A irrigação é uma das tecnologias cujo retorno econômico é mais facilmente percebido pelo usuário, apesar do seu alto custo de investimento inicial. Nos últimos anos, com o fim dos financiamentos subsidiados, o agricultor irrigante tem buscado alternativas tecnológicas que lhe permitam o retorno do investimento em prazos cada vez menores. As soluções encontradas vão desde o uso da agricultura intensiva com a semeadura de culturas em sucessão (cultivos anuais), ou de várias safras (perenes), como também a maximização do uso do equipamento de irrigação, que tem sido usado cada vez com maior freqüência para a aplicação de produtos químicos por meio da quimigação. Esse panorama tem gerado, em algumas situações, desequilíbrio no agroecossistema com várias conseqüências, entre as quais destacam-se a explosão de pragas e doenças e a compactação de solos. A quimigação, apresenta uma série de vantagens e desvantagens. Dentre as vantagens, destacam-se o seu baixo custo quando comparado a outros métodos, e o menor tráfego de máquina na área cultivada, o que também favorece o menor dano mecânico às culturas. Entre as limitações, destaca-se o fato de que nem todos os produtos podem ser utilizados, pois esses precisam apresentar determinadas características físico-químicas como baixa solubilidade em água e alta em solventes orgânicos. O presente trabalho visa gerar informações sobre o uso de óleos sem surfactantes para a dissolução de formulações comerciais de clorpirifós (concentrado emulsionável), e sua efetividade no controle da lagarta do cartucho (Spodoptera frugiperda) no milho quando comparado à formulação comercial sem mistura, aplicado por insetigação. Desse modo, espera-se viabilizar a quimigação foliar com possibilidade da redução de dose ou do número de aplicações além de propor uma metodologia para instalação de ensaios de quimigação a campo. Foram instalados durante os anos de 1996 e 1997, três ensaios em três safras distintas, todos com elevada pressão de infestação, em áreas comerciais, onde os tratamentos foram injetados diretamente na base do pivô por meio de uma bomba injetora-dosadora. As parcelas, em forma de cunha, tinham no mínimo 50 m no perímetro de modo a evitar contaminação entre os tratamentos, e uma área mínima de um hectare. Foram amostrados de quatro a seis pontos para cada parcela. Os resultados obtidos mostraram a viabilidade da técnica, uma vez que a metade da dose comercial dissolvida no óleo mostrou a mesma efetividade de controle que a dose comercial, o que sinaliza a possibilidade da redução da dose ou do número de aplicações minimizando os custos para o produtor com menor impacto ambiental, o que favorece o manejo integrado e a sustentabilidade da agricultura. As informações geradas viabilizam a quimigação como método de aplicação de baixo custo, e a metodologia sugerida comprovou-se eficiente para a instalação de ensaios a campo. / Irrigation is perceived by the farmers as the best payback technology, beside its initial investment cost. With the end of subsidized interests for agriculture in the last years, irrigated farmers are more and more looking for shorter paybacks of their investments. The alternatives presented are successive cropping (annual crops) or more that one harvest (perennial crops) per year, and to maximize the use of the irrigation equipment which is more frequently being used to apply chemical which is known as chemigation. This scenario generates in some situations an unbalance at the environment with several consequences like pest and diseases flare and soil compaction. Chemigation has advantages and limitations. Among the advantages is to be highlighted the lower application cost when compared to other methods, and less tracking in the area which reduce soil compaction and also crop losses due to crop damage. The main limitation is that not all products can be used in chemigation. They must attend some physical - chemical properties like low water solubility and high solubility in organic solvents. The present work has the intention to generate information about the feasibility of mixing commercial formulations (emulsifiable concentrate-EC) with oils and no addition of any surfactant and the effectiveness of this mixture in comparison to standard when insectigated to control fall armyworm (Spodoptera frugiperda) in corn. This way, it is expected to viabilize foliar chemigation, with the possibility of rate or applications number reduction, and also to propose a standardized methodology to install field trials. Three trials were installed during 1996 and 1997 in three different seasons, all of them with high infestation level, in commercial areas, where treatments were injected in the center of the pivots using an injection pump. The plots, in a wedge form, had a minimal lenght at the perimeter to avoid treatments contamination, which provided a minimal area of one hectare each plot. Sampling was done at four to six points for each plot. Data showed the viability of the techniques, since the half of commercial rate diluted in oil without surfactants had shown the same effectiveness that standard (full rate without oil). This sign up the possibility of a rate reduction or perhaps less applications, which will minimizes farmers’ costs with less environmental impact, being favorable to integrated pest management and agriculture sustainability. Generated information shows the workability of chemigation as a low cost application method, as well as the proposed methodology to install field trials had shown to be effective.

