O principal problema fitossanitário da citricultura atualmente é a doença conhecida como greening ou Huanglongbing (HLB), transmitida pelo inseto vetor Diaphorina citri. O presente trabalho objetivou desenvolver estratégias para incorporação dos fungos Beauveria bassiana ESALQ-PL63, registrado para o controle de outras pragas e/ou I. fumosorosea ESALQ-1296 em desenvolvimento como biopesticida especialmente para o manejo de D. citri, determinando a melhor concentração de adjuvantes e de conídios dos fungos a serem utilizados, sua eficácia em laboratório, semi-campo e campo; assim como sua compatibilidade com os agroquímicos utilizados na cultura. Na comparação dos dois fungos aplicados com seis adjuvantes no controle de D. citri revelaram que exceto Tween 80 a 0,001%, todos os outros adjuvantes contribuíram para aumentar a eficácia dos fungos. Os níveis de controle obtidos nos bioensaios em laboratório com B. bassiana e I. fumosorosea foram confirmados nos bioensaios em semi-campo e em campo. Realizaram-se 12 pulverizações com os dois fungos durante o período de dezembro de 2011 a janeiro de 2013 em campo nas concentrações de 2,5x106 conídios/mL em 2000 L/ha e 5x106 conídios/mL em 1000 L/ha ambos com 0,075% do adjuvante KBRAdj. Na comparação mês a mês, somente em janeiro de 2012 o controle químico foi superior a todos os outros tratamentos. No restante dos meses, as mortalidades causadas pelos tratamentos com fungos foram semelhantes entre si e semelhantes ao controle químico. As mortalidades de adultos de D. citri pelos fungos variaram de 96,4±3,8% (dezembro/2011) a 57,8±3,5% (outubro/2012). Os efeitos de 27 agroquímicos sobre o crescimento vegetativo e esporulação das colônias e a viabilidade dos conídios in vitro e a virulência dos fungos in vivo após mistura com cada um dos produtos foram determinados e revelaram que exceto os fungicidas a maioria dos produtos são compatíveis com os fungos e podem ser misturados na calda de aplicação. Estes resultados indicam que Beauveria bassiana ESALQ-PL63 e Isaria fumosorosea ESALQ-1296 são promissores para utilização no controle de D. citri em programas de manejo integrado de pragas de citros. / The main cause of the crop loss in citrus is currently the disease known as greening or Huanglongbing (HLB), transmitted by the insect vector Diaphorina citri. This study aimed to develop strategies for incorporating the fungi Beauveria bassiana ESALQ-PL63, registered for the control of other pests and Isaria fumosorosea ESALQ-1296 in development as biopesticide especially for the management of D. citri by determining the optimal concentration of adjuvants and of conidia to be used, their effectiveness in laboratory, semifield and field; as well as its compatibility with agrochemicals used in the crop. The comparison of the two fungi applied with six adjuvants for the control of D. citri revealed that except Tween 80 (0.001%), all other adjuvants contributed to increase the effectiveness of the fungi. Control levels obtained in the laboratory bioassays with B. bassiana and I. fumosorosea were confirmed in bioassays in semi-field and field. Both fungi were sprayed 12 times from December 2011 to January 2013 in the field at concentrations of 2.5x106 conidia/mL in 2000 L/ha and 5x106 conidia/mL in 1000 L/ha both with 0.075% of KBRAdj adjuvant. Only in January 2012 the chemical control was superior to all other treatments. In the remaining months, the mortalities caused by all the fungi treatments were similar to each other and similar to chemical control. The mortalities of adult D. citri by fungi ranged from 96.4 ± 3.8% (December/2011) to 57.8 ± 3.5% (October/2012). The effects of 27 agrochemicals on the vegetative growth and sporulation of colonies and conidia viability in vitro and in vivo virulence of the fungus after mixing it with each of the products were determined and showed that except the fungicides, the majority of products are compatible with the fungi and may be tank mixed. These results indicate that B. bassiana ESALQ-PL63 and I. fumosorosea ESALQ-1296 are promising for use in the control of D. citri in integrated pest management programs of citrus.

