One great issue of reforestation is the infestation of pest insects. The pine weevil (Hylobius abietis L.) is one of the economically most important pest insects in Europe and Asia. When insects feed on the trees the plants emit compounds that repel the insects. Methyl salicylate (MeSA) is one substance known to be emitted by conifer seedlings after herbivore feeding. Similar compounds are emitted by fungus isolated from pine weevil frass. Female pine weevils add frass to the egg laying site to protect the eggs from other pine weevils. The goal of the project was to cultivate the fungi Ophiostoma pluriannulatum and analyze the volatile substances. The fungus was cultivated using natural pine weevil frass (NF) as main substrate. Later new batches were prepared using lignin and potato dextrose broth (PDB), as feeding media to compare the results. Two sampling methods were used, SPME and adsorption with Porapak™ polymer column packing material. These samples were analyzed using GC-MS. Methyl salicylate was found in NF fungi cultures, peaking after ten days. Another, unexpected, product of the fungi was a sesquiterpene hydrocarbon. The lignin and the PDB samples did not produce MeSA. However, the sesquiterpene could be found in the lignin samples.

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Plants have evolved a variety of constitutive and induced chemical defense mechanisms against biotic stress. Emission of volatile compounds from plants facilitates interactions with both beneficial and pathogenic organisms. However, knowledge of the chemical defense in roots is still limited. In this study, we have examined the root-specific biosynthesis and function of volatile terpenes in the model plant Arabidopsis. When infected with the root rot pathogen Pythium irregulare, Arabidopsis roots release the acyclic C11-homoterpene (E)-4,8-dimethylnona-1,3,7-triene (DMNT), which is a common constituent of volatile blends emitted from insect-damaged foliage. We have identified a single cytochrome P450 monooxygenase of the CYP705 family that catalyzes a root-specific oxidative degradation of the C30-triterpene precursor arabidiol thereby causing the release of DMNT and a C19-degradation product named arabidonol. We found that DMNT shows inhibitory effects on P. irregulare mycelium growth and oospore germination in vitro, and that DMNT biosynthetic mutant plants were more susceptible to P. irregulare infection. We provide evidence based on genome synteny and phylogenetic analysis that the arabidiol biosynthetic gene cluster containing the arabidiol synthase (ABDS) and CYP705A1 genes possibly emerged via local gene duplication followed by de novo neofunctionalization. Together, our studies demonstrate differences and plasticity in the metabolic organization and function of terpenes in roots in comparison to aboveground plant tissues. Additionally, we demonstrated that the arabidiol cleavage product, arabidonol, is further modified by yet unknown enzymatic reactions into three products, which are found in root exudates. We suggested a pathway for their biosynthesis based on precursor feeding experiments and NMR analysis. Although DMNT biosynthetic genes are clustered on chromosome 4 along with several potential modification genes, we did not find a possible role of these genes in the derivatization of arabidonol. Preliminary experimental results using genetic and biochemical approaches for identifying genes involved in the modification steps are also presented. In summary, this study demonstrates an alternative route for volatile terpene formation belowground different from aboveground plant tissues via triterpene degradation and provides evidence for an unexplored triterpene catabolism pathway in Arabidopsis. / Ph. D.

Grape maturity is a critical attribute impacting potential wine quality. Maturity evaluation is difficult due to the many interrelated factors that impact physicochemical changes and limitations in the understanding of these factors. Current methods of assessing grape maturity are destructive, expensive, time consuming, subjective, and do not always strongly correlated to potential wine quality. This study evaluated the applicability of a conducting polymer-based electronic nose to monitor grape maturity by analyzing headspace volatiles. In the first part of the study, system and experimental parameters affecting the electronic nose operation were investigated to optimize detection of wine grape aroma. In the second part, the ability of an electronic nose to classify Cabernet Sauvignon (Vitis vinifera L.) grapes based on maturity was investigated. Maturity of samples collected at different weeks post-bloom was evaluated by measuring berry weight, pH, Brix, titratable acidity, total phenols, color intensity, hue, total anthocyanins, and total and phenol-free glycosides. Results were compared, using discriminant and canonical discriminant analysis, with analysis of headspace volatiles via the hand-held electronic nose. The electronic nose was able to determine the difference between the sample groups. Field measurements demonstrated the potential for the electronic nose as a rapid, non-destructive tool for evaluating grape maturity. / Master of Science

