Association of SO2-Generating Pads before Packaging and during Cold Storage to Extend the Conservation of 'Italia' Table Grapes.

The SO2-generating pads contain different concentrations of sodium metabisulfite, which absorbs water from the grapes' transpiration, releasing SO2 gas, and there are slow-(SlowSO2) and dual (DualSO2)-releasing pads (fast release in the first 48 h and slow for up to 60 days). The ultra-fast SO2-generating pad (FieldSO2) releases the SO2 quickly for up to 6 h, and it was designed to be used soon after the harvest and until the grapes' packaging. The goal was to study the effect of FieldSO2 associated with SlowSO2 and DualSO2 pads on gray mold incidence and physicochemical and appearance characteristics of 'Italia' table grapes. Grapes were harvested from a commercial vineyard in Parana, Brazil, in 2020 and 2021, and packaged in cardboard boxes, and the treatments were as follows: control (without SO2-generating pads); FieldSO2 + SlowSO2; and FieldSO2 + DualSO2. After 30, 45, 60, 75, and 90 days of cold storage (1 ± 1 °C), the grapes were assessed for gray mold incidence, mass loss, shattered berries, stem browning, and filamentous fungi on the surface. The use of FieldSO2 associated with SO2-generating pads is effective in controlling gray mold on 'Italia' table grapes, especially the treatment FieldSO2 + DualSO2, which provides the lowest incidence of the disease up to 90 days of cold storage, while the combination with SlowSO2 results in intermediate efficacy. Treatments combining these SO2-generating pads extend the postharvest shelf life of 'Italia' grapes, with few shattered berries, low mass loss and freshness of the rachis without impairing the bunch's appearance.

Control of Botrytis cinerea from Chilean grapevines by pydiflumetofen: baseline and carboxamide-mutant sensitivity.

The gray mold (Botrytis cinerea; Botrytis) is the main disease affecting grapevines production in Chile. Succinate Dehydrogenase Inhibitors (SDHI) belonging to the carboxamides fungicide family are a key tool for the control of Botrytis in grapevines from Chilean Central Valley. This study aimed to determine the sensitivity of Chilean Botrytis population to the new generation carboxamide pydiflumetofen. Conidial germination (CG) and germ-tube elongation (GTE) sensitivity assays were conducted on 200 single-spore isolates collected during the 2016-2017 season. The mean effective concentration that inhibited 50% (EC50) of CG in the Botrytis population was 0.0545 µg/mL, with mean values of 0.066 µg/mL and 0.042 µg/mL, for table and wine grapes, respectively. The mean EC50 value of GTE was 0.000245 µg/mL, 0.0003 µg/mL, and 0.0019 µg/mL for the total, table grape, and wine grape populations, respectively. The comparison between pydiflumetofen and fludioxonil, a highly-efficient fungicide carrying a different mode of action, showed the 87.5% and 97.5% of Botrytis control with an EC50 threshold of 0.1 µg/mL, in table grape, and wine grape populations, respectively. No cross-resistance between pydiflumetofen and fludioxonil was detected. For nine isolates with reduced pydiflumetofen sensitivity, we evaluated SdhB mutations using a qPCR-HRM diagnostic system. Two isolates carried the sdhBP225/H272R genotype and two the sdhBP225/H272Y. Additional analysis of SdhB mutant isolates determined that pydiflumetofen controls wild-type as well as sdhBP225/H272R and sdhBP225H/H272 mutants. Pydiflumetofen does not control CG in the sdhBP225/H272Y mutant but is effective in the GTE control. Pydiflumetofen significantly controls Botrytis independently of the SdhB genotype in wounded berry assays. This condition resembles the berry cracking due to heavy rainfall right before harvest, as seen in recent years in the Chilean Central Valley. The findings demonstrate that pydiflumetofen effectively controls the grapevine Botrytis population, suggest a moderate risk of pydiflumetofen resistance, and highlight the significance of incorporating genetic data into the design of control programs.

Microstructural and physicochemical quality maintenance in green bell pepper infected with Botrytis cinerea and treated with thyme essential oil combined with carnauba wax.

