Primeiro relato de atividade patogênica de Hirsutella thompsonii (Fischer) sobre o ácaro-da-erinose-da-lichia Aceria litchii (Keifer)

The litchi erineum mite, Aceria litchii (Keifer) is found in all producing regions attacking leaves and flowers of litchi plants. The mite attack young leaves and causes the erinea on leaf surface, which later become brown galls with velvety appearance. Severe attacks can cause leaf drop and destruction of branches end directly production affecting. In 2009 year it was registered a heavy infestation of the pest on litchi plants (Litchi chinensis Sonn.) in the municipality of Casa Branca, São Paulo, Brazil (2127'O; 4702'S; 679 m altitude). In June, many galls caused by mite infestation showed a mycelium of white coloration and many eriophyid dead. The fungus was identified as Hirsutella thompsonii (Fischer). The results suggest that galls may facilitate the fungus development and its permanence on the plants. Thus, the possibility of mite biological control with H. thompsonii should be investigated.

The role of native vegetation on infection rates of Calacarus heveae (Acari: Eriophyidae) by Hirsutella thompsonii (Ascomycota: Ophiocordycipitaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ácaros (Acari) de dois sistemas de cultivo da seringueira no noroeste do estado de São Paulo

A cultura da seringueira (Hevea brasiliensis Muell. Agr.) vem se expandindo consideravelmente no noroeste do estado de São Paulo. O objetivo deste trabalho foi avaliar a fauna de ácaros em dois sistemas comuns de cultivo nessa região, um em monocultivo e outro em consórcio com gariroba (Syagrus oleracea (Mart.) Becc. O estudo foi conduzido entre maio de 2002 e abril de 2003, tomando-se amostras mensais de 150 folíolos de seringueira de cada plantação e 150 folíolos de gariroba. As faunas dos dois sistemas foram semelhantes; os números de espécies, gêneros e famílias, assim como as famílias mais diversas e mais abundantes foram semelhantes entre eles. Dezoito espécies ocorreram exclusivamente em plantas de gariroba. O número de ácaros sobre estas foi consideravelmente menor, mas a riqueza de espécies foi maior que em seringueiras. Dentre as espécies mais freqüentes e abundantes em seringueira, Eutetranychus banksi (McGregor) e Pronematus sp. foram as únicas também freqüentes em gariroba, porém bem menos abundantes. Nenhum dos predadores abundantes em seringueira (Zetzellia aff. yusti, Euseius citrifolius Denmark & Muma, Pronematus sp. e Spinibdella sp.) foi abundante em gariroba. O fungo Hirsutella thompsonii Fisher foi o único patógeno de ácaros encontrado neste estudo, atacando Calacarus heveae Feres. Aparentemente, na forma como cultivada na região, a gariroba não constitui um reservatório adequado dos ácaros predadores mais importantes em seringueira.

Seasonality of pathogenic fungi in mites of rubber tree plantations adjacent to fragments of Cerrado

Fungos são os patógenos mais freqüentemente observados em populações de ácaros, auxiliando o controle destes em diferentes culturas. Vinte e cinco amostras de folhas foram coletadas de cultivos de seringueiras vizinhas a dois fragmentos de vegetação de Cerrado. em cada seringal, 25 plantas foram selecionadas para coletas e sete folhas ao redor da copa foram coletadas até a altura de sete a oito metros do solo. Aproximadamente 250 indivíduos de Calacarus heveae Feres, Phyllocoptruta seringueirae Feres e Tenuipalpus heveae Baker, coletados aleatoriamente, foram montados. Hirsutella thompsonii Fisher foi observado nas três espécies de ácaros e T. heveae foi a espécie mais infectada. Os maiores níveis de infecção ocorreram de novembro a fevereiro (estação chuvosa). Na estação seca, os níveis de infecção ficaram abaixo dos 5%. H. thompsonii tem potencial para ser usado como micoacaricida na estação chuvosa.