Mosquitoes, by transmitting vector-borne diseases through their saliva, impact nearly half of the world's population. Mosquito survival is dependent on their sense of smell, or olfaction, which allows a mosquito to differentiate between plant nectar, required for metabolic processes, and host odors, which will help them navigate towards hosts, source of the blood required for producing eggs. Mosquitoes interpret and respond to chemical volatiles very differently depending on their environment (temperature, humidity, time of day, etc.) and species-specific host preferences (humans, mammals, etc.). However, the impact of ecological factors on mosquito prevalence, sugar-feeding, and host seeking is relatively unknown. In an attempt to address this knowledge gap, we first investigate how several ecological factors (i.e., temporal, seasonal, and topographical) affect mosquito sugar feeding and population dynamics. Second, we focused on the anthropophilic mosquito species Aedes aegypti to investigate the role of plant associated compounds in host-seeking behavior, such as those which are commonly found in personal care products (i.e., body washes). Our results demonstrate that several ecological factors alter mosquito prevalence and behavior, including both sugar and host seeking behaviors. We anticipate these results to be a starting point for mosquito control strategies that depend not only on olfactory perception of plant odors, but also on the ecological and species-specific characteristics which shape the dynamics of vector-borne diseases. / Master of Science in Life Sciences / Most female mosquitoes must feed on a blood source in order to produce eggs, but in doing this they can transmit viruses and pathogens between hosts through their saliva, which are contracted by 700,000 people per year. In order to survive and fly to locate these host sources, both female and male mosquitoes require sugar and carbohydrates present in plant nectar. Mosquito identification of nectar and blood sources is primarily dependent on their sense of smell, or olfaction, which allows a mosquito to identify and locate chemical odors. The interpretation of these chemical odors is context dependent on both the mosquitoes' hunger status, ideal host (human, mammal, bird, etc.) and environmental cues (temperature, humidity, time of day, etc.). It is relatively unknown how the combination of these biological and environmental factors influences mosquito survival and biting frequency. In order to better understand this relationship, we first investigated mosquito species' habitat preferences (i.e., urban and forested) and ecological factors (i.e., season, weather). Second, we focused on Aedes aegypti mosquitoes, which almost exclusively feed on humans in urban habitats, to investigate mosquito behavior in the presence of combined plant and human odors caused by the usage of fragrant personal care products that are scented with plant odors. Our results demonstrate that mosquito prevalence, host-seeking, and plant-seeking are highly dependent on chemical and ecological factors. We anticipate that this research has avenues for both improving mosquito control strategies and better understanding the ecological dynamics of vector-borne diseases.

Effectiveness of a capture and return system for the partial retention of fermentation volatiles, as a means of improving white wine quality, was evaluated. Twenty-three aroma-active volatiles including ethyl esters, acetate esters, fusel alcohols, and fatty acids, were quantified using head-space solid phase microextraction with GC/MS. Volatile analysis of fermentations maintained at 15ºC demonstrated a trend of increased concentrations of ethanol, esters and ethyl esters of fatty acids and decreased concentrations of fusel alcohol acetates, fatty acids and higher alcohols in treatment wines. When fermentation temperature was maintained at 15ºC there was increased concentration and retention of fusel alcohols, fatty acids and higher alcohols compared to 15ºC. Sensory analysis of wines fermented at 15°C, using triangle difference testing, indicated variable differences in aroma among treatments. / Master of Science

Vineyard management practices are known to affect fruit composition and resultant wines, in part, by altering fruit volatiles. Methods currently used to evaluate the impact of vineyard practices on grape/wine composition include measuring physico-chemistry indices and performing wine sensory analyses. These activities are both time-consuming and destructive. Two electronic nose (ENose) systems: a hand-held conducting polymer-based and a portable surface acoustic wave-based systems were investigated as grape monitoring tools. Vineyard treatments included the effect of canopy side (East vs. West and North vs. South), cluster thinning (unthinned, 1 cluster/shoot, and 1 & 2 cluster/shoot) and ethanol spray (5% v/v) on Cabernet franc, Merlot and both varieties respectively. ENose data were obtained in the field (over two growing seasons for canopy side and in 2008 for cluster thinning and ethanol spray) and laboratory (2007 for canopy side), across different sampling dates and compared with nine-grape/eight-wine chemistry assays, GC/MS (cluster thinning) and wine aroma sensory evaluations (triangular difference testing). ENose results demonstrated 100% significant differences between all Cabernet franc and Merlot treatments. Grape/wine chemistry indices, for both Cabernet franc and Merlot, did not differ among treatments (except ethanol treatment) across sampling dates or growing seasons and vineyard management practices. Wine aroma sensory evaluations demonstrated only limited differences (3 out of 8 comparisons: East vs. West, 1 cluster/shoot vs. 1 & 2 clusters/shoot and 1 cluster/shoot and 1 & 2 clusters/shoot). The high level of discrimination by ENose systems may provide opportunities to enhance the understanding of vineyard management activities. / Master of Science in Life Sciences