Bell pepper presents rapid weight loss and is highly susceptible to gray mold caused by the fungus Botrytis cinerea. The most employed method to control this disease is the application of synthetic fungicides such as thiabendazole (TBZ); however, its continued use causes resistance in fungi as well as environmental problems. For these reasons, natural alternatives arise as a more striking option. Currently, bell pepper fruits are coated with carnauba wax (CW) to prevent weight loss and improve appearance. Moreover, CW can be used as a carrier to incorporate essential oils, and previous studies have shown that thyme essential oil (TEO) is highly effective against B. cinerea. Therefore, this study aimed to evaluate the effect of CW combined with TEO on the development of gray mold and maintenance of microestructural and postharvest quality in bell pepper stored at 13°C. The minimal inhibitory concentration of TEO was 0.5%. TEO and TBZ provoked the leakage of intracellular components. TEO and CW + TEO treatments were equally effective to inhibit the development of gray mold. On the quality parameters, firmness and weight loss were ameliorated with CW and CW + TEO treatments; whereas lightness increased in these treatments. The structural analysis showed that CW + TEO treatment maintained the cell structure reducing the apparition of deformities. The results suggest that CW + TEO treatment could be used as a natural and effective antifungal retarding the appearance of gray mold and maintaining the postharvest quality of bell pepper. PRACTICAL APPLICATION: CW and TEO are classified as generally recognized as safe (GRAS) by the US Food and Drug Administration (FDA). This combination can be employed on the bell pepper packaging system to extend shelf life and oppose gray mold developments. Bell pepper fruits are normally coated with lipid-base coatings such as CW before commercialization; therefore, TEO addition would represent a small investment without any changes on the packaging system infrastructure.

First Report of Gray Mold Caused by Botrytis cinerea on Dragon Fruit (Hylocereus undatus) in Mexico.

The dragon fruit is native of Mexico, and Puebla is the third-largest producing state (SIAP 2023). In June 2023, field sampling was conducted in El Paraíso, Atlixco (18° 49' 5.275" N, 98° 26' 52.353" W), Puebla, Mexico. The mean temperature and relative humidity were 20 °C and 75% for seven consecutive days. Dragon fruits cv. 'Delight' close to harvest with gray mold symptoms were found in a commercial area of 2 ha, with an incidence of 35 to 40% and an estimated severity of 75% on infected fruit. The symptoms included necrosis at the apex, which later spread throughout the fruit, along with a soft, black rot covered in abundant mycelium and sporulation. The fungus was isolated from 40 symptomatic fruits by disinfesting pieces of necrotic tissue with 3% NaClO for one minute, rinsing with sterile distilled water (SDW), plating on Petri dishes with potato dextrose agar, and incubating at 25 °C in the dark. One isolate was obtained from each diseased fruit by the hyphal-tip method. The colonies were initially white with a growth rate of 1.15-1.32 cm per day and turned gray after 10 days; the mycelium was dense and aerial. Spherical and irregular sclerotia were formed, measuring 0.9-1.4 × 0.6-1.1 mm (n = 100). Each Petri dish produced 56-278 sclerotia (n = 40) after 11 days; these were initially white and gradually turned dark brown. Brown to olive conidiophores were straight, septate, and branched, measuring 1075-1520 × 10-21 µm, with elliptical hyaline to light brown conidia of 6.6-11.5 × 5-8.1 µm (n=100). The isolates were tentatively identified as Botrytis cinerea based on morphological characteristics (Ellis 1971). Two representative isolates were chosen for molecular identification and genomic DNA was extracted by the CTAB protocol. The ITS region and the heat shock protein (HSP60), RNA polymerase binding II (RPB2) and glyceraldehyde 3-phosphate dehydrogenase (G3PDH) genes were sequenced (White et al. 1990; Staats et al. 2005). The sequences of a representative isolate (BcPh5) were deposited in GenBank (ITS-OR582337; HSP60-OR636622; RPB2-OR636623; and G3PDH-OR636621). BLAST analysis of the partial sequences of ITS (479 bp), HSP60 (1006 bp), RPB2 (1126 bp), and G3PDH (907 bp) showed 100% similarity to B. cinerea isolates (GenBank: KM840848, MH796663, MK919495, MF480679). Phylogenetic analysis confirmed that BcPh5 clustered with B. cinerea strains. Pathogenicity was confirmed by inoculating the non-wounded surface of 20 detached dragon fruits cv. 'Delight' using the BcPh5 isolate by depositing 20 µl of a 105 conidia/ml suspension with a sterile syringe. The fruits were placed on the rim of a plastic container and inserted in a moisture box with 2 cm of water at the bottom. The box was covered with a plastic sheet to maintain humidity. Control fruits were inoculated with SDW. The inoculated fruits became covered with abundant white to gray mycelium, and soft rot developed within eight days, while no symptoms were observed on the controls. The fungus was re-isolated from the inoculated fruits as described above, fulfilling Koch's postulates. The pathogenicity tests were repeated three times. Gray mold caused by B. cinerea was also recently reported in Mexico on pomegranate (Hernández et al. 2023) and rose apple (Isodoro et al. 2023). As far as we know, this is the first report of B. cinerea causing gray mold on dragon fruit in Mexico. This research is essential for designing integrated management strategies against gray mold on dragon fruits.

A site suitability analysis for castor (Ricinus communis L.) production during Brazil's second harvest incorporating disease prediction.