Aceria litchii (Keifer) em lichia: ocorrência sazonal, danos provocados e identificação de possíveis agentes de controle biológico

Pós-graduação em Agronomia - FEIS

Involvement of loop 5 lysine residues and the N-terminal β-hairpin of the ribotoxin hirsutellin A on its insecticidal activity

Ribotoxins are cytotoxic members of the family of fungal extracellular ribonucleases best represented by RNase T1. They share a high degree of sequence identity and a common structural fold, including the geometric arrangement of their active sites. However, ribotoxins are larger,with a well-defined N-terminal β-hairpin, and display longer and positively charged unstructured loops. These structural differences account for their cytotoxic properties.Unexpectedly, the discovery of hirsutellin A (HtA), a ribotoxin produced by the invertebrate pathogen Hirsutella thompsonii, showed how it was possible to accommodate these features into a shorter amino acid sequence. Examination of HtA N-terminal β-hairpin reveals differences in terms of length, charge, and spatial distribution. Consequently,four different HtA mutants were prepared and characterized. One of them was the result of deleting this hairpin [Δ(8-15)] while the other three affected single Lys residues in its close spatial proximity (K115E, K118E, and K123E). The results obtained support the general conclusion that HtA active site would show a high degree of plasticity,being able to accommodate electrostatic and structural changes not suitable for the other previously known larger ribotoxins, as the variants described here only presented small differences in terms of ribonucleolytic activity and cytotoxicity against cultured insect cells.

Biology and phenology of the eriophyid mite, Floracarus perrepae, on its native host in Australia, Old World climbing fern, Lygodium microphyllum

The biology and phenology of the eriophyid mite, Floracarus perrepae Knihinicki and Boczek,a potential biological control agent of Lygodium microphyllum (Cav.) R. Br., was studied in its native range - Queensland, Australia. F. perrepae forms leaf roll galls oil tile subpinnae of L. microphyllum. It has a simple biology, with females and males produced throughout the year. Tile Population was female biased at 10.5 to 1. The immature development time was 8.9 ± 0.1 and 7.0 ± 0.1 days; adult longevity was 30.6 ± 1.6 and 19.4 ± 1.2 days and mean fecundity per female was 54.5 ± 3.2 and 38.5 ± 1.6 eggs at 21 and 26 ° C, all respectively. Field studies showed that tile mite was active year round, with populations peaking when temperatures were cool and soil moisture levels were highest. Two species of predatory mites, Tarsonemus sp. and a species of Tydeidae, along with the pathogen Hirsutella thompsonii, had significant effects oil all life stages of F. perrepae. Despite high levels of predators and the pathogen, F. perrepae caused consistent damage to L. microphyllum at all the field sites over the entire 2 years of the study.

Characterization of a new toxin from the entomopathogenic fungus Metarhizium anisopliae: the ribotoxin anisoplin

Metarhizium anisopliae is an entomopathogenic fungus relevant in biotechnology with applications like malaria vector control. Studies of its virulence factors are therefore of great interest. Fungal ribotoxins are toxic ribonucleases with extraordinary efficiency against target ribosomes and suggested as potential insecticides. Here, we describe this ribotoxin characteristic activity in M. anisopliae cultures. Anisoplin has been obtained as a recombinant protein and further characterized. It is structurally similar to hirsutellin A, the ribotoxin from the entomopathogen Hirsutella thompsonii. Moreover, anisoplin shows the ribonucleolytic activity typical of ribotoxins and cytotoxicity against insect cells. How Metarhizium uses this toxin and possible applications are on perspective.

