Soybean [Glycine max (L.) Merr.] is an important crop worldwide. It has been widely consumed for protein, oil and other soy products. To develop soybean cultivars with greater resistance against pests and improved meal quality, it is important to elucidate the molecular bases of these traits. This dissertation aims to investigate the biochemical and biological functions of soybean genes from four gene families, which are hypothesized to be associated with soybean defense against pests and soybean meal quality. There are three specific objectives in this dissertation. The first one is to determine the function of components in the salicylic acid (SA) signaling pathway in soybean resistance against soybean cyst nematode (Heterodera glycines, SCN). The second one is to determine whether insect herbivory induce the emission of volatiles from soybean, and if so, how these volatiles are biosynthesized. The third objective is to identify and characterize soybean mannanase genes that can be used for the improvement of soybean meal quality. The soybean genome has been fully sequenced, which provides opportunities for cross-species comparison of gene families of interest and identification of candidate genes in soybean. The cloned cDNAs of putative genes were expressed in Escherichia coli to produce recombinant enzymes. Through biochemical assays, these proteins were proved to be soybean salicylic acid methyltransferase (GmSAMT1), methyl salicylate esterase (GmSABP2-1), α[alpha]-farnesene synthase (GmTPS1) and E-β[beta]-caryophyllene synthase (GmTPS2), and endo-β[beta]-mannanase (GmMAN1). Through a transgenic hairy root system harboring overexpression of GmSAMT1 and GmSABP2-1, both of these two genes were evaluated for their biological function related to resistance against SCN. The results showed that the over-expression of GmSAMT1 and GmSABP2-1 in the susceptible soybean background lead to enhanced resistance against SCN. Among four putative soybean mannanase genes, one gene was cloned and characterized. GmMAN1 showed the endo-β[beta]-mannanase hydrolyse activity and can hydrolyze cell walls isolated from soybean seeds. In summary, using comparative and functional genomics, a number of genes involved in soybean defense and meal quality were isolated and characterized. This study provides novel knowledge and molecular tools for the genetic improvement of soybean for enhanced resistance and improved meal quality.

Made available in DSpace on 2015-04-01T14:16:03Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2190162 bytes, checksum: 9763593619be7b764f8c70a928a7dba5 (MD5) Previous issue date: 2012-03-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Bornyl Salicylate (BS) is a salicylic derivative, obtained by sterification of salicylic acid and monoterpene (-)-borneol, and its topical use in inflammatory diseases has been described in the beginning of 20th century. It is also known that salicylic derivatives have anti-inflammatory activity and borneol have neuroprotective, genoprotective and analgesic activity. The goal of this study was to evaluate activity of BS in experimental models of acute inflammation. The toxicity of BS was analyzed by measuring water and food intake, weight, mortality and weight of major organs. To assess its anti-inflammatory effect, mice pre-treated with BS were subjected to carrageenan, prostaglandin E2, bradikynin or histamine-induced paw edema, zymosan-induced peritonitis and increased vascular permeability induced by acetic acid. NO production was analyzed in peritoneal macrophage cell culture. There were no signs of acute toxicity with the administration of 1000 mg/kg BS in male and female mice. Furthermore, pretreatment with BS was significantly (p<0.05) effective in reducing paw edema induced by carrageenin in early and late times; this effect is related to eicosanoid mediators and bradikynin, and independent of