Castor (Ricinus communis L.) is an important industrial crop with a wide range of industrial and pharmaceutical applications. Brazil is among the largest castor-producing countries. Between 2004 and 2010, castor cultivation was stimulated with an emphasis towards biodiesel production. However, this was not enough to leverage the production of castor in Brazil, mainly due to the lack of structured trade and the competition with other cheaper raw materials for the production of biodiesel. Despite this failure, the species presents itself as an excellent alternative for crop rotation in the second crop among soybean, corn, beans, and cotton cultivation areas as the oil is highly valuable for other products. Moreover, it has drawn the attention of producers and researchers in Brazil for this potential rotation as it is considered a plant tolerant of water-deficiency and is highly susceptible to gray mold, a disease favored by high humidity in the final stages of the crop. For instance, its cultivation in the second crop in Cerrado regions, where rains occur in the early stages of the crop and cease when the plants reach the final stage of production, has been successful and shows great promise. The current study aimed to evaluate the suitability of environments throughout Brazil to grow castor, incorporating variables associated with the incidence of gray mold and confirm these findings based on existing castor trial data obtained from the literature. The site suitability analysis determined that 74.99 million hectares - 8.8% of Brazilian territory - are highly suitable for castor production during second harvest, mostly located in the Northeastern and Midwestern regions. These results are surprising since Brazil currently has around 7.8% (∼66.81 million hectares) of its territory occupied with agriculture (grains, fruits, vegetables, and perennial crops). The findings of this study provide a method to perform site suitability for crops using data associated with agronomic and disease characteristics, as is the case with gray mold that often results in significant losses in castor production. Also, this analysis provides evidence for the great potential of Brazil to increase castor production and meet the world demand for its oil through utilization of second-crop cultivation.

Indigenous Yeasts for the Biocontrol of Botrytis cinerea on Table Grapes in Chile.

One hundred twenty-five yeast strains isolated from table grapes and apples were evaluated for the control Botrytis cinerea of in vitro and in vivo. Ten strains were selected for their ability to inhibit mycelial growth of B. cinerea in vitro. In the in vivo assays, these yeasts were tested at 20 °C on 'Thompson Seedless' berries for 7 days; only three were selected (m11, me99 and ca80) because they significantly reduced the incidence of gray mold. These three yeast strains were then evaluated at different concentrations (1 × 107, 1 × 108 and 1 × 109 cells mL-1) on 'Thompson Seedless' grape berries at 20 °C. The strains m11, me99 and ca80 reduced the incidence of B. cinerea to 11.9, 26.1 and 32.1%, respectively, when the berries were submerged in a yeast suspension at a concentration of 1 × 109 cells mL-1 24 h before inoculation with B. cinerea. The most favorable pH for antifungal activity was 4.6 in the three isolates. The three yeast strains secreted the hydrolytic enzymes chitinase and ß-1-glucanase, and two strains (me99 and ca80) produced siderophores. The three yeast strains exhibited low oxidative stress tolerance and only strain m11 had the ability to produce biofilms. The strains were identified using 5.8S-ITS rDNA PCR-RFLP and correspond to the Meyerozyma guilliermondii (m11) and Aureobasidium pullulans (me99 and ca80) species.

Identification of the main proteins secreted by Kluyveromyces marxianus and their possible roles in antagonistic activity against fungi.

Lytic enzymes secreted by Kluyveromyces marxianus can lyse Saccharomyces cerevisiae cells. Their ability to hydrolyze yeast cell walls can be used in biotechnological applications, such as the production of glucans and protoplasts, as well as a biological control agent against plant pathogenic fungi. Herein, 27 proteins secreted by K. marxianus were identified by mass spectrometry analyses. Importantly, 14 out of the 27 proteins were classified as hydrolases. Indeed, the enzyme extract secreted by K. marxianus caused damage to S. cerevisiae cells and reduced yeast cell viability. Moreover, K marxianus inhibited spore germination and mycelial growth of the phytopathogenic fungus Botrytis cinerea in simultaneous cocultivation assays. We suggest that this inhibition may be partially related to the yeast's ability to secrete lytic enzymes. Consistent with the in vitro antagonistic tests, K. marxianus was able to protect strawberry fruits inoculated with B. cinerea. Therefore, these findings suggest that K. marxianus possesses potential as a biocontrol agent against strawberry gray mold during the postharvest stage and may also have potential against other phytopathogenic fungi by means of its lytic enzymatic arsenal.

Synthesis of bis(ylidene) cyclohexanones and their antifungal activity against selected plant pathogenic fungi.

