Dans les thylakoïdes des végétaux supérieurs, la réaction photochimique de la photosynthèse implique un transfert d’électrons linéaire ayant lieu entre les photosystèmes II et I et aboutit à la production de pouvoir réducteur (NADPH) et d’ATP. En parallèle à ce transfert linéaire, le transfert cyclique d’électrons autour du PS I court-circuite le PS II et fait intervenir distinctement les complexes PGR (PGR5/PGRL1) et NADPH déshydrogénase (NDH). Ces deux voies alternatives permettent la production d’ATP sans accumulation de NADPH dans le stroma et particulièrement en conditions de stress environnemental. Ces voies sont essentielles pour la physiologie de la plante mais leur régulation demeure mal connue. Une hypothèse a été émise quant à leur régulation redox en étudiant plusieurs thiorédoxines chloroplastiques. Des expériences menées in vivo et in vitro sur des mutants d’Arabidopsis, de tabac et de tomate ont mis en évidence le rôle très spécifique de la thiorédoxine m4 dans la régulation négative des voies dépendantes de PGR et NDH. Une stratégie originale a été mise en place pour capturer les cibles putatives de cette thiorédoxine en mutant le site actif de la protéine chez Arabidopsis. Les résultats obtenus ont montré l’existence de plus de 160 cibles potentielles et certaines d’entre elles pourraient être impliquées dans la régulation des voies dépendantes des complexes PGR et NDH. / In plants thylakoids, the photochemical reaction of photosynthesis implies a linear electron flow between photosystems II and I which produces reducing power (NADPH) and ATP. In addition of linear electron flow, the cyclic electron flow around PS I bypasses PS II and involves distinctly the PGR (PGR5/PGRL1) and the NADPH dehydrogenase (NDH) complexes. These two alternative pathways lead to the production of ATP without accumulation of NADPH in the stroma especially under environmental stress conditions. Both pathways are essential for the plant physiology but their regulation remains unclear. An assumption concerning a redox regulation has been considered by surveying several chloroplastic thioredoxins. In vivo and in vitro studies of Arabidopsis, tobacco and tomato mutants revealed that the m4 thioredoxin plays a very specific role in the down-regulation of the PGR and NDH dependant pathways. An original strategy to capture putative targets of this thioredoxin has been established by mutating the active site of the protein in Arabidopsis. The results showed the existence of more than 160 potential targets and some of them could be involved in the regulation of PGR and NDH dependant pathways.

Plastid genomes of flowering plants are largely identical in gene order and content, but a few lineages have been identified with many gene and intron losses, genomic rearrangements, and accelerated rates of nucleotide substitutions. These aberrant lineages present an opportunity to understand the modes of selection acting on these genomes as well as their long-term stability. My research has focused on two areas within plastid genome evolution in Geraniaceae: first, an investigation of the diversity of unusual plastid genomes in a single genus, Erodium (Geraniaceae) for chapters one and three. Chapter two focuses on the evolution of subunits of the plastid-encoded RNA polymerase (PEP). The first chapter described the loss of plastid-encoded NADPH dehydrogenase (ndh) genes from a clade of 13 Erodium species. Divergence time estimates indicate this clade is less than 5 million years old. This recent loss of ndh genes in Erodium presents an opportunity to investigate changes in photosynthetic function through comparative biochemistry between Erodium species with and without plastid-encoded ndh genes. Second, I examined the evolution of the gene encoding the alpha subunit (rpoA) of PEP in three disparate angiosperm lineages—Pelargonium (Geraniaceae), Passiflora (Passifloraceae), and Annonaceae—in which this gene has diverged so greatly that it is barely recognizable. PEP is conserved in the plastid genomes of all photosynthetic angiosperms. I found multiple lines of evidence indicating that the genes remain functional despite retaining only ~30% sequence identity with rpoA genes from outgroups. The genomes containing these divergent rpoA genes have undergone significant rearrangement due to illegitimate recombination and gene conversion, and I hypothesized that these phenomena have also driven the divergence of rpoA. Third, I conducted a survey of plastid genome evolution in Erodium with the completion of 15 additional whole genomes. Except for Erodium and some legumes, all angiosperm plastid genomes share a quadripartite structure with large and small single copy regions (LSC, SSC) and two inverted repeats (IR). I discovered a species of Erodium that has re-formed a large inverted repeat. Demonstrating a precedent for loss and regain of the IR also impacts models of evolution for other highly rearranged plastid genomes. / text