histamine. Neutrophils migration and the release of pro-inflammatory cytokines (TNF-, IL-1 and IL-6) induced by zimosan, and the fluid leakage induced by acetic acid were also reduced in animals treated with BS. In vitro, BS reduced NO release in LPS-stimulated macrophage. These data suggest that BS has anti-inflammatory effects related to different mechanisms of inflammation, and further studies are needed to explore its potential. / O Salicilato de Borneol (SB) é um derivado salicílico, obtido pela esterificação do ácido salicílico e do monoterpeno (-)-borneol, e seu uso tópico em doenças inflamatórias foi descrito no início do século XX. Derivados salicílicos possuem excelente atividade anti-inflamatória e sabe-se que o borneol possui atividade neuroprotetora, genoprotetora e analgésica. Assim, o objetivo deste estudo foi avaliar a atividade do SB em modelos experimentais de inflamação aguda. A toxicidade do SB foi analisada, mensurando-se o consumo de água e ração, peso, letalidade e peso dos principais órgãos. Para avaliar seu efeito anti-inflamatório, camundongos pré-tratados com SB foram submetidos aos protocolos de edema de pata induzido por carragenina, prostaglandina E2, bradicinina ou histamina, peritonite induzida por zimosan e aumento da permeabilidade vascular induzido por ácido acético. A produção de óxido nítrico (NO) foi analisada em cultura de macrófagos peritoneais. Não foram observados sinais de toxicidade aguda com a administração de 1000 mg/kg de SB em camundongos machos e fêmeas. Além disto, o pré-tratamento com SB foi significativamente (p<0,05) eficaz na redução do edema de pata induzido por carragenina em tempos precoces e tardios, pela modulação de eicosanoides e bradicinina, e independente de histamina. A migração de neutrófilos e a liberação de citocinas pró-inflamatórias (TNF-, IL-1 e IL-6) provocadas pelo zimosan, bem como o extravasamento de fluidos mediado pelo ácido acético também foram reduzidos em animais tratados com SB. In vitro, o SB reduziu a liberação de NO em células estimuladas por lipopolissacarídeo de E. coli (LPS). Estes dados sugerem que SB tem efeitos anti-inflamatórios relacionados com diferentes mecanismos da inflamação, e novos estudos são necessários para explorar seu potencial.

Made available in DSpace on 2015-05-14T12:59:57Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1028566 bytes, checksum: 135624ca9af80cfe33db04d68975de22 (MD5) Previous issue date: 2014-02-07 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The pain is a serious public health problem, affects the quality of life of their patients, he brings harm to society. The science continues to seek new more potent analgesic drugs and with fewer side effects. Bornyl salicylate (BS) is a salicylic acid derivative obtained by esterification of salicylic acid with the monoterpene (-)-borneol, both with analgesic effects described in the literature. The aim of this study was to evaluate the antinociceptive effect of BS and its possible mechanisms in models of acute nociception in mice. For this purpose, we performed a behavioral pharmacological screening, test of acetic acid-induced writhing and formalin test. We used blockers pathways involved in pain in the formalin test to elucidate the possible mechanisms of action. BS (200 mg/kg, i.p.) showed behavioral changes indicative of CNS depressant drugs, no effect was observed on oral administration. Doses of 100 and 200 mg/kg (i.p.) reduced the number of writhings and increased the latency time to the first writhing. BS (100 and 200 mg/kg, i.p.) decreased paw licking time of animals only during the inflammatory phase of the formalin test. Pretreatment of animals with the precursor of nitric oxide (L-arginine, 600 mg/kg, i.p.), inhibitor of soluble guanylyl cyclase (methylene blue, 20 mg/kg, i.p.), non-selective muscarinic antagonist (atropine, 1 mg/kg, i.p.) and GABAA receptors