Previous studies have suggested that there may be species-specific differences in the metabolism of polybrominated diphenyl ethers (PBDEs) among different fish species. In this study, we investigated ...the in vitro hepatic metabolism of eleven individual PBDE congeners (tri- through decaBDEs) in three different fish species: rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio), and Chinook salmon (O. tschwatcha). In addition, we evaluated the influence of PBDE structural characteristics (i.e., bromine substitution patterns) on metabolism. Six of the eleven congeners we evaluated, BDEs 99, 153, 183, 203, 208, and 209, were metabolically debrominated to lower brominated congeners. All of the congeners that were metabolized contained at least one meta-substituted bromine. Metabolites were not detected for congeners without one meta-substituted bromine (e.g., BDEs 28, 47, and 100). Metabolite formation rates were generally 10 to 100 times faster in carp than in trout and salmon. BDEs 47, 49, 101, 154, and 183 were the major metabolites observed in all three species with the exception of BDE 47, which was only detected in carp. Carp demonstrated a preference toward meta-debromination, while trout and salmon debrominated meta- and para-bromine atoms to an equal extent. We compared glutathione-S-transferase (GST) and deiodinase (DI) activity among all three species as these enzyme systems have been hypothesized to play a role in PBDE debromination in teleosts. Carp exhibited a preference for meta-deiodination of the thyroid hormone thyroxine, which was consistent with the preference for meta-debromination of PBDEs observed in carp.
Fungal growth inhibition on solid media has been historically measured and calculated based on the average of perpendicular diameter measurements of growth on fungicide amended media. We investigated ...the sensitivity of the calculated area (DA) and the measured area (MA) for assessing fungicide growth inhibition of the ascomycete, Phyllosticta citricarpa on solid media. Both the calculated, DA and the actual measured area, MA were adequate for distinguishing significant treatment effects of fungicide on fungal growth, however MA was more sensitive at identifying significant differences between the controls and fungicide concentrations below 5 ppm.
Bacterial spot of pepper (BSP), caused by four different Xanthomonas species, primarily X. euvesicatoria (Xe), poses a significant challenge in pepper cultivation. Host resistance is considered the ...most important approach for BSP control, offering long-term protection and sustainability. While breeding for resistance to BSP for many years focused on dominant R genes, introgression of recessive resistance has been a more recent focus of breeding programs. The molecular interactions underlying recessive resistance remain poorly understood. In this study, transcriptomic analyses were performed to elucidate defense responses triggered by Xe race P6 infection by two distinct pepper lines: the Xe-resistant line ECW50R containing bs5, a recessive resistance gene that confers resistance to all pepper Xe races, and the Xe-susceptible line ECW. The results revealed a total of 3357 upregulated and 4091 downregulated genes at 0, 1, 2, and 4 days post-inoculation (dpi), with the highest number of differentially expressed genes (DEGs) observed at 2 dpi. Pathway analysis highlighted DEGs in key pathways such as plant-pathogen interaction, MAPK signaling pathway, plant hormone signal transduction, and photosynthesis - antenna proteins, along with cysteine and methionine metabolism. Notably, upregulation of genes associated with PAMP-Triggered Immunity (PTI) was observed, including components like FLS2, Ca-dependent pathways, Rboh, and reactive oxygen species (ROS) generation. In support of these results, infiltration of ECW50R leaves with bacterial suspension of Xe led to observable hydrogen peroxide accumulation without a rapid increase in electrolyte leakage, suggestive of the absence of Effector-Triggered Immunity (ETI). Furthermore, the study confirmed that bs5 does not disrupt the effector delivery system, as evidenced by incompatible interactions between avirulence genes and their corresponding dominant resistant genes in the bs5 background. Overall, these findings provide insights into the molecular mechanisms underlying bs5-mediated resistance in pepper against Xe and suggest a robust defense mechanism in ECW50R, primarily mediated through PTI. Given that bs5 provides early strong response for resistance, combining this resistance with other dominant resistance genes will enhance the durability of resistance to BSP.
