Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2014-12-18T10:55:48Z No. of bitstreams: 1 Dissertação - Juliana Camila Lopes Cavaion - 2012.pdf: 2928983 bytes, checksum: 908363ad8b87e3a821dd54fe80dcb4ce (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-12-18T14:24:31Z (GMT) No. of bitstreams: 1 Dissertação - Juliana Camila Lopes Cavaion - 2012.pdf: 2928983 bytes, checksum: 908363ad8b87e3a821dd54fe80dcb4ce (MD5) / Made available in DSpace on 2014-12-18T14:24:32Z (GMT). No. of bitstreams: 1 Dissertação - Juliana Camila Lopes Cavaion - 2012.pdf: 2928983 bytes, checksum: 908363ad8b87e3a821dd54fe80dcb4ce (MD5) Previous issue date: 2012-06-04 / Filamentous fungi have cytochrome P450 enzyme system similar to mammals whose application allows catalyze phase I oxidative reactions of metabolism. Obtaining functionalized derivatives by these microorganisms was the main purpose of this dissertation. Studies in silico to predict metabolism in the programs MetaPrint2D and SMARTCyp indicated S-oxidation, aromatic hydroxylation and dealkylation reactions, as the most likely S-oxidation in thiomorpholine ring of 2-[4-(1,4-thiazinan-4-ylsulfonyl)phenylcarbamoyl]benzoic acid (LASSBio-596). The filamentous fungi Absidia blakesleana ATCC 26617, Aspergillus candidus ATCC 1009, Beauveria bassiana ATCC 7159, Cunninghamella echinulata ATCC 9244, Cunninghamella echinulata ATCC 9245, Cunninghamella elegans ATCC 36112, Fusarium roseum ATCC 14717 and Streptomyces vendulae ATCC 8664 were used for biotransformation of LASSBio-596. In the reaction conditions applied, substrate dissolved in methanol with concentration of 0.25 mg/mL added to liquid culture medium PDSM and incubated at 27 °C with 200 rpm for 96 hours, the fungus Beauveria bassiana ATCC 7159 performed S-oxidation in the thiomorpholine ring resulting in sulfone derivative of LASSBio-596, called LaBioCon 214, yield of 5.84 %, and it was found the hydrolysis of amide reaction obtaining the compound 4-(thiomorpholinosulfonyl)aniline, LaBioCon 202, yield of 13.8 %. It was noted the solvent used affects the biotransformation process. The monitoring of the reactions and purification of the derivatives obtained were performed by high performance liquid chromatography with ultraviolet detector at 267 nm and preparative chromatography with column Microsorb 100-5 C18 (250 x 10 mm, 5 μm). Subsequently, sulfone derivative was obtained, called LaBioCon 223, by synthesis. Structural characterization was made by classical methods such as 1H NMR and mass spectrometry. / Os fungos filamentosos possuem o sistema enzimático citocromo P450 semelhante ao de mamíferos, cuja aplicação permite catalisar reações oxidativas de fase I do metabolismo. A obtenção de derivados funcionalizados por estes microrganismos foi o objetivo principal deste trabalho. Estudos in silico de previsão do metabolismo nos programas MetaPrint2D e SMARTCyp indicaram reações de S-oxidação, hidroxilação aromática e desalquilação, sendo mais provável a S-oxidação no anel tiomorfolina do ácido 2-[4-(1,4-tiazinan-4-ilsulfonil)fenilcarbamoil]benzóico (LASSBio-596). Os fungos filamentosos Absidia blakesleana ATCC 26617, Aspergillus candidus ATCC 1009, Beauveria bassiana ATCC 7159, Cunninghamella echinulata ATCC 9244, Cunninghamella echinulata ATCC 9245, Cunninghamella elegans ATCC 36112, Fusarium roseum ATCC 14717 e Streptomyces vendulae ATCC 8664 foram empregados para biotransformação do LASSBio-596. Nas condições reacionais aplicadas, substrato solubilizado em metanol com concentração de 0,25 mg/mL adicionado ao meio de cultura líquido PDSM e incubado a 27 °C com 200 rpm por 96 horas, a Beauveria bassiana ATCC 7159 promoveu a S-oxidação no anel tiomorfolina resultando no derivado sulfona do LASSBio-596, denominado LaBioCon 214, com rendimento de 5,84 %, e constatou-se a reação de hidrólise da amida obtendo o composto 4-(tiomorfolina-sulfonil)anilina, LaBioCon 202, com rendimento de 13,8 %. Verificou-se que o solvente utilizado interfere no processo de biotransformação. O monitoramento das reações e a purificação dos derivados obtidos foram realizados por cromatografia líquida de alta eficiência com detector ultravioleta em 267 nm e cromatografia preparativa com coluna Microsorb 100-5 C18 (250 x 10 mm, 5 μm). Posteriormente, foi obtido o derivado sulfona, LaBioCon 223, por via sintética. A caracterização estrutural foi realizada por métodos clássicos como a espectroscopia de ressonância magnética nuclear de hidrogênio e espectrometria de massas.

