</p> Part I The mass spectra of the seven isomeric monomethoxyquinolines, the seven monohydroxyquinolines, the two N-methylquinolones and six furoquinoline alkaloids have been recorded. Fragmentation mechanisms are proposed to account for the major peaks in all the spectra. Use has been made of intense metastable peaks found in most of the spectra to aid the interpretation of the fragmentation routes. In addition, deuterium and carbon-13 labelling have been employed in analyzing the fragmentation routes of the methoxyquinolines. </p> <p> Part II The Hofmann degradation of lobinaline is described. Examination of lobinaline, its derivatives and its Hofmann degradation products by mass spectrometry and other physical and chemical techniques has allowed a structure to be assigned to the alkaloid.</p> / Thesis / Doctor of Philosophy (PhD)

The present study was designed to evaluate the biological activity from the stem bark of Zanthoxylum rhoifolium and Zanthoxylum hyemale. The neutral methanol extract and fractions of this extract, the basic fractions that were obtained after acid-basic extraction from the methanolic extract, and pure compounds isolated from these fraction, were investigated in vitro for antimicrobial activity, antioxidant potential and for acetylcholinesterase inhibition. From the dichlorometane fraction of Zanthoxylum rhoifolium were isolated three furoquinoline alkaloids: skimianine, 8-hidroxy-4,7-dimetoxy-furoquinoline, and γ-fagarin, four benzophenantridine alkaloids: dihydrocheleritrine, dihydroavicine, zanthoxyline and cheleritrine. From the hexane fraction, the lignanes sesamine and lupeol were also isolated. From the neutral methanol extract were isolates de aporfinic alkaloid magnoflorine. The minimal inhibitory concentration of cheleritrine (MIC =1.50 μg/ml) for all tested bacteria, and between 3.12 and 12.5 μg/ml for the tested yeasts, showed this alkaloid have a excellent antimicrobial activity compared to chloramphenicol (between 3.5 to 6.25 μg/m for bacteria) and nistatin (between 5.5 to 10.3 μg/ml for yeasts). The study highlighted the antimicrobial activity of Z. rhoifolium stem bark. This activity is probably due their higher benzophenantridinic alkaloids level. The furoquinoline alkaloids, despite their insignificant antimicrobial activity, were the most active against the enzime AChE. In addition, in this work, the amide tembamide was isolated from the steam bark of Zanthoxylum hyemale, and their derivatives used for a structure-ativity analysis. / O presente estudo foi proposto para avaliar a atividade biológica da casca do caule de Zanthoxylum rhoifolium e Zanthoxylum hyemale. O extrato metanólico neutro e as frações deste extrato, as frações básicas obtidas após extração ácido-base do extrato metanólico e os compostos puros isolados dessas frações, foram avaliados in vitro quanto a atividade antimicrobiana, potencial antioxidante e inibição da enzima acetilcolinesterase. Da fração diclorometânica de Zanthoxylum rhoifolium foram isolados três alcalóides furoquinolínicos: skimianina, 8-hidroxi-4,7-dimetoxi-furoquinolinico e γ-fagarina, quatro alcalóides benzofenantridínicos: diidrocheleritrina, diidroavicina, zantoxilina e cheleritrina. Da fração hexânica foram isolados a lignana sesamina e o triterpeno lupeol. A partir do extrato metanólico neutro foi isolado o alcalóide aporfinico magnoflorina. A concentração inibitória mínima de cheleritrina (CIM = 1,50 μg/ml) para todas as bactérias testadas, e entre 3,12 e 12,5 μg/ml para as leveduras testadas, mostraram que este alcalóide possui uma excelente atividade antimicrobiana em comparação ao cloranfenicol (entre 3,5-6,25 μg/ml para as bactérias) e nistatina (entre 5,5-10,3 μg/ml para as leveduras). O estudo evidenciou a atividade antimicrobiana da casca do caule de Z. rhoifolium. Esta atividade é, provavelmente, devido ao seu alto nível de alcalóides benzofenantridínicos. Os alcalóides furoquinolinicos, mostraram pequena atividade antimicrobiana e foram os mais ativos frente a enzima AChE. Além disso, neste trabalho, a amida tembamida foi isolada da casca do caule de Zanthoxylum hyemale e seus derivados utilizados para uma análise de relação estrutura-atividade.