O principal problema fitossanitário da citricultura atualmente é a doença conhecida como greening ou Huanglongbing (HLB), transmitida pelo inseto vetor Diaphorina citri. O presente trabalho objetivou desenvolver estratégias para incorporação dos fungos Beauveria bassiana ESALQ-PL63, registrado para o controle de outras pragas e/ou I. fumosorosea ESALQ-1296 em desenvolvimento como biopesticida especialmente para o manejo de D. citri, determinando a melhor concentração de adjuvantes e de conídios dos fungos a serem utilizados, sua eficácia em laboratório, semi-campo e campo; assim como sua compatibilidade com os agroquímicos utilizados na cultura. Na comparação dos dois fungos aplicados com seis adjuvantes no controle de D. citri revelaram que exceto Tween 80 a 0,001%, todos os outros adjuvantes contribuíram para aumentar a eficácia dos fungos. Os níveis de controle obtidos nos bioensaios em laboratório com B. bassiana e I. fumosorosea foram confirmados nos bioensaios em semi-campo e em campo. Realizaram-se 12 pulverizações com os dois fungos durante o período de dezembro de 2011 a janeiro de 2013 em campo nas concentrações de 2,5x106 conídios/mL em 2000 L/ha e 5x106 conídios/mL em 1000 L/ha ambos com 0,075% do adjuvante KBRAdj. Na comparação mês a mês, somente em janeiro de 2012 o controle químico foi superior a todos os outros tratamentos. No restante dos meses, as mortalidades causadas pelos tratamentos com fungos foram semelhantes entre si e semelhantes ao controle químico. As mortalidades de adultos de D. citri pelos fungos variaram de 96,4±3,8% (dezembro/2011) a 57,8±3,5% (outubro/2012). Os efeitos de 27 agroquímicos sobre o crescimento vegetativo e esporulação das colônias e a viabilidade dos conídios in vitro e a virulência dos fungos in vivo após mistura com cada um dos produtos foram determinados e revelaram que exceto os fungicidas a maioria dos produtos são compatíveis com os fungos e podem ser misturados na calda de aplicação. Estes resultados indicam que Beauveria bassiana ESALQ-PL63 e Isaria fumosorosea ESALQ-1296 são promissores para utilização no controle de D. citri em programas de manejo integrado de pragas de citros. / The main cause of the crop loss in citrus is currently the disease known as greening or Huanglongbing (HLB), transmitted by the insect vector Diaphorina citri. This study aimed to develop strategies for incorporating the fungi Beauveria bassiana ESALQ-PL63, registered for the control of other pests and Isaria fumosorosea ESALQ-1296 in development as biopesticide especially for the management of D. citri by determining the optimal concentration of adjuvants and of conidia to be used, their effectiveness in laboratory, semifield and field; as well as its compatibility with agrochemicals used in the crop. The comparison of the two fungi applied with six adjuvants for the control of D. citri revealed that except Tween 80 (0.001%), all other adjuvants contributed to increase the effectiveness of the fungi. Control levels obtained in the laboratory bioassays with B. bassiana and I. fumosorosea were confirmed in bioassays in semi-field and field. Both fungi were sprayed 12 times from December 2011 to January 2013 in the field at concentrations of 2.5x106 conidia/mL in 2000 L/ha and 5x106 conidia/mL in 1000 L/ha both with 0.075% of KBRAdj adjuvant. Only in January 2012 the chemical control was superior to all other treatments. In the remaining months, the mortalities caused by all the fungi treatments were similar to each other and similar to chemical control. The mortalities of adult D. citri by fungi ranged from 96.4 ± 3.8% (December/2011) to 57.8 ± 3.5% (October/2012). The effects of 27 agrochemicals on the vegetative growth and sporulation of colonies and conidia viability in vitro and in vivo virulence of the fungus after mixing it with each of the products were determined and showed that except the fungicides, the majority of products are compatible with the fungi and may be tank mixed. These results indicate that B. bassiana ESALQ-PL63 and I. fumosorosea ESALQ-1296 are promising for use in the control of D. citri in integrated pest management programs of citrus.