Formulações a base de dispersões oleosas de conídios aéreos de fungos entomopatogênicos apresentam diversas vantagens em comparação à aplicação de produtos não formulados; entretanto, a complexidade e a falta de compreensão dos fatores que influenciam a persistência desse tipo de formulação fazem com que ainda seja pouco utilizada. O efeito de diferentes níveis de secagem de conídios, tipo de óleo, métodos de extração, estabilidade de blastosporos, compatibilidade de surfactantes e adição de aditivos foram avaliados para o desenvolvimento de micopesticidas formulados em óleo. Os melhores resultados obtidos com os conídios em óleo foram avaliados quanto à proteção conferida à radiação UV e a eficiência das formulações na mortalidade de Tenebrio molitor. Zeolita foi a argila mais eficiente na manutenção da viabilidade de blastosporos de M. anisopliae e B. bassiana, e apesar de resultados promissores a estabilidade desses propágulos foi muito inferior a de conídios aéreos. Vinte surfactantes foram avaliados quanto a inocuidade aos conídios, em dois ensaios, culminando com a seleção dos quatro mais compatíveis, dois deles E5 e E7, avaliados nos experimentos posteriores demonstraram bom potencial para compor a formulação como agentes emulsionantes. Dois aditivos foram avaliados na formulação e respostas distintas foram observadas. O A1 foi altamente incompatível, enquanto diferentes concentrações de A2 (1, 3 e 5%) incrementaram a sobrevivência dos conídios em até sete vezes. As vantagens da presença do A2 ficaram evidentes também em combinações de óleo com os quatro emulsionantes selecionados. As dispersões oleosas proporcionaram sobrevivência até sete vezes superior para conídios expostos a radiação UV em relação a conídios não formulados e dois experimentos comprovaram que as formulações oleosas testadas foram mais eficientes no controle de insetos, com reduções de até 2,3 vezes no tempo de sobrevivência de insetos. Os 12 bioensaios com mais de 20.000 leituras de viabilidade descritos neste trabalho permitiram um melhor entendimento dos fatores críticos que afetam a vida de prateleira de conídios, e indicam procedimentos que podem ser utilizados para melhorar a estabilidade de micopesticidas formulados em óleo. / Oil-based formulations of aerial conidia of entomopatogenic fungi have several advantages compared to the use of unformulated products; however, the complexity and lack of understanding of the factors that influence the persistence of this type of formulation make it still not widely used. The effect of different levels of drying conidia, oil type, extraction methods, blastospores stability, compatibility of surfactants and addition of additives were evaluated for the development of a mycopesticide oil formulation. The best results obtained with aerial conidia were evaluated regarding the protection against UV radiation and the efficacy of the formulations to control Tenebrio molitor. Zeolita was the most compatible clay in maintaining the viability of blastospores of M. anisopliae and B. bassiana, and despite promising results in the stability, the shelf life of these propagules was much lower than aerial conidia. Twenty surfactants were evaluated for safety to conidia in two trials resulting in the selection of the four most compatible, two of them E5 and E8, were evaluated in subsequent experiments demonstrating good potential as emulsifiers in oil-based formulations. Two additives were evaluated in the formulation and distinct responses were observed. A1 was highly toxic to the fungi, while different concentrations of A2 increased the survival of the conidia up to seven times. The advantages of A2 were evident also in combinations of oil with the four selected emulsifiers. Oil-based formulation provided survival up to seven times higher for conidia exposed to simulated UV radiation compared to unformulated conidia, and two experiments showed that oil formulations were more efficient in the insect control, with reductions of up to 2.3 times on the survival time of insects. Twelve bioassays and over 20.000 viability readings described in this work, allowed a better understanding of the critical factors that affect the shelf-life of conidia, and indicate methods that can be used to improve the stability of mycopesticides in oil-based formulations.