antagonist (bicuculline, 1 mg/kg, i.p.), do not reverted the antinociceptive effect of BS (200 mg/kg, i.p.) in the second phase of the formalin test. These data suggest that BS exerts peripheral antinociceptive effect (anti-inflammatory), which mechanism seems in principle not involve nitric oxide, cGMP, muscarinic cholinergic receptors or GABAA receptors. Further studies are required to clarify the effect produced by bornyl salicylate. / A dor é um grave problema de saúde pública, afeta a qualidade de vida de seus portadores e traz inúmeros prejuízos para a sociedade. A ciência continua buscando novos fármacos analgésicos mais potentes e com menos efeitos colaterais. O salicilato de bornila (SB) é um derivado salicílico obtido pela esterificação do ácido salicílico com o monoterpeno (-)-borneol, ambos com efeitos analgésicos já descritos na literatura. O objetivo deste estudo foi avaliar o efeito antinociceptivo do SB e seus possíveis mecanismos em modelos de nocicepção aguda em camundongos. Para tanto, realizou-se uma triagem farmacológica comportamental, teste das contorções abdominais induzidas por ácido acético e teste da formalina. Utilizou-se bloqueadores de vias envolvidas na dor no teste da formalina para evidenciar os possíveis mecanismos de ação. O SB (200 mg/kg, i.p.) promoveu alterações comportamentais indicativas de droga depressora do SNC, efeito não observado na administração por via oral. As doses de 100 e 200 mg/kg (i.p.) reduziram o número de contorções abdominais e aumentaram o tempo de latência para a primeira contorção. O SB (100 e 200 mg/kg, i.p.) diminuiu o tempo de lambida da pata dos animais somente na fase inflamatória do teste da formalina. O pré-tratamento dos animais com o precursor do óxido nítrico (L-arginina, 600 mg/kg, i.p.), inibidor da ciclase de guanilil solúvel (azul de metileno, 20 mg/kg, i.p.), antagonista muscarínico não-seletivo (atropina, 1 mg/kg, i.p.) e antagonista de receptores GABAA (bicuculina, 1 mg/kg, i.p.), não reverteram o efeito antinociceptivo do SB (200 mg/kg, i.p.) na segunda fase do teste da formalina. Estes dados sugerem que o SB apresenta efeito antinociceptivo periférico (anti-inflamatório), cujo mecanismo parece em princípio não envolver óxido nítrico, GMPc, receptores colinérgicos muscarínicos nem receptores GABAA. Novos estudos são necessários para melhor esclarecer o efeito produzido pelo salicilato de bornila.

Ziel der Dissertation war die Überprüfung der klinischen Wirksamkeit von Natriumsalicylat bei respiratorischen Erkrankungen von Mastputen unter Feldbedingungen. Bisher existieren keine konkreten Richtlinien bzw. Modelle zum Versuchsaufbau beim Geflügel, und explizit bei der Pute, zur Durchführung einer derartigen Studie. Daher stellten sowohl die Konzeption als auch die Implementierung eines Studiendesigns zur Untersuchung der Wirksamkeit eines Antiphlogistikums bei der Mastpute unter Feldbedingungen eine zusätzliche elementare Zielsetzung dieser Promotion dar. Die in dieser Dissertationsschrift entwickelten Feldstudien orientieren sich an den Leitlinien der good clinical practice sowie der guideline for the conduct of efficacy studies for non-steroidal anti-inflammatory drugs und entsprechen der Wertigkeitsklasse Ia der evidenzbasierten Veterinärmedizin. Jede der drei Feldstudien wurde als kontrollierte, randomisierte und verblindete Parallelgruppenstudie in verschiedenen Regionen Deutschlands durchgeführt und gliederte sich in eine viertägige experimentelle Phase (EP) und eine siebentägige postexperimentelle Phase (PoP). Die zunächst definierten Arbeitshypothesen bekamen entsprechende Beurteilungskriterien zugewiesen. Unter deren Nutzung erfolgte die spätere Evaluierung, inwieweit die entzündungshemmende Wirkung von Natriumsalicylat imstande war, den Heilungsprozess der respiratorischen Grunderkrankung zu beschleunigen und eine verbesserte restitutio ad integrum hervorzurufen.