O controle biológico de pragas é um importante serviço ecossistêmico em áreas agrícolas e de pastagem, podendo trazer tanto benefícios econômicos quanto ambientais. Em áreas de pastagens tropicais, as a cigarrinhas-das-pastagens podem trazer prejuízos da ordem de US$ 800 milhões por ano, sendo assim uma praga de grande importância. Para entendermos os fatores que potencialmente regulam o controle biológico destas cigarrinhas no nível da paisagem, nós avaliamos, através de experimentos de predação, o efeito da quantidade de cobertura florestal, do tamanho do fragmento e da distância na matriz ao fragmento no processo de predação. Além disso, testamos o efeito de diferentes procedimentos metodológico na detecção de padrões de relação entre o potencial controle de pragas e a estrutura da paisagem. Trabalhamos com nove paisagens inseridas na região de Mata Atlântica, e para cada paisagem utilizamos três transectos de 100 m de cumprimento para dentro das pastagens, ortogonalmente dispostos a partir de uma borda florestal. A taxa de predação apresentou uma relação positiva com a cobertura florestal no raio de 400 m e com o tamanho do fragmento, e uma relação negativa com a distância ao fragmento florestal. Os padrões de predação variaram com o tipo de presa utilizado e com o tempo de exposição do experimento. O experimento do tipo sentinel prey (com presas reais que são pragas comuns na região - Deois flavopicta) evidenciou padrões fracos com a cobertura florestal no raio de 3 km, enquanto o experimento com \"dummy caterpillars\" (com presas artificiais em formato de lagartas) evidenciou padrões mais claros associados, principalmente, com a cobertura florestal no raio de 400 m e com o tamanho do fragmento. O efeito negativo da distância ao fragmento florestal só ficou claro com tempos de exposição mais longos (96 horas). O processo de predação na matriz é assim influenciado pela estrutura da paisagem, particularmente em escala mais locais (400 m), e os diferentes tipos de presas e tempos de exposição de experimentos de predação influenciam nos padrões ecológicos que emergem. Nosso trabalho evidência a importância do manejo da estrutura da paisagem em áreas agrícolas para otimizar o oferecimento do serviço de controle de pragas, bem como a necessidade de se utilizar mais de um tipo de experimento para avaliar o processo de predação / Biological pest control is an important ecosystem service in agricultural and pastures areas, and can bring both economic and environmental benefits. In areas of tropical pastures, spittlebugs can bring losses of several millions per year, thus being a pest of great importance. To understand the factors that potentially regulate the biological control of these froghoppers, we evaluated through predation experiments the effects of forest cover amount, fragment size and distance in the matrix to fragment on predation process. In addition, we tested the effect of different methodological procedures in the detection of relationships between pest control and landscape structure. We worked with nine landscapes inserted in the Atlantic Forest region, and for each landscape used three transects of 100 m length inside pastures, orthogonally arranged from the forest edge. Predation rate showed a positive relationship with forest cover within 400 m and with fragment size, and a negative relationship with distance from forest fragment. Predation patterns varied with the type of prey used and the exposure time of the experiment. The sentinel prey\'s experiment (with real preys that are common pests in the region - Deois flavopicta) showed weak patterns with forest cover in 3 km, while the dummy caterpillars experiment (with artificial prey on caterpillars\' shape) showed clearer patterns mainly associated with forest cover within 400 m, and with fragment size. The negative effect of distance from the forest fragment became clear only with longer exposure times (96 hours). The predation process in the matrix is thus influenced by the landscape structure, particularly in more local scale (400 m), and different types of prey and exposures times of predation experiments influence the ecological patterns that emerge. Our work highlights the importance of landscape management structure in agricultural areas to optimize the offering of pest control service, as well as the need to use more than one type of experiment to access predation process

Gossypium hirsutum, le coton Upland, représente plus de 95% de la fibre de coton produite annuellement dans le monde et est cultivé dans environ 40 pays. La protection des champs de coton contre l’attaque des arthropodes herbivores nécessite des quantités importantes d’insecticides de synthèse, environ 18% de la consommation mondiale en 2000, bien que cela ait beaucoup diminué grâce au coton Bt et aux programmes d’éradication de certains ravageurs. Les composés organiques volatils (COV) naturellement émis par les plantes cultivées peuvent réduire les attaques d'insectes, les COV ayant une influence sur le comportement des arthropodes herbivores et des arthropodes auxiliaires. La recherche scientifique sur les COV des plantes a beaucoup augmenté depuis deux ou trois décennies. La stimulation des émissions de COV dans les champs de coton est désormais recommandée par les entomologistes du Cirad en tant que composante de la stratégie de protection. L’écimage (topping en anglais), c'est-à-dire couper la tête des cotonniers au cours de la floraison, stimule l'émission de COV de défense, une protection écologique qui limite les risques pour la santé dus aux insecticides synthétiques, pour les agriculteurs et les personnes des environs. L'objectif de la thèse était d'améliorer