Os fungos entomopatogênicos dos gêneros Metarhizium, Beauveria e Isaria (Ordem Hypocreales), são comumente encontrados em solo onde sobrevivem de maneira saprofítica ou como endofíticos do sistema radicular das plantas. Informações sobre a composição destas espécies bem como sua diversidade, distribuição e associação com diferentes tipos de cultivos e vegetação nativa são escassas no Brasil. O presente estudo foi desenvolvido para comparar a abundância de fungos entomopatogênicos e a diversidade genética de isolados de Metarhizium spp. em amostras de solo de cultivos anuais, perenes e vegetação nativa, em cinco estados brasileiros que representam os biomas Amazônia, Cerrado, Caatinga, Mata Atlântica e Pampa, em duas estações (seca e úmida) nos anos de 2012 e 2013. O isolamento dos fungos foi realizado com meio seletivo e \"Insect bait\" utilizando Galleria mellonella e Tenebrio molitor. Nos estudos de diversidade genética de Metarhizium spp. foram utilizadas sequências de DNA da região MzIGS3. Representantes dos haplótipos revelados nesta análise tiveram a região 5\'-TEF sequenciada para identificação específica. Fungos entomopatogênicos foram isolados de 86% das 1.056 amostras de solo sendo Metarhizium o gênero predominante (66% das amostras de solo), seguido por Beauveria (41,9%) e Isaria (10,8%). Em geral, as maiores densidades de fungos entomopatogênicos foram obtidas nos biomas Amazônia e Cerrado e as menores densidades detectadas no bioma Caatinga. Metarhizium spp. foi detectado em maior número de amostras de solo em vegetação nativa e cultivos anual e perene do Cerrado. A frequência de Isaria spp. foi baixa nas amostras de solo, sendo detectado em maior número de amostras nos solos com cultivos anuais e vegetação nativa na Amazônia e Caatinga. Metarhizium spp. foi geralmente encontrado em um maior número de amostras coletadas na estação úmida em comparação com as coletas da estação seca, por outro lado Beauveria spp. foi superior na estação seca. A diversidade dos isolados de Metarhizium spp. provenientes de áreas de vegetação nativa foi maior do que dos isolados de cultivos anuais e perenes. Seis linhagens foram encontradas neste estudo; M. robertsii, M. anisopliae, M. pingshaense e três espécies indeterminadas. M. robertsii foi a linhagem predominante (65% dos isolados) sendo encontrado em áreas com vegetação nativa e cultivos anual e perene dos cinco biomas. Metarhizium sp. indet. 1 apresentou a maior diversidade haplotípica dentre as linhagens estudadas. Uma nova linhagem, não caracterizada taxonomicamente, Metarhizium sp. indet. 