La cameline, Camelina sativa (Brassicaceae), fait aujourd'hui l'objet d'un regain d'intérêt. Il s'agit d'une plante aux vocations multiples (bioénergétique, industrielle, alimentaire), aux exigences agronomiques faibles et qui semble peu sujette aux attaques de ravageurs phytophages.Trois objectifs visant à appréhender la complexité des interactions multitrophiques autour de la cameline ont été définis au cours de ce travail de thèse. Le premier visait à déterminer le rôle de la cameline en tant que réservoir à pucerons ravageurs. Les deux autres reposaient sur les études des impacts des pratiques culturales (fertilisation azotée, culture en association) et des virus de plantes, sur les interactions tritrophiques cameline/pucerons/parasitoïdes. Les premiers résultats ont permis de montrer que la cameline pouvait être considérée comme plante réservoir pour différentes espèces de pucerons ravageurs présents dans les paysages agricoles du nord de la France, qu'il s'agisse d'espèces inféodées aux Brassicaceae (Brevicoryne brassicae), d'espèces polyphages (Myzus persicae, Aphis fabae) ou encore d'espèces spécialistes des Poaceae (Rhopalosiphum padi). En ce qui concerne les deux pratiques culturales étudiées, l'association de la cameline avec la fèverole a engendré des effets délétères sur le contrôle biologique du puceron Aphis fabae par le parasitoïde Aphidius matricariae. Le comportement des femelles A. matricariae ne semble pas satisfaire à la théorie de "l’optimal foraging". En parallèle, des effets de la fertilisation azotée ont été observés sur les trois niveaux trophiques et ont varié en fonction de la dose apportée à la plante et du degré de spécialisation du puceron. Ainsi, notre étude montre que les pratiques culturales peuvent avoir des répercussions importantes de type "bottom-up" sur les niveaux trophiques supérieurs. Enfin, nous avons montré un impact des deux virus de plantes étudiés qui s'est révélé variable non seulement en fonction du mode mais aussi de l'efficacité de la transmission par le puceron-vecteur. Ce dernier travail a également permis de mettre en évidence des effets "bottom-up" d'un virus sur le comportement de sélection de l'hôte par un parasitoïde de puceron mais aussi sur la physiologie des partenaires appartenant aux trois niveaux trophiques. En effet, les performances du puceron-vecteur sur la plante infectée semblent améliorées alors que celles du parasitoïde sont altérées, ce qui pourrait se traduire par une meilleure propagation du virus. Ces travaux soulignent l'importance de la prise en compte des relations multitrophiques avant la (ré-) introduction d’une espèce de plante dans les agro-écosystèmes, et en particulier l'intérêt d'identifier les risques associés en termes de pression de ravageurs et de pathogènes / Camelina (Camelina sativa, Brassicaceae) has recently received great attention as an alternative oil-seed crop presenting various (bioenergetic, industrial, alimentary) advantages, requiring low agronomic inputs and exhibiting few attacks by phytophagous insects.In this thesis, three objectives have been defined to understand the complexity of the multi-trophic interactions involving camelina. The first one was to determine the role of camelina as a reservoir of aphid pests. The other two were based on the study of the effects of cultural practices (nitrogen fertilization, mixed crops) and plant viruses on the tritrophic interactions involving camelina, aphids and parasitoids.Results have shown that camelina could be considered as a plant reservoir for various aphid pest species occurring in the agricultural landscape of northern France, among which were found not only species usually restricted to Brassicaceae (Brevicoryne brassicae), but also polyphagous species (Myzus persicae, Aphis fabae) and specialists of Poaceae (Rhopalosiphum padi). Regarding the two studied agricultural practices, the association of camelina with faba bean induced deleterious effects on the biological control of the aphid Aphis fabae by the parasitoid Aphidius matricariae. The behavior of parasitoid females did not seem in line with the "optimal foraging" theory. In addition, several effects of nitrogen fertilization have been observed on the three trophic levels depending on both the dose given to the plant and the degree of specialization of the aphid. Thus, our study has shown that cultural practices could have significant "bottom-up" impacts on higher trophic levels. Finally, we have shown the two studied plant viruses had an impact which not only varied depending on the virus mode of transmission by the vector but also according to its transmission efficiency. This work also revealed the existence of "bottom-up" effects of a virus on the host selection behavior by an aphid parasitoid as well as on the physiology of the partners belonging to three trophic levels. Indeed, the performance of the aphid vector was improved on the infected plant whereas that of the parasitoid was impaired, which could lead to a better spread of the virus.This work highlights not only the importance of taking into account the associated multi-trophic relations before the (re-) introduction of a plant species within agro-ecosystems but also the importance of identifying the risks in terms of pests and pathogens pressure