O objetivo do trabalho foi o desenvolvimento de tecnologias para melhoria da qualidade e vida de vaso em rosas (Rosa hybrida.) Vega. Os tratamentos précolheita foram realizados com aplicação de silício (0, 200, 400 e 800 mg L-1) por meio de fertirrigação via solo e via foliar. Foram realizadas seis aplicações semanais de silicato de potássio (12% Si) em rosas Vega, distribuídas em blocos casualisados com quatro repetições, e seis avaliações pós-colheita (dias 0, 2, 4, 6, 8 e 10). Após a colheita, foram avaliados: comprimento de haste, diâmetro de haste e de pedúnculo, comprimento de botão e a massa das hastes. As flores foram armazenadas em vasos com água deionizada a 20±1°C e 65±5% UR, sendo avaliadas: variação da massa fresca, conteúdo relativo de água (CRA) de pétalas e folhas, atividade de peroxidase, abertura floral, curvatura do pedúnculo, turgescência e escurecimento de pétalas, e coloração de pétalas. A aplicação de 400 mg L-1 de silício proporcionou maior comprimento de haste, maior CRA e cromaticidade das pétalas e menor atividade de peroxidase. A aplicação foliar também apresentou maior comprimento de botão, CRA de folhas, e luminosidade de pétalas, e a aplicação via solo também apresentou maior diâmetro de haste, maior massa inicial, menor variação de massa fresca e maior abertura floral, indicando que a aplicação de silício pode contribuir para melhorar a qualidade de rosas Vega. Os tratamentos pós-colheita foram realizados com aplicações de 0,1 mM de salicilato de metila (MeSa), 1 L L-1 de 1-metilciclopropeno (1-MCP), e 1 mM de ácido salicílico (AS). Após a aplicação dos tratamentos por 12 horas, as flores foram mantidas em vasos com água deionizada, armazenadas a 25±1°C e 65±5% U R. O delineamento experimental foi inteiramente casualisado, com quatro repetições por tratamento e seis avaliações (dias 0, 2, 4, 6, 8 e 10). Os tratamentos com AS e 1-MCP reduziram a atividade respiratória, produção de etileno, perda de massa e a curvatura do pedúnculo, tendo permitido maior manutenção do conteúdo relativo de água das pétalas, maior abertura floral, maior luminosidade das pétalas, e maior atividade de peroxidase. O AS também proporcionou melhor manutenção da turgescência, do conteúdo relativo de água das folhas, e menor escurecimento das pétalas. / The objective of this study was the development of technologies to increase the quality and vase life of cut roses (Rosa hybrida.) Vega. The pre-harvest treatments were the application of silicon (0, 200, 400 and 800 mg L-1) via foliar spray and root feeding, in quality of cut roses. Six weekly applications of potassium silicate (12% Si) were performed on roses Vega, distributed in a randomized block design with four replicates per treatment, and six evaluations (days 0, 2, 4, 6, 8 and 10). After harvest, the stem length, stem diameter, peduncle diameter, bud length and mass of the stems were evaluated. The flowers were kept in vases with deionized water, stored at 20±1°C and 65±5% RH, with evaluati ons of loss of fresh weight, leaf and petal relative water content (RWC), peroxidase activity, flower opening, bent neck, turgidity, petal darkening, and color of petals. The application of 400 mg L-1 of silicon provided increase in stem length, in petal RWC, petal chromaticity and reduced peroxidase activity. The application via foliar spray also increased bud length, the leaf RWC and the petal luminosity. The root feeding also increased the stem diameter, the floral opening, the initial mass, and reduced the fresh weight loss, indicating that the application of silicon can improve the quality of roses Vega. The postharvest treatments were the application of 0,1 mM of methyl salicylate (MeSa), 1 L L-1 of 1-methylcyclopropene (1-MCP), and 1 mM of salicylic acid (SA), in postharvest quality conservation of Vega roses. After the application of treatments for 12 hours, the flowers were kept in vases with deionized water, stored at 25±1°C and 65±5% RH. The experimental design was completely randomized, with four replications and six evaluations (days 0, 2, 4, 6, 8 and 10). The treatments with SA and 1-MCP reduced respiratory activity, ethylene production, loss of fresh weight and bent neck; and enable better maintenance of petal relative water content, largest flower opening and higher petal luminosity. SA also provided better maintenance of turgidity and leaf relative water content and less petal darkening.