nos connaissances sur les bases génétiques des émissions de COV après l’écimage. La combinaison de plusieurs disciplines telles que la bio-informatique, l'écologie chimique et la génétique moléculaire nous a permis de: 1) analyser les séquences génomiques des gènes des voies de biosynthèse des COVs terpènes et terpénoïdes et des facteurs de transcription (TF) liés à la réponse au stress, à l'aide des bases de données publiées sur les génomes de trois espèces de coton, G. raimondii, G. arboreum (cotons diploïdes) et G. hirsutum (coton tétraploïde), 2) étudier les émissions de COV par les feuilles de coton des plants de G. hirsutum en réponse à l’écimage, en capturant ces molécules en serre et en caractérisant leurs profils cinétiques par chromatographie en phase gazeuse-spectrométrie de masse (GC-MS), incluant des mesures quantitatives, et 3) étudier les modifications de l'expression ARN des plants de coton G. hirsutum après l’écimage, pour 44 gènes impliqués dans la biosynthèse des COV, et également par une comparaison du transcriptome complet au moyen d'une analyse RNA-seq. Les résultats des trois domaines scientifiques, bio-informatique, analyse chimique et expression des gènes, ont pu être liés dans notre thèse de recherche: par ex., deux des gènes initialement identifiés par la bio-informatique, correspondant à deux enzymes, TPS50 (EC: 4.2.3.106 - (E ) bêta-ocimène synthase) et TPS16 (EC: 4.2.3.111 - alpha-terpinéol synthase), ont montré une augmentation de leur expression après l’écimage, et l'analyse GC-MS montre une modification correspondante des profils d'émission de COV. Ces mêmes composés avaient déjà été caractérisés dans d'autres plantes en réponse aux dégâts d’arthropodes. Ce travail de thèse constitue une première exploration des bases génétiques des émissions défensives de COV par les cotonniers cultivés G. hirsutum. La variabilité des comportements d’expression génique observés entre les trois génotypes étudiés de coton Upland africain G. hirsutum, permet de supposer qu’une diversité génétique est présente pour les émissions défensives de COV, ce qui pourrait permettre d’améliorer et d’adapter ces mécanismes de défense naturels et leur réponse à l’écimage, dans la perspective d'une protection naturelle plus efficace des champs de coton. / Gossypium hirsutum, the Upland cotton, represents more than 95% of the cotton fiber annually produced worldwide and is grown in about 40 countries. The protection of cotton fields against the attack of herbivorous arthropods needs important quantities of synthetic insecticides, around 18% of the world consumption for the year 2000, although this decreased very much thanks to Bt cotton and eradication programs for some pests. Volatile organic compounds (VOCs) naturally emitted by crop plants can reduce insect attacks through the influence of VOCs on the behaviors of herbivorous arthropods and auxiliary arthropods. Scientific research about plant VOCs has been increasing much since two or three decades. The stimulation of VOCs emissions by cultivated cotton plants is now recommended by entomologists of CIRAD as a component of the cotton fields protection strategy. “Topping", that is, cutting the head of the cotton plants during the useful floriferous period, stimulates the emission of defense VOCs. It is an environmentally friendly method and it limits health hazards due to the use of synthetic insecticides for the farmers and the surrounding human populations. The objective of the thesis was to improve our knowledge about the genetic bases of VOCs emissions after topping. The combination of several disciplines such as bioinformatics, chemical ecology and molecular genetics allowed us to: 1) analyze the genomic sequences of VOCs genes of the terpene and terpenoid biosynthesis pathways and transcription factors (TF) related to stress response, using the published genome databases of three cotton species, G. raimondii, G. arboreum (both diploid cottons) and G. hirsutum (tetraploid cotton), 2) study the VOCs emissions by cotton leaves of G. hirsutum plants in response to topping, by capturing these molecules in greenhouse and then characterizing their kinetic profiles by means of gas-chromatography mass-spectrometry (GC-MS), with quantitative measurements, and, 3) study the modifications of the RNA expression of G. hirsutum cotton plants after topping, for genes involved in VOCs biosynthesis through quantitative PCR measurements on 44 targeted genes and also by means of a whole-transcriptome comparison through an RNA-seq analysis. The results from the three different fields, bioinformatics, chemical analysis and gene expression, could be interrelated in our research thesis: e.g., two of the genes initially identified by bioinformatics, corresponding to two enzymes, TPS50 (EC: 4.2.3.106 - (E) -beta-ocimene synthase) and TPS16 (EC: 4.2.3.111 - alpha-terpineol synthase), were shown to increase their expression after topping, while the GC-MS analysis showed an modification of the corresponding VOCs in emission profiles. These compounds have been already characterized in other organisms in response to wounds produced by herbivorous insects. This thesis work is a first exploration of the genetic bases of defensive VOCs emission by G. hirsutum cultivated cottons. The variability of genic expression behaviors observed amongst the three genotypes of African Upland cotton G. hirsutum that were studied permits to hypothesize that a genetic diversity is present for defensive VOCs emissions, that could permit to improve and adapt by breeding these natural defense mechanisms and the response to topping, in perspective of a more efficient natural protection of cotton fields.

