ABSTRACT
Phymastichus coffea LaSalle is an African endoparasitoid of adults of the coffee berry borer, Hypothenemus hampei (Ferrari) that was introduced into Mexico for biological control purposes. The objective of this study was to investigate the oviposition behavior of P. coffea on its host under laboratory conditions. The results indicate that P. coffea females exhibited 12 behavioral acts during and after oviposition, such as, walking, cleaning, flying, surface landing, resting, host landing, host antennation, touching the host with forelegs, host recognition, walking on the host, oviposition and cleaning after oviposition. The sequence of each behavior is described in an ethogram.
Subject(s)
Animals , Female , Behavior, Animal , Hymenoptera/anatomy & histology , Oviposition , Weevils/parasitology , Hymenoptera/physiologyABSTRACT
The aim of this study was to evaluate the coffee germplasm of the Paraná Agronomic Institute (IAPAR) for resistance to the coffee-berry-borer. Preliminary field evaluation was performed in August 2004 and the fruits of less damaged genotypes in the field were evaluated under controlled condition with obligated and free choice experiments established in a randomized complete design with three replications. The genotypes were evaluated fifteen days after infestation with one borer per fruit in Petri dishes. The data were analyzed by the Scott-Knott means test at 1 percent and by the χ2 test. Statistical analysis indicated that Coffea kapakata, Psilanthus bengalensis, C. eugenioides and genotypes with C. eugenioides genes were resistant. These genotypes presented low frequency of bored grains. C. eugenioides and C. kapakata could present resistance at epicarp level but not in the grain. P. bengalensis could present resistance also in the grains.
O objetivo deste trabalho foi identificar fontes de resistência genética a H. hampei em diferentes espécies de café do banco de germoplasma do Instituto Agronômico do Paraná (IAPAR), Londrina, PR. Foram realizadas avaliações preliminares de campo, para posterior testes de confinamento e de livre escolha, em laboratório, instalados em delineamento inteiramente casualizado com três repetições. Os genótipos foram avaliados quinze dias após a infestação com uma broca por fruto em placas de petri. Os dados foram analisados pelo teste de médias Scott-Knott a 1 por cento e pelo teste de χ2. Foi observado que C. eugenioides, C. kapakata e P. bengalensis constituem importantes fontes de resistência à broca, pois apresentaram menor freqüência de grãos brocados. Os dois primeiros podem apresentar substâncias voláteis antagônicas à broca na casca e a resistência de P. bengalensis pode estar também no grão.
ABSTRACT
La broca del café, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), es la plaga mas importante del cultivo de café en el mundo. Beauveria bassiana es un hongo entomopatógeno generalista ampliamente utilizado por los productores de café para controlar a H. hampei. Phymastichus coffea LaSalle es un endoparasitoide africano de adultos de la broca, recién importado a varios países de Latinoamérica y del Caribe para el control de esta plaga. En el presente estudio se realizaron bioensayos de laboratorio con el objetivo de determinar si B. bassiana es detrimental para el parasitoide P. coffea. La susceptibilidad del parasitoide fue evaluada en términos de supervivencia de adultos, concentración letal media (CL50), tiempo letal medio (TL50), reproducción y mortalidad de su progenie. El principal efecto del hongo resultó en la disminución de la longevidad de los adultos y en la mortalidad del 100 por ciento de los inmaduros del parasitoide. La CL50 para adultos de P. coffea fue de 0.11 por ciento equivalente a 9.53 x 10(7) conidias/ml de B. bassiana y una TL50 de 29.4h, equivalente a la reducción del 22 por ciento de su longevidad normal como adulto. P. coffea fue capaz de transmitir esporas de B. bassiana a brocas no infectadas con el hongo y originar indirectamente la muerte de su progenie. Este trabajo servirá de referencia cuando se contemple el empleo de ambos organismos a nivel de campo, especialmente dentro de un plan de manejo integral de la plaga.
The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is the most important coffee pest worldwide. Beauveria bassiana is a generalist entomopathogenic fungus widely used by coffee farmers to control this pest and Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) is an African endoparasitoid of H. hampei adults, recently imported to several Latin American and Caribbean countries to aid in the coffee berry borer control. The objective of this study was to determine if B. bassiana is detrimental to P. coffea. The susceptibility of the parasitoid was evaluated in terms of adult survivorship, mean lethal concentration (LC50), mean lethal time (LT50), reproduction and immature mortality. The main effect of the fungus resulted in reduction of adult longevity and mortality of 100 percent for immature stages of this parasitoid. The LC50 for adults was 0.11 percent equivalent to 9.53 x 10(7) conidia/ml of B. bassiana and a LT50 of 29.4h, equivalent to reduction of 22 percent of its normal longevity as an adult. P. coffea was capable of disseminating spores of B. bassiana to non-infected H. hampei adults, which could indirectly cause the death of its own progeny. These results could be valuable when considering the use of both organisms in the field, especially in an integrated pest management program.
Subject(s)
Animals , Beauveria , Hymenoptera/microbiology , Pest Control, BiologicalABSTRACT
A Brazilian isolate of Beauveria bassiana (CG425) that shows high virulence against the coffee berry borer (CBB) was examined for the production of subtilisin-like (Pr1) and trypsin-like (Pr2) cuticle-degrading proteases. Fungal growth was either in nitrate-medium or in CBB cuticle-containing medium under both buffered and unbuffered conditions. In unbuffered medium supplemented with cuticle, the pH of cultures dropped and Pr1 and Pr2 activities were detected in high amounts only at a pH of 5.5 or higher. In buffered cultures, Pr1 and Pr2 activities were higher in medium supplemented with cuticle compared to activities with nitrate-medium. The Pr1 and Pr2 activities detected were mostly in the culture supernatant. These data suggest that Pr1 and Pr2 proteases produced by strain CG425 are induced by components of CBB cuticle, and that the culture pH influences the expression of these proteases, indicating the occurrence of an efficient mechanism of protein secretion in this fungus. The results obtained in this study extend the knowledge about protease production in B. bassiana CG425, opening new avenues for studying the role of secreted proteases in virulence against the coffee berry borer during the infection process.
