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Abstract Background Cutaneous mycoses are common human infections among healthy and immunocompromised hosts, and the anthropophilic fungus Trichophyton rubrum is the most prevalent microorganism isolated from such clinical cases worldwide. The aim of this study was to determine the transcriptional profile of T. rubrum exposed to various stimuli in order to obtain insights into the responses of this pathogen to different environmental challenges. Therefore, we generated an expressed sequence tag (EST) collection by constructing one cDNA library and nine suppression subtractive hybridization libraries. Results The 1388 unigenes identified in this study were functionally classified based on the Munich Information Center for Protein Sequences (MIPS) categories. The identified proteins were involved in transcriptional regulation, cellular defense and stress, protein degradation, signaling, transport, and secretion, among other functions. Analysis of these unigenes revealed 575 T. rubrum sequences that had not been previously deposited in public databases. Conclusion In this study, we identified novel T. rubrum genes that will be useful for ORF prediction in genome sequencing and facilitating functional genome analysis. Annotation of these expressed genes revealed metabolic adaptations of T. rubrum to carbon sources, ambient pH shifts, and various antifungal drugs used in medical practice. Furthermore, challenging T. rubrum with cytotoxic drugs and ambient pH shifts extended our understanding of the molecular events possibly involved in the infectious process and resistance to antifungal drugs.

Florida's strawberry industry has been facing an emerging threat after several outbreaks of an aggressive Neopestalotiopsis sp. affecting fruit, leaf, and crown caused severe yield losses. Our studies found Neopestalotiopsis sp. can survive from one production season to the next on crop debris in soil, which led us to evaluate the effect of fumigants on resident inoculum in strawberry crowns and soil. Different rates of 1,3-dichloropropene, chloropicrin, and metam potassium were applied to pasteurized soil inside glass jars containing infected strawberry crowns. Soil samples were taken after broadcast and bed fumigation treatments with a combination of 1,3-dichloropropene/chloropicrin at ratios of 63:35 were applied at four commercial fields. Crowns and soil samples were processed and plated on a semi-selective medium for Neopestalotiopsis spp. and colony-forming units (CFU) were counted 5 days after plating. CFU counts in crowns treated with 1,3-dichloropropene, chloropicrin, and metam potassium decreased significantly as rates increased and were described by exponential decay models. CFUs were not recovered in most of the soil samples from fumigated strawberry beds after broadcast or bed fumigation. However, CFUs were found in non-fumigated row middles between fumigated beds which can serve as a source of inoculum to start new epidemics. Chloropicrin, 1,3-dichloropropene, and metam potassium were effective on reducing Neopestalotiopsis spp. inoculum in strawberry crowns and soil, providing new evidence on the fungicidal activity of 1,3-dichloropropene. Broadcast fumigation with 1,3-dichloropropene/chloropicrin at ratios of 63:35 could potentially be used to reduce inoculum of Neopestalotiopsis sp. in severely infested fields.

Caladium ( Caladium × hortulanum ) is an ornamental plant popular for its variable and colorful foliage. In 2020, plants showing leaf spots and blight, typical of anthracnose, were found in a field trial at the University of Florida's Gulf Coast Research and Education Center in Wimauma, Florida, U.S.A. Leaf samples consistently yielded a Colletotrichum- like species with curved conidia and abundant setae production in the acervuli. The internal transcribed spacer (ITS), partial sequences of the glyceraldehyde-3-phosphate dehydrogenase gene ( gapdh ), actin gene ( act ), chitin synthase 1 gene ( chs-1 ), beta-tubulin gene ( tub2 ), and histone3 gene ( his3 ) were amplified and sequenced. BLASTN searches in the NCBI GenBank database revealed similarities to species of the Colletotrichum truncatum species complex. Phylogenetic analyses using multilocus sequence data supports a distinct species within this complex, with the closest related species being C. curcumae . Based on morphological and phylogenetic analyses, a new species of Colletotrichum , named C. caladii , is reported. Pathogenicity assays and subsequent isolation confirmed that this species was the causal agent of the disease., Competing Interests: The author(s) declare no conflict of interest.

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.

In strawberry cultivation, precise disease management is crucial for maximizing yields and reducing unnecessary fungicide use. Traditional methods for measuring leaf wetness duration (LWD), a critical factor in assessing the risk of fungal diseases such as botrytis fruit rot and anthracnose, have been reliant on sensors with known limitations in accuracy and reliability and difficulties with calibrating. To overcome these limitations, this study introduced an innovative algorithm for leaf wetness detection systems employing high-resolution imaging and deep learning technologies, including convolutional neural networks (CNNs). Implemented at the University of Florida's Plant Science Research and Education Unit (PSREU) in Citra, FL, USA, and expanded to three additional locations across Florida, USA, the system captured and analyzed images of a reference plate to accurately determine the wetness and, consequently, the LWD. The comparison of system outputs with manual observations across diverse environmental conditions demonstrated the enhanced accuracy and reliability of the artificial intelligence-driven approach. By integrating this system into the Strawberry Advisory System (SAS), this study provided an efficient solution to improve disease risk assessment and fungicide application strategies, promising significant economic benefits and sustainability advances in strawberry production.

Devastating disease outbreaks with leaf spot symptoms and fruit rot caused by a fungus identified as a cryptic species of Neopestalotiopsis have generated concern in the Florida strawberry industry. Some Neopestalotiopsis species are saprobes in soil and plant debris, but the ability of the new Neopestalotiopsis sp. to survive in strawberry debris was unknown. Therefore, the objectives of this study were to develop a semiselective medium for isolation of Neopestalotiopsis spp. and to evaluate Neopestalotiopsis sp. survival in soil and strawberry plant debris over the summer in Florida. The Neopestalotiopsis semiselective medium (NSM) inhibited growth of most fungal species pathogenic to strawberry in Florida, except Neopestalotiopsis rosae , Neopestalotiopsis sp., and Phomopsis obscurans , in addition to Penicillium spp., which are nonpathogenic. However, Neopestalotiopsis species, P . obscurans , and Penicillium spp. could be distinguished in the medium through morphological characteristics. Soil samples arbitrarily collected from six commercial fields toward the end of the season (May) and before (July) and after (September) preplant soil fumigation in the following season were processed with NSM, and Neopestalotiopsis spp. populations were calculated as CFU per gram of soil. CFU ranged from 48.3 to 2,410.8 at the end of the season and from 1.7 to 630.8 before soil fumigation, but Neopestalotiopsis spp. were not recovered after soil treatment. However, 1.7 to 25 CFU were obtained from the nontreated areas in the row middles. Neopestalotiopsis sp. survival was also evaluated on diseased strawberry plants (leaves and crowns) exposed to the environment for 17 months. On leaves, Neopestalotiopsis spp. were recovered for 6 months before leaf decomposition, whereas the number of CFU on crowns declined monthly, but colonies were still recovered during the final evaluation months. A high-resolution melting analysis confirmed most of the colonies from soil and crowns were the new Neopestalotiopsis sp. Our results show Neopestalotiopsis sp. can survive in soil and strawberry debris under Florida summer conditions and may serve as a source of inoculum for the subsequent season., Competing Interests: The author(s) declare no conflict of interest.

Aggressive strains of Neopestalotiopsis sp. have recently emerged as devastating pathogens of strawberry (Fragaria × ananassa Duchesne ex Rozier), infecting nearly all plant parts and causing severe outbreaks of leaf spot and fruit rot in Florida and globally. The development of host resistance is imperative due to the absence of fungicides that effectively inhibit Neopestalotiopsis sp. growth on an infected strawberry crop. Here, we analyzed 1578 individuals from the University of Florida's (UF) strawberry breeding program to identify and dissect genetic variation for resistance to Neopestalotiopsis sp. and to explore the feasibility of genomic selection. We found that less than 12% of elite UF germplasm exhibited resistance, with narrow-sense heritability estimates ranging from 0.28 to 0.69. Through genome-wide association studies (GWAS), we identified two loci accounting for 7%-16% of phenotypic variance across four trials and 3 years. Several candidate genes encoding pattern recognition receptors, intra-cellular nucleotide-binding leucine-rich repeats, and downstream components of plant defense pathways co-localized with the Neopestalotiopsis sp. resistance loci. Interestingly, favorable alleles at the largest-effect locus were rare in elite UF material and had previously been unintentionally introduced from an exotic cultivar. The array-based markers and candidate genes described herein provide the foundation for targeting this locus through marker-assisted selection. The predictive abilities of genomic selection models, with and without explicitly modeling peak GWAS markers as fixed effects, ranged between 0.25 and 0.59, suggesting that genomic selection holds promise for enhancing resistance to Neopestalotiopsis sp. in strawberry., (© 2024 The Author(s). The Plant Genome published by Wiley Periodicals LLC on behalf of Crop Science Society of America.)

Phytophthora cactorum is the most common causal agent of Phytophthora crown rot and leather rot of strawberry, but P. nicotianae is also responsible for the disease in Florida. Studies of P. nicotianae populations have suggested that different groups of genotypes are associated with different hosts; however, it is not yet clear how many lineages exist globally and how they are related to different production systems. The aim of this study was to determine the genetic relationships of P. nicotianae isolates from Florida strawberry with genotypes reported from other hosts, quantify the genetic variation on strawberry, and test for an association with nursery source. A total of 49 isolates of P. nicotianae were collected from strawberry plants originating from multiple nursery sources during six seasons of commercial fruit production in Florida. Microsatellite genotyping identified 28 multilocus genotypes on strawberry that were distinct among 208 isolates originating from various hosts and locations. Based on STRUCTURE analysis, two genetic groups were identified: one consisting of isolates from strawberry, and the other comprising samples from different hosts. Multilocus genotypes were shared among nursery sources, and populations defined by nursery were not differentiated. Both mating types were found among the isolates from North Carolina- and California-origin plants and in most strawberry seasons; however, a predominance of A1 was observed, and regular sexual reproduction was not supported by the data. This study reveals a unique genetic population of P. nicotianae associated with strawberry and emphasizes the vital role of nursery monitoring in mitigating disease spread., Competing Interests: The author(s) declare no conflict of interest.