Submitted by Mario BC (mario@bc.ufrpe.br) on 2017-02-22T14:21:22Z No. of bitstreams: 1 Izanielle Batista dos Santos.pdf: 695252 bytes, checksum: 87a9486b9be20199fbb71b6da8e965e5 (MD5) / Made available in DSpace on 2017-02-22T14:21:22Z (GMT). No. of bitstreams: 1 Izanielle Batista dos Santos.pdf: 695252 bytes, checksum: 87a9486b9be20199fbb71b6da8e965e5 (MD5) Previous issue date: 2016-07-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Brazil is a country with enormous potential for the production of grain. However, the country faces serious problems in the control of stored grain pests. The Sitophilus zeamais Motsch. (Coleoptera, Curculionidae) is one of the most destructive pests of corn, whose control is done by insecticides, however, their indiscriminate use has led to the development of resistant populations. In this context, Diatomaceous Earth stands in the control of stored grain pests. In view of this, the objective of this study was to investigate whether there are differences among Brazilian populations of S. zeamais in its Diatomaceous Earth resistance capacity associated with some behavioral characteristics of locomotion and of S. zeamais control efficiency. We used four populations of S. zeamais from the states of the Federal District (DF), Minas Gerais (MG) and Pernambuco (PE). The bioassay control efficiency was structured in a factorial (3 doses of Diatomaceous Earth x 3 exposure times) with four replications. Each repetition was constituted by ten adult insects, placed in Petri dishes containing corn treated with diatomaceous earth. The number of live insects by repetition after 24, 48 and 72 hours of exposure to Diatomaceous Earth. The behavioral bioassay was structured in a factorial (4 people x 3 doses of Diatomaceous Earth), where each repetition consisted of a single insect housed in the Petri dish. The movement of the insects was recorded and their locomotor behavior was observed for ten minutes. The tested variables were distance (cm) traversal velocity (cm. S-1), moving time (s) and number of stops. The results demonstrate an increased mortality of insects S. zeamais with increasing doses. There was also different responses among populations of S. zeamais in their behavioral resilience associated with decrease in distance traveled, the number of stops of insects with increasing doses. Confirming the existence of behavioral resistance in populations of S. zeamais studied. In view of this, the elucidation of possible behavioral changes in populations of S. zeamais exposed to Diatomaceous Earth can provide new ways to increase the effectiveness in the management of this pest. / O Brasil é um país com enorme potencial para a produção de grãos. No entanto, o país enfrenta sérios problemas no controle de pragas de grãos armazenados. O Sitophilus zeamais Motsch. (Coleoptera, Curculionidae) é uma das pragas mais destrutivas do milho, cujo controle é feito por inseticidas, entretanto, seu uso indiscriminado tem ocasionado o desenvolvimento de populações resistentes. Neste contexto, a Terra Diatomácea destaca-se no controle de pragas de grãos armazenados. Em vista disto, o objetivo deste trabalho foi investigar se há diferença entre as populações brasileiras de S. zeamais na sua capacidade de resistência à Terra Diatomácea associada a alguma característica comportamental de locomoção e a eficiência de controle de S. zeamais. Foram utilizadas quatro populações de S. zeamais provenientes dos estados do Distrito Federal (DF), Minas Gerais (MG) e Pernambuco (PE). O bioensaio eficiência de controle foi estruturado em um fatorial (3 doses de Terra Diatomácea x 3 tempos de exposição) com quatro repetições. Cada repetição foi constituída por dez insetos adultos, colocados em placas de Petri contendo milho tratado com Terra Diatomácea. Foi contabilizado o número de insetos vivos por repetição após 24, 48 e 72 horas de exposição a Terra Diatomácea. O bioensaio comportamental foi estruturado em um fatorial (4 populações x 3 doses de Terra Diatomácea), onde cada repetição foi constituída por um único inseto alojado em placa de Petri. O movimento dos insetos foi gravado e seu comportamento locomotor foi observado durante dez minutos. As variáveis testadas foram distância percorrida (cm), velocidade de caminhamento (cm. s-1), tempo em movimento (s), e número de paradas. Os resultados demonstram uma mortalidade crescente de insetos de S. zeamais com o incremento das doses. Observou-se também, respostas diferentes entre as populações de S. zeamais em sua capacidade de resistência comportamental associada a diminuição da distância percorrida e do número de paradas dos insetos com o incremento das doses. Confirmando a existência de resistência comportamental nas populações de S. zeamais estudadas. À vista disso, a elucidação de possíveis modificações comportamentais de populações de S. zeamais expostas à Terra Diatomácea pode proporcionar novos caminhos para aumentar a efetividade no manejo dessa praga.