O objetivo do trabalho foi o desenvolvimento de tecnologias para melhoria da qualidade e vida de vaso em rosas (Rosa hybrida.) Vega. Os tratamentos précolheita foram realizados com aplicação de silício (0, 200, 400 e 800 mg L-1) por meio de fertirrigação via solo e via foliar. Foram realizadas seis aplicações semanais de silicato de potássio (12% Si) em rosas Vega, distribuídas em blocos casualisados com quatro repetições, e seis avaliações pós-colheita (dias 0, 2, 4, 6, 8 e 10). Após a colheita, foram avaliados: comprimento de haste, diâmetro de haste e de pedúnculo, comprimento de botão e a massa das hastes. As flores foram armazenadas em vasos com água deionizada a 20±1°C e 65±5% UR, sendo avaliadas: variação da massa fresca, conteúdo relativo de água (CRA) de pétalas e folhas, atividade de peroxidase, abertura floral, curvatura do pedúnculo, turgescência e escurecimento de pétalas, e coloração de pétalas. A aplicação de 400 mg L-1 de silício proporcionou maior comprimento de haste, maior CRA e cromaticidade das pétalas e menor atividade de peroxidase. A aplicação foliar também apresentou maior comprimento de botão, CRA de folhas, e luminosidade de pétalas, e a aplicação via solo também apresentou maior diâmetro de haste, maior massa inicial, menor variação de massa fresca e maior abertura floral, indicando que a aplicação de silício pode contribuir para melhorar a qualidade de rosas Vega. Os tratamentos pós-colheita foram realizados com aplicações de 0,1 mM de salicilato de metila (MeSa), 1 L L-1 de 1-metilciclopropeno (1-MCP), e 1 mM de ácido salicílico (AS). Após a aplicação dos tratamentos por 12 horas, as flores foram mantidas em vasos com água deionizada, armazenadas a 25±1°C e 65±5% U R. O delineamento experimental foi inteiramente casualisado, com quatro repetições por tratamento e seis avaliações (dias 0, 2, 4, 6, 8 e 10). Os tratamentos com AS e 1-MCP reduziram a atividade respiratória, produção de etileno, perda de massa e a curvatura do pedúnculo, tendo permitido maior manutenção do conteúdo relativo de água das pétalas, maior abertura floral, maior luminosidade das pétalas, e maior atividade de peroxidase. O AS também proporcionou melhor manutenção da turgescência, do conteúdo relativo de água das folhas, e menor escurecimento das pétalas. / The objective of this study was the development of technologies to increase the quality and vase life of cut roses (Rosa hybrida.) Vega. The pre-harvest treatments were the application of silicon (0, 200, 400 and 800 mg L-1) via foliar spray and root feeding, in quality of cut roses. Six weekly applications of potassium silicate (12% Si) were performed on roses Vega, distributed in a randomized block design with four replicates per treatment, and six evaluations (days 0, 2, 4, 6, 8 and 10). After harvest, the stem length, stem diameter, peduncle diameter, bud length and mass of the stems were evaluated. The flowers were kept in vases with deionized water, stored at 20±1°C and 65±5% RH, with evaluati ons of loss of fresh weight, leaf and petal relative water content (RWC), peroxidase activity, flower opening, bent neck, turgidity, petal darkening, and color of petals. The application of 400 mg L-1 of silicon provided increase in stem length, in petal RWC, petal chromaticity and reduced peroxidase activity. The application via foliar spray also increased bud length, the leaf RWC and the petal luminosity. The root feeding also increased the stem diameter, the floral opening, the initial mass, and reduced the fresh weight loss, indicating that the application of silicon can improve the quality of roses Vega. The postharvest treatments were the application of 0,1 mM of methyl salicylate (MeSa), 1 L L-1 of 1-methylcyclopropene (1-MCP), and 1 mM of salicylic acid (SA), in postharvest quality conservation of Vega roses. After the application of treatments for 12 hours, the flowers were kept in vases with deionized water, stored at 25±1°C and 65±5% RH. The experimental design was completely randomized, with four replications and six evaluations (days 0, 2, 4, 6, 8 and 10). The treatments with SA and 1-MCP reduced respiratory activity, ethylene production, loss of fresh weight and bent neck; and enable better maintenance of petal relative water content, largest flower opening and higher petal luminosity. SA also provided better maintenance of turgidity and leaf relative water content and less petal darkening.