3, foi encontrada predominantemente na Caatinga. Somente na Amazônia foram encontradas todas as linhagens. O conhecimento da composição das populações de fungos entomopatogênicos nativos bem como sobre a filogenia, diversidade e distribuição dos haplótipos de Metarhizium spp. em solos brasileiros, gerado neste estudo, poderá servir como subsídio para o desenvolvimento de estratégias de conservação e maximização do controle biológico natural de pragas. / Entomopathogenic fungi of the genera Metarhizium, Beauveria and Isaria (order Hypocreales) are associated to the soil where they survive saprofitically or as endophytes of the plants root system. Information on the species composition and its diversity, distribution and association of these fungi with different types of crops and native vegetation are scarce in Brazil. The present study was carried out to compare the abundance of entomopathogenic fungi and the genetic diversity of Metarhizium spp. Isolated from soil samples from annual and perennial crops and native vegetation in five Brazilian states that represent the biomes Amazon, Cerrado, Caatinga, Atlantic Forest and Pampa, in two seasons (wet and dry) in the years 2012 and 2013. The isolation of fungi was performed with selective medium and \"Insect bait\" using Galleria mellonella and Tenebrio molitor. DNA sequences of the region MzIGS3 were used in genetic diversity studies of Metarhizium spp. Representatives haplotypes revealed in the diversity analysis had the 5\'-TEF region sequenced for species identification. Entomopathogenic fungi were isolated from 86% of 1,056 soil samples and Metarhizium was the predominant genus (66% of soil samples), followed by Beauveria (41.9%) and Isaria (10.8%). In general, the highest densities of entomopathogenic fungi were obtained in the Amazon and Cerrado biomes and the lowest densities were detected in the Caatinga biome. Metarhizium spp. was detected in a greater number of soil samples from native vegetation and annual and perennial crops of Cerrado. The frequency of Isaria spp. was low in soil samples being detected in a greater number of soils with annual crops and native vegetation in the Amazon and Caatinga. Metarhizium spp. was usually found in a greater number of samples collected during the wet season compared to the collections in the dry season. On the other hand, Beauveria spp. was higher in the dry season. The diversity of isolates of Metarhizium spp. from areas of native vegetation was greater than that obtained from annual and perennial crops. Six lineages were found in this study; M. robertsii, M. anisopliae, M. pingshaense and three indeterminate species. M. robertsii was the predominant (65% of isolates) found in areas with native vegetation and in the annual and perennial crops of the five biomes. Metarhizium sp. indet. 1 showed the greatest haplotype diversity among the strains studied. A new strain, not characterized taxonomically, Metarhizium sp. indet. 3, was found predominantly in the Caatinga. Only in the Amazon, all lineages were found. The knowledge on species composition of entomopathogenic fungi as well as about phylogeny, diversity and distribution of haplotypes of Metarhizium spp. in Brazilian soils, generated in this study, may be useful for the development of strategies for conservation and maximization of natural biological control of pests.