Submitted by Neusa Fagundes (neusa.fagundes@unioeste.br) on 2017-08-30T18:44:32Z No. of bitstreams: 1 AnaPaula_Simonetti2016.pdf: 1680112 bytes, checksum: 8140e54ab237046b1634a3b33d12de36 (MD5) / Made available in DSpace on 2017-08-30T18:44:32Z (GMT). No. of bitstreams: 1 AnaPaula_Simonetti2016.pdf: 1680112 bytes, checksum: 8140e54ab237046b1634a3b33d12de36 (MD5) Previous issue date: 2017-02-07 / Maize has been of great importance as a cereal for the whole Brazilian, but not all harvested grains are used immediately, so much of the production is stored. So, pests are among the factors that affect the quality of stored grains. Sitophilus zeamais has been one of the biggest causes of losses, both in quantitative and qualitative for grains and seeds. Thus, pesticides usually applied to control this weevil can poison the applicators and cause pest resistance or even environmental damage. Therefore, there is an investigation for less aggressive alternatives as a control, and one of them is applying plants with insecticidal or repellent potential. So, this trial aimed at evaluating the effect powder, aqueous extracts and hydroalcoholic extracts of Crambe abyssinica on their survival, emergence, mortality and attractiveness of Sitophilus zeamais, besides its effects on corn seeds treated with this extract. This research was divided into three stages: the first one tested powder and aqueous extracts of crambe at 0%, 1%, 2%, 3% and 4% concentrations in survival, emergence, mortality, attractiveness, repellency and insecticidal action. During the second stage, the same concentrations of crambe extract were tested as in the other ones, but with a hydroalcoholic extract; in order to evaluate survival, emergence, mortality and insecticidal effect on Sitophilus Zeamais. The best effective results were observed with the application of crambe hydroalcohol extracts. The third step evaluated the use of these hydroalcoholic extracts in the following concentrations: 0%, 5%, 15% and 25% concerning attractiveness and insecticidal effect on insects, besides the physiological quality of corn seeds. The obtained results allowed the following conclusions: 1. aqueous extract of crambe grains provides less survival and emergence percentages of Sitophilus zeamais as well as lower consumption of treated corn grains, in relation to the use of crambe grain powder. 2. The hydroalcoholic extract of crambe has insecticidal potential on Sitophilus zeamais, and as concentration increases, from 0 to 4%, the results are more effective. It is also less attractive to the insect, since it provided a decrease in the number of corn grains infested by the pest and the number of insects that emerged from these grains in relation to its non-use. 3. 25% concentration of hydroalcoholic extract in crambe grains yields a high mortality rate (65%); 4. 15% concentration of hydroalcoholic extract in crambe grains stimulates germination percentage, normal seedlings and mass of corn seedlings; 5. 25% concentration hydroalcoholic extract of crambe grains does not negatively influence germination or initial development parameters of maize; and 6. Maize treated with hydroalcohol extract of crambe, stored until 60 days, stimulates germination, mass and length of maize seedlings. Therefore, new research is suggested to develop commercial product based on hydroalcoholic extract of crambe at 25% concentration. / O milho é um cereal de grande importância para todo o país, mas nem todos os grãos colhidos são utilizados imediatamente, assim, grande parte da produção é armazenada. Dentre os fatores que afetam a qualidade do grão armazenado estão as pragas, sendo o Sitophilus zeamais uma das maiores causadoras de perdas, tanto quantitativas como qualitativas para grãos e sementes. Para seu controle, via de regra são utilizados agrotóxicos, que podem causar intoxicação aos aplicadores, resistência da praga ou mesmo danos ambientais. Para evitar esse efeitos, é necessária a busca de alternativas de controle menos agressivas e, uma delas, é o uso de plantas com potencial inseticida ou repelente. O objetivo deste trabalho foi avaliar o efeito do uso de pó, extratos aquosos e hidroalcoólicos de Crambe abyssinica quanto à sobrevivência, emergência, mortalidade e atratividade do Sitophilus zeamais, além de seus efeitos sobre as sementes de milho tratadas com esse extrato. A pesquisa foi dividida em três etapas, a primeira testou pó e extratos aquosos de crambe nas concentrações de 0%, 1%, 2%, 3% e 4% na sobrevivência, emergência, mortalidade, atratividade, repelência e ação inseticida. Na segunda etapa foram testadas as mesmas concentrações do extrato de crambe que na primeira, porém, de extrato hidroalcoólico, nas quais foram avaliadas a sobrevivência, a emergência, a mortalidade e o efeito inseticida sobre o Sitophilus zeamais. Após observados resultados mais efetivos com o uso de extratos hidroalcoólicos de crambe, a terceira etapa avaliou o uso de extratos hidroalcoólicos nas concentrações: 0%, 5%, 15% e 25% na atratividade e efeito inseticida sobre os insetos; além da qualidade fisiológica das sementes de milho. Os resultados obtidos permitiram chegar as seguintes conclusões: 1. O extrato aquoso de grãos de crambe proporciona menor sobrevivência e emergência do Sitophilus zeamais e menor consumo dos grãos de milho tratados, em relação ao uso de pó de grãos de crambe; 2. O extrato hidroalcoólico de crambe tem potencial inseticida sobre o Sitophilus zeamais e à medida que aumenta a concentração, de 0 a 4%, mais efetivo é o resultado; além de ser menos atrativo ao inseto, já que proporcionou a diminuição do número de grãos de milho infestados pela praga e o número de insetos emergidos desses grãos, em relação ao não uso do mesmo; 3. O extrato hidroalcoólico de grãos de crambe na concentração de 25% proporciona taxa de mortalidade alta (65%); 4. O extrato hidroalcoólico de grãos de crambe a 15% estimula a porcentagem de germinação, plântulas normais e massa das plântulas de milho; 5. O extrato hidroalcoólico de grãos de crambe na concentração de 25% não influencia negativamente a germinação nem os parâmetros de desenvolvimento inicial do milho; 6. Milho tratado com extrato hidroalcoólico de crambe armazenado por até 60 dias, estimula a germinação, a massa e comprimento de plântulas de milho. Assim, sugerem-se novas pesquisas para desenvolvimento de produto comercial à base de extrato hidroalcoólico de crambe na concentração de 25%.