Botrytis cinerea, Rhizoctonia solani and Hemileia vastatrix are three species of phytopathogenic fungi behind major crop losses worldwide. These have been selected as target models for testing the fungicide potential of a series of bis(ylidene) cyclohexanones. Although some compounds of this chemical class are known to have inhibitory activity against human pathogens, they have never been explored for the control of phytopathogens until now. In the present work, bis(ylidene) cyclohexanones were synthesized through simple, fast and low-cost base- or acid-catalyzed aldol condensation reaction and tested in vitro against B. cinerea, R. solani and H. vastatrix. bis(pyridylmethylene) cyclohexanones showed the highest activity against the target fungi. When tested at 200 nmol per mycelial plug against R. solani., these compounds completely inhibited the mycelial growth, and the most active bis(pyridylmethylene) cyclohexanone compound had an IC50 of 155.5 nmol plug-1. Additionally, bis(pyridylmethylene) cyclohexanones completely inhibited urediniospore germination of H. vastatrix, at 125 µmol L-1. The most active bis(pyridylmethylene) cyclohexanone had an IC50 value of 4.8 µmol L-1, which was estimated as approximately 2.6 times lower than that found for the copper oxychloride-based fungicide, used as control. Additionally, these substances had a low cytotoxicity against the mammalian Vero cell line. Finally, in silico calculations indicated that these compounds present physicochemical parameters regarded as suitable for agrochemicals. Bis(ylidene) cyclohexanones may constitute promising candidates for the development of novel antifungal agents for the control of relevant fungal diseases in agriculture.

Inhibition of mycelial growth, conidial germination, and Botrytis cinerea Pers: Fr colonization in begonia with biocompatible products

This study evaluated the effects of potassium and sodium carbonate and bicarbonate, Bacillus subtilis (Cohn, 1872) QST-713, Bacillus pumilus (Meyer & Gottheil, 1901) QST-2808, and crude and roasted coffee oils on the inhibition of mycelial growth and conidial germination in Botrytis cinerea Pers.: Fr and the colonization of begonia (Begonia elatior Hort. ex Steud) leaf discs by B. cinerea inoculated before, simultaneously and after with these alternative products. The assays were carried out using the Baladin begonia cultivar. The inhibition of B. cinerea mycelial growth and conidial germination was proportional to increases in the concentration of all the products. The inhibition of conidial germination was directly proportional to the concentrations of B. pumilus QST-2808 and B. subtilis QST-713. Coffee oils were less efficient in inhibiting germination than the other products. The crude and roasted coffee oils, potassium and sodium carbonates and bicarbonates, and B. pumilus and B. subtilis sprayed 24 h before, simultaneously, or 24 h after pathogen inoculation inhibited the colonization of begonia leaf discs by B. cinerea. The positive results for the suppression of B. cinerea by the alternative products tested herein merit scrutiny. There is a pressing need to evaluate these products in the management of gray mold, as the severity of this disease is usually high under favorable conditions in greenhouses.

Mycofumigation of postharvest blueberries with volatile compounds from Trichoderma atroviride IC-11 is a promising tool to control rots caused by Botrytis cinerea.

Botrytis cinerea, the causal agent of the gray mold, is a filamentous fungus that infects blueberries and can cause important production losses in postharvest storage. Considering that the use of synthetic fungicides is not allowed on blueberries in postharvest conditions, alternative and natural strategies are needed to control gray mold. The objective of this work was to evaluate the capability of volatile organic compounds (VOCs) produced by Trichoderma atroviride IC-11 to control B. cinerea growth in blueberries after harvest. These VOCs inhibited almost completely B. cinerea growth in vitro. The most abundant volatile compound was 6-pentyl-α-pyrone (6PP). In vitro assays with pure 6PP confirmed its antifungal activity. The incidence of gray mold was evaluated in blueberries inoculated with B. cinerea and exposed to volatiles of T. atroviride IC-11. Gray mold incidence among those stored in air at 20 °C for 14 days was 100%, while the incidence among the volatile-treated fruit was 17%. Gray mold incidence among those stored in air at 4 °C for 31 days was 82%, while the incidence among the volatile-treated fruit was 11%. T. atroviride IC-11 VOCs inhibited mycelial growth and conidia germination of B. cinerea. The binding of VOCs to the surface of hyphae caused their vacuolation and deterioration. Selective cytotoxicity of 6PP on B. cinerea was observed but not on human intestinal cells at specific concentrations that controlled gray mold. The postharvest mycofumigation of blueberries with T. atroviride IC-11 VOCs is a promising approach to protect these fruits from gray mold.

Characterization of Botrytis cinerea and B. prunorum From Healthy Floral Structures and Decayed 'Hayward' Kiwifruit During Post-Harvest Storage.

