibitor named TS108, a plant phenolic acid derivative, in E. amylovora. We showed that adding TS108 to an in vitro culture of E. amylovora repressed the expression of several T3SS regulon genes, including the master regulator gene hrpL. Further studies demonstrated that TS108 negatively regulates CsrB, a global regulatory small RNA, at the posttranscriptional level, resulting in a repression of hrpS, which encodes a key activator of hrpL. Additionally, TS108 has no impact on the expression of T3SS in Dickeya dadantii or Pseudomonas aeruginosa, suggesting that its inhibition of the E. amylovora T3SS is likely species specific. To better evaluate the performance of T3SS inhibitors in fire blight management, we conducted five independent field experiments in four states (Michigan, New York, Oregon, and Connecticut) from 2015 to 2022 and observed reductions in blossom blight incidence as high as 96.7% compared with untreated trees. In summary, the T3SS inhibitors exhibited good efficacy against fire blight.

Many Gram-negative pathogenic bacteria rely on a functional type III secretion system (T3SS), which injects multiple effector proteins into eukaryotic host cells, for their pathogenicity. Genetic studies conducted in different host-microbe pathosystems often revealed a sophisticated regulatory mechanism of their T3SSs, suggesting that the expression of T3SS is tightly controlled and constantly monitored by bacteria in response to the ever-changing host environment. Therefore, it is critical to understand the regulation of T3SS in pathogenic bacteria for successful disease management. This review focuses on a model plant pathogen, Dickeyadadantii, and summarizes the current knowledge of its T3SS regulation. We highlight the roles of several T3SS regulators that were recently discovered, including the transcriptional regulators: FlhDC, RpoS, and SlyA; the post-transcriptional regulators: PNPase, Hfq with its dependent sRNA ArcZ, and the RsmA/B system; and the bacterial second messenger cyclic-di-GMP (c-di-GMP). Homologs of these regulatory components have also been characterized in almost all major bacterial plant pathogens like Erwiniaamylovora, Pseudomonassyringae, Pectobacterium spp., Xanthomonas spp., and Ralstonia spp. The second half of this review shifts focus to an in-depth discussion of the innovation and development of T3SS inhibitors, small molecules that inhibit T3SSs, in the field of plant pathology. This includes T3SS inhibitors that are derived from plant phenolic compounds, plant coumarins, and salicylidene acylhydrazides. We also discuss their modes of action in bacteria and application for controlling plant diseases.

Fire blight, caused by Erwinia amylovora, is a destructive disease of pome fruit trees. In the United States, apple and pear growers rely on applications of copper and antibiotics during bloom to control fire blight, but such methods have already led to regional instances of resistance. In this study, we used transcriptome analyses and field trials to evaluate the effectiveness of three commercially available plant defense elicitors and one plant growth regulator for fire blight management. Our data indicated that foliar applications of acibenzolar- S-methyl (ASM; Actigard 50WG) triggered a strong defense-related response in apple leaves, whereas applications of Bacillus mycoides isolate J (LifeGard WG) or Reynoutria sachalinensis extract (Regalia) did not. Genes upregulated by ASM were enriched in the biological processes associated with plant immunity, such as defense response and protein phosphorylation. The expression of several pathogenesis-related (PR) genes was induced by ASM as well. Surprisingly, many differentially expressed genes in ASM-treated apple leaves overlapped with those induced by treatment with prohexadione-calcium (ProCa; Apogee), a plant growth regulator that suppresses shoot elongation. Further analysis suggested that ProCa likely acts similarly to ASM to stimulate plant immunity because genes involved in plant defense were shared and significantly upregulated (more than twofold) by both treatments. Our field trials agreed with the transcriptome study, demonstrating that ASM and ProCa exhibit the best control performance relative to the other biopesticides. Taken together, these data are pivotal for the understanding of plant response and shed light on future improvements of strategies for fire blight management.

