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Made available in DSpace on 2020-12-10T20:00:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) 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 beta-tub, cytb, and cyp51b gene sequences did not reveal any mutations related to resistance to MBCs, QoIs, and DMIs, respectively. In mycelia] growth assays, mean EC50 values were 0.14, 0.11, and 0.21 mu g/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 mu g/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. Univ Sao Paulo, Escola Super Agr Luiz Queiroz, BR-13418900 Piracicaba, SP, Brazil Univ Florida, Gulf Coast Res & Educ Ctr, Wimauma, FL 33598 USA Sao Paulo State Univ, Campus Expt Registro, Registro, SP, Brazil Fundo Defesa Citricultura Res & Dev, BR-14807040 Araraquara, SP, Brazil Sao Paulo State Univ, Campus Expt Registro, Registro, SP, Brazil CNPq: 2015/06007-2

Rhizopus rot, caused by Rhizopus stolonifer, is a major postharvest disease of stone fruits. The disease is related to the occurrence of mechanical and physical damage; however, observations at a Brazilian wholesale market suggest that direct penetration can occur. Therefore, the penetration mechanisms of R. stolonifer in stone fruits were evaluated. To identify the production of enzymes that help with direct penetration by the pathogen, esterase activity, both in mycelial discs and in spore suspensions of the fungus in water and in modified Van Etten nutrient solution, was measured. Assays were also conducted to evaluate the growth of R. stolonifer on glucose or cutin as a sole carbon source. The pathogen grew on both media, and higher esterase activity was observed in the cutin medium. Wounded and unwounded peaches and nectarines were inoculated with R. stolonifer spore suspensions in water or in modified Van Etten nutrient solution. Wounded fruit inoculated with either of the R. stolonifer spore suspensions developed rhizopus rot, whereas unwounded fruit developed the rot only in the presence of spores in the modified Van Etten nutrient solution. Scanning electron and light microscopic examination showed the fungus can directly penetrate the nectarine cuticle. Diisopropyl fluorophosphate, a serine hydrolase inhibitor, prevented rot development in peaches. The results provide valuable evidence for the ability of R. stolonifer to directly penetrate unwounded stone fruits, probably due to the production of esterase enzymes.

Brown rot, caused by Monilinia spp. , is one of the most important peach (Prunuspersica (L.) Batsch) diseases and the main cause of postharvest losses. Currently, alternative methods for postharvest disease control, such as the use of volatiles, are under investigation. The objective of this study was to determine the effects of hexanal on the in vitro development of Monilinia fructicola and M. laxa and on monocyclic components of brown rot on peaches. To evaluate the effect on pathogen development in vitro, a single dose of 215 µL of liquid hexanal was placed on glass jars in closed plastic containers (4.3 L) at the moment of fungi transfer, 24 or 48 h after transferring to Petri dishes. After hexanal application, the Petri dishes were kept inside the containers that were closed for 24 h at 20 ºC. Mycelial growth was measured seven days after hexanal removal. For in vivo assays, inoculated fruits were kept in closed plastic containers, and hexanal was applied at the moment of fruit inoculation or 24 hours thereafter. The monocyclic components infection frequency, expressed as brown rot incidence, lesion diameter and lesion sporulation, were assessed daily for seven days. Overall, hexanal was more effective in inhibiting mycelial growth when applied at the moment of pathogen transfer. Hexanal did not prevent pathogen infection, but reduced lesion diameter and completely inhibited spore production on the fruit for both treatments. Hexanal provides a promising alternative for chemical control and can be used in postharvest handling systems.