Aims: To identify whether microorganisms isolated from blueberry flowers can inhibit the growth of Colletotrichum, an opportunistic plant pathogen that infects flowers and threatens yields, and to assess the impacts of floral microbes and Colletotrichum pathogens on artificial nectar sugars and honey bee consumption., Methods and Results: The growth inhibition of Colletotrichum (Colletotrichum acutatum, Colletotrichum fioriniae, and Colletotrichum gloeosporioides) was screened using both artificial nectar co-culture and dual culture plate assays. All candidate nectar microbes were screened for antagonism against a single C. acutatum isolate. Then, the top four candidate nectar microbes showing the strongest inhibition of C. acutatum (Neokomagataea thailandica, Neokomagataea tanensis, Metschnikowia rancensis, and Symmetrospora symmetrica) were evaluated for antagonism against three additional C. acutatum isolates, and single isolates of both C. fioriniae and C. gloeosporioides. In artificial nectar assays, single and three-species cultures inhibited the growth of two of four C. acutatum isolates by ca. 60%, but growth of other Colletotrichum species was not affected. In dual culture plate assays, inhibition was observed for all Colletotrichum species for at least three of four selected microbial antagonists (13%‒53%). Neither honey bee consumption of nectar nor nectar sugar concentrations were affected by any microbe or pathogen tested., Conclusions: Selected floral microbes inhibited growth of all Colletotrichum species in vitro, although the degree of inhibition was specific to the assay and pathogen examined. In all microbial treatments, nectar sugars were preserved, and honey bee preference was not affected., (Published by Oxford University Press on behalf of Applied Microbiology International 2023.)

Botrytis fruit rot (BFR) and anthracnose fruit rot (AFR) are diseases of concern to strawberry growers. Both diseases are managed mainly by fungicide applications from the nursery (plant production) to the end of the growing season (fruit production). In Florida, captan is the main broad-spectrum fungicide used to control BFR and AFR. It has been tested in many trials over the years in various programs in alternation with single-site fungicides or weekly applications. Due to its broad-spectrum activity, captan is a pivotal tool in fungicide resistance management, because resistance to several fungicides has been reported in populations causing BFR and AFR. Our objective was to determine the effectiveness and profitability of applications of captan for controlling BFR and AFR based on a univariate meta-analysis considering data from 25 field trials conducted from 2005 to 2021 in Florida. Captan applications significantly improved marketable yields and reduced BFR and AFR incidence during late- and total-season periods. Marketable yields were significantly improved even when the effect on disease control was not significant. Our results indicate 100% probability that weekly captan applications would return the investment during low, medium, and high strawberry pricing regimes, although the magnitude of the return will depend on strawberry market prices. However, the probabilities of reducing BFR, AFR, and culls were lower. Results from our meta-analysis demonstrate the value of captan as an important tool for strawberry growers. Captan applications, in addition to effectively controlling BFR and AFR and improving marketable yields, will result in investment returns at any strawberry price level., Competing Interests: The author(s) declare no conflict of interest.

Ultraviolet light at wavelengths from 254 to 283 nm/has been reported to effectively suppress powdery mildews in several crops, including some cucurbits. Its use to suppress powdery mildew ( Podosphaera xanthii ) specifically in cantaloupe has not been previously reported. We evaluated the foregoing technology in cantaloupe fields for suppression of powdery mildew and possible effects on plant growth and yield. In a controlled laboratory study, greenhouse-grown cantaloupe plants were exposed to a gradient of UV-C (254 nm) doses during darkness, and the effects upon powdery mildew development and the plant were evaluated. We also evaluated the efficacy of nighttime applications of UV-C at 100 and 200 J/m 2 against powdery mildew on adaxial leaf surfaces in greenhouse, high-tunnel, and open-field plantings. UV-C at the foregoing doses reduced sporulation and germination of P. xanthii conidia without damaging plants. On cantaloupe seedlings in the greenhouse, disease severity was equivalently suppressed at all doses and frequencies of applications of the light. In high-tunnel and open-field experiments, the most effective control of powdery mildew was provided by UV-C applied at 200 J/m 2 twice every week, where suppression provided by UV-C was generally equal to and sometimes better than the fungicide treatment. The foregoing UV-C dose and frequency of application also provided the highest yield under field conditions, indicating that UV-C treatment is a promising technology for commercially relevant suppression of powdery mildew on cantaloupe in a variety of growing systems., Competing Interests: The author(s) declare no conflict of interest.

Recently, the Florida strawberry industry faced unprecedented outbreaks of an emerging disease caused by the fungus Neopestalotipsis spp. Currently, there are no fungicides labeled to control this disease in the United States and the efficacy of single- and multisite fungicides is unknown. Therefore, this study aimed to determine the in vitro sensitivity of Neopestalotiopsis spp. isolates to fungicides with different modes of action and to evaluate the efficacy of these products on detached fruit and in the field. In preliminary in vitro tests, 30 commercially available fungicides were screened using discriminatory doses. The effective concentration that inhibited mycelial growth by 50% was determined for the most effective single-site fungicides. Four field experiments were conducted during the 2019-20, 2020-21, and 2021-22 seasons to determine product efficacy in managing the disease. The single-site fungicides fludioxonil, fluazinam, and sterol demethylation inhibitors, and the multisite fungicides captan, thiram, and chlorothalonil were the most effective in inhibiting pathogen growth and suppressing disease development. Conversely, products in Fungicide Resistance Action Committee (FRAC) groups 1 (methyl benzimidazole carbamate) and 7 (succinate-dehydrogenase inhibitors), except for benzovindiflupyr, were not effective against Neopestalotiopsis spp. Resistance to fungicides from FRAC group 11 (e.g., azoxystrobin) was confirmed by the presence of the G143A mutation in the cytochrome b gene together with inoculation tests and field trials. Our results provide information to support or discourage the registration of fungicides to manage Neopestalotiopsis fruit rot and leaf spot in strawberry production., Competing Interests: The author(s) declare no conflict of interest.

Phytophthora crown rot (PhCR) caused by Phytophthora cactorum is one of the most damaging diseases of strawberry worldwide. Mefenoxam is one of the major fungicides currently used to manage PhCR. However, the emergence and spread of resistant isolates have made controlling the pathogen in the field problematic. In the present study, using whole genome sequencing analysis, mutations associated with mefenoxam-resistant isolates were identified in six different genomic regions of P. cactorum. The 95.54% reads from a sensitive isolate pool and 95.65% from a resistant isolate pool were mapped to the reference genome of P. cactorum P414. Four point mutations were in coding regions while the other two were in noncoding regions. The genes harboring mutations were functionally unknown. All mutations present in resistant isolates were confirmed by sanger sequencing of PCR products. For the rapid diagnostic assay, SNP-based high-resolution melting (HRM) markers were developed to differentiate mefenoxam-resistant P. cactorum from sensitive isolates. The HRM markers R3-1F/R3-1R and R2-1F/R2-1R were suitable to differentiate both sensitive and resistant profiles using clean and crude DNA extraction. None of the mutations associated with mefenoxam resistance found in this study were in the RNA polymerase subunit genes, the hypothesized target of this compound in oomycetes. Our findings may contribute to a better understanding of the mechanisms of resistance of mefenoxam in oomycetes since serves as a foundation to validate the candidate genes as well as contribute to the monitoring of P. cactorum populations for the sustainable use of this product., (© 2023. The Author(s).)

A new Neopestalotiopsis sp. was recently reported causing outbreaks of leaf spot and fruit rot on strawberry in Florida, Georgia, and South Carolina. In contrast to other Pestalotiopsis pathogens, the new species appears more aggressive and destructive on strawberry. Current chemical options for management are disease suppressive at best, and affected growers have been experiencing major yield losses. In this study, we developed a molecular method based on polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) for identification of the new Neopestalotiopsis sp. from strawberry. Isolates of the new Neopestalotiopsis sp. collected in Florida; isolates of N. rosae , N. honoluluana , N. ellipsopora , N. saprophytica , N. samarangensis , and P. rhododendri ; and isolates from South Carolina suspected to be the new Neopestalotiopsis sp. were included in this study. This method is based on PCR amplification of a β-tubulin gene fragment using a previously published set of primers (Bt2a and Bt2b), followed by use of the restriction enzyme BsaWI. The enzyme cuts the PCR product from the new Neopestalotiopsis sp. twice, yielding fragments of 290 base pairs (bp) and 130 and 20 bp in size, whereas fragments from other species are only cut once, yielding fragments of 420 and 20 bp. This method will aid research labs and diagnostic clinics in the accurate and fast identification of the aggressive Neopestalotiopsis sp. variant from strawberry., Competing Interests: The author(s) declare no conflict of interest.

