A Semi-Selective Medium to Evaluate Over-Summering Survival of Neopestalotiopsis sp. in Florida Strawberry Fields.

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.

Identification of sequence mutations in Phytophthora cactorum genome associated with mefenoxam resistance and development of a molecular assay for the mutant detection in strawberry (F. × ananassa).

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.

Old but Gold: Captan Is a Valuable Tool for Managing Anthracnose and Botrytis Fruit Rots and Improving Strawberry Yields Based on a Meta-Analysis.

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.

Resistance of Strawberry Cultivars and the Effects of Plant Ontogenesis on Phytophthora cactorum and P. nicotianae Causing Crown Rot.

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.

Efficacy of Single- and Multi-Site Fungicides Against Neopestalotiopsis spp. of Strawberry.

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.

Phosphite Is More Effective Against Phytophthora Crown Rot and Leather Rot Caused by Phytophthora cactorum than P. nicotianae.

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% (EC50) 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, EC50 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.

A reassessment of the fungicidal efficacy of 1,3-dichloropropene, chloropicrin, and metam potassium against Macrophomina phaseolina in strawberry.

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.

Sensitivity of Colletotrichum acutatum Species Complex from Strawberry to Fungicide Alternatives to Quinone-Outside Inhibitors.

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.

Phytophthora Crown Rot of Florida Strawberry: Inoculum Sources and Thermotherapy of Transplants for Disease Management.

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.

Cultivar Selection Is an Effective and Economic Strategy for Managing Charcoal Rot of Strawberry in Florida.

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.

Physical, Cultural, and Chemical Alternatives for Integrated Management of Charcoal Rot of Strawberry.

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.

Outbreak of Leaf Spot and Fruit Rot in Florida Strawberry Caused by Neopestalotiopsis spp.

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, ß-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.

Sensitivity of Colletotrichum acutatum Isolates from Citrus to Carbendazim, Difenoconazole, Tebuconazole, and Trifloxystrobin.

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 ß-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 EC50 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 EC50 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 EC50 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.

Evaluation of ethanedinitrile (EDN) as a preplant soil fumigant in Florida strawberry production.

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.

Sources of Inoculum and Survival of Macrophomina phaseolina in Florida Strawberry Fields.

Macrophomina phaseolina, the causal agent of charcoal rot, affects strawberry crowns, inducing plant collapse. The fungus survives in the soil through the production of microsclerotia and is usually controlled by preplant fumigation of soil. However, in the 2016 to 2017 Florida strawberry season, even after soil fumigation, about 30% plant mortality still occurred in plastic-covered beds that were used for a second season and where crop residue (mainly old strawberry crowns) was disposed of between beds. Therefore, this study was conducted to determine if M. phaseolina can survive on strawberry debris over summer in Florida and if so, verify whether strawberry debris might act as a source of inoculum for new transplants. Crowns from the previous season were collected from commercial farms where charcoal rot had been reported, and M. phaseolina was recovered from all samples. In a research field, infected crowns were buried in the soil at different depths and retrieved every 2 weeks during the summer. After 8 weeks, M. phaseolina could be recovered at all depths. Moreover, inoculation of strawberry plants by drenching the soil, dipping roots, or spraying leaves with a M. phaseolina microsclerotial suspension from pure cultures or infected crowns produced symptoms with differences in incubation periods depending on cultivar susceptibility. Furthermore, infected crowns disposed of in the aisles between beds or buried next to new transplants of cultivars Strawberry Festival, Florida Beauty, and Winterstar induced charcoal rot, with the level of aggressiveness depending on the cultivar susceptibility and inoculum placement in the field.

Effect of Formulations of Allyl Isothiocyanate on Survival of Macrophomina phaseolina from Strawberry.

Management of Macrophomina phaseolina, causal agent of charcoal rot in many crops worldwide, including strawberry, has become more challenging since the phase out of methyl bromide (MeBr). The search for a fumigant equally effective as MeBr to control soilborne pathogens has been extensive. Allyl isothiocyanate (AITC), a biofumigant recently registered in the United States, was evaluated at different rates, formulations, fumigant combinations, and application methods in the fall of 2014 and 2015 at two research facilities in Balm and Dover, FL. The efficacy of treatments was determined by evaluating the survival of M. phaseolina inoculum on infested corn-cob litter buried in bags 7.6 and 20.3 cm deep in the center, and 7.6 cm deep in the side, of plastic mulched raised beds. The biofumigant was shown to be more or as effective in reducing populations of M. phaseolina in the soil compared with standard fumigants, such as chloropicrin and 1,3-dichloropropene with chloropicrin. Thus, AITC is a promising biofumigant alternative for managing charcoal rot of strawberry, particularly in organic production systems, and should be evaluated for the management of other soilborne pathogens.

Sensitivity of Botrytis cinerea Isolates from Conventional and Organic Strawberry Fields in Brazil to Azoxystrobin, Iprodione, Pyrimethanil, and Thiophanate-Methyl.

Botrytis fruit rot, caused by Botrytis cinerea, is one of the most important strawberry diseases worldwide, and fungicide applications are often used to manage the disease in commercial production. Isolates of B. cinerea were collected from conventional and organic strawberry fields in four Brazilian States from 2013 to 2015 and their sensitivity to the main single-site mode-of action fungicides used in Brazil was tested. Resistance to azoxystrobin, iprodione, pyrimethanil, and thiophanate-methyl was found and values for effective concentration that inhibited mycelial growth by 50% were higher than 71.9, 1.2, 5.0, and 688 µg/ml, respectively, regardless the production system. Resistance to these fungicides was observed in 87.5, 76.6, 23.4, and 92.2% of isolates from conventional fields and 31.4, 22.9, 14.3, and 51.4% of isolates from organic fields, respectively. Moreover, frequencies of isolates with multiple fungicide resistance to the four active ingredients were 20.6 and 2.8% whereas 6.3 and 27.8% were sensitive to the four fungicides for conventional and organic areas, respectively. Molecular analyses of the cytochrome b, ß-tubulin, and bos1 genes revealed the presence of G143A; E198A; and I365 N/S, Q369P, or N373S mutations, respectively, in resistant isolates of B. cinerea. Field rates of fungicides sprayed preventively to inoculated strawberry fruit failed to control disease caused by the respective resistant isolates.