Made available in DSpace on 2016-04-28T20:16:21Z (GMT). No. of bitstreams: 1 2007-Everton Kort Kamp Fernandes.pdf: 2844359 bytes, checksum: a56e9f806dc8f1f0e7cc6b4641b4a78c (MD5) Previous issue date: 2007-03-30 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Beauveria bassiana is an entomopathogenic fungus used worldwide as a biological control agent of pest arthropods. The present study aimed at characterizing several isolates of B. bassiana and selects those with major potential to control ticks population and other arthropods under natural conditions. Thus, 53 isolates of B. bassiana and 7 isolates including 5 other Beauveria spp. and 1 Engyodontium albus (=Beauveria alba) were investigated. These isolates were originally from different regions of Brazil and other nations, and different hosts or substrate. Initially, all the isolates were morphologically analyzed and investigated about the potential of conidial production. Afterwards, experiments were conducted to investigate the genetic diversity of Brazilian isolates of B. bassiana, to evaluate the spatial distribution of genotypes, and to understand the genetic relationship among isolates of different species of Beauveria. Thus, all isolates were analyzed by isozymes electrophoresis, AFLP-PCR and sequencing of ITS1 and ITS2 regions. The results have permitted the taxonomic identification of isolates and the selection of those with major potential to conidial production. The results have also showed the detection of high genetic variability among B. bassiana isolates, indicating that this fungus represents a species aggregate. Furthermore, the genetic distance among isolates was directly related to the geographic distances. In addition, the isolates were evaluated to UV-B tolerance, heat tolerance, and cold activity. Heat and cold are two abiotic factors of the environment that restrict the use of entomopathogenic fungi as agents for biological control of insects. In addition, solar radiation, particularly the UV-B wavelength, is a major environmental factor that can negatively affect this entomopathogenic fungus in the field. High variability in UV-B tolerance was detected among the isolates after 2 h of radiation exposure. Conidial relative germination ranged from 0% to almost 80%, and a delayed germination was detected. Moreover, isolates of B. bassiana originating from lower latitudes tended to have lower UV-B tolerance than isolates from higher latitudes. A high variability in thermotolerance was also observed among the isolates after 2 h of heat exposure at 45 ?C, ranging from low, to medium, to high viability. The thermal death point of most resistant isolates was between 44 ?C and 45 ?C after 6 h of heat exposure. At cold temperatures most of the isolates grew at 5 ?C. An attempt to correlate the latitude of origin with thermotolerance or cold activity indicated that isolates from higher latitudes were more cold-active than isolates from nearer the equator. The present study also evaluated the virulence of the isolates toward Boophilus microplus larvae, and virulence ranged from very low to very high. It was observed that different populations of B. microplus may present different susceptibility to B. bassiana infection. Therefore, the present study has selected isolates of this entomopathogenic fungus with major effective potential to control B. microplus and other pest arthropods under environmental conditions. / Beauveria bassiana ? um fungo entomopatog?nico utilizado em v?rias partes do mundo como agente de controle biol?gico de artr?podes. Baseado nisso, o presente estudo buscou caracterizar diversos isolados de B. bassiana e selecionar aqueles com maior potencial para controlar popula??es de carrapatos e outros artr?podes em condi??es naturais. Neste sentido, 53 isolados de B. bassiana e mais sete isolados incluindo cinco esp?cies de Beauveria spp. e uma de Engyodontium albus (=Beauveria alba) foram estudadas. Esses isolados foram oriundos de v?rias regi?es geogr?ficas brasileiras e estrangeiras, e de diferentes hospedeiros ou substratos. Os isolados foram inicialmente avaliados morfologicamente e quanto ao potencial de produ??o de con?dios. Em seguida, buscou-se investigar a