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Fusarium wilt of watermelon (Citrullus lanatus) caused by Fusarium oxysporum f. sp. niveum (Fon), has become an increasing concern of farmers in the southeastern USA, especially in Florida. Management of this disease, most often through the use of resistant cultivars and crop rotation, requires an accurate understanding of an area's pathogen population structure and phenotypic characteristics. This study improved the understanding of the state's pathogen population by completing multilocus sequence analysis (MLSA) of two housekeeping genes (BT and TEF) and two loci (ITS and IGS), aggressiveness and race-determining bioassays on 72 isolates collected between 2011 and 2015 from major watermelon production areas in North, Central, and South Florida. Multilocus sequence analysis (MLSA) failed to group race 3 isolates into a single large clade; moreover, clade membership was not apparently correlated with aggressiveness (which varied both within and between clades), and only slightly with sampling location. The failure of multilocus sequence analysis using four highly conserved housekeeping genes and loci to clearly group and delineate known Fon races provides justification for future whole genome sequencing efforts whose more robust genomic comparisons will provide higher resolution of intra-species genetic distinctions. Consequently, these results suggest that identification of Fon isolates by race determination alone may fail to detect economically important phenotypic characteristics such as aggressiveness leading to inaccurate risk assessment. [ABSTRACT FROM AUTHOR]

Bacterial spot disease caused by Xanthomonas euvesicatoria is one of the major constraints for pepper production in Florida and worldwide. The common prevalence of copper (Cu)-tolerant X. euvesicatoria strains has reduced the efficacy of Cu based bactericides against bacterial spot of pepper. In this study, we evaluated antimicrobial activity of three Cu-based nanocomposites, namely, core-shell Cu (CS–Cu), fixed-quat Cu (FQ-Cu) and mixed-valence Cu (MV-Cu), along with Kocide® 3000, which is a micron size metallic Cu bactericide and is commercially available, both in vitro and in greenhouse trials, against a Cu-tolerant strain of X. euvesicatoria. In repeated in vitro tests, X. euvesicatoria growth was completely inhibited 24 h after exposure of bacterial cells to 500 and 1000 μg/ml of MV-Cu and all concentrations of FQ-Cu and CS-Cu (100, 200, 500, and 1000 μg/ml) treated for 1 h. Micron size copper, even at 1000 μg/ml, had no statistical difference in growth in comparison to the untreated control (P = 0.05; LSD). In the repeated greenhouse experiments, plants treated with all the copper composites exhibited less bacterial spot severity than untreated control and similar or significantly less than Cu-mancozeb (P < 0.05). However, MV-Cu was the only Cu composite with no phytotoxicity on plants under controlled conditions. This study shows potential to use Cu-based nanoparticles for efficient management of bacterial spot on pepper. [ABSTRACT FROM AUTHOR]

Summer squash (Cucurbita pepo L.) is a major vegetable crop produced in Georgia and Florida during the fall season. This production is vulnerable to whitefly (Bemisisia tabaci Genn.)-transmitted viruses that lead to severe yield losses. Over the past several years, whitefly populations have increased during the fall, thus leading to an increase in whitefly-transmitted viruses such as Cucurbit leaf crumple virus (CuLCrV) and Cucurbit yellow stunting disorder virus (CYSDV). Whitefly management for summer squash relies on the use of insecticides and can be costly without providing adequate management of the viruses. Deployment of host resistance to whiteflies and their transmitted viruses (CuLCrV and CYSDV) is the best strategy for mitigating yield loss of summer squash; however, no resistant cultivars are commercially available. In the current study, resistance or tolerance to whiteflies, CuLCrV, and CYSDV was determined for squash germplasm from the U.S. Department of Agriculture (USDA) Germplasm Resources Information Network (GRIN), university breeding programs, and commercial companies in Georgia and Florida across 2 years. In both locations and years, visual virus symptom severity scores were collected and a quantitative polymerase chain reaction (qPCR) was used to determine the CuLCrV viral load and CYSDV presence in Georgia. Whitefly-induced feeding damage was evaluated by directly assessing the intensity of silverleaf symptoms and visual counts of whitefly adults on the foliage in the field or in photographs. Virus symptom severity was lower in C. moschata Duchesne ex Poir. genotypes, namely, PI 550689, PI 550692, PI 550694, PI 653064, and Squash Betternut 900, than in other evaluated genotypes. Two C. pepo accessions were common between both locations for viral severity (PI 442294) or viral severity and viral load (PI 171625). Lower CuLCrV loads were identified in C. ecuadorensis Cutler & Whitaker (PI 540895), and C. okeechobeensis (Small) L.H.Bailey (PI 540900) than other evaluated genotypes. Four genotypes tested negative for CYSDV during both years: C. pepo (PI 507882), C. moschata (PI 483345), C. ecuadorensis (PI 390455), and C. okeechobeensis (PI 540900); they are potential sources of resistance. Six C. moschata accessions (PI 211999, PI 550690, PI 550692, PI 550694, PI 634982, and PI 653064) showed high tolerance to silverleaf disorder and had the lowest adult whitefly counts. Collectively, the accessions identi- fied in the current study are potential sources of resistance or tolerance to whitefly and whitefly-transmitted viruses (CuLCrV and CYSDV [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The invasive mite Phyllocoptes fructiphilus Keifer (Acari: Eriophyidae) feeds on plants in the genus Rosa. Phyllocoptes fructiphilus is associated with the rose rosette emaravirus (rose rosette virus) and acts as the only known vector of rose rosette virus, the causal agent of rose rosette disease (Emaravirus). The mite P. fructiphilus is reported for the first time in the state of Florida, USA. No roses showed signs or symptoms of viral infection, and current molecular methods were unable to detect the virus. Phyllocoptes fructiphilus represents a potential threat to the Florida rose industry if rose rosette disease becomes established. [ABSTRACT FROM AUTHOR]