Brassica carinata (A.) Braun and Camelina sativa (L.) Crantz are two re-emerging oilseed crops of the Brassicaceae family that are being adapted for cultivation in western Canada. Both seeds of these species reportedly accumulate considerable amounts of sulfur-containing secondary metabolites called glucosinolates. The purpose of the current work was to gain knowledge of the occurrence and distribution of glucosinolates during primary processing of these oilseeds, including during pressing and dehulling. In the first study, a reversed phase HPLC method was developed for the analysis of sinigrin, the major glucosinolate in B. carinata. Both C18 columns selected were able to separate the compound with an isocratic eluent containing 100% tetramethylammonium bromide (10 mM, pH 5) delivered at 1 mL/min at a column temperature of 25oC. These chromatographic conditions were applied and sinigrin concentration of whole B.carinata seed was estimated to be 29 μg/mg. Average matrix effect was estimated to be 104% that was caused by other components in the B. carinata seed matrix. In the second study, high concentrations of glucosinolates were detected and identified in fractions of C. sativa seeds using HPLC-ESI-MS. Methods for extraction, isolation, and purification of three individual glucosinolates from these fractions are reported. Quantitation of total glucosinolates was performed on proton NMR using DMF as an internal standard. Quantitation of individual glucosinolates was achieved by using MS extracted ion chromatogram data. Total glucosinolates were found in C. sativa whole seed at a concentration of 14 μg/mg, and glucocamelinin, the major glucosinolate, constituted 65% of the total amount. In addition, a dehulling treatment was applied to C. sativa seeds, from which both oil content and crude protein content increased after dehulling of the seeds.

Master of Science / Biochemistry and Molecular Biophysics Interdepartmental Program / Timothy P. Durrett / Using vegetable oils directly as an alternative biofuel presents several problems as such oils typically possess poor fuel qualities including high viscosity, low volatility, and poor cold temperature properties. The ornamental shrub Euonymus alatus produces unusual acetyl-1,2-diacyl-sn-glycerols (acetyl-TAGs) that have an acetyl group in the sn-3 position instead of a long chain fatty acid. The presence of this sn-3 acetyl-group give acetyl-TAGs properties desirable for biofuels, such as reduced viscosity, comparted to the normal long chain triacyglycerols found in most vegetable oils. Acetyl-TAGs are synthesized by the Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) and Euonymus fortunei diacylglycerol acetyltransferase (EfDAcT) enzymes. Both enzymes catalyze the transfer of an acetyl group from acetyl-CoA to diaclglycerol (DAG) to produce acetyl-TAGs. Previous work demonstrated that expression of EaDAcT combined with the suppression of a diacylglycerol aceyltransferase (DGAT1) in Camelina sativa led to seeds with 85 mol % acetyl-TAGs. Increasing acetyl-TAG levels further was explored using two strategies. Over expression of citrate lyase to increase the pool of acetyl-CoA to be used as a substrate for the acetyltransferase enzymes failed to increased levels of acetyl-TAGs. A second approach involved expressing EfDAcT in Camelina sativa. EfDAcT has demonstrated higher activity in vitro and in vivo and its expression in yeast leads to approximately 50 % higher levels of acetyl-TAGs compared to EaDAcT. The expression of EfDAcT coupled with the suppression of DGAT1 in Camelina sativa resulted in 90 mol % acetyl-TAGs in the transgenic seeds. Levels of EfDAcT protein analyzed in developing transgenic Camelina sativa seeds across a 40 day time period were highest at 15 and 20 days after flowering. Following these time points acetyl-TAG accumulation increased rapidly, coinciding with the higher enzyme expression levels. The optimization of additional promoters to ensure expression of EfDAcT in the last half of seed development could represent another way to further increase acetyl-TAGs in the future.

