Lime sulfur to control citrus flat mite and its interactions with the entomopathogenic fungus Lecanicillium muscarium

Lime sulfur is one of the few products indicated to control Brevipalpus yothersi in Brazilian organic citrus orchards. Other strategies, such as the use of entomopathogenic fungi should be evaluated, and Lecanicillium muscarium is one of the basic choices for pest management. Knowledge of the interactions between lime sulfur and this entomopathogen is critical for developing control strategies. With this goal, it was conducted the toxicological characterization of lime sulfur to B. yothersi and the compatibility evaluation with L. muscarium. Finally, the effects of L. muscarium and lime sulfur mixtures on B. yothersi control were evaluated. Product evaluation for B. yothersi was done through direct and residual contact bioassay, and different concentrations of lime sulfur mixed in potato dextrose agar culture medium were used to evaluate compatibility with L. muscarium. Lime sulfur was effective against adults of B. yothersi and caused eggs unviability of up to 71.0%, at a dose of 80 L per 2,000 L of H2O. The lethal concentration (LC50 and LC99) of lime sulfur estimated for mite adults were 246.62 and 858.5 µg of sulfur per mL of H2O (ppm a.i.). Lime sulfur concentrations of 180 to 560 ppm a.i. showed promise for use in combination with L. muscarium. However, concentrations of 1,000 and 5,600 ppm significantly reduced colony size and the number of spores/colony. The mixture of 100 and 180 ppm a.i. of lime sulfur with L. muscarium (108 conidia·mL­1) was not able to reduce the lethal time of entomopathogen on B. yothersi.

Lethal and sublethal effects of pesticides used in Brazilian citrus groves on Panonychus citri (Acari: Tetranychidae) / Efeitos letais e subletais de agrotóxicos utilizados em citros sobre o ácaro Panonychus citri (Acari: Tetranychidae)

Pesticides have been blamed as the principal factor responsible for biological disequilibrium favoring the population increase of Panonychus citri (McGregor) in Brazilian citrus groves. In order to generate subsidies for the integrated management of this pest, we evaluated the lethal and sublethal effects of pesticides used in citrus on P. citri. We tested 18 pesticides by evaluating mortality of adult female, egg viability and adult avoidance. For imidacloprid, pyriproxyfen, dimethoate, deltamethrin, lambda-cyhalothrin and lime sulfur, the principal insecticides used in Brazilian orchards, the oviposition behavior and the incidence of adult females on discontinuous pesticide residues were evaluated. The pesticides which caused adult P. citri mortality were: abamectin (94%), dimethoate (86%), lime sulfur (69%), fenpropathrin (44%), diafenthiuron (25%) and lambda-cyhalothrin (23%). The pesticides affecting egg viability were: fenpropathrin (53%), dimethoate (30%) and lime sulfur (22%). Fenpropathrin, lambda-cyhalothrin and lime sulfur caused repellent effects of 55, 45 and 22%, respectively. P. citri preferred areas untreated with deltamethrin, lambda-cyhalothrin and lime sulfur for oviposition and permanence, but the species could not distinguish areas treated with imidacloprid, pyriproxyfen and dimethoate. The insecticides abamectin, dimethoate and diafenthiuron affected survival and did not cause changes in P. citri behavior. These pesticides should be used mainly in periods P. citri occurs. On the other hand, fenpropathrin, lambda-cyhalothrin, lime sulfur and deltamethrin affect dispersal behavior, oviposition and P. citri incidence and their use should be avoided.(AU)