Phytophthora crown rot (PhCR) and leather rot (LR) caused by Phytophthora spp. are major threats to strawberry production worldwide. In the United States, these diseases are mainly caused by Phytophthora cactorum ; however, P. nicotianae has also been recently reported causing PhCR. Growers have relied on three different chemical products (i.e., mefenoxam and phosphites for PhCR and LR, and azoxystrobin for LR). Because resistance to mefenoxam and azoxystrobin has been reported, this study aimed to assess the in vitro sensitivity of Phytophthora spp. isolates from strawberry to phosphites and investigate its efficacy on in vivo assays. In vitro sensitivity of P. cactorum ( n = 128) and P. nicotianae ( n = 24) isolates collected from 1997 to 2018 was assessed for phosphite at 10, 50, 100, 150, and 300 µg/ml. Regardless of the Phytophthora sp. and isolation organ, most of the isolates (75% for P. cactorum and 54.2% for P. nicotianae ) had effective concentration that inhibits pathogen growth by 50% (EC 50 ) values ranging from 50 to 100 µg/ml. In vivo tests with strawberry fruit and plants revealed that commercial formulations of phosphite applied at the highest field rate controlled P. cactorum isolates but failed to control PhCR and LR caused by some isolates of P. nicotianae . In this study, EC 50 results from in vitro assay did not truly translate the efficacy of phosphites on controlling LR and PhCR caused by P. cactorum and P. nicotianae . Our findings support the hypothesis that the product acts in a dual way: direct on the pathogen and stimulating the plant immune system. Moreover, this has important implications for disease management, highlighting the importance of a correct diagnosis before phosphite recommendations, because its efficacy varies within Phytophthora spp., Competing Interests: The author(s) declare no conflict of interest.

Phytophthora crown rot (PhCR) is an important disease of strawberry worldwide. Phytophthora cactorum is the most common causal agent, however, P. nicotianae was also recently reported causing PhCR in the U.S. Therefore, the goals of this study were to evaluate the resistance of strawberry cultivars from Florida and California, and to study the etiology of the two Phytophthora species causing PhCR. Sixteen strawberry cultivars were evaluated over three Florida seasons for susceptibility to P. cactorum , and P. nicotianae . Inoculations at different days after transplanting (DAT) were also carried out to evaluate the ability of both species to cause PhCR at different phenological stages of the plant. Plant wilting and mortality were assessed weekly, and disease incidence, and the area under the disease progress curve were calculated. Cultivars Sensation 'Florida127', 'Winterstar FL 05-107', and 'Florida Radiance' were susceptible, whereas 'Florida Elyana', 'Camarosa', 'Fronteras', 'Sweet Charlie', and 'Strawberry Festival' were highly resistant to both Phytophthora species. However, some cultivars exhibited stronger resistance to one species over the other. P. cactorum caused more PhCR when plants were inoculated at transplanting, 45, and 60 DAT, whereas P. nicotianae only caused disease when inoculated at transplanting. These results emphasize the importance of screening for disease resistance to guide management recommendations in commercial strawberry production as well as the need for proper pathogen identification since cultivar susceptibility might differ. Varying susceptibility to P. cactorum and P. nicotianae at different growth stages emphasizes the importance of considering both plant and pathogen biology when making management recommendations.

Background: Major pests of Florida strawberry (two-spotted spider mites, Tetranychus urticae Koch; chilli thrips, Scirtothrips dorsalis Hood; and western flower thrips, Frankliniella occidentalis Pergande) are difficult to manage using only conventional chemical control. Previous research has demonstrated high mortality of T. urticae eggs after exposure to ultraviolet B (UV-B) light. Additionally, ultraviolet C (UV-C) light has been shown to suppress powdery mildew in strawberry at doses between 85 and 200 J m -2 with no damage to the plant. Therefore, UV-C may also have the potential to be used as an integrated pest management tool for arthropod pests of strawberry. The objectives of this study were to: (i) determine the effect of UV-C on T. urticae, S. dorsalis, and F. occidentalis natural populations in open-field strawberry; and (ii) determine the effect of UV-C on T. urticae egg hatch after application in open-field strawberry. Field studies were conducted during the 2019-2020 and 2020-2021 strawberry seasons in Florida, USA. Four treatments were compared: (i) foliar application of spinetoram in response to natural pest pressure; (ii) application of UV-C 200 J m -2 twice a week; (iii) application of UV-C 350 J m -2 twice a week; and (iv) a non-treated control., Results: In the field trials, suppression of T. urticae was observed at 350 J m -2 in 2020-2021. In the other field trials, no effect was observed due to low natural infestations. No negative impact on yield was observed from UV-C applications., Conclusion: UV-C shows promise as a component of an integrated pest management program for T. urticae in strawberry. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Powdery mildew (PM) is a common fungal disease in many important crops. The PM caused by Podosphaera xanthii has been the most challenging problem in commercial Gerbera ( Gerbera hybrida ) production globally, often leading to severe losses of crop yield and quality. A small number of PM-resistant breeding lines and cultivars have been reported in Gerbera , but the underlying genetics for PM resistance in Gerbera is largely unknown. Scarcity of genomic resources such as genetic linkage maps and molecular markers has severely hindered the effort to understand the genetic basis and locate loci controlling PM resistance in Gerbera . This study aimed to construct a genome-wide genetic linkage map, identify quantitative trait loci (QTL), and molecular markers for PM resistance in Gerbera . A segregating mapping population was developed by crossing PM-resistant and -susceptible Gerbera breeding lines, genotyped by sequencing, and phenotyped for PM resistance. A genome-wide genetic linkage map constructed with 791 single polymorphic site (SNP) markers spans 1912.30 cM across 27 linkage groups (LG) and reaches a density of 1 marker per 2.42 cM. One major consistent QTL was discovered in LG16, explaining more than 16.6% of the phenotypic variance for PM resistance. The QTL was tagged with two flanking SNP markers. The availability of this genetic linkage map will be very useful for locating and tagging QTLs for other important traits in Gerbera , and the newly discovered QTL and SNP markers will enable development of molecular markers for improving Gerbera for resistance to PM., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Bhattarai, Sharma, Verma, Peres, Xiao, Clark and Deng.)

Blackberry ( Rubus L. subgenus Rubus Watson) is a deciduous berry crop that is the fourth most economically important berry crop, and its production is expanding in the southeastern United States. However, since most commercially available cultivars were bred under temperate conditions, they are not always well adapted and could be threatened by new pathogen populations inhabiting subtropical areas. In 2017, plants showing purple or brown leaf spots and angular-to-irregular lesions on both leaf surfaces, with clusters of black conidiophores at the center, were observed in a field trial at the University of Florida's Gulf Coast Research and Education Center (UF/GCREC) in Wimauma, FL. A fungus resembling Cercospora / Pseudocercospora was isolated from the lesions. The ribosomal DNA internal transcribed spacers, the translation elongation factor 1-alpha, and the actin genes were amplified and sequenced. Based on the phylogenetic analysis, the closest related species was Pseudocercospora pancratii . Pathogenicity assays and subsequent reisolation confirmed that this species is the causal agent of the disease. Among eight cultivars screened, no complete resistance was found. However, 'Osage' was the least susceptible, and 'Kiowa' was the most susceptible. This study is the first report of P . pancratii causing leaf spots on blackberry worldwide, and it may help shape future research into disease epidemiology and management for a crop that is rapidly expanding but has very limited disease information currently available for Florida growers.

The Strawberry Advisory System (SAS) is a tool developed to help Florida strawberry growers determine the risk of common fungal diseases and the need for fungicide applications. Leaf wetness duration (LWD) is one of the important parameters in SAS disease risk modeling. By accurately measuring the LWD, disease risk can be better assessed, leading to less fungicide use and more economic benefits to the farmers. This research aimed to develop and test a more accurate leaf wetness detection system than traditional leaf wetness sensors. In this research, a leaf wetness detection system was developed and tested using color imaging of a reference surface and a convolutional neural network (CNN), which is one of the artificial-intelligence-based learning methods. The system was placed at two separate field locations during the 2021-2022 strawberry-growing season. The results from the developed system were compared against manual observation to determine the accuracy of the system. It was found that the AI- and imaging-based system had high accuracy in detecting wetness on a reference surface. The developed system can be used in SAS for determining accurate disease risks and fungicide recommendations for strawberry production and allows the expansion of the system to multiple locations.

Strawberry powdery mildew, caused by Podosphaera aphanis , can be particularly destructive in glasshouse and plastic tunnel production systems, which generally are constructed of materials that block ultraviolet (UV) solar radiation (about 280 to 400 nm). We compared epidemic progress in replicated plots in open fields and under tunnels constructed of polyethylene, which blocks nearly all solar UV-B, and two formulations of ethylene tetrafluoroethylene (ETFE), one of which contained a UV blocker and another that transmitted nearly 90% of solar UV-B. Disease severity under all plastics was higher than in open-field plots, indicating a generally more favorable environment in containment structures. However, the foliar severity of powdery mildew within the tunnels was inversely related to their UV transmissibility. Among the tunnels tested, incidence of fruit infection was highest under polyethylene and lowest under UV-transmitting ETFE. These effects probably transcend crop, and the blocking of solar UV transmission by glass and certain plastics probably contributes to the widely observed favorability of greenhouse and high-tunnel growing systems for powdery mildew.

Colletotrichum acutatum is a species complex that causes anthracnose fruit rot and root necrosis on strawberry. The major and minor species within the complex that affect strawberry production are C. nymphaeae and C. fioriniae , respectively. The disease can significantly reduce yield under conducive weather, and its management has greatly relied on quinone-outside inhibitor fungicides (QoI). However, due to the emergence of resistant isolates, such products are no longer effective. Therefore, alternative fungicides were investigated. C. nymphaeae and C. fioriniae isolates were collected from multiple strawberry fields in the United States from 1995 to 2017. The sensitivity of benzovindiflupyr, penthiopyrad, pydiflumetofen, fluazinam, fludioxonil, and cyprodinil was assessed by in vitro and in vivo assays. Both Colletotrichum species were sensitive to benzovindiflupyr, penthiopyrad, fluazinam, and fludioxonil based on mycelial growth assays. Interestingly, of these products, only penthiopyrad did not inhibit conidial germination at 100 µg/ml. For cyprodinil, C. nymphaeae was sensitive based on the mycelial growth, whereas C. fioriniae was not inhibited. Neither species was inhibited by pydiflumetofen in mycelial growth, conidial germination, nor detached fruit assays. The prepackaged mixtures fludioxonil + cyprodinil and fludioxonil + pydiflumetofen were effective in a field trial; however, their use should be carefully considered because of the lack of efficacy of one of the compounds in the mixture. This study sheds light on the potential registration of products alternative to QoIs, such as benzovindiflupyr and fluazinam, which could improve the management of strawberry anthracnose.