To better understand the nature and function of N-acylethanolamines (NAEs) in Camelina sativa, we used mass spectrometry analysis to identify and quantify NAE types in developing seeds, desiccated seeds and seedlings. Developing seeds showed a differential increase in individual NAE species and an overall increase in NAE content with seed development and maturation. The NAE composition in mature, desiccated seeds mostly reflected the total fatty acid composition in the seed tissues, except for a noted absence of 11-eicosenoic (20C monounsaturated) fatty acid in the NAE pool. During seed stratification and seedling growth, individual NAE species were depleted at similar rates. Simulated drought treatments during seedling development resulted in a significant rise in NAE levels for the major 18C NAE types compared with untreated seedlings. Arabidopsis and Camelina mutants with reported altered fatty acid profiles were analyzed for their NAE compositions; both Arabidopsis and Camelina had relatively similar changes between compositions of total seed fatty acids and NAEs. Furthermore, seeds were analyzed from transgenic Arabidopsis and Camelina with engineered, non-native, long-chain polyunsaturated fatty acids (18C, 20C and 22C), and the results showed the production of novel N-acylphosphatidylethanolamines (presumed precursors of NAEs) and NAEs with the same long acyl chains. These results demonstrate i) that NAE levels change dramatically with seed and seedling development in Camelina, ii) that NAE levels in Camelina seedlings can be elevated in response to environmental stress, and iii) that the pool of NAEs can be manipulated by making alterations to seed fatty acid compositions, some of which may have promise for the production of novel, bioactive NAEs.

Master of Science / Biochemistry and Molecular Biophysics Interdepartmental Program / Timothy P. Durrett / Many Euonymus species express an acetyltransferase enzyme in their seeds which catalyzes the transfer of an acetyl group from acetyl-CoA to the sn-3 position of diacylglycerol (DAG) producing unusual acetyl-1,2-diacyl-sn-glycerols (acetyl-TAG). The presence of the sn-3 acetate group gives acetyl-TAG with unique physical properties over regular triacylglycerol (TAG) found in vegetable oils. The useful characteristics of acetyl-TAG oil offer advantages for its use as emulsifiers, lubricants, and 'drop-in' biofuels. One enzyme, Euonymus alatus diacylglycerol acetyltransferase (EaDAcT), responsible for acetyl-TAG synthesis in nature was previously isolated from the seeds of Euonymus alatus (burning bush) and expressed in the oilseed crop Camelina sativa. Expression of EaDAcT successfully led to production of high levels of acetyl-TAG in camelina seeds. To further increase acetyl-TAG accumulation in transgenic camelina seeds, multiple strategies were examined in this study. Expression of a new acetyltransferase enzyme (EfDAcT) isolated from the seeds of Euonymus fortunei, which was previously shown to possess higher in vitro activity and in vivo acetyl-TAG levels compared to EaDAcT, increased acetyl-TAG accumulation by 20 mol%. Suppression of the endogenous competing enzyme DGAT1 further enhanced acetyl-TAG accumulation to 90 mol% in selected transgenic line. Studying the regulation of EfDAcT transcript, protein, and acetyl-TAG levels during seed development further provided new insights on the factors limiting acetyl-TAG accumulation.