O Brasil é o país líder no uso do fungo entomopatogênico Metarhizium anisopliae para o controle pragas agrícolas. Pouco se conhece sobre a diversidade e o impacto dos micovírus em M. anisopliae sensu stricto. Este trabalho mostra a riqueza dos micovírus associados a isolados de M. anisopliae da coleção de entomopatógenos da Universidade de São Paulo, usinas sucroalcooleiras e produtos microbianos à base do entomopatógeno. RNAfd foram encontrados em 55% dos 36 isolados de Metarhizium e apresentaram 16 diferentes padrões eletroforéticos consistindo de 3 a 18 bandas de RNAfd em gel de poliacrilamida. RNAfd não foram detectados em nenhum dos produtos comerciais utilizados no presente estudo. Os diferentes padrões de vírus encontrados nos isolados aqui estudados, aparentemente, não têm relação com os locais onde foram coletados. O inibidor de síntese protéica ciclohexamida não foi eficiente na eliminação dos vírus de RNAfd em nenhum dos isolados de fungos testados. Colônias dos isolados de M. anisopliae ESALQ 866, M. anisopliae ESALQ 1256 e M. anisopliae CTC F8 foram curadas por meio do cultivo monoconidial ou isolamento de ponta de hifas. Alguns segmentos do isolado M. anisopliae PL26 foram perdidos após o subcultivo de ponta das hifas. Colônias de M. anisopliae ESALQ PL26 obtidas por meio do cultivo monoconidial ou subcultivo de ponta de hifas apresentaram grande variabilidade morfológica, no entanto, essa variação não foi correlacionada com a presença de micovírus. Colônias isogênicas de M. anisopliae ESALQ 1256 mostraram diferenças no crescimento, na produção de conídios e na virulência, no entanto, essas diferenças não foram associadas à presença de RNAfd. Não foram observadas diferenças na tolerância aos raios ultravioleta e ao calor entre as colônias de M. anisopliae ESALQ 1256, com e sem vírus de RNAfd. Colônias oriundas de setores formados no isolado M. anisopliae PL26 produziram menor quantidade de conídios e apresentaram menor quantidade de vírus RNAfd em comparação com as colônias oriundas de outras regiões da colônias originais com características normais. A repicagem sucessiva dos isolados de M. anisopliae ESALQ PL26, ESALQ 1256, CTC F8 e CTC F15, infectados por diferentes vírus de RNAfd, nos meios de cultura BDA, SDAY e meio de arroz, bem como a passagem dos fungos em larvas de Tenebrio molitor, em geral, não afetaram a replicação dos micovírus. / Brazil is the leading country in the use of the entomopathogenic fungus Metarhizium anisopliae against agricultural pests. Little is known about the diversity and the impact of mycovirus in M. anisopliae sensu stricto. This study shows the richness of mycovirus associated with M. anisopliae isolates from the collections of entomopathogens of the University of São Paulo, from sugar-alcohol factories and microbial based products. dsRNA were found in 55% of the 36 Metarhizium isolates and showed 16 different electrophoretic patterns consisting of 3 to 18 dsRNA bands in polyacrylamide gels. dsRNA was not detected in any of the commercial products used in this study. The different viral patterns found in the isolates studied here apparently have no relation to the locations where they were collected. The inhibitor of protein synthesis cycloheximide culture was efficient in eradicating dsRNA virus from fungi. Colonies of isolates of M. anisopliae ESALQ 866, M. anisopliae ESALQ 1256 and M. anisopliae F8 were cured by monoconidial or by hyphal tip isolation of. Some segments of the M. anisopliae PL26 isolate were lost following hyphal tip subculture. Colonies both from monoconidial culture and hyphal tip subculture of M. anisopliae ESALQ PL26 showed great morphological variability; however, this variation was not correlated with the presence of mycoviruses. Isogenic colonies of M. anisopliae ESALQ 1256 showed differences in growth, conidia production and virulence; however, these differences were not associated to the presence of the dsRNA. No difference was observed regarding tolerance to ultraviolet rays and heat among the colonies of M. anisopliae ESALQ 1256, with and without the dsRNA virus. Sectors formed in the isolate M. anisopliae PL26 produced a smaller number of conidia and fewer dsRNA virus in comparison to the original colonies with normal characteristics. The repeated subculture of isolates of M. anisopliae ESALQ PL26, ESALQ 1256, CTC F8 and CTC F15, infected by different virus of dsRNA, in the PDA, SDAY culture media and in rice, as well as the fungi passage in larvae of Tenebrio molitor, in general, did not affect the replication of the mycovirus.