Background: The effectiveness of metam potassium, 1,3-dichloropropene, chloropicrin, and different ratios of 1,3-dichloropropene and chloropicrin on the reduction of natural and artificial inoculum of Macrophomina phaseolina were investigated in laboratory and field experiments. Additionally, a multivariate meta-analysis with data from six field trials conducted in Florida from 2012 to 2018 was performed., Results: In small-plot field experiments using drip stakes, the highest rate (468 L ha -1 ) of metam potassium was most effective in controlling M. phaseolina in infected crowns buried at 15.2 cm from the point of fumigant injection, whereas none of the rates was able to reduce inoculum buried at 30.5 cm. In closed-container experiments, use of the highest rate of 1,3-dichloropropene (168 kg ha -1 ) resulted in the highest level of pathogen control. Different rates of chloropicrin also reduced inoculum when compared to the non-treated control. 1,3-dichloropropene + chloropicrin at different ratios were also highly effective in controlling M. phaseolina. Results from the meta-analysis of open-field experiments indicated that metam potassium and 1,3-dichloropropene + chloropicrin (63:35, v:v) treatments were significantly more effective in reducing M. phaseolina than the 1,3-dichloropropene + chloropicrin (39:60, v:v) treatment; however, metam potassium was not as effective at the side of the beds., Conclusion: 1,3-dichloropropene alone and in mixture with chloropicrin were more effective in reducing inoculum of M. phaseolina than chloropicrin alone, indicating the fungicidal efficacy of 1,3-dichloropropene. Formulation with higher 1,3-dichloropropene concentration performed better than the formulation with higher chloropicrin concentration in field trials. Metam potassium was effective when applied at the highest rate, but with limited lateral movement perpendicular to the drip irrigation line. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Aphelenchoides besseyi and A. pseudogoodeyi are foliar nematodes associated with commercial strawberry production in Florida, United States. The reproductive and feeding habits of these two nematode species were assessed on Florida isolates of the fungi Botrytis cinerea , Colletotrichum gloeosporioides , Macrophomina phaseolina , and Neopestalotiopsis rosae , which are pathogenic to strawberry, and nonpathogenic isolates of Fusarium oxysporum and Monilinia fructicola grown on potato dextrose agar in Petri dishes. Each culture was inoculated with six specimens of mixed life stages of A. besseyi or A. pseudogoodeyi and incubated at 24°C under axenic and nonaxenic conditions 23 and 31 days after inoculation, respectively. A. besseyi reproduction rates were greater on strawberry-pathogenic isolates of B. cinerea , C. gloeosporioides , and N. rosae than on the nonpathogenic isolates of F. oxysporum and M. fructicola . In contrast, reproductive rates of A. pseudogoodeyi did not vary among fungi cultures. For both nematode species, M. phaseolina was a poor host because it did not produce mycelium on the media used. Our findings indicate that A. besseyi is more selective in its fungal-feeding preference than A. pseudogoodeyi . Additionally, A. pseudogoodeyi eggs and juveniles were significantly more numerous than adults. Yet, for A. besseyi , adult stages were more abundant. Fungi aid in the maintenance of soil-dwelling populations of these two nematode species. Removing fungus-infected strawberry plant residues is both a desirable and effective management practice to limit A. besseyi in central Florida commercial strawberry fields.

Phytophthora cactorum and P . nicotianae cause leather rot (LR) of fruit and Phytophthora crown rot (PhCR) in strawberry. LR occurs sporadically but can cause up to 70% fruit loss when weather is conducive. In Florida's annual strawberry winter production system, PhCR can be severe, resulting in plant stunting, mortality, and severe yield loss. Azoxystrobin is labeled for control of LR but not for PhCR. The aims of this research were to determine the sensitivity of P . cactorum and P . nicotianae isolates from strawberry to azoxystrobin and to investigate mechanisms of quinone-outside-inhibitor resistance present in P . cactorum and P . nicotianae based on the known point mutations within the cytochrome b ( cytb ) gene. Isolates of both Phytophthora spp. causing LR and PhCR were collected from multiple strawberry fields in Florida between 1997 and 2020. Isolates were tested for sensitivity to azoxystrobin at 0, 0.01, 0.1, 1.0, 10, and 50 μg/ml on potato dextrose agar amended with salicylhydroxamic acid (100 μg/ml). Isolates were separated into two groups - sensitive isolates with the 50% effective concentration ( EC 50 ) values <1.0 μg/ml, and resistant isolates having EC 50 values >50 μg/ml. P . cactorum and P . nicotianae resistance to azoxystrobin was found for isolates collected after 2010. The first 450 nucleotides of the mitochondrial cytb gene were sequenced from a selection of resistant and sensitive isolates of both species. The G143A mutation reported to confer resistance to azoxystrobin was found in all resistant P . cactorum isolates. However, in P . nicotianae , qualitative resistance was observed, but the isolates lacked all the known mutations in the cytb gene. This is the first report of resistance to azoxystrobin in P . cactorum and P . nicotianae .

Colletotrichum gloeosporioides is the causal agent of Colletotrichum crown rot of strawberry in the southern United States. Recent multigene studies defined C. gloeosporioides as a complex species comprised of 37 species. In our study, we phylogenetically characterized C. gloeosporioides isolates from strawberry and other noncultivated plants around strawberry fields. One hundred fifteen strawberry isolates and 38 isolates from noncultivated hosts were sequenced for five genomic regions: internal transcribed spacer, actin, calmodulin, chitin synthase, and glyceraldehyde-3-phosphate dehydrogenase. Phylogenetic analysis using the maximum likelihood and Bayesian inference methods, based on partition-specific models, revealed that most of the isolates in Florida (86%) were closely related to C. siamense , whereas 14 isolates were closely related to C. theobromicola (syn. C. fragariae ), four isolates were C. fructicola , and three isolates were C. clidemiae . However, only the first three species were pathogenic to strawberry. Morphological characteristics evaluated show that mycelial growth of all species is approximately 5 mm/day, but colony morphology varies by species and incubation conditions. In vitro mating of the isolates demonstrated that C. fructicola is homothallic whereas C. siamense and C. theobromicola isolates are heterothallic. The biological importance of these different Colletotrichum species is currently being investigated to determine whether different management strategies are needed in strawberry production fields.

During the fall of 2020 and summer of 2021, symptoms of leaf rust were observed on blackberry plants of 'Kiowa', and breeding line 1734 (progeny of 'Natchez' and Arapaho') in a field trial at the University of Florida, Wimauma, FL. Symptoms consisted of small chlorotic spots (1 to 3 mm) on the upper side of the leaf, while the underside had yellow-orange pustules. Disease incidence was up to 100% on both 'Kiowa' and the breeding line 1734, and severity was up to 20% with most of the symptoms observed on older leaves. Two isolates were collected from 'Kiowa' and one from the breeding line 1734 for further investigation. Isolates were maintained and multiplied on healthy 'Kiowa' plants in growth chambers (25 ºC and 12-12 h photoperiod). Uredinia (n=30) were erumpent and ranged from 90 to 320 µm (Average=285 µm, SD=5.3 µm) in diameter. Urediniospores (n=50) were obovoid, yellow, and ranged from 24 to 36 μm long (Average=32 µm, SD=3.2 µm) and 22 to 30 μm wide (Average=28 µm, SD=2.5 µm). Based on morphology and literature, the pathogen was tentatively identified as Kuehneola uredinis (Link) Arth (Arthur 1906; Shands et al., 2018). Spores from a single uredinium of each isolate were collected with a needle and suspended in 50 µL of molecular biology-grade water yielding a final concentration of approximately 5 x 104 spores/mL. Two µL of each spore suspension was used for the PCR reactions. Two DNA fragments were amplified using the primers Rust2inv and LR6, and Rust18S-R and NS1 for the 5.8S-ITS2-28S gene region of rDNA (1,755 bp) and partial 18S gene region of rDNA (2,684 bp), respectively. The amplified products of the partial 28S gene region were sequenced with the primers LR3 and LR0R, and the 18S gene region with NS5, NS6, and NS4 (Aime 2006). DNA sequences were deposited in GenBank (accession nos. OK509845 - OK509848). BLASTn searches revealed that the isolates were 100% identical to K. uredinis reported causing leaf rust on blackberry in California (1044/1044bp, and 1540/1540bp for accession numbers MF158087, and MF158088, respectively). To test for pathogenicity, blackberry cultivars Kiowa, Natchez, Osage, Ouachita, Ponca, Prime-Ark® 45, Prime-Ark® Freedom, Prime-Ark® Traveler, and Prime-Ark® Horizon were inoculated. Five plants of each cultivar were inoculated with a mixture of spores of the three isolates, and two plants of each cultivar were used as controls. Spores were washed from leaves of 'Kiowa' exhibiting sporulation using a suspension of 1% Tween 20 in deionized water. The final concentration of the inoculum was adjusted to 104 spores/mL. Plants were inoculated in the greenhouse with a spray bottle until run-off and kept inside clear plastic boxes for 48 h. Controls were sprayed with sterile deionized water. Plants were watered by mists of 3 s every 10 min twice a week. Disease incidence and severity were evaluated weekly on five leaves per plant that had been tagged before inoculation. The experiment was repeated once. Symptoms identical to the original were only observed in 'Kiowa' and 'Prime-Ark® Freedom'. One week after inoculation, disease incidence was already 100% in both cultivars, with at least one pustule on all the inoculated leaves, and six weeks later disease severity was up to 50% (Average= 35%, SD=2.4%). To our knowledge, this is the first report of K. uredinis causing leaf rust on blackberry in Florida. This disease was reported on Rubus spp. in several U.S. states, and recently in California on Rubus ursinus (Farr and Rossman 2021; Shands et al. 2018). Blackberry is an emerging crop in Florida and efforts should be implemented to monitor the occurrence and spread of leaf rust considering that urediniospores disperse long distances by wind, especially if growers choose the susceptible cultivars 'Kiowa' and 'Prime-Ark Freedom'. The apparent resistance observed in other commercial cultivars such as 'Osage', 'Ouachita', and 'Ponca' may serve as valuable breeding parents for developing new blackberry cultivars with resistance to leaf rust.