Made available in DSpace on 2014-06-11T19:32:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-01-29Bitstream added on 2014-06-13T20:44:14Z : No. of bitstreams: 1 mochi_da_dr_jabo.pdf: 1918684 bytes, checksum: f29aa98fefb34604c991782bab720e3b (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A mosca-dos-chifres Haematobia irritans é um dos principais ectoparasitos associados a bovinos em pastagens. O presente trabalho objetivou analisar a ação patogênica dos isolados E9, IBCB425 e IBCB159 do fungo Metarhizium anisopliae, JAB06, JAB07 e AM09 de Beauveria bassiana, IBCB133 e CB75 de Isaria fumosorosea (=Paecilomyces fumosoroseus) e CG189 e CG195 de Isaria farinosa (=Paecilomyces farinosus) para a mosca-dos-chifres H. irritans em laboratório e campo. Avaliou-se em laboratório, a ação de todos os isolados dos fungos sobre ovos, larvas, pupas e adultos. Grupos de 30 ovos foram inoculados com suspensões fúngicas, e colocados sobre papel de filtro esterilizado, em 5 g de fezes bovinas frescas, acondicionadas em tubos de plástico. No ensaio com larvas, pedaços de papel filtro esterilizado contendo na superfície grupos de 30 ovos foram colocados sobre 5 g de fezes bovinas frescas, contidas em tubos de plástico, às quais incorporaram-se diversas concentrações de conídios mg fezes-1. No ensaio com ovos e larvas, o término do experimento ocorreu com a emergência dos adultos. Grupos de 20 pupas foram imersas por 60 segundos em suspensão do fungo, e colocadas em placas de Petri. Após a emergência, os adultos foram transferidos para gaiolas de isopor pequenas. Para o ensaio com adultos, grupos de 30 adultos foram separados e pulverizados com 0,3 mL das suspensões de conídios. Doze horas após, as moscas foram transferidas para gaiolas de isopor e avaliadas diariamente até o 15o dia. Para todos os ensaios, foram utilizadas suspensões contendo 106, 107 e 108 conídios mL-1. O isolado que promoveu os melhores resultados nos ensaios anteriores, foi usado em um experimento de campo para avaliar a ação fúngica em larvas e adultos de H. irritans em bovinos naturalmente infestados. Para o controle de larvas, foi oferecido aos bovinos conídios de M. anisopliae isolado... / Horn fly Haematobia irritans is one of the main ectoparasites associated to cattle on pastures. The present work had the objective of analyzing the pathogenic action of the isolates E9, IBCB425 and IBCB159 of the fungus Metarhizium anisopliae, JAB06, JAB07 and AM09 of Beauveria bassiana, IBCB133 and CB75 of Isaria fumosorosea (=Paecilomyces fumosoroseus) and CG189 and CG195 of Isaria farinosa (=Paecilomyces farinosus) for horn fly H. irritans, in laboratory and field conditions. The action of all the fungi isolates over the eggs, larvae, pupae and adults were evaluated in laboratory. Groups of 30 eggs were inoculated with fungal suspensions and placed over sterilized filter paper on top of 5 g of fresh bovine feces, put in plastic tubes. In the larvae assay, peaces of sterilized filter paper, containing groups of 30 eggs on the surface, were placed over 5 g of fresh bovine feces, contained in plastic tubes, in which different concentrations of conidia mg feces-1 were incorporated. In the eggs and larvae assay, the end of the experiment matched the emergence of the adults. Groups of 20 pupae were immersed for 60 seconds in the fungus suspension and placed on Petri dishes. After the emergence, the adults were transferred to small Styrofoam cages. In the adults assay, groups of 30 adults were separated and pulverized with 0,3 ml of the conidia suspensions. After twelve hours, the adults were transferred to Styrofoam cages and the evaluations were done daily until the 15th day. Suspensions containing 106, 107 and 108 conidia ml-1 were used for all the assays. The isolate that promoted best results on the previous assays was used in a field experiment, where the fungal action on larvae and adults of H. irritans, on naturally infested bovines, was evaluated. For the larvae control conidia of the E9 isolate of M. anisopliae, microencapsulated... (Complete abstract click electronic access below)

This thesis is focused on evaluation of the compatibility of entomopathogenic fungus Isaria fumosorosea with different species of entomopathogenic fungi and mycoparasitic fungi at different temperatures. The strains of species Beauveria bassiana, Metarhizium anisopliae, Lecanicillium muscarium, Coniothyrium minitans and Clonostachys rosea f. catenulata were selected for experiments base on compatibility. The results showed that combination of I. fumosorosea with species L. muscarium is compatible. The species do not limit to each other in the environment at all temperatures (15, 23 and 25 °C). On the other side, fungus I. fumosorosea in combination with other species have affected their growth and spore production. The efficacy of entomopathogenic fungi against larvae Tenebrio molitor was evaluated. The most effective species against larvae were species I. fumosorosea, B. bassiana and M. anisopliae. On the contrary, the smallest effective was observed after infection larvae by L. muscarium. Mycoparasitic fungus C. rosea f. catenulata was not able to directly infect larvae of T. molitor. This species did not infect healthy larvae. However it is able to infect weakened individuals or is growing as saprotrophs on the cadavers.