京都大学 / 新制・課程博士 / 博士(理学) / 甲第23048号 / 理博第4725号 / 新制||理||1677(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 鹿内 利治, 准教授 竹中 瑞樹, 教授 長谷 あきら / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第20210号 / 理博第4295号 / 新制||理||1617(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 鹿内 利治, 教授 長谷 あきら, 准教授 小山 時隆 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM

This thesis deals with phylogenetic relationships at different taxonomic levels. All the plants studied are or have been included in the order Gentianales, which comprises about 17 200 species. The phylogenies are based on parsimony analyses of nucleotide sequence data (DNA) from different regions in the chloroplast genome, but morphological characteristics have also been studied. Analyses of sequence data from the genes rbcL and ndhF provide the interfamilial structure of the order Gentianales, shown to comprise the families Apocynaceae (incl. Asclepiadaceae), Gelsemiaceae, Gentianaceae, Loganiaceae, and Rubiaceae. Exclusion of certain genera from the Loganiaceae is confirmed and their phylogenetic positions are clarified. Some of these genera remain within the Gentianales, while others belong to other orders. Exclusion of the tribe Buddlejeae from the Loganiaceae is confirmed, and a monophyletic group formed by Buddleja, Emorya, Gomphostigma, and Nicodemia is recognized and placed in the Lamiales. The Loganiaceae s.str. forms, after these exclusions, a strongly supported monophyletic group comprising 13 genera. The tribe Paederieae in the family Rubiaceae is analysed using sequence data from the regions rbcL gene, rps16 intron, and the regions trnT-F, and is shown to be paraphyletic. These results lead to a new circumscription of Paederieae comprising the genera Leptodermis, Paederia, Serissa, and Spermadictyon. The tribe Putorieae is reestablished with the single genus Plocama, including 34 species. Aitchisonia, Choulettia, Crocyllis, Gaillonia, Jaubertia, Pseudogaillonia, Pterogaillonia, and Putoria are reduced to synonyms of Plocama based on the molecular analyses and morphological studies. The Mediterranean species of the re-circumscribed Plocama, previously segregated as the genus Putoria, are revised. Two species, Plocama calabrica and P. brevifolia, are recognized, their synonymics are established, and seven lectotypes are selected.

Palladium-catalysed carbonylative multicomponent reactions have proven useful for the synthesis of structurally diverse compounds. Carbon monoxide serves as an atom-efficient, one-carbon building block, which allows for further structural elaboration of the carbonyl compound. By varying the components of the carbonylative multicomponent reaction, considerable product diversity can readily be attained. However, due to the reluctance to use toxic CO gas, considerable efforts have been directed at exploring non-gaseous approaches. The work described in this thesis has mainly focused on the development of palladium(0)-catalysed, carbonylative multicomponent synthetic methodology, using the non-gaseous CO source molybdenum hexacarbonyl, in the synthesis of heterocycles and other biologically relevant functional groups. The first part of this work describes the development of a non-gaseous carbonylative Sonogashira cross-coupling of bifunctional ortho-iodoanilines and terminal alkynes. Where 4-quinolones were synthesised via a carbonylation/cyclisation sequence. Using a similar synthetic strategy, three different N-cyanobenzamide intermediates were prepared by palladium-catalysed carbonylative couplings of various aryl halides and bromides and cyanamide. The formed intermediates provided a basis for further chemical transformations. First, ortho-iodoanilines were carbonylatively coupled with cyanamide and subsequently cyclised to yield heterocyclic 2-aminoquinazolinones. Next, building on those findings, the same synthetic strategy was applied to ortho-halophenols to provide a highly convenient domino carbonylation/cyclisation method for the preparation of benzoxazinones. The developed method was used to evaluate the efficiency of various non-gaseous CO sources. Third, the palladium-catalysed carbonylative synthesis of N-cyanobenzamides, was used to produce biologically relevant N-acylguanidines with considerable product diversity. Finally, one of the developed carbonylative methodologies was used in the preparation of potential NDH-2 inhibitors based on a quinolinyl pyrimidine scaffold. The prepared compounds were biologically evaluated in terms of inhibition of oxidoreductase NDH-2 and antibacterial activity on Gram-negative bacteria, S. aureus and Mtb. The biological evaluation revealed that some of the quinolinyl pyrimidines exerted inhibitory activity on the NDH-2 enzyme and possessed antibacterial properties. The work described in this thesis has been devoted to the development of non-gaseous one-pot, multicomponent carbonylation/cyclisation and carbonylation/amination reactions. The described methods offer highly attractive synthetic strategies that can be of great value to synthetic and medicinal chemists.