Gray mold is the primary postharvest disease of 'Hayward' kiwifruit (Actinidia deliciosa) in Chile, with a prevalence of 33.1% in 2016 and 7.1% in 2017. Gray mold develops during postharvest storage, which is characterized by a soft, light to brown watery decay that is caused by Botrytis cinerea and B. prunorum. However, there is no information on the role of B. prunorum during the development and storage of kiwifruit in Chile. For this purpose, asymptomatic flowers and receptacles were collected throughout fruit development and harvest from five orchards over two seasons in the Central Valley of Chile. Additionally, diseased kiwifruits were selected after storage for 100 days at 0°C and 2 days at 20°C. Colonies of Botrytis sp. with high and low conidial production were consistently obtained from apparently healthy petals, sepals, receptacles, and styles and diseased kiwifruit. Morphological and phylogenetic analysis of three partial gene sequences encoding glyceraldehyde-3-phosphate dehydrogenase, heat shock protein 60, and DNA-dependent RNA polymerase subunit II were able to identify and separate B. cinerea and B. prunorum species. Consistently, B. cinerea was predominantly isolated from all floral parts and fruit in apparently healthy tissue and diseased kiwifruit. During full bloom, the highest colonization by B. cinerea and B. prunorum was obtained from petals, followed by sepals. In storage, both Botrytis species were isolated from the diseased fruit (n = 644), of which 6.8% (n = 44) were identified as B. prunorum. All Botrytis isolates grew from 0°C to 30°C in vitro and were pathogenic on kiwifruit leaves and fruit. Notably, B. cinerea isolates were always more virulent than B. prunorum isolates. This study confirms the presence of B. cinerea and B. prunorum colonizing apparently healthy flowers and floral parts in fruit and causing gray mold during kiwifruit storage in Chile. Therefore, B. prunorum plays a secondary role in the epidemiology of gray mold developing in kiwifruit during cold storage.

Diversity, prevalence and phylogenetic positioning of Botrytis species in Brazil.

Botrytis is a necrotrophic fungal genus of great economic importance worldwide. Together, the Botrytis species are able to infect over one thousand host plant species, including dicotyledons and monocotyledons. As the identification of Botrytis species in Brazil has mostly been based only on morphological characterization and comparisons of the rDNA ITS region, which is not informative in the genus, its diversity remains unknown. Thus, in this study we determined the diversity and prevalence of Botrytis spp. in Brazil by multilocus phylogeny. Phylogenetic reconstruction of the genus was performed using the nuclear genes glyceraldehyde-3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60) and RNA polymerase II second largest subunit (RPB2). From analyses of 56 Botrytis isolates obtained from different hosts and geographical regions in Brazil, we found that Botrytis cinerea is the most prevalent species with considerable intraspecific genetic diversity detected by nuclear genes. Two new hosts to B. cinerea and eight host never previously reported in Brazil were found. We also reported for the first time the occurrence of Botrytispseudocinerea associated with Accasellowiana (Myrtaceae). Due to the new phylogenetic positioning of Botrytispelargonii and Botrytiseucalypti, a taxonomic review of these species was suggested.

Reproducibility of the Development and Validation Process of Standard Area Diagram by Two Laboratories: An Example Using the Botrytis cinerea/Gerbera jamesonii Pathosystem.

Standard area diagrams (SADs) are plant disease severity assessment aids demonstrated to improve the accuracy and reliability of visual estimates of severity. Knowledge of the sources of variation, including those specific to a lab such as raters, specific procedures followed including instruction, image analysis software, image viewing time, etc., that affect the outcome of development and validation of SADs can help improve standard operating practice of these assessment aids. As reproducibility has not previously been explored in development of SADs, we aimed to explore the overarching question of whether the lab in which the measurement and validation of a SAD was performed affected the outcome of the process. Two different labs (Lab 1 and Lab 2) measured severity on the individual diagrams in a SAD and validated them independently for severity of gray mold (caused by Botrytis cinerea) on Gerbera daisy. Severity measurements of the 30 test images were performed independently at the two labs as well. A different group of 18 raters at each lab assessed the test images first without, and secondly with SADs under independent instruction at both Lab 1 and 2. Results showed that actual severity on the SADs as measured at each lab varied by up to 5.18%. Furthermore, measurement of the test image actual values varied from 0 to up to 24.29%, depending on image. Whereas at Lab 1 an equivalence test indicated no significant improvement in any measure of agreement with use of the SADs, at Lab 2, scale shift, generalized bias, and agreement were significantly improved with use of the SADs (P ≤ 0.05). An analysis of variance indicated differences existed between labs, use of the SADs aid, and the interaction, depending on the agreement statistic. Based on an equivalence test, the interrater reliability was significantly (P ≤ 0.05) improved at both Lab 1 and Lab 2 as a result of using SADs as an aid to severity estimation. Gain in measures of agreement and reliability tended to be greatest for the least able raters at both Lab 1 and Lab 2. Absolute error was reduced at both labs when raters used SADs. The results confirm that SADs are a useful tool, but the results demonstrated that aspects of the development and validation process in different labs may affect the outcome.