Characterization of a new toxin from the entomopathogenic fungus Metarhizium anisopliae: the ribotoxin anisoplin

Metarhizium anisopliae is an entomopathogenic fungus relevant in biotechnology with applications like malaria vector control. Studies of its virulence factors are therefore of great interest. Fungal ribotoxins are toxic ribonucleases with extraordinary efficiency against target ribosomes and suggested as potential insecticides. Here, we describe this ribotoxin characteristic activity in M. anisopliae cultures. Anisoplin has been obtained as a recombinant protein and further characterized. It is structurally similar to hirsutellin A, the ribotoxin from the entomopathogen Hirsutella thompsonii. Moreover, anisoplin shows the ribonucleolytic activity typical of ribotoxins and cytotoxicity against insect cells. How Metarhizium uses this toxin and possible applications are on perspective.

Aislamiento, producción y evaluación de la patogenicidad de cepas de Hirsutella citriformis Speare para el control de Diaphorina citri Kuwayama, vector de la enfermedad Huanglongbing de los cítricos

La industria citrícola es una de las más importantes industrias con las que cuenta el país y en los últimos años se encuentra amenazada por una plaga que está ocasionando estragos en todas las áreas citrícolas en México, el Huanglongbing o greening la cual es transmitida por el psílido asiático de los cítricos, Diaphorina citri Kuwayama, insecto que ocasiona daño en las plantas al alimentarse de los brotes tiernos y que además es el vector de la bacteria Candidatus Liberibacter asiaticus para la cual no hay cura y por lo que es necesario controlar al insecto que la transmite. Por lo que el presente trabajo consistió en aislar hongos de D. citri micosados, identificarlos en base a sus características morfométricas y usando técnicas moleculares, cultivarlos en diferentes medios de cultivo comerciales de agar para seleccionar aquel en el cual crecen y conidian mejor, se determinó también el tipo de exudados producidos por Hirsutella citriformis en los medios de agar, se cultivaron en dos diferentes medios de cultivo líquido y un sustrato vegetal para obtener mayor cantidad de conidios; también se realizaron bioensayos de laboratorio para determinar la virulencia de las cepas y seleccionar aquellas que pudieran controlar mejor al insecto; se evaluó la patogenicidad que pudieran presentar sobre insectos depredadores de D. citri, y con los resultados obtenidos se seleccionó las cepas que presentaron mayor patogenicidad sobre el insecto vector del HLB para evaluar su actividad sobre el mismo en bioensayo de campo. Se aislaron cinco cepas de insectos micosados provenientes de diferentes regiones citrícolas, se trabajó con ellos además de tres cepas con las cuales ya contaba el INIFAP de General Terán de Nuevo León. Se identificaron morfométrica y molecularmente como Hirsutella citriformis Speare. Los medio comerciales de cultivo en los cuales crecen y presentan mejor conidiación son los medios agar papa dextrosa enriquecidos con extracto de levadura al 0.5 y 1 % en los cuales se obtuvo un crecimiento radial de 3.2 a 3.8 cm para las cepas IB-Hir-2 e INIFAP-Hir-1 respectivamente, a los 34 días de cultivo a 25 ± 1 °C. La conidiación obtenida para las ocho cepas no correspondió al crecimiento radial, pero si concordó en presentarse en los medios enriquecidos con extracto de levadura, los valores variaron desde 1.22 x 10⁶ a 2.79 x 10⁷ para las cepas 2 INIFAP-Hir-2 en agar papa dextrosa e IB-Hir-2 en agar papa dextrosa con extracto de levadura al 1% respectivamente. En medios líquidos la mayor producción de blastosporas obtenida fue 4.3 x 108 blastosporas /ml a los 9 días en el medio de casaminoácidos para la cepa INIFAP-Hir-1, y en caldo papa dextrosa la mayor producción la presentó a los 11 días la cepa IB-Hir-2 (7.7 x 107 blastosporas/ml). La mayor producción de conidios en el sustrato vegetal (arroz) se obtuvo a los 14 días por la cepa INIFAP-Hir-4 (1.97 x 107 conidios/gr). Se analizaron los dos tipos de exudados observados en las cepas usando los métodos de Bradford para proteínas y el del Fenol Sulfúrico para carbohidratos y con estos métodos se determinó que todas las cepas producen ambos tipos de compuestos, además de que las proteínas son los exudados amarillo claro y oscuro, mientras que los carbohidratos corresponden a las gotas cristalinas producidas por las cepas en los diferentes medios de agar. En los diferentes bioensayos de laboratorio las cepas presentaron mayor mortalidad cuando se inoculó por contacto, donde las cepas INIFAP-Hir-1, INIFAP-Hir-2, IB-Hir-1 e IB-Hir-2 mostraron los mayores porcientos de mortalidad media. Al evaluar la patogenicidad de cuatro cepas sobre dos depredadores de D. citri, los resultados fueron similares para tratamientos y testigo y no se presentó micosamiento en los insectos muertos. Bajo condiciones de laboratorio la CL50 y CL90 obtenidas para el aislado INIFAP-Hir-1 fueron, 34 x 105 y 26.7 x 107 respectivamente. En el bioensayo de campo se usaron las cepas INIFAP.Hir-2, INIFAP.Hir-4, IB-Hir-1 e IB-Hir-2 y el mayor porciento de mortalidad obtenida fue 51.049 y el menor 35.7 para las cepas INIFAP-Hir-4 e IB-Hir-1 respectivamente, mientras que el testigo con adherente mostro 6.059 y en el testigo absoluto 4.84 % de mortalidad promedio. Además logramos aislar la cepa que los micosaba de 20 insectos colectados en diferentes puntos de muestreo, identificando en base a sus características morfológicas de crecimiento colonial a tres de las cepas evaluadas y a la cepa nativa de Martínez de la Torre.