Choisya ternata est une plante ornementale souvent touchée par la maladie de la pourriture racinaire provoquée par Phytophthora. Cette maladie peut induire de pertes allant jusqu’à 80 %, ce qui implique l’utilisation intensive de produits phytosanitaires. Afin de limiter l’utilisation de ces produits toxiques, une meilleure connaissance des acteurs de la défense des plantes est nécessaire. Pour cela, les objectifs de cette thèse sont I) d’identifier les espèces du genre Phytophthora pathogènes de C. ternata, II) d’étudier des acteurs de défense au niveau racinaire et III) de développer une approche de lutte alternative aux phytosanitaires. L’identification des espèces de Phytophthora par l’amplification et le séquençage de la région ITS, montre la présence de Phytophthora parasitica dans la production de C. ternata en pépinière. Ils mettent aussi en évidence, pour la première fois en France et chez Choisya, la présence de Phytophthora tropicalis. Deux cultivars de C. ternata sont utilisés pour étudier les acteurs de la défense racinaire contre P. parasitica, Aztec pearl (moins sensible à la pourriture racinaire) et Goldfinger (plus sensible). Les arabinogalactane-protéines (AGPs) sont des glycomolécules de la paroi cellulaire qui constitue une barrière physique face aux pathogènes. Des études ont montré le rôle des AGPs dans l’interaction plante-pathogène et plus particulièrement avec les oomycètes. Toutefois, le rôle des AGPs racinaires de Choisya dans l’interaction avec P. parasitica n’est pas étudié. Nos résultats montrent des différences biochimiques au niveau de la composition monosaccharidique des AGPs racinaires d’Aztec pearl en comparaison avec les AGPs de feuilles de ce cultivar et les AGPs de feuilles et de racines de Goldfinger. Contrairement aux autres fractions, ces AGPs n’ont pas augmenté la croissance du mycélium de P. parasitica. Ces résultats suggèrent que l’oomycète n’est pas capable de dégrader cette fraction pour l’utiliser comme une source d’énergie. Pendant l’infection de la plante par P. parasitica, ces AGPs peuvent freiner la pénétration du pathogène, et par conséquent diminuer la sévérité de la maladie. Les plantes ont aussi développé des molécules chimiques nommées métabolites secondaires (MS) capables de les protéger contre leurs agresseurs. Des études ont montré que la partie foliaire de Choisya est riche en MS tels que les alcaloïdes dont plusieurseffets pharmacologiques sont connus. Cependant, leur composition racinaire et leur rôle dans la protection de la plante contre P. parasitica ne sont pas élucidés. Nos résultats montrent que les racines des deux cultivars sont riches en alcaloïdes furoquinoliques. Certains alcaloïdes sont présents en plus grande quantité chez Aztec pearl, mais suite à l’inoculation de zoospores de P. parasitica cette différence n’est plus détectable. De plus, l’extrait contenant les alcaloïdes totaux d’Aztec pearl ont été capables d’inhiber la croissance du mycélium de l’oomycète, contrairement à l’extrait issu de Goldfinger. Ces résultats montrent le rôle potentiel des alcaloïdes furoquinoliques dans la protection de la plante contre cet oomycète. / Choisya ternata is an ornamental plant that suffers from root rot disease due to Phytophthora. This disease can lead to severe production losses (up to 80 %), which require intensive use of phytosanitary products. A better understanding of plants defenses in required in order to reduce the use of these products. Therefore, the objectives of this thesis are I) identifying Phytophthora spp. pathogens of C. ternata, II) studying roots defense actors and III) developing an alternative control approach. Phytophthora spp. identification by ITS region amplification and sequencing highlighted the presence of Phytophthora parasitica on Choisya. Furthermore, Phytophthora tropicalis was identified for the first time in France and on Choisya culture. Two C. ternata cultivars were used to study the plants root defense actors against P. parasitica, Aztec pearl (less susceptible to root rot) and Goldfinger (more susceptible to root rot). Arabinogalactan proteins (AGPs) are glycomolecules of the cell wall which constitutes a physical barrier to pathogens. Studies showed the role of AGPs in the plant-pathogen interaction and more specifically in the case of oomycetes. However, Choisya root AGPs role in the interaction with P. parasitica is not studied. Our results showed biochemical differences in the monosaccharide composition of Aztec pearl root AGPs and the other AGPs fractions (Aztec pearl leaves and Goldfinger roots and leaves). Contrary to other fractions Aztec pearl root AGPs did not increase P. parasitica mycelium growth. These results suggest that oomycete was not able to degrade this fraction and use it as energy source. When P. parasitica infects the plant, these AGPs may be able to slow the infection and reduce disease severity. Plants have also developed chemical molecules known as secondary metabolites (SM) capable of protecting them against attackers. Studies showed that C. ternata leaves are rich with SM as alkaloids that have many pharmacological activities. Nevertheless, roots alkaloids composition and role in plant protection are not studied. Our results showed that roots of the two cultivars are riche with furoquinoline alkaloids. Some these alkaloids were more concentrated in Aztec pearl. But after inoculation with P. parasitica zoospores, the difference was not detectable anymore. Moreover, the total alkaloids extract of Aztec pearl inhibited P. parasitica mycelium growth, unlike the extract of Goldfinger. Another strategy of plants protection is the use of beneficial soil microorganisms that limits pathogens development and stimulate plant defenses. These microorganisms are known as biological control agents (BCA) and are sometimes used in horticulture as an alternative control strategy. In our study, treatments of C. ternata by different BCA were evaluated by a developed real time PCR (qPCR) targeting ypt1 and by symptoms annotation. This evaluation showed that combined treatments by Glomus intraradices with Gliocladium catenulatum and G. intraradices with Trichoderma atroviridae (respectively mycorrhizal fungi and filamentous fungi) offer a better protection against P. parasitica.