Priming of Defense Systems and Upregulation of MYC2 and JAZ1 Genes after Botrytis cinerea Inoculation in Methyl Jasmonate-Treated Strawberry Fruits.

Several attempts have been made to study the effects of methyl jasmonate (MeJA) on plants in the past years. However, the comparative effects of the number and phenological time of MeJA applications on the activation of defense systems is currently unknown in strawberries. In the present research, we performed three field treatments during strawberry (Fragaria× ananassa 'Camarosa') fruit development and ripening which consisted of differential MeJA applications at flowering (M3), and the large green (M2 and M3) and red ripe (M1, M2, and M3) fruit stages. We also checked changes in gene expression related to plant defense against Botrytis cinerea inoculation post-harvest. In M3 treatment, we observed an upregulation of the anthocyanin and lignin contents and the defense-related genes, encoding for chitinases, ß-1,3-glucanases and polygalacturonase-inhibiting proteins, after harvest (0 hpi), along with the jasmonate signaling-related genes FaMYC2 and FaJAZ1 at 48 h after B. cinerea inoculation (48 hpi) during postharvest storage. Although we did not find differences in gray mold incidence between the MeJA treatments and control, these results suggest that preharvest MeJA treatment from the flowering stage onwards (M3) primes defense responses mediated by the upregulation of different defense-related genes and retains the upregulation of MYC2 and JAZ1 at 48 hpi.

In vitro control of Botrytis cinerea and Penicillium italicum by antagonistic yeasts / Controle in vitro de Botrytis cinerea e Penicillium italicum por leveduras antagonistas

Gray mold in grapes (caused by Botrytis cinerea) and blue mold in citrus (caused by Penicillium italicum) are post-harvest diseases that cause major losses in these crops. The control of these diseases is based on the use of synthetic chemical fungicides. The increase in regulatory policies and demand to reduce the application of pesticides, due to harmful effects on the environment and humans, have led to the search for more ecofriendly alternatives, such as biological control agents. Thus, the present work aims to verify the antagonistic potential of four yeast strains, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762), and Lachancea thermotolerans (CCMA0763), against of B cinerea and P. italicum. To assess the antagonism of volatile compounds, Petri plates with two divisions containing potato-dextrose-agar (PDA) were used by placing a fungal mycelial disc and yeast suspension (3.0 × 106 cells mL-¹) on opposite sides of the plate. The colony diameter and mycelial growth rate index of the fungi were evaluated via comparisons with the control plate without yeast. For the evaluation of the antagonism of diffusible substances in the medium, yeasts were striated 3 cm from the center of the plates containing PDA. After 48 h, a mycelial disc of each phytopathogen was placed in the center of the plates. The colony growth, inhibition halo, and mycelial growth rate index were evaluated via comparisons with the control plate. All yeast strains showed an antagonistic effect on the mycelial growth of B. cinerea in both tests. In the volatile compounds test, H. opuntiae, L. thermotolerans, P. caribbica, and P. manshurica inhibited mycelial growth by approximately 82%, 75%, 72%, and 50%, respectively. In the antagonism test of the diffusible substances in the medium, [...].

O mofo cinzento na uva (causada por Botrytis cinerea) e o bolor azul nos citros (causada por Penicillium italicum) são doenças de pós-colheita que causam perdas importantes nestas culturas. O controle destas doenças é baseado na utilização de fungicidas químicos sintéticos. O aumento das políticas regulatórias e a demanda para reduzir a aplicação de agrotóxicos, devido a efeitos prejudiciais ao meio ambiente e aos seres humanos, levam à procura de alternativas mais ecológicas como o uso de agentes de controle biológico. Desta forma, o presente trabalho teve por finalidade analisar o potencial antagônico de quatro cepas de leveduras, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762) e Lachancea thermotolerans (CCMA 0763) no controle de B. cinerea e P. italicum. Para avaliar o antagonismo por compostos voláteis, foram utilizadas placas bipartidas contendo meio batata-dextrose-ágar (BDA) onde foram colocados um disco micelial dos fungos e uma suspensão de 3,0x106 cél. mL-¹ das leveduras em lados opostos da placa. Avaliou-se o diâmetro da colônia e índice de velocidade de crescimento micelial dos fungos, comparando-a com a placa controles em a levedura. Para a avaliação do antagonismo de substâncias difusíveis no meio, as leveduras foram estriadas a 3 cm do centro de placas contendo meio BDA e após 48 h colocou-se um disco micelial de cada fitopatógeno no centro das mesmas. Avaliou-se o crescimento da colônia, a formação do halo de inibição e o índice de velocidade de crescimento micelial comparando-os com a placa controle. Todos os isolados apresentaram efeito antagônico no crescimento micelial de B. cinerea nos dois testes realizados. H. opuntiae, L. thermotolerans, P. caribbica e P. manshurica inibiram o crescimento micelial em aproximadamente 82%, 75%, 72% e 50%, respectivamente, no teste de compostos voláteis. P. caribbica e P. manshurica inibiram o crescimento micelial em 58% e 33%, [...].