The potential synergism between chosen species of entomopathogenic nematodes (Steinernema arenarium, Steinernema feltiae) and entomopathogenic fungi (Beauveria bassiana, Isaria fumosorosea, Lecanicillium lecanii) is investigated in this study. It is theoretically possible to expect increasing of their efficiency at the collective introduction into environment. Creating of uniform laboratory method which will be possible to use at the research of this interaction system is a part of this study.

My diploma thesis deals with genetic variability of entomopathogenic fungi of the Isaria genus encountered in the Czech Republic. Individual representative of the genus can be found in soil where they attack all developmental stages of insects, giving preference to larvae and pupae. The Isaria fungi find application first and foremost where plants have to be provided biological protection. In case of mitosporic fungi is the precise identification very difficult, taxonomy is often unclear in many genera, including the genus Paecilomyces/Isaria to demonstrate their polyphyletic nature. The fungi are classified primarily with reliance on morphological studies. The most common markers used to identify fungi are the shapes and sizes of their conidia and the biological properties (germination of spores, tests of biological efficiency). Identification made in consideration of the morphological markers is inaccurate and very variable. To overcome those accuracies, there are very useful molecular DNA markers, which can be relevant in ecology, biology and in fungi genetics. This paper relies on applying the ITS region (Internal Transcribed Spacer) as a molecular marker. ITS regions are partial constituent rDNA carrying no code - that is why the regions are likely to accumulate evolutionary changes in the DNA sequence, which makes them suitable for extensive use in taxonomic analyses of many organisms. The study results in a phylogenetic trees constructed by comparing different sequences of ITS regions obtained from the samples of entomopathogenic fungi of the Isaria genus gathered in the Czech Republic during the monitoring stage 2013 to 2014. Thereunder detection of Isaria sp. occurring in the Czech Republic.

This diploma theses focuses on comparison of natural and intentionally inducated supressiveness of environment induced by application of entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, Isaria fumosorosea a Lecanicillium muscarium. In tests were evaluated the in vitro parameters as well as the effectiveness in vivo biotests on insect host larva Tenebrio molitor. The species of entomopatoghenic fungi were applied in suspensions, single strains and also in combination of two strains. In the in vitro conditions the possibilities of objective evaluation of the supresivity level were tested by using the CFU test (Colony Forming Units) on three different nutrient media (PDA, PDA + A , PDA + D), as one of the basic evaluation parameters. Further the germination tests were evaluated according to GI (Germination Index), determination of radial growth (comparison of median cultures) and interaction of strain suspensions on nutrient media PDA. In the in vivo biotests were watched the epizooties from suspensions of these entomopathogenic fungi on insect larva Tenebrio molitor in competitive test of strains according to FDI (Fungl development index) evaluation scala. Chosen larva covered by fully sporulating mycelium from epizootie were further evaluated in CFU test. The result were dominant strain/s on the larva from applied suspensions.