Citrus black spot (CBS) caused by Phyllosticta citricarpa, is the most recent introduction of an exotic citrus pathogen into Florida and has been a challenge to control to date. Understanding the ...dispersal pattern of the disease within affected groves is vital in developing effective control strategies to limit the spread of the disease. The spatial pattern of CBS-affected trees was studied in two commercial 'Valencia' orange groves over three consecutive citrus seasons. Cluster analyses based on nearest-neighbor distance (F, G and J-functions) and pairwise distances between points (Ripley's K function, Besag's L function and the pair correlation function, g) were used to test the hypothesis of complete spatial randomness (CSR) of CBS infected trees within the groves. In both groves, the hypothesis of CSR was rejected for all tests performed including quadrats testing (2 × 2 trees up to 10 × 10 trees). The relationship between tree age and disease was assessed at one experimental site. Citrus trees bearing fruit for the first time accounted for approximately 13% of trees positive for disease and were located within areas of heavy disease pressure. These findings support short distance movement of inoculum as the main spread of disease in the groves studied.
BACKGROUND: Huanglongbing (HLB) or citrus greening is a bacterial disease vectored by the Asian citrus psyllid (ACP) causing tree decline, and yield loss. Vector control and foliar nutrition are used ...in Florida to slow the spread of HLB and mitigate debilitating effects of the disease. A four year replicated field study was initiated February 2008 in a 5.2‐ha commercial block of young ‘Valencia’ orange trees employing a factorial design to evaluate individual and compound effects of vector management and foliar nutrition. Insecticides were sprayed during tree dormancy and when psyllid populations exceeded a nominal threshold. A mixture consisting primarily of micro‐ and macro‐nutrients was applied three times a year corresponding to the principal foliar flushes. RESULTS: Differences in ACP numbers from five‐ to 13‐fold were maintained in insecticide treated and untreated plots. Incidence of HLB estimated by polymerase chain reaction (PCR), rose from 30% at the beginning of the study to 95% in only 18 months. Highest yields all four years were seen from trees receiving both foliar nutrition and vector control. Production for these trees in the fourth year was close to the pre‐HLB regional average for 10 year old ‘Valencia’ on ‘Swingle’. Nevertheless, at current juice prices, the extra revenue generated from the combined insecticide and nutritional treatment did not cover the added treatment costs. CONCLUSIONS: This experiment demonstrated that vector control, especially when combined with enhanced foliar nutrition, could significantly increase yields in a citrus orchard with high incidence of HLB. Economic thresholds for both insecticide and nutrient applications are needed under different market and environmental conditions.
Citrus black spot (CBS) caused by the fungus Phyllosticta citricarpa occurs in tropical and sub-tropical citrus production regions and affects all varieties of citrus. In Florida, the disease cycle ...is unique, having only the asexual spore. This work examines incidence and severity of CBS (hard spot symptoms) on fruit in two citrus groves during 2013-2014, 2014-2015 (Grove III) and 2015-2016 (Grove II) citrus seasons. Disease incidence and severity on fruit were analyzed based on citrus season, side of tree evaluated, height within the canopy, tree health, and tree age. Results indicate an increase in CBS incidence in Grove III between 2013-2014 and 2014-2015 seasons, with more infected or symptomatic fruit on the road side of the canopy and a higher incidence above 2 meters. Tree health status affected incidence but not severity and tree age had a significant effect on severity of CBS in Grove II. Analysis of weather data conducive for infection, between 2010 and 2017, indicated an average of 172 days per year (range: 104-261 days) when the temperature (15-35 °C) and relative humidity (RH ≥ 90% for 8 consecutive hours) were conducive for infection of fruit and an average of 98 days per year (range: 72-123 days) when the fruit were susceptible to infection.
Recombination is a major driver of evolution in bacterial populations, because it can spread and combine independently evolved beneficial mutations. Recombinant lineages of bacterial pathogens of ...plants are typically associated with the colonization of novel hosts and the emergence of new diseases. Here we show that recombination between evolutionarily and phenotypically distinct plant-pathogenic lineages generated recombinant lineages with unique combinations of pathogenicity and virulence factors.
and
are two closely related lineages causing bacterial spot disease on tomato and pepper worldwide. We sequenced the genomes of atypical strains collected from tomato in Nigeria and observed recombination in the type III secretion system and effector genes, which showed alleles from both
and
Wider horizontal gene transfer was indicated by the fact that the lipopolysaccharide cluster of one strain was most similar to that of a distantly related
pathogen of barley. This strain and others have experienced extensive genomewide homologous recombination, and both species exhibited dynamic open pangenomes. Variation in effector gene repertoires within and between species must be taken into consideration when one is breeding tomatoes for disease resistance. Resistance breeding strategies that target specific effectors must consider possibly dramatic variation in bacterial spot populations across global production regions, as illustrated by the recombinant strains observed here.