In vitro control of Botrytis cinerea and Penicillium italicum by antagonistic yeasts / Controle in vitro de Botrytis cinerea e Penicillium italicum por leveduras antagonistas

Gray mold in grapes (caused by Botrytis cinerea) and blue mold in citrus (caused by Penicillium italicum) are post-harvest diseases that cause major losses in these crops. The control of these diseases is based on the use of synthetic chemical fungicides. The increase in regulatory policies and demand to reduce the application of pesticides, due to harmful effects on the environment and humans, have led to the search for more ecofriendly alternatives, such as biological control agents. Thus, the present work aims to verify the antagonistic potential of four yeast strains, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762), and Lachancea thermotolerans (CCMA0763), against of B cinerea and P. italicum. To assess the antagonism of volatile compounds, Petri plates with two divisions containing potato-dextrose-agar (PDA) were used by placing a fungal mycelial disc and yeast suspension (3.0 × 106 cells mL-¹) on opposite sides of the plate. The colony diameter and mycelial growth rate index of the fungi were evaluated via comparisons with the control plate without yeast. For the evaluation of the antagonism of diffusible substances in the medium, yeasts were striated 3 cm from the center of the plates containing PDA. After 48 h, a mycelial disc of each phytopathogen was placed in the center of the plates. The colony growth, inhibition halo, and mycelial growth rate index were evaluated via comparisons with the control plate. All yeast strains showed an antagonistic effect on the mycelial growth of B. cinerea in both tests. In the volatile compounds test, H. opuntiae, L. thermotolerans, P. caribbica, and P. manshurica inhibited mycelial growth by approximately 82%, 75%, 72%, and 50%, respectively. In the antagonism test of the diffusible substances in the medium, [...].(AU)

O mofo cinzento na uva (causada por Botrytis cinerea) e o bolor azul nos citros (causada por Penicillium italicum) são doenças de pós-colheita que causam perdas importantes nestas culturas. O controle destas doenças é baseado na utilização de fungicidas químicos sintéticos. O aumento das políticas regulatórias e a demanda para reduzir a aplicação de agrotóxicos, devido a efeitos prejudiciais ao meio ambiente e aos seres humanos, levam à procura de alternativas mais ecológicas como o uso de agentes de controle biológico. Desta forma, o presente trabalho teve por finalidade analisar o potencial antagônico de quatro cepas de leveduras, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762) e Lachancea thermotolerans (CCMA 0763) no controle de B. cinerea e P. italicum. Para avaliar o antagonismo por compostos voláteis, foram utilizadas placas bipartidas contendo meio batata-dextrose-ágar (BDA) onde foram colocados um disco micelial dos fungos e uma suspensão de 3,0x106 cél. mL-¹ das leveduras em lados opostos da placa. Avaliou-se o diâmetro da colônia e índice de velocidade de crescimento micelial dos fungos, comparando-a com a placa controles em a levedura. Para a avaliação do antagonismo de substâncias difusíveis no meio, as leveduras foram estriadas a 3 cm do centro de placas contendo meio BDA e após 48 h colocou-se um disco micelial de cada fitopatógeno no centro das mesmas. Avaliou-se o crescimento da colônia, a formação do halo de inibição e o índice de velocidade de crescimento micelial comparando-os com a placa controle. Todos os isolados apresentaram efeito antagônico no crescimento micelial de B. cinerea nos dois testes realizados. H. opuntiae, L. thermotolerans, P. caribbica e P. manshurica inibiram o crescimento micelial em aproximadamente 82%, 75%, 72% e 50%, respectivamente, no teste de compostos voláteis. P. caribbica e P. manshurica inibiram o crescimento micelial em 58% e 33%, [...].(AU)

Control of Gray Mold on Clamshell-Packaged 'Benitaka' Table Grapes Using Sulphur Dioxide Pads and Perforated Liners.