O objetivo desse trabalho foi avaliar o efeito in vitro e in vivo dos sanificantes cloreto de benzalcônio (Fegatex®), biomassa cítrica (Ecolife40®) e ozônio no controle da podridão parda (Monilinia fructicola) e da podridão mole (Rhizopus stolonifer) em pêssegos das cultivares Aurora, Dourado e Flor da Prince. Cloreto de benzalcônio e biomassa cítrica, aplicados in vitro, ambos na concentração de 1000 mL L-1, inibiram totalmente o crescimento radial (micelial) de M. fructicola, porém nenhum deles foi eficiente no controle de R. stolonifer. Cloreto de benzalcônio aplicado de forma preventiva, na concentração de 3000 mL L-1, reduziu a podridão parda em frutos inoculados sem ferimentos. Quando aplicado de forma curativa em frutos não feridos esse produto foi eficiente em todas as concentrações testadas. Nenhum produto aplicado nos frutos de forma curativa foi eficiente no controle da podridão parda, quando a inoculação do fungo foi realizada através de ferimentos. Nenhum dos produtos foi eficiente no controle da podridão mole. O ozônio (0,1 mL L-1) não foi eficiente no controle das podridões parda e mole. The purpose of this work was to study the in vitro and in vivo effects of the sanitizing products benzalkonium chloride (Fegatex®), citric biomass (Ecolife40®) and ozone gas on the control of brown rot (Monilinia fructicola) and soft rot (Rhizopus stolonifer) in peaches cultivars "Aurora", "Dourado" and "Flor da Prince". Benzalkonium chloride and citric biomass, both at 1000 mL L-1, completely inhibited the radial (mycelial) growth of M. fructicola, although none of them controlled R. stolonifer in vitro. The preventive use of benzalkonium chloride at 3000 mL L-1 reduced brown rot in non-injured peaches. Benzalkonium chloride applied as curative in non-injured peaches was effective against M. fructicola at all concentrations. None of the products tested curatively was effective in preventing the development of brown rot when the fungus was inoculated over fruit injuries. None of the products was effective in controlling soft rot. Ozone gas at 0,1 mL L-1 was not effective in controlling brown or soft rots in peaches.

Este trabalho teve por objetivo avaliar o efeito da irradiação UV-C no controle in vitro de Monilinia fructicola e Rhizopus stolonifer e no controle das doenças causadas por estes fungos em pêssegos inoculados com ferimento. No experimento in vitro, avaliou-se o crescimento micelial dos fungos em meio BDA após a exposição nas doses de UV-C de 0, 0,26, 0,52, 1,04, 3,13, 5,22, 10,44, 15,66, e 31,32 kJ.m-2 num equipamento com quatro lâmpadas com taxa de fluência de 1,74 mW.cm-2. Nos experimentos in vivo, os frutos foram tratados com irradiação UV-C de forma protetora e curativa. No tratamento protetor, os frutos foram expostos a 1,04 kJ.m-2 por 1 min. e foram inoculados imediatamente após e 16, 24 e 40 h após. No tratamento curativo, os frutos foram inoculados, incubados e irradiados com doses de UV-C de 0, 1,04, 5,22, 10,44, 15,66 e 31,32 kJ.m². Avaliou-se a incidência das doenças e a severidade da podridão parda. No experimento in vitro, apenas as doses aplicadas durante 1 e 10 min. de exposição reduziram o crescimento micelial de M. fructicola enquanto que a aplicação da luz UV-C entre 10-15 minutos reduziu o crescimento micelial de R. stolonifer e a dose aplicada durante 30 minutos inibiu completamente o crescimento micelial deste fungo. Não houve efeito protetor da luz UV-C no controle das doenças. Não houve controle curativo da podridão parda. A irradiação UV-C foi eficiente no controle curativo da podridão mole e o tempo de exposição de 10 min. foi o que apresentou melhor resultado. The purpose of this work was to evaluate the effect of UV-C irradiation in vitro to control M. fructicola and R. stolonifer and the diseases caused by them in peach fruits. Mycelial growth was evaluated after exposure of the pathogens on PDA medium to different UV-C irradiation concentrations of 0, 0.26, 0.52, 1.04, 3.13, 5.22, 10.44, 15.66 and 31.32 kJ.m-2. The UV cabinet was equipped with four lamps with the fluency rate of 1.74 mW.cm-2. Peach fruits were treated with UV-C irradiation in a protective and curative way. In the protector treatment, fruits were exposed to 1.04 kJ.m-2 for 1 min. and were inoculated immediately and after 16, 24 and 40 hours. In the curative treatment, peach fruits were inoculated, incubated and irradiated with 0, 1.04, 5.22, 10.44, 15.66 and 31.32 kJ.m-2 UV-C concentrations. Disease incidence and severity of brown rot were evaluated. In vitro, only the concentrations applied during 1 and 10 min. of exposure reduced the mycelial growth of M. fructicola, while the application of UV-C for 10-15 minutes reduced the mycelial growth of R. stolonifer and the concentration applied for 30 minutes completely inhibited the mycelial growth of this pathogen. There was no effect caused by UV-C light in the protective control, or by UV-C light in the curative control of brown rot. The irradiation of fruits with UV-C was effective in the curative control of soft rot, and the best results were observed with an exposure time of 10 minutes.