The pathogens that cause bacterial spot of tomato and pepper are extensively studied models of plant-microbe interactions and cause problematic disease worldwide. Atypical bacterial spot strains collected from tomato in Nigeria, and other strains from Italy, India, and Florida, showed evidence of genomewide recombination that generated genetically distinct pathogenic lineages. The strains from Nigeria and Italy were found to have a mix of type III secretion system genes from
and
, as well as effectors from
These genes and effectors are important in the establishment of disease, and effectors are common targets of resistance breeding. Our findings point to global diversity in the genomes of bacterial spot pathogens, which is likely to affect the host-pathogen interaction and influence management decisions.
Cucurbit yellow stunting disorder virus (CYSDV) is a single-stranded positive-sense RNA virus that produces devastating disease in watermelon and squash. Foliar symptoms of CYSDV consist of ...interveinal yellowing, brittleness, and thickening of older leaves leading to reduced plant vigor. A rapid diagnostic method for CYSDV would facilitate early detection and implementation of best viral-based management practices. We developed a rapid isothermal reverse transcription-recombination polymerase amplification (exo RT-RPA) assay for the detection of CYSDV. The primers and a 6-fluorescein amidite (6-FAM) probe were developed to target the nucleocapsid gene. The real-time assay detected CYSDV at 2.5 pg purified total RNA extracted from CYSDV-infected leaf tissue and corresponded to 10 copies of the target molecule. The assay was specific and did not cross-react with other common cucurbit viruses found in Florida and Georgia. The performance of the exo RT-RPA was evaluated using crude extract from 21 cucurbit field samples and demonstrated that the exo RT-RPA is a rapid procedure, thus providing a promising novel alternative approach for the detection of CYSDV.
•An exo RT-RPA assay developed for simple and rapid detection of Cucurbit yellow stunting disorder virus (CYSDV).•The exo RT-RPA primers/probe were highly specific and sensitive in the detection of CYSDV.•The assay detected CYSDV from crude extract of watermelon and squash leaves.
Genotyping-by-sequencing (GBS) was performed on 257 Phytophthora infestans isolates belonging to four clonal lineages to study within-lineage diversity. The four lineages used in the study were US-8 ...(n = 28), US-11 (n = 27), US-23 (n = 166), and US-24 (n = 36), with isolates originating from 23 of the United States and Ontario, Canada. The majority of isolates were collected between 2010 and 2014 (94%), with the remaining isolates collected from 1994 to 2009, and 2015. Between 3,774 and 5,070 single-nucleotide polymorphisms (SNPs) were identified within each lineage and were used to investigate relationships among individuals. K-means hierarchical clustering revealed three clusters within lineage US-23, with US-23 isolates clustering more by collection year than by geographic origin. K-means hierarchical clustering did not reveal significant clustering within the smaller US-8, US-11, and US-24 data sets. Neighbor-joining (NJ) trees were also constructed for each lineage. All four NJ trees revealed evidence for pathogen dispersal and overwintering within regions, as well as long-distance pathogen transport across regions. In the US-23 NJ tree, grouping by year was more prominent than grouping by region, which indicates the importance of long-distance pathogen transport as a source of initial late blight inoculum. Our results support previous studies that found significant genetic diversity within clonal lineages of P. infestans and show that GBS offers sufficiently high resolution to detect sub-structuring within clonal populations.
Bacterial spot disease was first reported from South Africa by Ethel M. Doidge in 1920. In the ensuing century after the initial discovery, the pathogen has gained global attention in plant pathology ...research, providing insights into host-pathogen interactions, pathogen evolution, and effector discovery, such as the first discovery of transcription activation-like effectors, among many others. Four distinct genetic groups, including
(proposed name:
pv.
),
(proposed name:
pv.
),
(proposed name:
pv.
), and
, are known to cause bacterial spot disease. Recently, a new race of a bacterial spot pathogen, race T5, which is a product of recombination between at least two
species, was reported in Nigeria. In this review, our focus is on the progress made on the African continent,
progress made in the global bacterial spot research community to provide a body of information useful for researchers in understanding the diversity, evolutionary changes, and management of the disease in Africa.