Thesis (Ph. D. Agriculture (Plant Protection)) -- University of Limpopo, 2013 / Global withdrawal of synthetic fumigant and non-fumigant nematicides due to their ecounfriendly impacts and high toxicity to non-target organisms, respectively, increased the research and development of alternatives for managing population densities of plantparasitic nematodes, particularly the root-knot (Meloidogyne species) nematodes. Although Meloidogyne species had been managed using genotypes that are resistant to plant-parasitic nematodes in various crops, various challenges negate the available or introgressed nematode resistance. In tomato (Solanum lycopersicum) production, nematode races and instability of nematode resistant genotypes under certain conditions necessitated the continued research and development of alternatives since most of the existing commercial tomato cultivars are highly susceptible to various biological races of Meloidogyne species. The aim of the study was to research and develop appropriate dosages of two phyto- nematicides which could be applied through drip irrigation system in open field tomato production systems, while the specific objectives were to: (1) determine whether a computer-based model could provide nonphytotoxic concentrations to tomato plants using fresh fruits of wild watermelon (Cucumis africanus) and wild cucumber (C. myriocarpus) under greenhouse conditions, (2) determine whether computer-based concentrations from the two plant species when using dried fruits would be less phytotoxic and more suppressive to nematodes, (3) investigate application time intervals for the two products, (4) determine responses of plant growth in tomato and nematode suppression in respect to the derived dosages, and and (5) validate dosages of fermented crude extracts from the two plant species with respect to plant growth of tomato and suppression of nematode numbers. xxxiii Greenhouse, microplot and field studies were set to test the hypotheses intended to achieve the stated objectives, with reliability of measured variables being ensured by using statistical levels of significance (P ≤ 0.05) and coefficients of determination (R2), while validity was ensured by conducting experiments at the same location over two seasons and/or by setting up factorial treatments. Firstly, fermented plant extracts of fresh fruits from C. africanus and C. myriocarpus consistently reduced population densities of Meloidogyne species by 80-92% and 50-90%, respectively. Tomato plants were highly sensitive to the two products as shown by the total degree of sensitivities (Σk) and biological index of 0 and 3, respectively. Also, the mean concentration stimulation range (MCSR) of 11% and 7% concentrations, respectively, attested to this phytotoxicity. Secondly, fermented crude extracts of dried fruits from C. africanus and C. myriocarpus also reduced population densities of Meloidogyne species by 78-97% and 87-97%, respectively. Tomato plants were highly tolerant to the two products in dried form as shown by the total degree of sensitivities (Σk) and biological index of 4 and 3, respectively. The MCSR values for C. africanus and C. myriocarpus dried fruits on tomato were 2.64% and 2.99%, respectively, which for the purpose of this study were individually adjusted to 3%, which translated to 36 L undiluted material/ha of 4 000 tomato plants. In subsequent studies, 3% concentration was used as the standard, along with double strength concentration, namely, 6% concentration. Thirdly, the MCSR values derived in Objective 4, namely 3% and 6% concentration for both Cucumis species using the CARD model were used in the optimisation of application time interval using the innovative concept of weeks (0, 1, 2, 3 and 4) in a 30-day month period. Application time interval for 3% and 6% concentrations of C. africanus fruits was xxxiv optimised at 2.40 and 2.61 weeks in a 30-day month period, respectively, which translated to 18 days [(2.4 weeks/4 weeks) × 30 days] and 20 days [(2.6 weeks/4 weeks) × 30 days], respectively. In contrast, for both concentrations from fermented crude extracts of C. myriocarpus fruits, application time interval was optimised at 16 days for 2.2 and 2.1 weeks, respectively. During optimisation of application frequencies, fermented crude extracts from C. africanus and C. myriocarpus reduced final population densities of M. incognita race 2 by 70-97% and 76-96%, respectively. Fourthly, optimum application intervals (time), allowed computation of dosage, which is a product of concentration and application frequency (dosage = concentration × application frequency). Fifthly, validation of the dosages under open field conditions suggested that 6% × 16-day dosage under crude extracts from C. myriocarpus fruit significantly (P ≤ 0.05) improved growth of tomato plants when compared with those of either 0% (untreated control) or 3% at 16 days. In contrast, dosages of C. africanus fruit at two application frequency had no effect on growth of tomato plants – suggesting that either of the dosages was suitable for use in tomato production since both reduced nematode numbers. During validation, the materials reduced nematode numbers by margins similar to those observed previously under other environments. In conclusion, crude extracts of the two Cucumis species have stimulatory concentrations which have potential similar reductive effects on population densities of Meloidogyne species and could serve as botanical nematicides. However, since plant responses to the two products differed in terms of their respective dosages and active ingredients, it implied that for further improvement of the two, the overriding focus should be on their interaction with the protected plants and nematode numbers. Ideally, future research xxxv should include environmental impact studies, especially on the influence of the products fruit quality of tomato, earthworms, fish and bees.