diversidade gen?tica dos isolados brasileiros de B. bassiana, avaliar a distribui??o espacial de gen?tipos, e conhecer a rela??o gen?tica entre os isolados das diferentes esp?cies. Para esta finalidade os isolados foram submetidos as t?cnicas de eletroforese de isoenzimas, AFLP-PCR e seq?enciamento de regi?es ITS1 e ITS2. Os resultados permitiram a confirma??o taxon?mica dos isolados e a sele??o daqueles com maior capacidade de produ??o de con?dios. Permitiram tamb?m detectar grande variabilidade gen?tica entre os isolados de B. bassiana indicando que este fungo representa um agregado de esp?cies. Al?m disso, a dist?ncia gen?tica detectada entre os isolados foi fortemente relacionada a dist?ncia geogr?fica de origem dos isolados. Posteriormente, os isolados tamb?m foram avaliados quanto a toler?ncia ? radia??o UV-B, toler?ncia ao calor e atividade ao frio. Tanto o calor quanto o frio s?o fatores abi?ticos ambientais que restringem o uso de fungos como agentes de biocontrole, enquanto a radia??o solar, particularmente UV-B, ? o maior fator ambiental que pode afetar negativamente a a??o deste entomopat?geno. Neste sentido, foi observada alta variabilidade entre os isolados analisados em rela??o ? toler?ncia a UV-B ap?s duas horas de exposi??o ? radia??o, obtendose percentuais de germina??o de con?dios que variaram entre 0% e aproximadamente 80%. Al?m disso, foi observado atraso na germina??o dos con?dios, e revelado que os isolados originados de baixas latitudes tenderam a apresentar menor toler?nc ia a UV-B do que aqueles provenientes de latitudes mais elevadas. Foi tamb?m encontrada alta variabilidade em termotoler?ncia dos con?dios ap?s duas horas de exposi??o a 45 ?C, variando entre baixa, m?dia e alta viabilidade. Os con?dios mais resistentes n?o toleraram exposi??o entre 44 ?C e 45 ?C por seis horas. Quando submetidos a 5 ?C a maioria dos isolados apresentaram atividade. Os isolados de B. bassiana obtidos de latitudes mais elevadas foram mais ativos ao frio do que aqueles originados pr?ximos ? linha do equador. Al?m disso, os isolados tamb?m foram avaliados quanto ao potencial virulento sobre larvas de Boophilus microplus, e neste sentido foi poss?vel diferenci?-los em baixa, m?dia e alta virul?ncia. Foi detectado tamb?m que diferentes popula??es desta esp?cie de carrapato podem manifestar diferentes n?veis de suscetibilidade ? infec??o por B. bassiana. Neste sentido, o presente estudo permitiu selecionar isolados deste entomopat?geno com maior potencial para o controle biol?gico de B. microplus e outros artr?podes em condi??es de campo.

Made available in DSpace on 2014-06-12T15:55:40Z (GMT). No. of bitstreams: 2 arquivo9374_1.pdf: 2655923 bytes, checksum: 4a8cd8dfda72129aa8308dbf088c4d10 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2004 / Beauveria bassiana, Metarhizium anisopliae var. anisopliae e Metarhizium flavoviride var. flavoviride são fungos entomopatogênicos com ação comprovada contra várias espécies, embora não tenham sido testados em Chrysomya albiceps, um díptero de importância na saúde pública. Assim, objetivou-se avaliar a ação de B. bassiana, M. anisopliae var. anisopliae e M. flavoviride var. flavoviride em ovos, larvas e adultos de C. albiceps, utilizando as concentrações 104, 105, 106, 107, 108 conídios.mL-1, considerando o percentual de eclosão de larvas, período de pré-pupa, pupa, percentual de emergência de adultos, ritmo de emergência, morte acumulativa, longevidade, período de postura e percentual de eclosão a partir de fêmeas infectadas. O comportamento dos fungos reisolados também foi avaliado através dos parâmetros biológicos: percentual de germinação, número de conídios, número e diâmetro de colônias, bem como a citologia no que se refere à descrição das estruturas vegetativas e reprodutivas. De acordo com a metodologia empregada, verificou-se que os três fungos apresentaram ação contra ovos, larvas e adultos de C. albiceps. Já em relação ao comportamento, foi observado que os fungos reisolados de larva apresentaram o melhor desempenho em relação ao controle e os aspectos citológicos não diferiram quando comparados ao controle. Esses resultados sugerem a possibilidade do emprego desses fungos no controle de C. albiceps.