O intenso uso de pesticidas na citricultura tem sido apontado como o principal fator responsável por desequilíbrios biológicos, favorecendo o aumento populacional de Panonychus citri (McGregor) na citricultura brasileira. Para gerar subsídios ao manejo integrado dessa praga, avaliaram-se os efeitos letais e subletais de agrotóxicos utilizados em citros sobre P. citri. Avaliaram-se 18 agrotóxicos na mortalidade de fêmeas adultas, viabilidade de ovos e repelência de P. citri. Foram também avaliados o comportamento de oviposição e a incidência de P. citri em superfícies com áreas tratadas e não tratadas com imidacloprid, piriproxifem, dimetoato, deltametrina, lambda-cialotrina e calda sulfocálcica, por serem os principais inseticidas utilizados nos pomares brasileiros. Os pesticidas que causaram mortalidade em adultos de P. citri foram abamectina (94%), dimetoato (86%), calda sulfocálcica (69%), fempropatrina (44%), diafentiurom (25%) e lambda-cialotrina (23%). Os pesticidas que afetaram a viabilidade de ovos foram fempropatrina (53%), dimetoato (30%) e calda sulfocálcica (22%). Quanto aos efeitos subletais, fempropatrina, lambda-cialotrina e calda sulfocálcica ocasionaram repelências de 55, 45 e 22%, respectivamente. P. citri preferiu áreas não tratadas com deltametrina, lambda-cialotrina e calda sulfocálcica para oviposição e permanência. Em contraste, P. citri não foi capaz de distinguir superfícies tratadas com imidacloprid, piriproxifem e dimetoato. Os inseticidas abamectina, dimetoato e diafentiurom afetaram a sobrevivência e não causaram mudanças no comportamento de P. citri, podendo ser utilizados principalmente em períodos que P. citri ocorre. Em contrapartida, fempropatrina, lambda-cialotrina, calda sulfocálcica e deltametrina afetaram o comportamento de dispersão e oviposição de P. citri e deveriam ser evitados.(AU)

An antifungal peptide from Coffea canephora seeds with sequence homology to glycine-rich proteins exerts membrane permeabilization and nuclear localization in fungi.

BACKGROUND: The superfamily of glycine-rich proteins (GRPs) corresponds to a large and complex group of plant proteins that may be involved in many developmental and physiological processes such as RNA biogenesis, stress tolerance, pollen hydration and plant-pathogen interactions, showing defensive activity against fungi, bacteria and viruses. METHODS: In this study, the peptides from Coffea canephora seeds were extracted according to the methods of Egorov et al. (2005). The purified peptide was submitted for amino acid sequencing and antimicrobial activity measurement. RESULTS: The purified peptide with a molecular weight of 7kDa, named Cc-GRP, was observed to display homology to GRPs. The Cc-GRP-fungi interaction led to morphological changes and membrane permeability, including the formation of pseudohyphae, which were visualized with the aid of SYTOX green dye. Additionally, Cc-GRP also prevented colony formation by yeasts. Antifungal assays of Fusarium oxysporum and Colletotrichum lindemuthianum, observed by light microscopy, showed that the two molds exhibited morphological changes after the growth assay. Cc-GRP coupled to FITC and its subsequent treatment with DAPI revealed the presence of the peptide in the cell wall, cell surface and nucleus of F. oxysporum. CONCLUSIONS AND GENERAL SIGNIFICANCE: In this work we purified, characterized and evaluated the in vitro effect on fungi of a new peptide from coffee, named Cc-GRP, which is involved in the plant defense system against pathogens by acting through a membrane permeabilization mechanism and localized in the nuclei of fungal cells. We also showed, for the first time, the intracellular localization of Cc-GRP during antimicrobial assay.

[Resistance of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) to acaricides that inhibit cellular respiration in citrus: cross-resistance and fitness cost]. / Resistência de Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) a acaricidas inibidores da respiração celular em citros: resistência cruzada e custo adaptativo.