Master of Science / Department of Biological & Agricultural Engineering / Donghai Wang / Camelina sativa (L). Crantz that belongs to Brassicaceae family has been grown as a dicotyledonous oilseed crop in the cold places like America and Canada. Camelina seeds are widely used for the extraction of oil and protein. Recently, research found that camelina gum is an excellent candidate for food and industrial uses as thickener or stabilizer. The objectives of this research were 1) to increase camelina gum isolation efficiency using spray drying technology, and 2) to develop an innovative method to remove gum from seed bran to increase protein and oil extraction efficiency and quality. The camelina gums isolated using ethanol precipitation and spray drying method from the whole camelina seeds were compared. Effects of spray drying temperature on yield, gum morphology, and gum rheological and thermal properties were studied. The representative sample dried at 165°C was chosen to study the effects of concentration, temperature, pH and additives (NaCl, CaCl₂, sucrose, and ethanol) on viscosity and viscoelastic properties of the isolated gum. The gum showed a shear thinning behavior when shear rate increased gradually, higher concentrations of additives only slightly affect the rheological properties. Results showed that spray drying is an effective method in terms of saving time and energy, and provided positive rheology benefits on camelina gum isolation. Pre-removal of gum from camelina seeds can increase protein and oil yield and their quality. Decortication can separate 10-17% of the total camelina seed as bran. A wind tunnel was used to separate lighter bran particles from heavier endosperm and unbroken seeds. Camelina gum isolation from the separated seed bran using the traditional ethanol precipitation method was optimized using response surface methodology where the simultaneous effect of the three independent variables (seed bran to water ratio, isolation temperature, and isolation time) were investigated for gum yield, purity, and optimum rheological properties. Three independent quadratic modules were developed and the original data fitted the models fitted (R² = 0.995, 0.877, and 0.804). The optimal isolation conditions were seed bran to water ratio of 1:39, isolation temperature of 35 ºC, and isolation time of 1.5 h and 0.839 desirability was obtained by the rigorous statistics analysis. The protein yield and quality extracted from decorticated endosperm were improved significantly compared with that extracted from whole seeds meal without decortication. In addition, the degumming step can be eliminated before protein and oil extraction that increase protein and oil extraction efficiency.

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Aviation greenhouse gas (GHG) emissions have risen faster than any other transport sector to double between 1990 and 2005. Such emissions from aviation could increase another 700 percent globally, and at least 150 percent in the European Union (EU), by 2050 due to continuously increasing consumer demand. To reverse the trend of rising emissions writ large, the EU has set 2030 climate goals of reducing its GHG emissions by 40 percent (relative to 2005) and having 32 percent of gross final energy consumption from renewables. The EU’s recast Renewable Energy Directive (RED-II) calls for 14 percent of transport energy from renewables, gives multipliers to advanced biofuels, and restricts biomass that is from ecologically valuable lands or that causes land use change. Energy security and energy independence are also long-term EU goals. Many of these goals and targets have also been adopted by the European Free Trade Area (EFTA). Despite these efforts, options are limited to reduce aviation emissions compared to other transport sectors, leaving aviation biofuels, also known as renewable jet fuels (RJFs), as currently the only commercialized option. Against this backdrop, in this thesis scenario analyses were conducted to produce biomass from EU+EFTA lands, project RJF yields from this biomass, and estimate emissions savings of these RJFs compared to petroleum jet fuel. Particular effort was devoted to identifying biomass, biofuels, and EU+EFTA lands that comply with RED-II criteria. The two RJF pathways selected were hydroprocessed esters and fatty acid (HEFA) conversion of Camelina sativa vegetable oil and Fischer-Tropsch (FT) synthesis of forestry residue lignocellulosic biomass. Over 117 million hectares in the EU+EFTA was identified as available for Camelina sativa cultivation, which could yield over 64 Mt of RJF each year, or 113 percent of the total jet fuel consumed in the EU+EFTA in 2017. Conversely, if 50 percent of the forestry residues generated as by-products from EU+EFTA roundwood harvesting operations in 2017 were extracted from harvest sites, 40 Mt of forestry residues would be available as biomass, which would yield almost 7.6 Mt of RJF annually (13% of 2017 jet fuel consumption). If all 144 million hectares of EU+EFTA forest lands deemed available for wood supply were logged, 1,772 Mt of forestry residues would be produced in total (at 50 percent extraction), which could result in almost 337 Mt of RJF, or 590% of the jet fuel consumed in the region in 2017. Hence, RJF can be feasibly produced from biomass from EU+EFTA lands, in amounts that meet or exceed the annual jet fuel consumption of the EU+EFTA, and in ways that meet or exceed RED-II sustainability criteria. However, the proportion of these RJF yields to total annual EU+EFTA jet fuel consumption will decrease over time as the number of flights and their resulting emissions increase. The two RJFs also emit 67 percent and 91 percent fewer GHG emissions, respectively, than petroleum-based jet fuel, showing them to be important tools for the EU to meet its 2030 renewables and emissions reductions targets. Producing the biomass feedstocks and RJFs in these quantities will require the EU to make serious decisions on land use trade-offs, such as whether livestock production is more important than biofuel production.

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