Phytophthora cactorum and Phytophthora nicotianae cause leather rot on fruit and crown rot (PhCR) of strawberry plants. Leather rot is not a common disease in Florida; however, up to 50% yield loss has been reported in harvests after intense rainfall events. PhCR is an important disease worldwide and is characterized by a sudden wilting and collapse of plants. Mefenoxam is the most effective and widely used fungicide to control both diseases. P. cactorum and P. nicotianae isolates from leather rot and PhCR have been collected from multiple strawberry fields in Florida since 1997, and the sensitivity of 185 isolates was tested at 0, 0.05, 0.5, 5, and 100 µg/ml. The 50% effective concentration (EC 50 ) values of sensitive isolates ranged from 0.05 to 1 µg/ml. Resistance to mefenoxam (EC 50 values >100 µg/ml) was found among P. cactorum isolates collected after 2015, but no resistance was found in P. nicotianae isolates. During the 2015 to 2016, 2016 to 2017, 2017 to 2018, and 2018 to 2019 seasons, resistance was detected on 9, 10, 21, and 23% of the isolates collected, respectively. Mefenoxam-resistant isolates originated from 3 of the 24 strawberry nurseries monitored. This is the first report of the occurrence of P. cactorum resistance to mefenoxam in Florida, suggesting that alternative control strategies are needed to avoid the increase of mefenoxam-resistant populations of P. cactorum in Florida fields.

Phytophthora crown rot, caused mainly by Phytophthora cactorum but also by P. nicotianae , reported in 2018, is an important disease in the Florida strawberry annual production system. Mefenoxam is the most effective and widely used fungicide to manage this disease. However, because of pathogen resistance, alternatives to chemical control are needed. Phytophthora spp. were rarely recovered during the summer from soil of commercial farms where the disease was observed during the season. In a more detailed survey on research plots, neither of the two species was recovered 1 month after the crop was terminated and water was shut off. Therefore, Phytophthora spp. does not seem to survive in the soil over summer in Florida. In a field trial, asymptomatic nursery transplants harboring quiescent infections were confirmed as the major source of inoculum for these pathogens in Florida. Heat treatment of P. cactorum zoospores at 44°C for as little as 5 min was effective in inhibiting germination and colony formation; however, oospore germination was not inhibited by any of the tested temperatures in vitro. In the field, thermotherapy treatment of inoculated plants was shown to have great potential to serve as a nonchemical approach for managing Phytophthora crown rot in production fields and reducing mefenoxam-resistant populations in nursery transplants.

We designed and deployed an apparatus to apply UV light for suppression of powdery mildew in open field production of strawberry. The unit was evaluated in a commercial production field for one season, and for two additional seasons in open field research plots at the University of Florida Gulf Coast Research and Education Center. The apparatus contained two 180-cm-long hemicylindrical arrays of twenty 55-W low-pressure discharge UV-C lamps (operated at 30 W; peak wavelength = 254 nm) backed by polished aluminum reflectors covering two adjacent beds of the strawberry planting. The lamp arrays were suspended within a steel carriage that was tractor-drawn through the planting at 2.3, 4.6, and 5.6 km h -1 . Nighttime applications of UV-C at doses ranging from 65 to 170 J⋅m -2 either once or twice weekly provided suppression of foliar and fruit disease that was consistently equal to or better than that provided by a commercial calendar-based fungicide spray program.

Anthracnose fruit rot (AFR) and Botrytis fruit rot (BFR) are primary diseases affecting strawberry ( Fragaria × ananassa ), which typically drive fungicide applications throughout the growing season. The Strawberry Advisory System (StAS), a disease forecasting tool, was originally developed in Florida to better time the fungicide sprays by monitoring AFR and BFR infection risk based on leaf wetness and temperature input in real-time. Thirteen field trials were conducted in Maryland and Virginia between 2017 and 2019 to evaluate the StAS performance in the Mid-Atlantic region. As a result, 55, 18, and 31% fewer sprays were recorded on average in the model-based StAS treatment compared with the grower standard treatment in 2017, 2018, and 2019, respectively. Marketable yield, as well as AFR and BFR incidence, were largely comparable between the two treatments. However, poor disease control occurred during the StAS treatment in four trials in 2017, presumably because of a missed fungicide spray during a high-risk infection event and attributable to heavy rainfall that led to impassable fields. The implementation of the StAS may be further challenged by the employment of floating row covers that are essential for growing strawberries in plasticulture systems in open fields in the Mid-Atlantic region. Preliminary results indicated that row covers can alter canopy-level microclimatic conditions, possibly increasing the risk for disease occurrence. Overall, the StAS can be a valuable tool for Mid-Atlantic growers to control AFR and BFR, but sprays may need to be promptly applied when consecutive or heavy rainfalls are predicted, especially for highly susceptible cultivars. Complications in disease forecasting and management arising from the use of row covers need to be further addressed in this region because of its highly diverse climate.

Macrophomina phaseolina , the causal agent of charcoal rot, is a soilborne pathogen that affects strawberry crowns leading to plant wilt and collapse. Disease management involves a combination of physical, cultural, and chemical methods. Field trials were conducted for 10 consecutive Florida seasons (2010-11 to 2019-20) to determine the susceptibility of strawberry cultivars to charcoal rot and the effect of cultivar selection on disease and to estimate the economic impact of cultivar selection on disease management. Six cultivars grown commercially in Florida were chosen and grouped as highly susceptible (HS) ('Strawberry Festival' and 'Treasure'), susceptible (S) ('Florida Radiance' and 'Florida Beauty'), and moderately resistant (MR) (Sensation 'Florida127' and Winterstar 'FL05-107') according to their susceptibility levels. After a primary analysis of the individual trials, a network meta-analysis was conducted to estimate and compare the final disease incidence and the disease progress rate of each susceptibility group. The economic impact of charcoal rot on strawberry production and gross revenue was estimated based on plant production functions, weekly fruit prices, and disease progress over time with parameters obtained via the meta-analytical models. Disease incidence was reduced by 91.5 and 77.3%, respectively, when the MR and S cultivar groups were adopted instead of the HS group. There was a 62.5% reduction in the disease incidence when the MR group was used instead of the S group. Significant differences in disease progress rates were also observed when the MR and S groups were adopted instead of the HS group. Therefore, the adoption of more resistant cultivars is an effective strategy when incorporated into a charcoal rot integrated management program and can significantly impact growers' revenue by reducing disease incidence, preventing yield loss, and, consequently, minimizing economic losses.

Rapid and accurate disease diagnosis is a prerequisite for an effective disease management program in strawberry production. In Florida, Colletotrichum spp., Phytophthora spp., and Macrophomina phaseolina are the primary microorganisms causing strawberry crown rot. Even though the diseases can be caused by different pathogens, symptoms are indistinguishable and equally devastating. To inform strawberry growers in a timely fashion of diagnostic results for effective deployment of chemical control practices, we developed a multiplex high-resolution melting (HRM) assay to rapidly and accurately detect the abovementioned pathogens. The multiplex HRM assays using three predesigned primer pairs showed high specificity for individual species by generating specific melting peaks without cross-reaction between primers or with other common strawberry pathogens. The amplification limit of the assay was 1 pg of Colletotrichum and Phytophthora and 100 pg of M. phaseolina DNA per 10-μl reaction. However, the presence of different melting peaks was observed in mixed DNA samples and was concentration and target DNA dependent. A crude DNA extraction protocol was developed to allow high-throughput screening by minimizing the inhibitory effects. Moreover, we applied the HRM assay to 522 plant samples and found high correlations between conventional pathogen isolation and HRM and between singleplex and multiplex assays. Altogether, this multiplex HRM assay is specific, cost effective, and reliable for the timely detection of strawberry crown rot pathogens.

Blueberry is an increasingly important crop in Florida. Anthracnose fruit rot (AFR), caused mostly by Colletotrichum gloeosporioides , is favored by long wetness periods and temperatures from 15 to 27°C. Currently, the model in the Strawberry Advisory System (StAS) guides fungicide applications targeting strawberry AFR. Given the similarity between blueberry and strawberry AFR, we hypothesized that the model used in StAS could be used in a decision support system (DSS) built for blueberry AFR. There is no information on inhibition posed by fungicides on C. gloeosporioides isolates from blueberry. Our objectives were to demonstrate that the model used in the StAS could be used for blueberry AFR management in Florida and to assess the sensitivity of isolates to fungicides. Four trials were undertaken in blueberry fields in Florida during two seasons to compare the effectiveness of fungicide applications according to the model with that of the growers' standard calendar. Sensitivity of blueberry C. gloeosporioides isolates to azoxystrobin, benzovindiflupyr, penthiopyrad, pydiflumetofen, boscalid, thiophanate-methyl, fluazinam, and fludioxonil was evaluated. AFR incidence and yield were compared between treatments. Following recommendations from the model resulted in disease control as effective as the standard program and in some cases with fewer applications. All isolates were sensitive to benzovindiflupyr, penthiopyrad, fluazinam, and fludioxonil. Low frequency of in vitro inhibition of isolates by azoxystrobin, pydiflumetofen, boscalid, and thiophanate-methyl should raise concern about fungicide resistance. Our results indicate that the model used in StAS could be used in a DSS to help Florida growers to manage AFR in blueberry.