A taxonomic guide to the mysids of the South Atlantic Bight.

Following the examination of extensive collections from the National Museum of Natural History (NMNH), the Southeastern Regional Taxonomic Center (SERTC), and other regional institutions, 18 species of the family Mysidae are recognized and described from the South Atlantic Bight (Cape Lookout, North Carolina to Cape Canaveral, Florida). This report includes synonymies of previous records, as well as new species distribution records. Previous regional accounts of Metamysidopsis munda and Metamysidopsis mexicana are attributed to Metamysidopsis swifti. New regional records are established for Amathimysis brattegardi, Heteromysis beetoni, and Siriella thompsonii. Two other species tentatively identified as Amathimysis sp. (nr. serrata) and Mysidopsis sp. (cf. mortenseni) may represent new taxa. Neobathymysis renoculata is included and discussed as a potential regional species. An illustrated key to the species currently known from the South Atlantic Bight is presented. Relevant taxonomic, distributional, and ecological information is also included for each species. (PDF file contains 45 pages.)

Controle biologico do percevejo-de-renda (Leptopharsa heveae Drake & Poor) em seringais de cultivo.

A mosca-de-renda ou percevejo-de-renda (Leptopharsa heveae - Hemiptera: Tingidae) e o mais importante inseto-praga da seringueira no Brasil. Essa praga de origem amazonica invadiu os seringais do Estado de Mato Grosso por volta de 1985 e, de 1992 a 1998, chegou aos seringais do Estado Sao Paulo e de Goias. Acredita-se que breve chegara aos Estados da Bahia, Espirito Santo e Minas Gerais. O principal agente disseminador desse inseto-praga entre estados e seringais tem sido os veiculos transportadores de coagulos e latex semiprocessado (GEB). O controle quimico dessa praga, alem de muito caro, coloca em risco os aplicadores e o meio ambiente. Em 1986, no Amazonas, o fungo Sporothrix insectorum foi constatado parasitando individuos do percevejo-de-renda. Com base nessa constatacao foram feitos alguns estudos em campo e laboratorio onde se determinaram a eficiencia biologica (capacidade do inimigo natural para infectar e destruir a praga) e a eficiencia tecnica (capacidade do inimigo natural para reduzir os danos causados na planta pela acao da praga). Os resultados desses testes evidenciaram o elevado potencial do S. insectorum para o controle do percevejode-renda. Imediatamente apos os testes, esse fungo foi enviado para os Estados de Mato Grosso, Acre, Rondonia, e mais recentemente para Sao Paulo e Goias onde vem sendo utilizado com o fungo Hirsutella verticillioides por diversos produtores e empresas. O fungo H. verticillioides, outro inimigo natural do percevejo-de-renda, isolado em 1990, na Guiana Francesa, vem sendo bem mais agressivo nos seringais da regiao Centro-Oeste e de Sao Paulo que o S. insectorum. Os resultados obtidos de avaliacoes efetuadas em alguns seringais de Goias, Mato Grosso e Sao Paulo indicam eficiencia biologica (porcentagem de insetos colonizados e fixados na folha por ambos os fungos em fevereiro), variando de 53% a 100%. Por outro lado, a eficiencia tecnica, medida pelo nivel de descoloracao da folha ou % de danos causados pela praga em fevereiro, tendo como referencia folhas nao atacadas, variou de 16% a 63%.

