L’objectif de ce projet de thèse était de caractériser le métabolisme de terpénoïdes (ou isoprénoïdes) chez les plantes supérieures. L’essentiel du travail a consisté à caractériser des triterpènes synthases (TTPS) d’Arabidopsis thaliana, un modèle végétal, ainsi que celles de l’épurge (Euphorbia lathyris), une euphorbe pour laquelle des applications agronomiques sont envisagées. Au cours de ma thèse, j’ai aussi contribué à l’étude du métabolisme et des fonctions des précurseurs de triterpènes et de stérols, ainsi qu’à leurs fonctions biologiques.Les triterpènes synthases, ou 2,3-oxydosqualène cyclases (OSCs), convertissent le substrat 2,3-oxydosqualene (SqO) en une multitude d'alcools triterpéniques, et ainsi amorcent la biosynthèse de dérivés triterpénoïdes (triterpènes oxydés, conjugués, etc ..). Arabidopsis thaliana contient 13 OSCs produisant divers squelettes triterpéniques, de type stéroïdien ou non-stéroïdien. Les produits de cyclisation du SqO ont été élucidés structuralement (GC-MS, RMN) après expression hétérologue des enzymes en levure erg7. Cette levure est déficiente en lanostérol synthase (ERG7), ce qui permet d'accumuler le SqO, substrat des cyclases. Lorsque le mutant est transformé avec un ADNc codant une triterpène synthase, il est capable de convertir le SqO en un ou plusieurs triterpènes. Cependant, la caractérisation des 13 OSCs d'Arabidopsis réalisée de façon hétérologue en levure n’a pas été établie inplanta. De façon surprenante, certains des composés produits dans les levures erg7 transformées n'ont jamais été détectés chez Arabidopsis. C'est pourquoi il a été nécessaire de reconsidérer les fonctions biochimiques exactes de ces enzymes dans un contexte végétal. / The subject of this PhD thesis is part of a research project entitled "Production of energy-rich triterpenoids in Euphorbia lathyris, a potential crop for third generation biofuels," whose acronym is EULAFUEL. This project is funded by a multipartner program ANR-KBBE and has been extended until December 2013. The aim of this PhD project is to get new insights into the aspects related with the biosynthesis and accumulation of latex triterpenoids. In addition, for comparison, a major objective of the thesis is to characterize functionally the enzymes involved in the synthesis of triterpenes in the model plant Arabidopsis thaliana. Triterpene synthases, also named oxidosqualene cyclases (OSCs), convert 2,3-oxidosqualene (OS) into a multitude of triterpene alcohols and there by initiate triterpene biosynthesis. Arabidopsis thaliana for instance has 13 OSCs producing diverse skeletons of steroidal or non-steroidal triterpenes. Cyclization products of a given enzyme have been characterized biochemically using a yeast heterologous expression system. However, for the majority of Arabidopsis triterpene synthases, inplanta studies are lacking. In fact, most of the compounds produced in yeast expressing such enzymes have never been detected in wild-type Arabidopsis. This is a reason why we should reconsider the exact biochemical function of triterpene synthases in the plant context. Then, in a comparative approach in E. lathyris, we project to study the specific triterpene accumulation in the laticifers, a specialized cell type where high amounts of lanosterol, an unusual OS cyclization product for plants, accumulate.

Rhizoctonia solani, Pythium aphanidermatum, and Macrophomina phaseolina were isolated from Euphorbia lathyris grown in fields near Tucson, Arizona. R. solani occurred as a damping-off organism in the fall. P. aphanidermatum infected seeds, seedlings, and mature plants in laboratory and greenhouse tests. Although P. aphanidermatum was infrequently isolated from field plants in Arizona, it was a major pathogen of greenhouse plants growing at high temperatures in nonsterile soil. M. phaseolina was a major pathogen. Infection of E. lathyris roots occurred within 1 mo of an October 1980 planting, but symptoms were not significant until June. Sclerotia of this fungus ranged in numbers from 1 to 246 sclerotia/g field soil. Population densities of 0.2 sclerotium/g soil were sufficient to cause more than 90% plant death in field plots. Some plants infected with M. phaseolina were growing in an area newly cleared of native desert vegetation. Subsequently, M. phaseolina was found in uncultivated soils from four vegetative communities in southern Arizona at elevations from 600 to 2,000 m; the fungus also was recovered from roots of several symptomless native plants.

An evaluation was made of soil solarization to control Macrophomina phaseolina and Sclerotium rolfsii under the climatic conditions of the Sonoran Desert region of Arizona. Tarping of moist soil with clear polyethylene in the summer was most effective in raising soil temperatures, while tarping during the fall and spring were less efficient. In one summer trial, the maximum temperatures achieved were 7-8 C higher than control plots at 1, 15, and 30 cm depths. In none of the tests was the application of tarp effective in reducing M. phaseolina populations to non-detectable levels. After a six-week summer solarization treatment, S. rolfsii was controlled at the 15-cm but not the 30-cm depth. During a fall treatment control of S. rolfsii was achieved only at the 1-cm depth. After solarization, seeds of Euphorbia lathyris were planted in solarized and control plots. The incidence of M. phaseolina-associated mortality among seedlings planted in solarized plots was always the same as the control plot with the highest disease incidence. The utility of this technique may be limited by the heat tolerance of the target organisms. Studies of bacterial antagonists were initiated to determine their utility in enhancing pathogen control after the solarization treatment. Screening 43 bacterial isolates showed that seven of Pseudomonas fluorescens and one of Serratia marcescens exhibited some form of antagonism toward M. phaseolina, S. rolfsii and Verticillium dahliae in vitro. Antagonism was manifested as a complete inhibition of fungal development or reduced hyphal development coupled with suppression of sclerotial development. The action of the antagonists was found to be fungitoxic or fungistatic rather than fungicidal. None of the four P. fluorescens isolates tested were effective in preventing M. phaseolina infection of E. lathyris seedlings. The ability of these bacteria to prevent sclerotial formation while still permitting hyphal growth may be a useful technique for studying the two phases of the fungal life cycle separately.

Thesis (Ph. D. - Arid Lands Resource Sciences)--University of Arizona, 1983. / Includes bibliographical references (leaves 164-170).

The objective of this study is to develop a model to evaluate the economic feasibility of biofuels production, and in particular to isolate the variables crucial to feasibility. The model constructed to define these variables is unique in its ability to accommodate a variety of plants and to integrate all portions of the production process; it was tested on a case study of a Euphorbia lathyris industry. The model minimizes costs of production to determine the best configuration for the industry. Total cost equals the sum of costs incurred in each segment of the process: growth, harvest, transport, and extraction. The solution is determined through a non-linear transportation- transshipment algorithm which describes production as a series of nodes and links. Specific application of the model was analysis of E. lathyris biofuel production in Arizona. Simulations were run examining the sensitivity of biocrude cost to changes in input parameters. Conclusions are summarized as follows. * No change in any single element can reduce final cost sufficiently to enable competitive production in the near future. * The major factor necessary to bring cost into range is improvement in biological yield. Two components of equal importance are tonnage produced per acre and percentage extractables recovered in processing. * Lowering cropping costs provided the most effective improvements of economic inputs. Perennial crops significantly reduced farm costs. * Transportation costs outweighed economies of scale in extraction; extractor location close to crops is more efficient than centralized. The cost minimization model was successfully used to isolate the critical factors for an E. lathyris industry in an arid region. Results determine that this industry would not be competitive in Arizona without dramatic improvements in yields and moderate changes in a combination of input costs. Viability is critically dependent on improvements in tonnage yield produced per acre and percent extractables recovered.