Hemp is an annual herbaceous plant that is used for its fiber and oil in a variety of commercial and industrial products. In Florida, it is currently being explored as a new specialty crop. During a field trial from October to January 2019 in Wimauma, FL, a stem canker was observed on up to 60% of three-month-old plants of 'Eletta Campana', 'Carmagnola Selezionata', and 'Tygra'. Symptoms started on the main stems with light-to-dark brown lesions of different sizes and shapes. Over time, the lesions coalesced into large necrotic areas and bore pycnidia. Isolations were made from diseased stem tissues on General Isolation medium (Amiri et al. 2018) after surface disinfestation (Marin et al. 2020). The plates were placed in a growth chamber at 25°C under a 12/12 photoperiod. A fungus with white, floccose, aerial mycelium and pycnidia producing alpha and beta conidia was consistently isolated. Three single spore isolates were chosen for identification and pathogenicity tests. Pycnidia on PDA were globose to irregular and ranged from 170 to 250 μm long (210 ± 2.5, n = 50) and 140 to 220 μm wide (180 ± 2.7, n = 50). The alpha conidia were unicellular, hyaline, ellipsoidal to fusiform and ranged from 5.3 to 7.7 μm long (6.5 ± 1.6, n = 50) and 1.5 to 4.6 μm wide (2.8 ± 1.8, n = 50). The beta conidia were hyaline, elongated, filiform, straight or curved and ranged from 10.2 to 17.7 μm long (16.1 ± 2.2, n = 50) and 0.5 to 1.8 μm wide (0.8 ± 0.2, n = 50). Perithecia were not observed. Based on morphological features, the fungus was similar to anamorphs of Diaporthe spp. (Santos et al. 2011; Udayanga et al. 2015). DNA from the same three isolates was extracted using the FastDNA kit, and the ribosomal internal transcribed spacer (ITS), β-tubulin (TUB), and calmodulin (CAL) regions were amplified following Udayanga et al. (2014), and Sanger sequenced by Genewiz. Sequences were deposited in GenBank (accession no. MT497039 to MT497047 for ITS, TUB, and CAL). BLASTn searches revealed isolates 20-58, 20-59, and 20-60 were 96.34% identical to the epitype isolate D. phaseolorum AR4203 for ITS (KJ590738.1, 527 bp out of 547 bp), 100% for TUB (KJ610893.1, 459 bp out of 459 bp), and 100% for CAL (KJ612135.1, 522 bp out of 522 bp) (Udayanga et al. 2015). Their identity was confirmed by phylogenetic analyses using maximum likelihood and Bayesian inference methods. To complete Koch's postulates, pycnidia of the same three isolates were harvested and crushed in 2 mL Eppendorf tubes containing 0.01% Tween 20. Conidia suspensions were adjusted to 106 spores/mL. Three 5-week-old potted plants of 'Eletta Campana' and 'Carmagnola Selezionata' per isolate were inoculated using a 1 mL syringe with a needle by injecting 200 µL of the suspension into the stem. Plants were placed inside clear plastic bags for 48 h and maintained in the greenhouse. Control plants were injected with sterile deionized water and kept under the same conditions. The pathogenicity test was repeated once. Four weeks after inoculation, inoculated plants developed stem cankers from which the same pathogen was isolated, whereas controls remained healthy. To our knowledge, this is the first report of D. phaseolorum causing stem canker on hemp. This pathogen has been reported causing canker on sunflower and Phaseolus spp. (Gomzhina and Gannibal 2018; Udayanga et al. 2015; Vrandecic et al. 2009). This discovery may help shape future research into disease epidemiology and management for a crop in which very limited disease information is available at the moment.

Macrophomina phaseolina , the causal agent of charcoal rot in strawberry, induces plant wilting and collapse. The pathogen survives through the production of microsclerotia in the soil and in strawberry debris. However, its management is difficult, and the disease has become an increasing problem for the strawberry industry. Physical, cultural, and chemical alternatives for integrated management of charcoal rot were evaluated in laboratory and field trials during the 2017-18 and 2018-19 strawberry seasons. In a laboratory trial, M. phaseolina microsclerotia were subjected to heat treatment and germination was inhibited at 52, 56, 80, and 95°C after 30, 10, 1, and 0.5 min of exposure, respectively. In infected strawberry crowns, microsclerotial viability was reduced after 5 min, regardless of temperature, whereas in the field, reduction was observed after 1 min. In field trials, charcoal rot incidence of inoculated strawberry plants transplanted into white-striped plastic-mulched beds was reduced to 20.8%, compared with 60.8% for plants grown in the black plastic mulch. On commercial farms, crop residue removal from infested areas reduced the M. phaseolina population in the soil but did not decrease charcoal rot incidence. Moreover, M. phaseolina propagule densities in the soil and in strawberry debris was reduced by fumigant application at crop termination but surviving propagules allowed the population to increase over the summer. Furthermore, preplant fumigation with metam potassium reduced soil population and charcoal rot incidence. Overall, the adoption of integrated approaches such as physical, chemical, and/or cultural methods played a significant role in reducing M. phaseolina inoculum and contributed to control of the disease in areas with high disease pressure.

Pestalotiopsis -like species have been reported affecting strawberry worldwide. Recently, severe and unprecedented outbreaks have been reported in Florida commercial fields where leaf, fruit, petiole, crown, and root symptoms were observed, and yield was severely affected. The taxonomic status of the fungus is confusing because it has gone through multiple reclassifications over the years. Morphological characteristics, phylogenetic analyses, and pathogenicity tests were evaluated for strawberry isolates recovered from diseased plants in Florida. Phylogenetic analyses derived from the combined internal transcribed spacer, β-tub , and tef1 regions demonstrated that although there was low genetic diversity among the strawberry isolates, there was a clear separation of the isolates in two groups. The first group included isolates recovered over a period of several years, which was identified as Neopestalotiopsis rosae . Most isolates recovered during the recent outbreaks were genetically different and may belong to a new species. On potato dextrose agar, both groups produced white, circular, and cottony colonies. From the bottom, colonies were white to pale yellow for Neopestalotiopsis sp. and pale luteous to orange for N. rosae . Spores for both groups were five-celled with three median versicolored cells. Mycelial growth and spore production were higher for the new Neopestalotiopsis sp. isolates. Isolates from both groups were pathogenic to strawberry roots and crowns. However, the new Neopestalotiopsis sp. proved more aggressive in fruit and leaf inoculation tests, confirming observations from the recent outbreaks in commercial strawberry fields in Florida.

Powdery mildew ( Podosphaera aphanis ) is a destructive and widespread disease of strawberry ( Fragaria × ananassa ), especially when susceptible cultivars are grown in high plastic tunnels or glasshouses. Many powdery mildews thrive in humid environments but free water films on plant surfaces can inhibit conidial germination of some species. We hypothesized that P. aphanis might be directly suppressed by rain through the action of water films and meteoric water. In repeated experiments, the hydrophobic conidia of P. aphanis collected on the surface of water droplets, resulting in their removal when the droplets rolled over the leaf surfaces and fell to the ground. Meteoric water and water films also damaged conidiophores. Brief midday water mists applied in pulses lasting 1 min each four times per day were as effective as multiple fungicide treatments in suppressing powdery mildew. Rapid drying of the pulsed mists resulted in effective suppression of powdery mildew without consequent increases of fungal pathogens that might benefit from water films. The timing and duration of water sprinkling has been refined to the point where it can provide a commercially relevant degree of suppression of powdery mildew on strawberry in a high-tunnel production system.

Background: RNA sequencing has been widely used to profile genome-wide gene expression and identify candidate genes controlling disease resistance and other important traits in plants. Gerbera daisy is one of the most important flowers in the global floricultural trade, and powdery mildew (PM) is the most important disease of gerbera. Genetic improvement of gerbera PM resistance has become a crucial goal in gerbera breeding. A better understanding of the genetic control of gerbera resistance to PM can expedite the development of PM-resistant cultivars., Results: The objectives of this study were to identify gerbera genotypes with contrasting phenotypes in PM resistance and sequence and analyze their leaf transcriptomes to identify disease resistance and susceptibility genes differentially expressed and associated with PM resistance. An additional objective was to identify SNPs and SSRs for use in future genetic studies. We identified two gerbera genotypes, UFGE 4033 and 06-245-03, that were resistant and susceptible to PM, respectively. De novo assembly of their leaf transcriptomes using four complementary pipelines resulted in 145,348 transcripts with a N50 of 1124 bp, of which 67,312 transcripts contained open reading frames and 48,268 were expressed in both genotypes. A total of 494 transcripts were likely involved in disease resistance, and 17 and 24 transcripts were up- and down-regulated, respectively, in UFGE 4033 compared to 06-245-03. These gerbera disease resistance transcripts were most similar to the NBS-LRR class of plant resistance genes conferring resistance to various pathogens in plants. Four disease susceptibility transcripts (MLO-like) were expressed only or highly expressed in 06-245-03, offering excellent candidate targets for gene editing for PM resistance in gerbera. A total of 449,897 SNPs and 19,393 SSRs were revealed in the gerbera transcriptomes, which can be a valuable resource for developing new molecular markers., Conclusion: This study represents the first transcriptomic analysis of gerbera PM resistance, a highly important yet complex trait in a globally important floral crop. The differentially expressed disease resistance and susceptibility transcripts identified provide excellent targets for development of molecular markers and genetic maps, cloning of disease resistance genes, or targeted mutagenesis of disease susceptibility genes for PM resistance in gerbera.

