As sessile organisms, plants are unable to escape from attack by herbivorous insects. To cope with this pressure, plants have evolved several defense strategies, including the production of secondary metabolites, specialized chemicals with ecological functions. Most studies have focused on the role of secondary metabolites in plant defense against chewing insects. Little is known about what compounds are present in phloem sap and how they affect phloem feeding insects. Therefore, I investigated the effects of phenolic compounds on phloem feeders, using Chaitophorus aphids in bioassays with wildtype and transgenic poplar overexpressing the transcription factor MYB 134, which results in elevated levels of tannins and reduced levels of phenolic glycosides. Aphids produced significantly more offspring on MYB 134 plants but showed a significant preference for lower tannin leaf tissue. Analysis of poplar phloem exudates and aphid extracts provides direct evidence that the phenolic glycosides salicin, salicortin and tremulacin are present in poplar phloem and are ingested by aphids. These results are discussed in relation to what is driving the differences in aphid fecundity and choice between plant types. / Graduate / 0309 / 0307 / 0329 / alphareghanwong@gmail.com

Made available in DSpace on 2015-04-11T13:38:49Z (GMT). No. of bitstreams: 1 Claudia Comandolli.pdf: 4289017 bytes, checksum: 2bef35ae167e04ceccfedeedc54fbb31 (MD5) Previous issue date: 2010-12-15 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The objective of the present study was to evaluate the antimicrobial, antioxidant and cytotoxicity potentials of extracts of leaves and branches of Casearia javitensis, and to purify and identify their secondary metabolites. Leaves and branches were collected in Reserva Adolpho Ducke, Manaus, AM. The plant material was dried, grinded and the extracts were prepared with DCM, MeOH and water using ultrasound for 20 minutes each. The extracts were concentrated and tested for antibacterial, cytotoxicity and antioxidant activity. Antibacterial activity assays were done using the agar diffusion method by well technique, against Aeromonas hydrophila, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were measured for the most active extracts. The cytotoxicity assay employed brine shrimp Artemia salina and the evaluation of antioxidant activity used a method with DPPH and ascorbic acid. DCM branches extract was fractioned in chromatographic column (CC), and recrystallization. Gas Chromatography-Mass Spectrometry (GC-MS) and Nuclear Magnetic Resonance (NMR) analyses afforded steroids, as β-sitosterol. DCM extracts of leaves and branches were analyzed by GC-MS in appropriated conditions to steroids and triterpenes analyses, and triterpene friedelin was detected in branches extracts. Antimicrobial assay showed high activity for branches MeOH extract and leaves MeOH extract against A. hydrophila, MIC value < 2 mg/mL and showed no cytotoxicity in Artemia salina. These MeOH extracts were dissolved in water and partitioned between dichloromethane (DCM), ethyl acetate (AcOEt) and n-buthanol (n-BuOH), and these phases were tested for antibacterial activity. The AcOEt phases were the most active, and showed high antimicrobial activity. The DCM phase from branches MeOH extract was fractioned by CC and afforded β-friedelanol. The AcOEt fraction of branches MeOH extract was fractioned by CC and some fractions were purified by HPLC, and afforded five phenolic glycosides, which structures are not yet determined. 1D and 2D-NMR and mass spectrometry analyses are still on progress. The n BuOH phase from branches MeOH extract was fractioned by exclusion chromatography (Sephadex LH-20) and subjected to CC over a Poliamide-6 column, and NMR analyses of one fraction suggest the presence of a phenolic glycoside. / O presente trabalho teve por objetivo isolar e identificar os metabólitos secundários presentes nos extratos de Casearia javitensis, assim como avaliar o potencial antimicrobiano, citotóxico e antioxidante destes extratos. Para o trabalho, foram realizadas quatro coletas na Reserva Adolpho Ducke, Manaus-AM. Os materiais coletados foram secos, moídos e extraídos em ultrassom com diclorometano (DCM), metanol (MeOH) e H2O. Para os testes antimicrobianos, utilizou-se o método de difusão em ágar, pela técnica de