Acaricides that inhibit cellular respiration play an important role in the control of Brevipalpus phoenicis (Geijskes) in citrus groves in Brazil. Studies were conducted to evaluate: (a) the variability in the susceptibility among B. phoenicis populations collected from citrus groves to cyhexatin, azocyclotin, propargite and sulphur; (b) cross-resistance relationships between propargite and azocyclotin, cyhexatin, dinocap, pyridaben and sulphur; and (c) the fitness cost associated with propargite resistance in B. phoenicis under laboratory conditions. A residual-type contact bioassay was used to characterize the susceptibility. The susceptibility was estimated with diagnostic concentrations based LC(95) of each acaricide. The cross-resistance was evaluated by characterizing the concentration-mortality responses of susceptible (S) and propargite-resistant (Propargite-R) strains. The fitness cost was evaluated by measuring the biological parameters of S and Propargite-R strains on citrus fruits at 25 +/- 1 degrees C and fotophase of 14h. Significant differences in the susceptibility of B. phoenicis were detected at diagnostic concentration of cyhexatin (survivorship from 16.3% to 80.5%), azocyclotin (from 3.0% to 15.0%), propargite (from 1.0% a 71.6%) and sulphur (from 9.0% to 82.6%). A low intensity of cross-resistance was detected between propargite and the acaricides azocyclotin (1.8-fold), cyhexatin (4.6-fold), dinocap (3.5-fold) and pyridaben (3.5-fold). On the other hand, the intensity of cross-resistance to sulphur (> 111-fold) was very high. There was no fitness cost associated with B. phoenicis resistance to propargite, based on biological parameters evaluated. Therefore, the use of these acaricides should also be done very carefully in resistance management of B. phoenicis to acaricides.

Resistência de Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) a acaricidas inibidores da respiração celular em citros: resistência cruzada e custo adaptativo / Resistance of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) to acaricides that inhibit cellular respiration in citrus: cross-resistance and fitness cost

Acaricidas inibidores da respiração celular são importantes para o controle de Brevipalpus phoenicis (Geijskes) na cultura dos citros no Brasil. Foram conduzidos estudos para avaliar: (a) a variabilidade na suscetibilidade de populações de B. phoenicis coletadas em pomares de citros para cihexatim, azociclotim, propargite e enxofre; (b) relações de resistência cruzada entre propargite e os acaricidas azociclotim, cihexatim, dinocape, piridabem e enxofre; e (c) o custo adaptativo associado à resistência de B. phoenicis a propargite em condições de laboratório. O método de bioensaio adotado foi o contato residual. A suscetibilidade foi estimada com concentrações diagnósticas baseadas na CL95 de cada acaricida. A resistência cruzada foi avaliada pela caracterização da curva de concentração-mortalidade das linhagens suscetível (S) e resistente a propargite (Propargite-R). O custo adaptativo foi avaliado mediante comparação de parâmetros biológicos das linhagens S e Propargite-R em frutos de laranja a 25 ± 1°C e fotofase de 14h. Diferenças significativas na sobrevivência de B. phoenicis foram verificadas aos acaricidas cihexatim (de 16,3 por cento a 80,5 por cento), azociclotim (de 3,0 por cento a 15,0 por cento), propargite (de 1,0 por cento a 71,6 por cento) e enxofre (de 9,0 por cento a 82,6 por cento). Baixa intensidade de resistência cruzada foi verificada entre propargite e os acaricidas azociclotim (1,8 vezes), cihexatim (4,6 vezes), dinocape (3,5 vezes) e piridabem (3,5 vezes). Por outro lado, a intensidade de resistência cruzada a enxofre (> 111 vezes) foi bastante alta. Não foi verificada presença de custo adaptativo associado à resistência de B. phoenicis a propargite. Portanto, o uso desses acaricidas também deve ser feito de maneira criteriosa no manejo da resistência de B. phoenicis a acaricidas.