Aerated steam-based thermotherapy was developed and evaluated for its efficacy in managing three nematode species ( Aphelenchoides besseyi , Meloidogyne hapla , and Pratylenchus penetrans ) that are often transported as quiescent passengers on strawberry transplants shipped to Florida from out-of-state nurseries. Initial studies were focused on evaluating the intrinsic temperature sensitivity of each nematode species to hot water in laboratory conditions. Each nematode species was exposed to hot water at 40, 44, 48, and 52°C for 1, 5, 10, 30, 60, 120, and 240 min. Exposure for 60 min or higher at 40°C paralyzed all three nematode species when examined immediately after heat treatment. Examination of the nematodes 24 hr post-treatment suggested that 100% mortality of all three nematode species was achieved when nematodes were exposed to hot water at a minimum temperature of 44°C for 120 min. Further studies were conducted to evaluate the efficacy of aerated steam to kill all three nematode species by exposing nematode-infested strawberry transplants at 44°C for 60, 120, and 240 min. Exposure of nematode inoculated plants to steam for 60 or 120 min reduced the populations of all three nematode species, but this was not enough to completely eradicate any of the three nematode species. Exposure for 240 min, however, was the most effective in reducing the populations of the three nematode species. A 240 min of exposure to aerated steam completely eradicated A. besseyi and M. hapla while P. penetrans populations were reduced only by 85%. Furthermore, the aerated steam had minimal to no adverse effect on plant biomass. Results from both the laboratory and greenhouse studies indicated that M. hapla was more sensitive to heat treatment followed by A. besseyi and P. penetrans . Results from this study suggested that aerated steam-based thermotherapy has good potential as a non-chemical method of management of nematodes of strawberry transplants., (© 2020 Authors.)

The current management of Colletotrichum crown rot (CCR) of strawberry, caused by Colletotrichum gloeosporioides sensu lato, relies on the use of a few fungicide classes, particularly QoI fungicides. Since resistance to QoI fungicides has recently been detected, alternative fungicide groups are needed to control this disease. Our objective was to evaluate the efficacy of succinate-dehydrogenase-inhibitor (SDHI) fungicides in managing CCR. Five SDHI fungicides, fluopyram, isofetamid, penthiopyrad, fluxapyroxad, and benzovindiflupyr, were applied 2 days before or 1 day after inoculation of cultivar Strawberry Festival. SDHI treatments were compared with the most common fungicides used for CCR management, i.e., thiophanate-methyl, pyraclostrobin, and captan. Benzovindiflupyr applied 1 day after inoculation was effective in reducing plant mortality and disease development. The baseline sensitivity of C. gloeosporioides isolates was determined in vitro using a spiral gradient dilution assay. The EC 50 for benzovindiflupyr and penthiopyrad varied from 0.08 to 1.11 and 0.45 to 3.17 µg/ml, respectively, whereas the other SDHI fungicides did not inhibit fungal growth. If registered, benzovindiflupyr could serve as an alternative to manage CCR in Florida.

In a number of pathosystems involving the powdery mildews (Erysiphales), plant stress is associated with decreased disease susceptibility and is detrimental to pathogen growth and reproduction. However, in strawberry, anecdotal observations associate severe powdery mildew ( Podosphaera aphanis ) with water stress. In a 2017 survey of 42 strawberry growers in Norway and California, 40 growers agreed with a statement that water-stressed strawberry plants were more susceptible to powdery mildew compared with nonstressed plants. In repeated in vitro and in vivo experiments, we found that water stress was consistently and significantly unfavorable to conidial germination, infection, and increases in disease severity. Deleterious effects on the pathogen were observed from both preinoculation and postinoculation water stress in the host. Soil moisture content in the range from 0 to 50% was correlated ( R 2 = 0.897) with germinability of conidia harvested from extant colonies that developed on plants growing at different levels of water stress. These studies confirm that P. aphanis fits the norm for biotrophic powdery mildews and hosts under stress. Mild water stress, compared with a state of optimal hydration, is likely to decrease rather than increase susceptibility of strawberry to P. aphanis . We believe it is possible that foliar symptoms of leaf curling due to diffuse and inconspicuous infection of the lower leaf surfaces by P. aphanis could easily be mistakenly attributed to water stress, which we observed as having a nearly identical leaf curling symptom in strawberry.

During the 2019-2020 Florida strawberry season (October to April), a strawberry ( Fragaria × ananassa ) fruit rot was observed in two fields (Plant City and Wimauma, FL) with up to 5% incidence on short-day cultivars Sensation TM Florida127, and Florida Brilliance. Symptoms on pink and ripe fruit consisted of softening, discoloration, watery rot with white fuzzy mycelium, and initial sclerotium formation. Diseased tissue was placed on General Isolation (GI) medium (Amiri et al. 2018) and incubated at 25°C under a 12-h photoperiod. A fungus producing spreading cottony white colonies with dark sclerotia near the outer edges of the plates was consistently isolated. One isolate from each cultivar field (20-51 and 20-55) was selected for identification and pathogenicity tests. Apothecial production was induced following the protocol of Li and Rollins (2009), and apothecia (n=30) had an average diameter of 4.5 (3.5 to 7.2) mm. Sclerotia were 3.6 (2.5 to 6.2) mm by 4.5 (3.1 to 5.9) mm (n=30) in size. Dimensions of asci were 130.2 (115.1 to 160.5) μm by 8.5 (6.2 to 13.1) μm (n=30), and those of ascospores were 12.2 (10.8 to 14.6) μm by 6.8 (5.7 to 8.1) μm (n=30). Based on these morphological features, the pathogen was tentatively identified as Sclerotinia sclerotiorum (Lib.) de Bary (Maas 1998). DNA was extracted from the same two isolates using the FastDNA kit (MP Biomedicals, Solon, OH), and the ribosomal internal transcribed spacer (ITS) region was amplified using the primers ITS1 and ITS4 (White et al. 1990). Sequences were deposited in GenBank (accession nos. MT378215 and MT378216). BLASTn searches revealed that isolates 20-51 and 20-55 were 99.62% identical (526 / 528 bp) to S. sclerotiorum CBS 499.50 (MH856725.1). Immature pink fruit of Sensation TM 'Florida127' were harvested, surface disinfested in bleach solution (0.08% NaClO) for 90 sec, rinsed twice with deionized water, then placed into styrofoam egg cartons inside clean plastic boxes (30x24x7 cm) containing 150 ml of sterile deionized water to maintain moisture, and kept at 25°C with a 12-h photoperiod. Ascospores and sclerotia were used for inoculation tests with three repetitions in an egg carton containing 12 fruit each per isolate and inoculation method. The experiment was repeated once. Fruit were inoculated by placing 20 μl of a 1 × 10 6 ascospores/mL suspension or a single sclerotium on the upper half of the fruit. Controls were included, by placing 20 μl of sterile DI water or fruit with no sclerotia. Evaluations were done 6, 10, and 15 days after inoculation (DAI). Control fruit remained healthy, while inoculated fruit developed symptoms of softening and discoloration. For ascospore inoculation, disease incidence was 55 (± 5) and 78% (± 4), for 6 and 15 DAI, respectively, whereas for sclerotia inoculation incidence was 100% 6 DAI. Morphologically identical fungi to the original isolates were re-isolated from the diseased fruit. No other fruit decay fungi were observed. S. sclerotiorum has been previously reported causing strawberry fruit rot in Washington state in the United States, England, Israel, and Scotland (Alcorn 1966; Maas 1998; McLean 1957). It has also been listed in the indices of plant diseases from Florida, North Carolina, and California as causing crown rot (Farr and Rossman 2020). To our knowledge, this is the first report of S. sclerotiorum causing strawberry fruit rot in Florida. The pathogen is an aggressive necrotroph with prolonged survival and affects several vegetable crops grown in Florida (Paret et al. 2018). Because only the strawberry beds are fumigated, sclerotia remaining in the alleys could serve as inoculum sources. Currently, the disease is rare and of minor significance to strawberry production. However, efforts should be implemented to monitor its occurrence and spread.

Strawberry ( Fragaria x ananassa ) is an important crop in the U.S., and Florida is the second major producer in the country. In January 2020, an unknown fruit rot was observed in two strawberry fields in Dover (seedling selection) and Plant City (cultivar Florida127), Florida. Disease incidence varied from less than 1% in one field to up to 15% in the second field during some harvests where over-ripe fruit were present. Affected fruit had a water-soaked soft rot with a sour smell, and sometimes with white mycelium on the fruit surface. Direct isolation was performed from symptomatic fruit from each area by touching the surface of a lesion with a sterile needle and streaking the fungus over general isolation medium (Amiri et al. 2018). The fungus was incubated at 25°C and 12-h photoperiod for five days. Four single-spore isolates (20-46 and 20-47 from Plant City; 20-49 and 20-50 from Dover) obtained from different colonies were grown on potato dextrose agar (PDA). Colonies were white to cream, flat, with a powdery surface, and had a characteristic sour odor. Hyphae were hyaline, septate, growing in a dichotomous pattern and often disarticulating into arthroconidia, which were unicellular, spherical to cylindrical, measuring 4.8 to 9.5 × 3.5 to 5.6 µm (n = 50). Based on the morphological characteristics, the fungus was identified as Geotrichum candidum (De Hoog et al. 1986). To confirm the identity of the isolates, the ribosomal internal transcribed spacer (ITS-rDNA) and the translation elongation factor 1-alpha (TEF1-α) gene regions were amplified and sequenced using the primers ITS1/ITS4 (White et al. 1990), and EF1-728F/EF1-1567R (Carbone and Kohn 1999), respectively. Consensus sequences were deposited in the GenBank (accession numbers MT353975 to MT353978 for ITS-rDNA, and MT346367 to MT346370 for TEF1-α). The BLASTn analysis revealed 99% identity with reference sequences of G. candidum for ITS-rDNA (KU373122) and TEF1-α (MK397513). Phylogenetic analysis, including ITS sequences of G. candidum and other spp., obtained from GenBank, was performed using maximum likelihood and Bayesian inference methods, implemented in MEGA-X and MrBayes, respectively. The isolates were grouped within the G. candidum clade. The pathogenicity of the fungus (isolate 20-46) was evaluated in ripe and green strawberry fruit (cultivar Florida127). Non-wounded and superficially wounded (with a sterilized needle) fruit were inoculated with 20 µl of a spore suspension (1 × 10 7 spore/ml), prepared by washing the surface of a 5-day-old colony on PDA, with sterile deionized water (SDW) plus 0.01% (v/v) Tween 20. Wounded and non-wounded control fruit were treated with SDW. Fruit were maintained in moist containers at 25°C. Each treatment consisted of 12 fruit (three replicates, each containing four fruit) and was evaluated daily for 10 days. Symptoms of sour rot were visible as soon as 48 hr after inoculation, but only on ripe (100% incidence) and green (58% incidence) wounded fruit. Symptoms progressed to an intense water-soaking with the presence of a typical white mycelium on the surface. Control fruit remained symptomless. The fungus was successfully recovered from symptomatic fruit, fulfilling Koch's postulates. Although strawberry fruit rots caused by G. candidum have already been reported in Pakistan and China (Hussain et al. 2016; Ma et al. 2018), this is the first report of this species causing sour rot on strawberry in the U.S. The disease may be a minor problem on damaged or over-ripe fruit, but further studies might be needed to determine its importance, distribution, and potential strategies for control.