Thesis (Ph. D. Agriculture (Plant Protection)) -- University of Limpopo, 2013 / Global withdrawal of synthetic fumigant and non-fumigant nematicides due to their ecounfriendly impacts and high toxicity to non-target organisms, respectively, increased the research and development of alternatives for managing population densities of plantparasitic nematodes, particularly the root-knot (Meloidogyne species) nematodes. Although Meloidogyne species had been managed using genotypes that are resistant to plant-parasitic nematodes in various crops, various challenges negate the available or introgressed nematode resistance. In tomato (Solanum lycopersicum) production, nematode races and instability of nematode resistant genotypes under certain conditions necessitated the continued research and development of alternatives since most of the existing commercial tomato cultivars are highly susceptible to various biological races of Meloidogyne species. The aim of the study was to research and develop appropriate dosages of two phyto- nematicides which could be applied through drip irrigation system in open field tomato production systems, while the specific objectives were to: (1) determine whether a computer-based model could provide nonphytotoxic concentrations to tomato plants using fresh fruits of wild watermelon (Cucumis africanus) and wild cucumber (C. myriocarpus) under greenhouse conditions, (2) determine whether computer-based concentrations from the two plant species when using dried fruits would be less phytotoxic and more suppressive to nematodes, (3) investigate application time intervals for the two products, (4) determine responses of plant growth in tomato and nematode suppression in respect to the derived dosages, and and (5) validate dosages of fermented crude extracts from the two plant species with respect to plant growth of tomato and suppression of nematode numbers. xxxiii Greenhouse, microplot and field studies were set to test the hypotheses intended to achieve the stated objectives, with reliability of measured variables being ensured by using statistical levels of significance (P ≤ 0.05) and coefficients of determination (R2), while validity was ensured by conducting experiments at the same location over two seasons and/or by setting up factorial treatments. Firstly, fermented plant extracts of fresh fruits from C. africanus and C. myriocarpus consistently reduced population densities of Meloidogyne species by 80-92% and 50-90%, respectively. Tomato plants were highly sensitive to the two products as shown by the total degree of sensitivities (Σk) and biological index of 0 and 3, respectively. Also, the mean concentration stimulation range (MCSR) of 11% and 7% concentrations, respectively, attested to this phytotoxicity. Secondly, fermented crude extracts of dried fruits from C. africanus and C. myriocarpus also reduced population densities of Meloidogyne species by 78-97% and 87-97%, respectively. Tomato plants were highly tolerant to the two products in dried form as shown by the total degree of sensitivities (Σk) and biological index of 4 and 3, respectively. The MCSR values for C. africanus and C. myriocarpus dried fruits on tomato were 2.64% and 2.99%, respectively, which for the purpose of this study were individually adjusted to 3%, which translated to 36 L undiluted material/ha of 4 000 tomato plants. In subsequent studies, 3% concentration was used as the standard, along with double strength concentration, namely, 6% concentration. Thirdly, the MCSR values derived in Objective 4, namely 3% and 6% concentration for both Cucumis species using the CARD model were used in the optimisation of application time interval using the innovative concept of weeks (0, 1, 2, 3 and 4) in a 30-day month period. Application time interval for 3% and 6% concentrations of C. africanus fruits was xxxiv optimised at 2.40 and 2.61 weeks in a 30-day month period, respectively, which translated to 18 days [(2.4 weeks/4 weeks) × 30 days] and 20 days [(2.6 weeks/4 weeks) × 30 days], respectively. In contrast, for both concentrations from fermented crude extracts of C. myriocarpus fruits, application time interval was optimised at 16 days for 2.2 and 2.1 weeks, respectively. During optimisation of application frequencies, fermented crude extracts from C. africanus and C. myriocarpus reduced final population densities of M. incognita race 2 by 70-97% and 76-96%, respectively. Fourthly, optimum application intervals (time), allowed computation of dosage, which is a product of concentration and application frequency (dosage = concentration × application frequency). Fifthly, validation of the dosages under open field conditions suggested that 6% × 16-day dosage under crude extracts from C. myriocarpus fruit significantly (P ≤ 0.05) improved growth of tomato plants when compared with those of either 0% (untreated control) or 3% at 16 days. In contrast, dosages of C. africanus fruit at two application frequency had no effect on growth of tomato plants – suggesting that either of the dosages was suitable for use in tomato production since both reduced nematode numbers. During validation, the materials reduced nematode numbers by margins similar to those observed previously under other environments. In conclusion, crude extracts of the two Cucumis species have stimulatory concentrations which have potential similar reductive effects on population densities of Meloidogyne species and could serve as botanical nematicides. However, since plant responses to the two products differed in terms of their respective dosages and active ingredients, it implied that for further improvement of the two, the overriding focus should be on their interaction with the protected plants and nematode numbers. Ideally, future research xxxv should include environmental impact studies, especially on the influence of the products fruit quality of tomato, earthworms, fish and bees.

Orientador: Leonardo Fernandes Fraceto / Resumo: Para o controle de pragas, contínuamente se faz o uso de elevadas quantidades de agroquímicos que muitas vezes possuem efeitos adversos tanto para o ambiente quanto para a saúde humana. Este fato tem motivado a busca por alternativas menos impactantes. Neste cenário, diversos mecanismos estão sendo estudados a fim de minimizar estes danos, como por exemplo, o desenvolvimento de sistemas carreadores utilizando polímeros biodegradáveis e proteínas. Aliado a isto, a utilização de inseticidas botânicos também tem demonstrado potencialidade para o combate a pragas, devido aos possíveis menores impactos causados por esses produtos de origem natural. Portanto, o presente trabalho descreve o preparo e caracterização de um nanobiopesticida baseado em nanopartículas de zeína carreadas com óleo de neem junto com a avaliação da sua toxicidade contra organismos não alvos (usando Allium cepa, microbiota do solo envolvida no ciclo do nitrogênio e Caenorhabditis elegans) e da sua fitotoxicidade (usando Phaseolus vulgaris). Ademais, a atividade biológica desse novo sistema em pragas agrícolas (Acanthoscelides obtectus, Bemisia tabaci and Tetranychus urticae) também foi investigada. As nanopartículas esféricas apresentaram diâmetro médio de 278 ± 61,5 nm, alta eficiência de encapsulação (> 80%) e boa estabilidade durante os experimentos. Nos ensaios de toxicidade com A. cepa, as nanopartículas de zeína carregadas com óleo de neem mitigaram o aumento do índice de danos relativo ao DNA causado p... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

Graduation date: 1995