Erwinia amylovora is a plant pathogen causing necrotrophic fire blight disease of apple, pear, and other rosaceous plants. This bacterium colonizes host vascular tissues via the production of exopolysaccharides (EPSs) including amylovoran. It is well-established that the nearly ubiquitous plasmid pEA29 of E. amylovora is an essential virulence factor, but the underlying mechanism remains uncharacterized. Here, we demonstrated that pEA29 was required for E. amylovora to produce amylovoran and to form a biofilm, and this regulation was dependent on the thiamine biosynthesis operon thiOSGF. We then conducted carbohydrate and genetic analyses demonstrating that the thiamine-mediated effect on amylovoran production was indirect, as cells lacking thiOSGF produced an EPS that did not contain glucuronic acid, one of the key components of amylovoran, whereas the transcriptional activity and RNA levels of the amylovoran biosynthesis genes were not altered. Alternatively, addition of exogenous thiamine restored amylovoran production in the pEA29-cured strain of E. amylovora and positively impacted amylovoran production in a dose-dependent manner. Individual deletion of several chromosomal thiamine biosynthesis genes also affected amylovoran production, implying that a complete thiamine biosynthesis pathway is required for the thiamine-mediated effect on amylovoran production in E. amylovora. Finally, we determined that an imbalanced tricarboxylic acid cycle negatively affected amylovoran production, which was restored by addition of exogenous thiamine or overexpression of the thiOSGF operon. In summary, our report revealed a novel signaling pathway that impacts E. amylovora virulence in which thiamine biosynthesis enhances bacterial respiration that provides energetic requirements for the biosynthesis of EPS amylovoran.[ ] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Erwinia amylovora is a bacterial pathogen that causes fire blight, an important disease in apples and pears. Applying the antibiotic streptomycin during the phenological bloom stage is considered the most effective management tactic for fire blight. Although streptomycin-resistant (SmR) E. amylovora populations have emerged in major U.S. apple-producing regions, antibiotic resistance data for medium- to small-sized apple-producing regions such as the Midwest are still lacking. This Short Communication article collected symptomatic fire blight samples from Iowa apple orchards during 2022 and 2023, where recent fire blight outbreaks persisted despite streptomycin use. Among E. amylovora isolates from seven counties in Central and Eastern Iowa, approximately 90% of them were SmR. All SmR isolates exhibited a single base pair mutation in codon 43 of the rpsL gene, conferring resistance to streptomycin levels exceeding 1,000 μg/ml. Through clustered regularly interspaced short palindromic repeat (CRISPR) analysis, we characterized two E. amylovora genotypes unique to our region. Whole genome sequencing of one representative SmR isolate, IA01, confirmed its CRISPR genotype, and subsequent phylogenetic analysis suggested that IA01 is genetically similar to Michigan isolates and distinct from those in eastern and western regions of North America. Furthermore, the disease-causing ability of IA01 was comparable to that of the highly virulent Ea110 strain, a streptomycin-sensitive strain isolated from Michigan, in immature pears. Overall, this study underscores the urgent need for regional strategies to address antibiotic resistance and provides insights into its genetic basis and geographic distribution, which are crucial for sustainable orchard management.

The secondary shoot blight phase of fire blight is a critical component of disease epidemics in apples, pears, and other Rosaceae family plants with infection occurring at the tips of vigorously growing branches. Shoot blight infections are exacerbated in modern high-density apple plantings, where growers emphasize maximizing tree growth to recapture planting costs and increase yields of high-quality fruit. The overarching goal of this study was to develop new strategies for shoot blight management that do not impact the growth and yield of young apple trees. ‘Gala’ apple trees of various ages were inoculated with the fire blight pathogen Erwinia amylovora. Being treated with a combination of reduced rate mixtures of prohexadione calcium (ProCa; 6-12× rate reduction) with acibenzolar-S-methyl (ASM; 2× reduction) resulted in a significant decrease in shoot blight incidence and severity without significant impacts on branch growth. The systemic spread of E. amylovora was significantly reduced in trees sprayed with these lower-rate mixtures. Comparable rates of either treatment alone were not as effective in reducing lesion length. A transcriptomic analysis revealed a synergistic effect in which the expression of marker genes associated with systemic acquired resistance was higher in apple trees sprayed with the low-rate mixture of ProCa + ASM than with either compound alone. We conclude that the combination of ProCa + ASM at reduced rates is an effective treatment for the shoot blight phase of fire blight without impacting horticultural practices associated with high-density apple production.