The use of vented clamshells has become popular in the packaging of grapes for local and international markets. The aim of this study is to evaluate the postharvest preservation of 'Benitaka' table grapes individually packaged in vented clamshells using different types of SO2-generating pads and perforated plastic liners during cold storage. A completely randomized design with four replications in a two-factor arrangement with an additional treatment [(4 × 3) + 1] was used. The trials were carried out under two situations: Artificial or natural infections with Botrytis cinerea, which is the causal agent of gray mold on table grapes. The incidence of gray mold, shattered berries, and stem browning were evaluated at 30 and 45 days of cold storage at 1 ± 1 °C and 3 days of shelf-life at 22 ± 1 °C after the period of cold storage. Mass loss and berry firmness were also examined at the end of the cold storage period. The use of dual-release SO2-generating pads containing 5 or 8 g of a.i. and slow-release pads with 7 g of a.i. was effective in controlling the incidence of gray mold in grapes packaged in vented clamshells and kept under cold storage for up to 45 days. Under these storage conditions, perforated plastic liners with 0.3% ventilation area or micro-perforated liners with 1.0% ventilation area reduced the percentage of mass loss and shattered berries.

Natural Compounds That Modulate the Development of the Fungus Botrytis cinerea and Protect Solanum lycopersicum.

Botrytis cinerea is the causal agent of gray mold disease and is responsible for the loss of millions of dollars in crops in worldwide. Currently, this pathogen exhibits increasing resistance to conventional fungicides; therefore, better control methods and novel compounds with a more specific mechanism of action but without biocidal effects, are required. In this work, several natural compounds to control B. cinerea were analyzed in vitro. Detected effects were dependent on the stage of fungus development, and 3-phenyl-1-propanol displayed the most potent inhibition of in vitro germination, germ tube development, and sporulation. However, it had lower protection of leaves and postharvest fruit in plant infection. Isoeugenol and 1-phenylethanol exhibited lower inhibition of in vitro germination and sporulation, but at the highest concentrations, they inhibited germ tube elongation. Although the lowest rates of foliage infection were recorded using isoeugenol and 3-phenyl-1-propanol, 1-phenylethanol significantly decreased the disease in postharvest tomato fruit, with an efficacy like Mancozeb, but at 18 times lower micromolar concentration. All compounds resulted in high cell viability after spores were removed from the treatment solution exhibited high cell viability, suggesting a non-biocidal effect. The diversity of in vitro and in-plant effects seems to indicate a different mechanism of action.

Evaluation of Rhodosporidium fluviale as biocontrol agent against Botrytis cinerea on apple fruit.

The aim of the present work was to evaluate the ability of the native yeast Rhodosporidium fluviale to control Botrytis cinerea on apple fruit and to study the possible mechanisms of action with the goal of improving the control of gray mold. For this, the influence of application time of the yeast was studied simulating preventive and curative effects. Also, the effect of nonviable cells of the yeast in the biocontrol was assessed. According to the results obtained, the following mechanisms of action of R. fluviale could be proposed: 1- competition for space, 2- direct interaction between antagonist and pathogen, 3- induction of ß-1,3-glucanase in apple tissue, 4- Probable production of glucanase in the apple wounds and 5- antifungal action of cellular components, probably chitin, present in the wall of yeast cells that could be the explanation for the activity of nonviable cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Significance and Impact of the Study: Botrytis cinerea Pers: Fr, which causes gray mold of fruits and vegetables around the world, is difficult to control successfully because it is genetically variable and rapidly develops resistance to the chemicals commonly used for its control. This study is a contribution to the biocontrol of this phytopathogen fungus. The evaluation of the native yeast Rhodosporidium fluviale as biocontrol agent and the elucidation of possible mechanisms of action, including the participation of nonviable cells of this yeast, have not been reported up to date.

The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of Botrytis cinerea Produced by Pseudomonas aeruginosa LV Strain.

One of the most important postharvest plant pathogens that affect strawberries, grapes and tomatoes is Botrytis cinerea, known as gray mold. The fungus remains in latent form until spore germination conditions are good, making infection control difficult, causing great losses in the whole production chain. This study aimed to purify and identify phenazine-1-carboxylic acid (PCA) produced by the Pseudomonas aeruginosa LV strain and to determine its antifungal activity against B. cinerea. The compounds produced were extracted with dichloromethane and passed through a chromatographic process. The purity level of PCA was determined by reversed-phase high-performance liquid chromatography semi-preparative. The structure of PCA was confirmed by nuclear magnetic resonance and electrospray ionization mass spectrometry. Antifungal activity was determined by the dry paper disk and minimum inhibitory concentration (MIC) methods and identified by scanning electron microscopy and confocal microscopy. The results showed that PCA inhibited mycelial growth, where MIC was 25 µg mL-1. Microscopic analysis revealed a reduction in exopolysaccharide (EPS) formation, showing distorted and damaged hyphae of B. cinerea. The results suggested that PCA has a high potential in the control of B. cinerea and inhibition of EPS (important virulence factor). This natural compound is a potential alternative to postharvest control of gray mold disease.