Acaricides that inhibit cellular respiration play an important role in the control of Brevipalpus phoenicis (Geijskes) in citrus groves in Brazil. Studies were conducted to evaluate: (a) the variability in the susceptibility among B. phoenicis populations collected from citrus groves to cyhexatin, azocyclotin, propargite and sulphur; (b) cross-resistance relationships between propargite and azocyclotin, cyhexatin, dinocap, pyridaben and sulphur; and (c) the fitness cost associated with propargite resistance in B. phoenicis under laboratory conditions. A residual-type contact bioassay was used to characterize the susceptibility. The susceptibility was estimated with diagnostic concentrations based LC95 of each acaricide. The cross-resistance was evaluated by characterizing the concentration-mortality responses of susceptible (S) and propargite-resistant (Propargite-R) strains. The fitness cost was evaluated by measuring the biological parameters of S and Propargite-R strains on citrus fruits at 25 ± 1°C and fotophase of 14h. Significant differences in the susceptibility of B. phoenicis were detected at diagnostic concentration of cyhexatin (survivorship from 16.3 percent to 80.5 percent), azocyclotin (from 3.0 percent to 15.0 percent), propargite (from 1.0 percent a 71.6 percent) and sulphur (from 9.0 percent to 82.6 percent). A low intensity of cross-resistance was detected between propargite and the acaricides azocyclotin (1.8-fold), cyhexatin (4.6-fold), dinocap (3.5-fold) and pyridaben (3.5-fold). On the other hand, the intensity of cross-resistance to sulphur (> 111-fold) was very high. There was no fitness cost associated with B. phoenicis resistance to propargite, based on biological parameters evaluated. Therefore, the use of these acaricides should also be done very carefully in resistance management of B. phoenicis to acaricides.

Mecanismos de dispersão de Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) em pomares de citros / Dispersal mechanisms of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) in citrus groves

O conhecimento da dispersão de pragas é de grande importância para a compreensão da dinâmica da resistência a pesticidas. Desta forma, o principal objetivo do presente trabalho foi estudar a capacidade de dispersão de Brevipalpus phoenicis (Geijskes) em condições de laboratório, casa de vegetação e pomares de citros. Estudos de dispersão por caminhamento em condições de casa de vegetação numa superfície constituída de areia fina mostraram que esse mecanismo de dispersão é relativamente limitado. Com a liberação de 6.000 ácaros em um determinado ponto, aproximadamente 3 por cento atingiram distâncias de 40 a 50 cm. As distâncias percorridas pelo ácaro foram inferiores a 1 cm.dia-1. Em condições de laboratório, verificou-se que ventos de 23 km.h-1 não foram capazes de incitar o processo de dispersão de ácaros da superfície de frutos. Velocidades de 30 e 40 km.h-1 foram capazes de incitar a dispersão de menos de 1 por cento da população de ácaros presentes nos frutos. Trabalhos de dispersão em condições de campo mediante a utilização de armadilhas adesivas também comprovaram que a dispersão de B. phoenicis é limitada quando comparada à de outras espécies de ácaros que ocorrem nos pomares de citros. Nesse experimento, de um total de 2.420 e 661 ácaros coletados em cada um de dois talhões de citros na região de Descalvado, SP, 0,4 e 11,8 por cento dos ácaros pertenciam à família Tenuipalpidae. A limitada dispersão de B. phoenicis exige muito cuidado na implementação de estratégias de manejo da resistência a acaricidas.

The knowledge of pest dispersal is crucial for understanding the dynamics of resistance to pesticides. Therefore, the main objective of this research was to study the dispersal of Brevipalpus phoenicis (Geijskes) under laboratory, greenhouse and citrus groves conditions. Dispersal studies of B. phoenicis on sandy surface conducted under greenhouse conditions showed that dispersal capacity by crawling is relatively limited. After the release of 6,000 mites in one point, approximately 3 percent reached distances of 40 to 50 cm from the release point. This mite moved less than 1 cm.day-1. Wind speed of 23 km.h-1 was not enough to trigger mite dispersal from the surface of citrus fruits under laboratory conditions. Wind speeds of 30 and 40 km.h-1 were capable to trigger the dispersal of less than 1 percent of mites on fruits. Mite dispersal studies with the use of sticky traps under field conditions also showed that B. phoenicis dispersal is limited when compared to other mite species from citrus groves. In this experiment, from a total of 2,420 and 661 mites collected from each of two citrus fields in Descalvado, region, State of São Paulo, Brazil, 0.4 and 11.8 percent of the mites belonged to the family Tenuipalpidae. The limited dispersal of B. phoenicis requires considerable attention in the establishment of acaricide resistance management strategies.