This dissertation describes efforts to improve the oxidative capacity of n-alkane- induced Beauveria bassiana; a fungus and a versatile whole cell biocatalyst used in the biotransformation of steroids. n-Hexadecane was used as the carbon source during the growth of B. bassiana, presumably to induce the expression of oxidative enzymes, thus enhancing the oxidation of unactivated carbons. Dehydroepiandrosterone (DHEA) is an essential endogenous male-hormone and serves as a metabolic intermediate in the production of more potent androgens. Using DHEA as a substrate also provides the opportunity to study the hydroxylation of an unfunctionalized carbon, an attractive reaction that produces valuable intermediates for chemical synthesis. Results showed that exposing and inducing cells in n-hexadecane improves the synthesis of 11α-hydroxy derivatives. Reactions were carried out with cells grown on n-hexadecane, resulting in 65 ± 6.3 % conversion of DHEA to androstenediol (40.3% mM) and 3β,11∝,17β- trihydroxyandrost-5-ene (22.8% mM), as determined by HPLC, NMR and LCMS analyses. However, experiments with non-induced cells resulted in a poor substrate conversion (17%). To extend use of B. bassiana to pharmaceutical applications, it was necessary to optimize reaction conditions such as biocatalyst preparation, substrate concentration, agitation reaction temperature and pH. Higher substrate conversion, selectivity and yield of desired product were achieved with the reactor arrangement of “Resting Cells”. The apparent rate of reaction fits a Michaelis-Menten kinetic model with a maximum reaction rate of 4.45 mM/day, revealing that the transformation of intermediate androstenediol to desired 3β,11∝,17β-trihydroxyandrost-5-ene is the limiting step in the reaction. Interestingly, when a diluted amount of substrate was used, a higher yield of 11∞-hydroxy steroid was achieved. Also, reactions at 26°C with pH ranges between 6.0 and 7.0, resulted in the highest conversion (70%) and the higher product yield (45.8%). The maximum conversion of DHEA (71%) was achieved in experiments with high biomass loading, and the increment of desired product yield (11∝-hydroxy) was directly proportional to the amount of biomass used. Moreover, a high VMax/KM value was achieved with high biomass yields. Interestingly, the changes in biomass yield did not have a considerable effect on reaction selectivity. The main drawbacks of biocatalysis for production of steroids were addressed and approaches to minimize the drawbacks have been presented. The production of desired product (11∝-DHEA) was significantly improved using cells previously adapted to n-hexadecane.

Echinacea purpurea is a native herbaceous perennial with substantial economic value for its medicinal and ornamental qualities. Arbuscular mycorrhizae are symbiotic fungi that form relationships with plant roots and are known to enhance growth in the host. Mycorrhizae and other fungal endophytes often affect stress resistance and secondary metabolism in the host, as well as the ecology of other endophytes in the plant. A newly emerging paradigm in sustainable biotechnique is the targeted use of fungal endophytes to enhance growth and secondary metabolism in crops. Many of the therapeutic compounds in E. purpurea could be affected by fungal colonization. In this research the effects of inoculation of Echinacea purpurea with two classes of fungal endophytes: the arbuscular mycorrhizal fungi Glomus intraradices and Gigaspora margarita and the entomopathogenic endophyte Beauveria bassiana were evaluated . Endophyte colonization and impacts on plant growth and phytochemistry were tested in multiple greenhouse experiments. Arbuscular mycorrhizae and B. bassiana effectively colonized E. purpurea with some significant interactive effects. Consistent, substantial, and significant increases in all growth parameters were observed in mycorrhizal plants; mycorrhizal plants produced up to four times the biomass of controls in 12 weeks. Broad spectrum changes in fertilization were necessary to produce mycorrhizal and nonmycorrhizal samples of equal size, and severely nutrient-limited mycorrhizal E. purpurea seedlings maintained growth rates comparable to well fertilized samples. Treatment with B. bassiana had minor and inconsistent effects on some plant growth parameters, and there were significant interactive effects with arbuscular mycorrhizae. Phytochemical concentrations in all metabolite classes tested responded significantly to inoculation with both classes of fungal endophytes. Changes were observed in various pigments, caffeic acid derivatives, alkylamides, and terpenes. Many of the affected compounds have important roles in metabolism or have bioactive value as natural products. When considered from a net production perspective (concentration X dry weight), compared to controls, plants inoculated with endophytes produced as much as 30 times the content of some compounds in 12 weeks. This work effectively demonstrates that fungal endophytes can enhance the bioactivity of plant tissues and the production of natural products in E. purpurea.