A highly sensitive one-step multiplex RT-PCR (OMRT-PCR) system with an internal plant gene control was developed to simultaneously detect single and mixed RNA and DNA viral pathogens affecting watermelon and squash in the Southeastern United States. Current methods rely on a singleplex detection system for several viruses which is time-consuming for large-scale field monitoring. The gene-specific primers for one DNA virus, cucurbit leaf crumple virus (CuLCrV), and three RNA viruses, cucurbit yellow stunting disorder virus (CYSDV), squash vein yellowing virus (SqVYV), and cucurbit chlorotic yellows virus (CCYV), were used for the development of OMRT-PCR. A plant gene, rubisco from cucurbits was used as an internal control in the standardization of the assay and to monitor false amplification. In addition, we examined the use of total nucleic acid as a template in OMRT-PCR for simultaneously detection of mixed viral infection. The OMRT-PCR demonstrated high sensitivity and detected all four virus infections from 1 ng of total plant nucleic acid in a single reaction. The multiple virus detection approach was successful in the diagnosis of 16 watermelon and 18 squash samples collected from commercial fields in Florida. The OMRT-PCR system is specific, rapid, sensitive, and cost-efficient for the detection of multiple viruses. This technique is expected to improve the ease in the detection of CuLCrV, CYSDV, SqVYV and CCYV and assist in the monitoring of the viral pathogens in commercial cucurbit production fields. • The OMRT-PCR assay is developed for simultaneous detection of viruses infecting cucurbits: CuLCrV, CYSDV, SqVYV and CCYV. • The newly developed OMRT-PCR assay is highly specific and sensitive for detection of these mixed viruses. • The detection limit of this OMRT-PCR system is up to 1 ng of total nucleic acid. • The plant gene internal control was used to monitoring false amplification. • This assay have the potential to reduce cost, time and labor for the diagnosis of a large number of samples. [ABSTRACT FROM AUTHOR]

In Florida, angular leaf spot, caused by Xanthomonas fragariae , was the only known bacterial disease in strawberry, which is sporadic and affects the foliage and calyx. However, from the 2019-2020 to 2023-2024 Florida strawberry seasons, unusual bacterial-like symptoms were observed in commercial farms, with reports of up to 30 % disease incidence. Typical lesions were water-soaked and angular in early stages that later became necrotic with a circular-ellipsoidal purple halo, and consistently yielded colonies resembling Pseudomonas on culture media. Strains were pathogenic on strawberry, fluorescent, oxidase- and arginine-dihydrolase-negative, elicited a hypersensitive reaction on tobacco, and lacked pectolytic activity. Although phenotypic assays, such as fatty acid methyl profiles and Biolog protocols, placed the strains into the Pseudomonas group, there was a low similarity at the species level. Further analysis using 16S rRNA genes, housekeeping genes, and whole genome sequencing showed that the strains cluster into the Pseudomonas group but do not share more than 95 % average nucleotide identity compared to representative members. Therefore, the genomic and phenotypic analysis confirm that the strains causing bacterial spot in strawberry represent a new plant pathogenic bacterial species for which we propose the name Pseudomonas fragariae sp. nov. with 20-417 T (17 T =LMG 32456 T =DSM 113340 T ) as the type strain, in relation to Fragaria × ananassa , the plant species from which the pathogen was first isolated. Future work is needed to assess the epidemiology, cultivar susceptibility, chemical sensitivity, and disease management of this possible new emerging strawberry pathogen.