poço, contra Aeromonas hydrophila, Staphylococcus aureus, Escherichia coli e Pseudomonas aeruginosa. A Concentração Inibitória Mínima (CIM) e Concentração Mínima Bactericida (CMB) foram determinadas para os extratos mais ativos. O ensaio citotóxico utilizou Artemia salina como organismo-teste e o ensaio de atividade antioxidante empregou o método quantitativo utilizando o radical DPPH e equivalência com o ácido ascórbico. O extrato DCM dos galhos foi fracionado por coluna cromatográfica (CC) e uma fração foi purificada através de recristalização. Frações analisadas por Cromatográfo à Gas acoplado à Espectrometria de Massas (CG-EM) e Ressonância Magnética Nuclear (RMN) mostraram presença de esteroides, possuindo o β-sitosterol como majoritário nas frações. Os demais extratos DCM foram analisados por CG-EM em uma condição para análise de esteroides e triterpenos, e o triterpeno friedelina foi detectado nos extratos dos galhos. O extrato metanólico dos galhos da primeira coleta e o extrato metanólico das folhas da terceira coleta apresentaram halos de inibição superiores a 10 mm no teste contra Aeromonas hydrophila, CIM abaixo de 2 mg/mL e ausência de atividade citotóxica em Artemia salina. Estes extratos foram submetido à partição com diclorometano (DCM), acetato de etila (AcOEt) e n-butanol (n-BuOH), e as fases obtidas foram avaliadas para atividade antimicrobiana. As fases AcOEt destes extratos mostraram-se como as mais ativas, concentrando a atividade antimicrobiana. A fase DCM do extrato MeOH dos galhos foi fracionada por sucessivas CCs e uma fração mostrou a presença de β-friedelanol por análise em CG-EM. A fase AcOEt foi fracionada por CC e algumas frações foram purificadas por CLAE, fornecendo cinco fenólicos glicosilados, os quais a determinação estrutural ainda não está concluída, pois espectros de RMN mono e bidimensionais são necessários para a conclusão desta etapa. A fase n -BuOH foi fracionada em CC utilizando Sephadex LH-20 e CC de Poliamida-6, e o espectro de RMN de uma fração semipurificada sugere a presença também de fenólicos glicosilados.

Quaking aspen (Populus tremuloides) clones within close proximity to one another can exhibit drastically different levels of browsing by ungulates. The objectives of this study were to (1) determine interclonal differences in plant chemistry between adjacent clones exhibiting different degrees of herbivory which may influence the browsing behavior and patterns of ungulates, and (2) determine if correlation exists in the levels of salicortin and tremulacin between current year's suckers and current year's growth on older trees. This second objective was meant to indicate a protocol for land managers for identifying clones meriting increased protection from herbivory after treatment and wildfire. In July of 2005, 6 pairs of clones were identified on the Dixie National Forest, Utah, and on Cedar Mountain, east of Cedar City, Utah. Pairs consisted of 2 clones within the same pasture and/or grazing allotment and within a minimal distance from one another; one clone displaying moderate to high levels of ungulate utilization of aspen suckers, and one exhibiting minimal to no ungulate utilization of aspen suckers. Soil samples were taken at each clone and leaf tissues were sampled to determine genet. Aspen suckers were sampled for nutrient content, combined phenolic glycoside concentration (salicortin and tremulacin), condensed tannins, and the presence of extra floral nectaries (EFNs), at intervals throughout the growing season (August 3-6, August 31-September 2, and October 12-14). Current year's growth from representative mature trees was sampled for phenolic glycoside concentration at these times as well. All tests demonstrated high levels of insignificance for both leaves and stems.Sucker nitrogen values may have been elevated during portions of the sampling year in clones displaying moderate to high levels of ungulate utilization, possibility indicating an ungulate preference for nitrogen, but due to missing values, this is far from conclusive. P-values for forest floor factors were also highly non-significant with the exception of forest floor C (0.04) in the regenerating clones. Two post-project hypotheses are postulated in an attempt to explain the differences of forest floor carbon in terms of factors that may be influencing ungulate herbivory.