Echinacea purpurea is a native herbaceous perennial with substantial economic value for its medicinal and ornamental qualities. Arbuscular mycorrhizae are symbiotic fungi that form relationships with plant roots and are known to enhance growth in the host. Mycorrhizae and other fungal endophytes often affect stress resistance and secondary metabolism in the host, as well as the ecology of other endophytes in the plant. A newly emerging paradigm in sustainable biotechnique is the targeted use of fungal endophytes to enhance growth and secondary metabolism in crops. Many of the therapeutic compounds in E. purpurea could be affected by fungal colonization. In this research the effects of inoculation of Echinacea purpurea with two classes of fungal endophytes: the arbuscular mycorrhizal fungi Glomus intraradices and Gigaspora margarita and the entomopathogenic endophyte Beauveria bassiana were evaluated . Endophyte colonization and impacts on plant growth and phytochemistry were tested in multiple greenhouse experiments. Arbuscular mycorrhizae and B. bassiana effectively colonized E. purpurea with some significant interactive effects. Consistent, substantial, and significant increases in all growth parameters were observed in mycorrhizal plants; mycorrhizal plants produced up to four times the biomass of controls in 12 weeks. Broad spectrum changes in fertilization were necessary to produce mycorrhizal and nonmycorrhizal samples of equal size, and severely nutrient-limited mycorrhizal E. purpurea seedlings maintained growth rates comparable to well fertilized samples. Treatment with B. bassiana had minor and inconsistent effects on some plant growth parameters, and there were significant interactive effects with arbuscular mycorrhizae. Phytochemical concentrations in all metabolite classes tested responded significantly to inoculation with both classes of fungal endophytes. Changes were observed in various pigments, caffeic acid derivatives, alkylamides, and terpenes. Many of the affected compounds have important roles in metabolism or have bioactive value as natural products. When considered from a net production perspective (concentration X dry weight), compared to controls, plants inoculated with endophytes produced as much as 30 times the content of some compounds in 12 weeks. This work effectively demonstrates that fungal endophytes can enhance the bioactivity of plant tissues and the production of natural products in E. purpurea.

No description available.

Tomato (Solanum lycopersicum L.) produces an array of volatile secondary metabolites that act as constitutive and induced defenses against a variety of insect pests and diseases. We studied the effect of beneficial microorganisms, an arbuscular mycorrhiza fungus (AM), Glomus intraradices, an entomopathogenic fungus (Bb), Beauveria bassiana and a combination of both (AM+Bb), as well as, pests, such as rootknot nematode (RKN) (Meloidogyne incognita) and beet armyworm (BAW) (Spodoptera exiguae H.) on the production of volatile compounds in the leaves and roots of tomato. Benzyl alcohol, 3-hexenoic acid, total other compounds and β-myrcene were significantly increased (P<0.05) in the leaves by AM and Bb colonization. Upon herbivory, control plants showed a trend for high volatile contents among all four treatment groups contrary to the trend observed without herbivory, when control showed low volatile contents. Herbivory significantly decreased all the volatiles in the leaves compared to those without herbivory. Pest bioassay revealed that these two fungi together can significantly induce resistance against BAW. In the experiment with RKN, a trend was observed with low production of most volatile compounds in the leaves from the RKN plants. Varying durations of herbivory exposure had significant effects on many leaf volatile compounds compared to those without herbivory. Effect of RKN was significant (P<0.05) on the production of methyl salicylate, and (Z)-geraniol in the roots. Interaction effect of RKN with 18 hours of herbivory was significant for (Z)-geraniol, and with 42 hours of herbivory, it was significant for (Z)-geraniol, benzyl alcohol, and total volatiles in the roots. Beet armyworm preferred RKN plants and caused greater damage to them compared to the control plants. Semi-quantitative RT-PCR showed higher expression of the SlSAMT gene in the roots colonized with RKN, compared to the control roots and those from a resistant line. The experiments demonstrated that interaction with these organisms can change the volatile compounds in the leaves and roots of tomato plant, can alter herbivore preference, and can upregulate defense genes such as SlSAMT.