Influência de fragmentos de cerrado vizinhos à seringal (Hevea brasiliensis Muell. Arg.) na distribuição e infecção fúngica de ácaros (Acari, Arachnida)

Pós-graduação em Biologia Animal - IBILCE

Distribution of planktonic foraminifera during the Mid-Pleistocene of ODP Site 181-1123

Planktonic foraminiferal assemblages and artificial neural network estimates of sea-surface temperature (SST) at ODP Site 1123 (41°47.2'S, 171°29.9'W; 3290 m deep), east of New Zealand, reveal a high-resolution history of glacial-interglacial (G-I) variability at the Subtropical Front (STF) for the last 1.2 million years, including the Mid-Pleistocene climate transition (MPT). Most G-I cycles of ~100 kyr duration have short periods of cold glacial and warm deglacial climate centred on glacial terminations, followed by long temperate interglacial periods. During glacial-deglacial transitions, maximum abundances of subantarctic and subtropical taxa coincide with SST minima and maxima, and lead ice volume by up to 8 kyrs. Such relationships reflect the competing influence of subantarctic and subtropical surface inflows during glacial and deglacial periods, respectively, suggesting alternate polar and tropical forcing of southern mid-latitude ocean climate. The lead of SSTs and subtropical inflow over ice volume points to tropical forcing of southern mid-latitude ocean-climate during deglacial warming. This contrasts with the established hypothesis that southern hemisphere ocean climate is driven by the influence of continental glaciations. Based on wholesale changes in subantarctic and subtropical faunas, the last 1.2 million years are subdivided into 4-distinct periods of ocean climate. 1) The pre-MPT (1185-870 ka) has high amplitude 41-kyr fluctuations in SST, superimposed on a general cooling trend and heightened productivity, reflecting long-term strengthening of subantarctic inflow under an invigorated Antarctic Circumpolar Current. 2) The early MPT (870-620 ka) is marked by abrupt warming during MIS 21, followed by a period of unstable periodicities within the 40-100 kyr orbital bands, decreasing SST amplitudes, and long intervals of temperate interglacial climate punctuated by short glacial and deglacial phases, reflecting lower meridional temperature gradients. 3) The late MPT (620-435 ka) encompasses an abrupt decrease in the subantarctic inflow during MIS 15, followed by a period of warm equable climate. Poorly defined, low amplitude G-I variations in SSTs during this interval are consistent with a relatively stable STF and evenly balanced subantarctic and subtropical inflows, possibly in response to smaller, less dynamic polar icesheets. 4) The post-MPT (435-0 ka) is marked by a major climatic deterioration during MIS 12, and a return to higher amplitude 100 kyr-frequency SST variations, superimposed on a long term trend towards cooler SSTs and increased mixed-layer productivity as the subantarctic inflow strengthened and polar icesheets expanded.