Postbloom fruit drop (PFD) of citrus is caused by the Colletotrichum acutatum and C. gloeosporioides species complexes. The disease is important when frequent rainfall occurs during the flowering period of citrus trees. In Brazil, until 2012, PFD was mainly controlled by preventive applications of the methyl-benzimidazole carbamate (MBC) carbendazim and demethylation-inhibitor (DMI) fungicides such as difenoconazole. Since then, mixtures containing the DMI tebuconazole and the quinone-outside inhibitor (QoI) trifloxystrobin have been commonly used. Fungicides are often applied preventively, sometimes even when conditions are not conducive for PFD development. Excessive fungicide applications may favor the selection of resistant populations of Colletotrichum spp. In this study, we assessed the fungicide sensitivity of C. acutatum isolates collected during the two distinct periods of PFD management in Brazil: before and after the trifloxystrobin and tebuconazole mixture became widely employed. The sensitivity of 254 C. acutatum isolates to carbendazim and difenoconazole and of 164 isolates to tebuconazole and trifloxystrobin was assessed. Mycelial growth inhibition of these isolates was evaluated for all the fungicides using either serial dilution of fungicide rates or the spiral gradient dilution method. In addition, inhibition of conidial germination was also assessed for trifloxystrobin. Analysis of partial β-tub , cytb , and cyp51b gene sequences did not reveal any mutations related to resistance to MBCs, QoIs, and DMIs, respectively. In mycelial growth assays, mean EC 50 values were 0.14, 0.11, and 0.21 μg/ml for difenoconazole, tebuconazole, and trifloxystrobin, respectively. The conidial germination inhibition by trifloxystrobin was similar among the tested isolates, and the mean EC 50 value was 0.002 μg/ml. All isolates had similar mean mycelial growth inhibition for carbendazim, regardless of the fungicide concentrations. Therefore, based on similar EC 50 values and molecular analyses, no shift in the sensitivity of isolates has been observed to the fungicides commonly used in different citrus-producing areas in Brazil.

Succinate dehydrogenase inhibitors (SDHIs) are the fungicides most commonly used to control Botrytis fruit rot on commercial strawberry in Florida. The medium-to-high risk of selection of resistance in the causal agent Botrytis cinerea is a threat to the efficacy of this fungicide group. In this study, we characterized the sensitivity of B. cinerea to the SDHI isofetamid, evaluated the SdhB gene mutation associated with resistance, and monitored resistance frequencies to five SDHI fungicides for two consecutive seasons. EC 50 values of 70 isolates were obtained using the spiral gradient dilution (SGD) method and averaged 0.098 µg/ml of isofetamid. EC 50 averages of 3.04 and >500.00 µg/ml were obtained for isolates with the N230I and P225F mutations indicating moderate and high resistance to isofetamid, respectively. A total of 565 B. cinerea isolates collected during 2015-2016 and 2016-2017 seasons from strawberry nurseries and Florida production fields were evaluated using conidial germination assays. Results for the first season showed resistance frequencies of 95, 33, 21, 25, and 0% to boscalid, penthiopyrad, fluopyram, benzovindiflupyr, and isofetamid, respectively. The respective resistance frequencies for the following season were 91, 95, 44, 27, and 1.3%. Only three isolates were found to be moderately resistant to isofetamid during the second season, and the mutation N230I was identified after sequence analysis. These isolates were confirmed to be resistant to isofetamid in fruit assays with disease incidence of 55.6 to 77.0%; however, the conidial production of the isolates was inhibited by an average of 83.9%. In general, isofetamid efficacy was higher than the other evaluated SDHIs, but a slight increase in resistance frequencies was observed in our study.

Background: Ethanedinitrile (EDN) has shown promising efficacy against weeds, plant-parasitic nematodes and soil-borne pathogens. Field experiments were conducted for three strawberry (Fragaria× ananassa D.) seasons in Balm and Dover, FL, USA to evaluate the control efficacy of EDN applied through drip tape on various weed species and the fungus Macrophomina phaseolina, the causal agent of charcoal rot of strawberry., Results: Results revealed that 224, 336, 448 or 560 kg ha -1 EDN, and 104 kg ha -1 1,3-dichloropropene (1,3-D) + 176 kg ha -1 chloropicrin (Pic) applied through drip tapes under totally impermeable film were safe for strawberry production, with no adverse effect on strawberry growth and yield. EDN rates at 224, 336, 448 and 560 kg ha -1 were highly efficacious and equally effective in reducing purple nutsedge (Cyperus rotundus L.) in two of three growing seasons. Compared with the efficacy on purple nutsedge, the evaluated EDN rates were generally less effective on various broadleaf and grass weeds emerging on the bare ground and planting holes, but at 560 kg ha -1 consistently exhibited similar levels of control as 1,3-D + Pic. For M. phaseolina, the evaluated EDN rates were more efficacious than 1,3-D + Pic and significantly reduced inoculum buried in different bed locations in plastic-mulched beds., Conclusion: EDN has potential to be an efficacious tool for soil disinfestation and weed control in plasticulture strawberry production. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Succinate dehydrogenase inhibitors (SDHIs) are an essential group of fungicides for managing gray mold, caused by Botrytis cinerea , in numerous crops. Resistance to boscalid, an early-generation SDHI, is widespread worldwide and was linked to mutations in the iron-sulfur protein encoding the SdhB subunit of the SDH complex. Herein, we report on four simultaneous dependent mutations at codons 85 (G85A), 93 (I93V), 158 (M158V), and 168 (V168I) of the membrane-anchored SdhC subunit of B. cinerea . Isolates without and with mutations in SdhC were referred to as C - and C + genotypes, respectively. The C + genotype was found in all the five surveyed hosts from different U.S. regions but its frequency was higher, 25 to 40%, in the tree fruit isolates compared with 12 to 25% in the small fruit populations. The four SdhC mutations were found in isolates without mutations in SdhB or with mutations known to confer resistance to the SDHIs in SdhB. However, the frequency of C + isolates was significantly higher in the SdhB wild-type isolates, which suggests that SDHI sprays may have played a role in selecting for the C - over the C + genotype. Field C + isolates exhibited reduced sensitivity to fluopyram and increased sensitivity to boscalid and penthiopyrad in vitro and on detached fruit. Homology modeling confirmed the positioning of the four mutations in the ubiquinone-binding pocket. The SdhC G85A is found in the proximal ubiquinone binding site and SdhC M158V is positioned in the iron sulfur protein interface next to the [3Fe-4S] cluster, whereas SdhC I93V is positioned next to the heme b with vital functions in the SDH enzyme. Beside the differential sensitivity to the SDHIs, these mutations caused a significant fitness cost in the C + isolates including sporulation and increased sensitivity to reactive oxygen species. The presence of Botrytis populations differentially sensitive to the SDHIs suggests increased risks for resistance development but also opens up new perspective for future gray mold management using different SDHI fungicides.

Anthracnose fruit rot and leaf blight caused by Colletotrichum species are important diseases of pomegranate in the southeastern United States. In this study, 26 isolates from pomegranate were identified based on pathological and molecular characterization. Isolates were identified to species based on multilocus sequence analysis with the internal transcribed spacer region, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin, and chitin synthase genomic genes. Pomegranate isolates grouped within the C. acutatum and C. gloeosporioides species complexes, with more than 73% belonging to the latter group. Three species were identified within the C. acutatum species complex ( C. nymphaeae [ n = 5], C. fioriniae [ n = 1], and C. simmondsii [ n = 1]), and three other species were identified within the C. gloeosporioides species complex ( C. theobromicola [ n = 11], C. siamense [ n = 6], and C. gloeosporioides [ n = 2]). Inoculations of pomegranate fruit showed that isolates from the C. acutatum species complex were more aggressive than isolates from the C. gloeosporioides species complex. Interestingly, opposite results were observed when leaves of rooted pomegranate cuttings were inoculated. In addition, Colletotrichum isolates from pomegranate, strawberry, blueberry, mango, and citrus were cross-pathogenic when inoculated to fruit. This is the first study identifying six different species of Colletotrichum causing pomegranate leaf blight and fruit anthracnose in the southeastern United States and the potential cross-pathogenic capability of pomegranate isolates to other commercially important crops.