O isolado brasileiro de Beauveria bassiana (CG425) que apresenta alta virulência contra a broca do café (CBB) foi analisado quanto à produção de proteases degradadoras de cutícula, tipo-subtilisina (Pr1) e tipo-tripsina (Pr2). O crescimento fúngico foi realizado em meio contendo nitrato e em meio contendo cutícula da broca em condições de pH tamponado e não tamponado. Em meio não tamponado, suplementado com cutícula, o pH da cultura caiu e as atividades de Pr1 e Pr2 foram detectadas somente em valores de pH igual ou superior a 5,5. Em culturas tamponadas, as atividades Pr1 e Pr2 foram superiores em meio suplementado com cutícula, comparativamente as atividades em meio contendo nitrato. As atividades Pr1 e Pr2 ocorreram predominantemente no sobrenadante de cultivo. Os dados obtidos sugerem que Pr1 e Pr2 produzidas pelo isolado CG425 são induzidas por componentes da cutícula da broca do café (CBB), e que o pH da cultura influencia a expressão destas proteases, indicando a ocorrência de um mecanismo eficiente de secreção por este fungo. Os resultados obtidos neste estudo aumentam o conhecimento a respeito da produção de proteases por B. bassiana CG425, abrindo novos caminhos para o estudo do papel de proteases na virulência contra a broca do café durante o processo de infecção.
Subject(s)
Beauveria/growth & development , Beauveria/isolation & purification , Clinical Enzyme Tests , Culture Media , Environmental Microbiology , Fungi/growth & development , Fungi/isolation & purification , In Vitro Techniques , Peptide Hydrolases/analysis , Coffee , Methods , VirulenceABSTRACT
The coffee berry borer Hypothenemus hampei Ferrari (Coleoptera: Scolytidae) was first reported infecting Costa Rican coffee plantations in the year 2000. Due to the impact that this plague has in the economy of the country, we were interested in seeking new alternatives for the biological control of H. hampei, based on the entomopathogenic bacteria Bacillus thuringiensis. A total of 202 B. thuringiensis isolates obtained from Costa Rican coffee plantations infested with H. hampei were analyzed through crystal morphology of the crystal inclusions and SDS-PAGE of 6-endotoxins, while 105 strains were further evaluated by PCR for the presence cry, cyt and vip genes. Most of the Bt strains showed diverse crystal morphologies: pleomorphic (35%), oval (37%), bipyramidal (3%), bipyramidal and oval (12%), bipyramidal, oval and pleomorphic (10%) and bipyramidal, oval and cubic (3%). The SDS-PAGE analyses of the crystal preparations showed five strains with delta-endotoxin from 20 to 40 kDa, six from 40 to 50 kDa, seven from 50 to 60 kDa, 19 from 60 to 70 kDa, 29 from 70 to 100 kDa and 39 from 100-145 kDa. PCR analyses demonstrated that the collection showed diverse cry genes profiles having several genes per strain: 78 strains contained the vip3 gene, 82 the cry2 gene, 45 the cry1 and 29 strains harbored cry3-cry7 genes. A total of 13 strains did not amplified with any of the cry primers used: cry1, cry2, cry3-7, cry5, cry11, cry12 and cry14. Forty-three different genetic profiles were found, mainly due to the combination of cry1A genes with other cry and vip genes. The genetic characterization of the collection provides opportunities for the selection of strains to be tested in bioassays against H. hampei and other insect pests of agricultural importance.
Subject(s)
Animals , Genetic Variation , Bacillus thuringiensis/genetics , Coleoptera , Coffee , Endotoxins/genetics , Hemolysin Proteins/genetics , Bacterial Proteins/genetics , Bacterial Toxins/genetics , Bacillus thuringiensis/chemistry , Bacillus thuringiensis/ultrastructure , Coffee/microbiology , Coffee/parasitology , Pest Control, Biological , Electrophoresis, Polyacrylamide Gel , Microscopy, Electron, Scanning , Polymerase Chain ReactionABSTRACT
In a study conducted to determine natural enemies of coffee berry borer (Hypothenemus hampei, Ferrari) in the state of Rondônia, the occurrence of the fungus Beauveria bassiana (Bals.) Vuill. was identified at Embrapa Rondônia. Coffee fruits with coffee borer containing the fungus were collected in Conilon coffee plantations (Coffea canephora), in Rolim de Moura, Ouro Preto do Oeste and Machadinho do Oeste, state of Rondônia, Brazil, from February to March, 2000. The infection percentage in fruits with coffee berry borer varied from 0.23% to 0.47%, less than reported from other regions of Brazil.
Em trabalho efetuado visando o levantamento de inimigos naturais da broca-do-café (Hypothenemus hampei, Ferrari) no estado de Rondônia, verificou-se a ocorrência do fungo Beauveria bassiana (Bals.) Vuill. conforme identificação da Embrapa Rondônia. Os frutos de café foram coletados em cafezais da cultivar Conilon (Coffea canephora), nos municípios de Rolim de Moura, Ouro Preto do Oeste e Machadinho do Oeste, Rondônia, nos meses de fevereiro a março de 2000. A percentagem média de infecção de B. bassiana em frutos broqueados variou de 0,23% a 0,47%, sendo menor que em outras regiões do Brasil.