Experiments were conducted to assess whether a concept termed 'attract and reward' (A&R) could enhance conservation biological control (CBC). In A&R, a synthetically-produced herbivore induced plant volatile (HIPV) ('attract') is combined with a floral resource ('reward'). It is anticipated that the two will work synergistically attracting natural enemies into the crop ('attract') and maintaining them within the crop ('reward'). The study system consists of brassica, the most commonly occurring brassica herbivores, their natural enemies and higher order natural enemies. The HIPV deployed is methyl salicylate (MeSA) and the floral resource is buckwheat Fagopyrum esculentum. The aim of the first two field experiments, in 2007 and 2008, was to evaluate the effects of MeSA and MeSA combined with buckwheat (A&R) on the abundance of arthropods from three trophic levels. In 2007, a field experiment was conducted using MeSA alone. The mean abundance of the leafmining fly Scaptomyza flava (trophic level 2), the diamondback moth (trophic level 2)(DBM) parasitoid Diadegma semiclausum (trophic level 2) and the hoverfly Melangyna novaezealandiae (trophic level 3) was increased in MeSA-treatments by up to 300% and for the brown lacewing parasitoid Anacharis zealandica a maximum mean increase of 600% was recorded. Significantly more females of the D. semiclausum and M. novaezealandiae were attracted to MeSA than males. When A&R was deployed in 2008, were arthropods from the third and fourth trophic levels affected. For none of the species was there a synergistic effect between 'attract' and 'reward' on their abundance. The brown lacewing Micromus tasmaniae (trophic level 3), two parasitoids of DBM and one of cabbage white butterfly Pieris rapae (trophic level 2) increased significantly in treatments with buckwheat. The hoverfly Melanostoma fasciatum (trophic level 3) was significantly more abundant in treatments with MeSA, but significantly less abundant in treatments with buckwheat. The effect of MeSA on the fourth trophic level parasitoid Anacharis zealandica (trophic level 4) was inconsistent between years. Here it significantly decreased its abundance, while treatments with buckwheat increased it. Significantly fewer male than female D. semiclausum were attracted to MeSA only treatments. These experiments show that MeSA and buckwheat can have unwanted effects on arthropod abundance which may disrupt CBC. To assess the effect of A&R on CBC a further field experiment evaluating herbivore densities, predation, parasitism and hyper-parasitism rates was conducted. The only effect was significantly higher aphid parasitism in treatments with MeSA. Based on the results from the field experiments it remained unclear whether it was MeSA or a blend of volatiles produced by MeSA-induced host plants that were attractive to the arthropods. An olfactory experiment was conducted to evaluate if the aphid parasitoid Aphidius colemani can be attracted to two different concentrations of MeSA diluted in Synertrol oil. Significantly more parasitoids were attracted to 2.0% MeSA than to air while the parasitoid did not respond to the 0.5% concentration. These results indicate that A&R has potential as a CBC technique, as long as any unwanted side effects can be managed. Although there were no synergistic effects between 'attract' and 'reward' on the abundance of individual natural enemies, combining MeSA and buckwheat could still be beneficial because the two techniques increase the abundance of different natural enemies.