Cold hardy citrus is an emerging industry in north Florida. However, it is under the threat of Candidatus Liberibacter asiaticus (CLas), the agent of the citrus disease huanglongbing. Distribution and phenology of the Asian citrus psyllid, Diaphorina citri (Kuwayama), the vector of CLas, was investigated over a 2-year sampling period in north Florida. Diaphorina citri was only found in backyard and ornamental citrus along the Gulf of Mexico, and was not observed in cultivated citrus groves during the 2 years (2017-2018) of the survey. Diaphorina citri population peaks occurred approximately 2 mo later than in central Florida with major population peaks occurring in July. The number of D. citri adults was significantly higher on CLas infected than uninfected citrus trees, whereas more nymphs were found on uninfected trees. Most D. citri were negative for CLas except in Franklin county where both infected trees and psyllids were found. We were able to find adult D. citri during all winter months, despite temperatures as low as -5.5°C. During two consecutive winters, we conducted experiments to determine D. citri cold hardiness by caging D. citri under ambient conditions in mid-November and assessing survivors in the following spring. In 2018, approximately 21%, of D. citri adults survived overwintering whereas 16% survived in 2019 despite lower temperature in 2018 than in 2019. As we are at the earliest stage of HLB infestation, management of D. citri and CLas in north Florida should focus on removal of CLas-infected trees to reduce the reservoir of pathogen., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Gummy stem blight, caused by Stagonosporopsis spp., is a major disease of cucurbits in the United States and China that is managed primarily through the use of fungicides. The objective of this study was to monitor and compare the recent fungicide resistance profiles of Stagonosporopsis spp. in Florida open-field and East China protected-structure production systems. Isolates of Stagonosporopsis spp. were evaluated for sensitivity to the commonly used fungicides azoxystrobin, boscalid, tebuconazole, and thiophanate-methyl at discriminatory rates of 0.096, 0.034, 0.128, and 100 mg/liter, respectively. Isolates were collected from Jiangsu, Jiangxi, Zhejiang, and Anhui provinces in China ( n = 69), 12 counties in Florida ( n = 89), and one county in Georgia ( n = 6). More than 50% of isolates from Florida and East China were resistant to thiophanate-methyl. Boscalid resistance was detected in both isolate collections but was two times more frequent in China. Resistance to azoxystrobin was detected in 66% of isolates in Florida but only 7% in China. Tebuconazole was effective in controlling the mycelia growth of Stagonosporopsis spp. in both collections. The results indicate that both production systems currently face similar challenges related to the development of fungicide resistance in Stagonosporopsis spp. However, the resistance profiles are unique for both production systems.

Multispectral imaging is increasingly used in specialty crops, but its benefits in assessment of disease severity and improvements in conventional scouting practice are unknown. Multispectral imaging was conducted using an unmanned aerial vehicle (UAV), and data were analyzed for five flights from Florida and Georgia commercial watermelon fields in 2017. The fields were rated for disease incidence and severity by extension agents and plant pathologists at randomized locations (i.e., conventional scouting) followed by ratings at locations that were identified by differences in normalized difference vegetation index (NDVI) and stress index (i.e., UAV-assisted scouting). Diseases identified by the scouts included gummy stem blight, anthracnose, Fusarium wilt, Phytophthora fruit rot, Alternaria leaf spot, and cucurbit leaf crumple disease. Disease incidence and severity ratings were significantly different between conventional and UAV-assisted scouting ( P < 0.01, Bhapkar/exact test). Higher severity ratings of 4 and 5 on a scale of 1 to 5 from no disease to complete loss of the canopy were more consistent after the scouts used the multispectral images in determining sampling locations. The UAV-assisted scouting locations had significantly lower green, red, and red edge NDVI values and higher stress index values than the conventional scouting areas ( P < 0.05, ANOVA/Tukey), and this corresponded to areas with higher disease severity. Conventional scouting involving human evaluation remains necessary for disease validation. Multispectral imagery improved watermelon field scouting owing to increased ability to identify disease foci and areas of concern more rapidly than conventional scouting practices with early detection of diseases 20% more often using UAV-assisted scouting.

Field trials were conducted at two locations in Florida to evaluate transgenic tomato expressing the ELONGATION FACTOR TU RECEPTOR (EFR) gene from Arabidopsis thaliana, the Bs2 gene from pepper, or both Bs2 and EFR (Bs2/EFR) for managing bacterial wilt caused by Ralstonia solanacearum and bacterial spot caused by Xanthomonas perforans. Expression of EFR or Bs2/EFR in the susceptible genotype Fla. 8000 significantly reduced bacterial wilt incidence (50 to 100%) and increased total yield (57 to 114%) relative to lines expressing only Bs2 or the nontransformed Fla. 8000 control, although the marketable yield was not significantly affected. Following harvest, surviving symptomatic and nonsymptomatic plants were assessed for colonization by R. solanacearum. There were no significant differences in the population at the lower stem. Interestingly, in the middle stem, no bacteria could be recovered from EFR or Bs2/EFR lines but viable bacterial populations were recovered from Bs2 and nontransformed control lines at 10 2 to 10 5 CFU/g of stem tissue. In growth-chamber experiments, the EFR transgenic tomato lines were found to be effective against seven different R. solanacearum strains isolated from the southeastern United States, indicating utility across the southeastern United States. In all of the bacterial spot trials, EFR and Bs2/EFR lines had significantly reduced disease severity (22 to 98%) compared with the Fla. 8000 control. The marketable and total yield of Bs2/EFR were significantly higher (43 to 170%) than Fla. 8000 control in three of four field trials. These results demonstrate for the first time the potential of using the EFR gene for field management of bacterial wilt and bacterial spot diseases of tomato.