Traumatic brain injury (TBI) is a major cause of mortality and disability. As common sequelae in survivors of TBI are disabling functional, emotional and cognitive disturbances, improved treatment of TBI patients is urgently needed. At present, no neuroprotective pharmacological treatment exists. The formation of oxygen-centered free radicals, reactive oxygen species (ROS), is considered an important event in the pathophysiology of TBI. In the present thesis, the fluid percussion (FPI) and controlled cortical contusion injury models of TBI in rats were used. Two nitrone radical scavengers, α-Phenyl-N-tert -butyl nitrone (PBN) and the sulfonated analogue of PBN, 2-sulfophenyl-N-tert-butyl nitrone (S-PBN), were used as tools to study the role of ROS in TBI. Pre-treatment with PBN (30 mg/kg) improved morphological and cognitive outcome after severe controlled cortical contusion injury. Treatment with equimolar doses of PBN and S-PBN administered 30 min after FPI followed by a 24 h intravenous infusion improved morphological outcome. Only S-PBN improved cognitive outcome as assessed in the Morris Water Maze. Surprisingly, pre-treatment with PBN increased the number of apoptotic neurons at 24 hours after injury despite a reduced lesion volume. FPI resulted in an early increase in glucose uptake and a reduction in regional cerebral blood flow (rCBF) assessed by fluoro-2-deoxyglucose (FDG) and hexamethylpropylene amine oxime (HMPAO) autoradiography. At 12 h, a marked reduction in glucose uptake and rCBF ensued. These TBI-induced changes were attenuated by PBN and S-PBN pre-treatment. A method for ROS detection using 4-hydroxybenzoate in conjunction with microdialysis was evaluated. The results showed a marked increase in ROS formation as assessed by an increase in the single adduct 3,4-DHBA, lasting 90 min after injury. In a separate study, PBN and S-PBN equally reduced 3,4-DHBA formation despite no detectable brain concentrations of S-PBN at 30 or 60 min post-injury. In conclusion, ROS play an important role in the injury process after TBI. We report a method for ROS detection with potential clinical utility. Nitrones increased ROS elimination and improved functional and morphological outcome. Nitrone treatment may have a clinical potential as a neuroprotective concept in TBI.

Experiments were conducted to assess whether a concept termed 'attract and reward' (A&R) could enhance conservation biological control (CBC). In A&R, a synthetically-produced herbivore induced plant volatile (HIPV) ('attract') is combined with a floral resource ('reward'). It is anticipated that the two will work synergistically attracting natural enemies into the crop ('attract') and maintaining them within the crop ('reward'). The study system consists of brassica, the most commonly occurring brassica herbivores, their natural enemies and higher order natural enemies. The HIPV deployed is methyl salicylate (MeSA) and the floral resource is buckwheat Fagopyrum esculentum. The aim of the first two field experiments, in 2007 and 2008, was to evaluate the effects of MeSA and MeSA combined with buckwheat (A&R) on the abundance of arthropods from three trophic levels. In 2007, a field experiment was conducted using MeSA alone. The mean abundance of the leafmining fly Scaptomyza flava (trophic level 2), the diamondback moth (trophic level 2)(DBM) parasitoid Diadegma semiclausum (trophic level 2) and the hoverfly Melangyna novaezealandiae (trophic level 3) was increased in MeSA-treatments by up to 300% and for the brown lacewing parasitoid Anacharis zealandica a maximum mean increase of 600% was recorded. Significantly more females of the D. semiclausum and M. novaezealandiae were attracted to MeSA than males. When A&R was deployed in 2008, were arthropods from the third and fourth trophic levels affected. For none of the species was there a synergistic effect between 'attract' and 'reward' on their abundance. The brown lacewing Micromus tasmaniae (trophic level 3), two parasitoids of DBM and one of cabbage white butterfly Pieris rapae (trophic level 2) increased significantly in treatments with buckwheat. The hoverfly Melanostoma fasciatum (trophic level 3) was significantly more abundant in treatments with MeSA, but significantly less abundant in treatments with buckwheat. The effect of MeSA on the fourth trophic level parasitoid Anacharis zealandica (trophic level 4) was inconsistent between years. Here it significantly decreased its abundance, while treatments with buckwheat increased it. Significantly fewer male than female D. semiclausum were attracted to MeSA only treatments. These experiments show that MeSA and buckwheat can have unwanted effects on arthropod abundance which may disrupt CBC. To assess the effect of A&R on CBC a further field experiment evaluating herbivore densities, predation, parasitism and hyper-parasitism rates was conducted. The only effect was significantly higher aphid parasitism in treatments with MeSA. Based on the results from the field experiments it remained unclear whether it was MeSA or a blend of volatiles produced by MeSA-induced host plants that were attractive to the arthropods. An olfactory experiment was conducted to evaluate if the aphid parasitoid Aphidius colemani can be attracted to two different concentrations of MeSA diluted in Synertrol oil. Significantly more parasitoids were attracted to 2.0% MeSA than to air while the parasitoid did not respond to the 0.5% concentration. These results indicate that A&R has potential as a CBC technique, as long as any unwanted side effects can be managed. Although there were no synergistic effects between 'attract' and 'reward' on the abundance of individual natural enemies, combining MeSA and buckwheat could still be beneficial because the two techniques increase the abundance of different natural enemies.

El objetivo de esta Tesis Doctoral es estudiar diferentes herramientas para el manejo de plagas y enfermedades del cultivo de la platanera. Entre las herramientas que vamos a desarrollar, se van a analizar los compuestos orgánicos volátiles (COVs) fúngicos derivados de hongos entomopatógenos (HE) y nematófagos, como fuente de metabolitos con actividad antagónica contra el picudo negro (PN) de la platanera, Cosmopolites sordidus, para su control y manejo en el campo. Así mismo, vamos a estudiar las respuestas de cultivares de plataneras a quitosano, un polisacárido biodegradable, para evaluar su posible uso en el campo como estimulante y protector de las plantas frente a plagas y patógenos, como Fusarium oxysporum f. sp. cubense. El conjunto de capítulos de esta tesis pretende sentar las bases de una estrategia de manejo sostenible de plagas y enfermedades del cultivo de la platanera, basada en el uso de COVs derivados de hongos presentes de forma natural en los cultivos, en combinación con la suplementación de quitosano en el riego, para un efecto de protección y activación de las defensas de las plataneras antes de cualquier infección de plagas y/o enfermedades. El objetivo principal de esta Tesis Doctoral es encontrar nuevas fórmulas para la gestión integrada de plagas como Cosmopolites sordidus y enfermedades de la platanera en condiciones de campo. En esta Tesis Doctoral hemos ideado enfoques sostenibles para la gestión de las plagas y enfermedades de las plataneras. Nuestros objetivos son: a) Cosmopolites sordidus (picudo negro de la platanera, PN), la principal plaga de los cultivos de plátano y, b) el hongo del marchitamiento Fusarium oxysporum f. sp. cubense Raza Tropical 4 (FocTR4), agente causante de una nueva variante extremadamente virulenta de la enfermedad del “Mal de Panamá”, que se está extendiendo rápidamente por todo el mundo. Nuestras herramientas de gestión sostenible son: a) los hongos entomopatógenos (HE, conocidos por su uso como agentes de control biológico, ACBs) aislados de campos comerciales de plátanos, b) sus compuestos orgánicos volátiles (COVs) y, c) el quitosano, un compuesto biodegradable y elicitor de la inmunidad de las plantas con actividad antimicrobiana. Damos evidencia de que los COVs de los hongos agentes de control biológico son repelentes del PN. Pueden utilizarse en los cultivos de platanera mediante estrategias de push and pull para gestionar la plaga de forma sostenible. El quitosano puede utilizarse en el riego para prevenir las defensas de la platanera local y sistémicamente. Por lo tanto, este polímero, con probada actividad antimicrobiana frente a otros patógenos de marchitamiento de Fusarium spp., podría utilizarse contra la actual pandemia en las plataneras causada por FocTR4. La capacidad de inducir reguladores del crecimiento de las plantas sostiene también el papel fertilizante del quitosano. La inducción de compuestos relacionados con la respuesta sistémica adquirida (RSA) hace que el riego con quitosano sea una herramienta para manejar también las plagas de las plataneras sobre el suelo (PN) y las enfermedades (Sigatoka). De esta manera, los COVs y el quitosano podrían ayudar a reducir el uso de agroquímicos tóxicos en los cultivos de platanera en todo el mundo.