Central and storage carbon metabolism of the brown alga Ectocarpus siliculosus: insights into the origin and evolution of storage carbohydrates in Eukaryotes

International audienceBrown algae exhibit a unique carbon (C) storage metabolism. The photoassimilate d‐fructose 6‐phosphate is not used to produce sucrose but is converted into d‐mannitol. These seaweeds also store C as β‐1,3‐glucan (laminarin), thus markedly departing from most living organisms, which use α‐1,4‐glucans (glycogen or starch).Using a combination of bioinformatic and phylogenetic approaches, we identified the candidate genes for the enzymes involved in C storage in the genome of the brown alga Ectocarpus siliculosus and traced their evolutionary origins.Ectocarpus possesses a complete set of enzymes for synthesis of mannitol, laminarin and trehalose. By contrast, the pathways for sucrose, starch and glycogen are completely absent.The synthesis of β‐1,3‐glucans appears to be a very ancient eukaryotic pathway. Brown algae inherited the trehalose pathway from the red algal progenitor of phaeoplasts, while the mannitol pathway was acquired by lateral gene transfer from Actinobacteria. The starch metabolism of the red algal endosymbiont was entirely lost in the ancestor of Stramenopiles. In light of these novel findings we question the validity of the ‘Chromalveolate hypothesis’

Development and physiology of the brown alga Ectocarpus siliculosus: two centuries of research

International audienceBrown algae share several important features with land plants, such as their photoautotrophic nature and their cellulose‐containing wall, but the two groups are distantly related from an evolutionary point of view. The heterokont phylum, to which the brown algae belong, is a eukaryotic crown group that is phylogenetically distinct not only from the green lineage, but also from the red algae and the opisthokont phylum (fungi and animals). As a result of this independent evolutionary history, the brown algae exhibit many novel features and, moreover, have evolved complex multicellular development independently of the other major groups already mentioned. In 2004, a consortium of laboratories, including the Station Biologique in Roscoff and Genoscope, initiated a project to sequence the genome of Ectocarpus siliculosus, a small filamentous brown alga that is found in temperate, coastal environments throughout the globe. The E. siliculosus genome, which is currently being annotated, is expected to be the first completely characterized genome of a multicellular alga. In this review we look back over two centuries of work on this brown alga and highlight the advances that have led to the choice of E. siliculosus as a genomic and genetic model organism for the brown algae

Role and Evolution of the Extracellular Matrix in the Acquisition of Complex Multicellularity in Eukaryotes: A Macroalgal Perspective

Multicellular eukaryotes are characterized by an expanded extracellular matrix (ECM) with a diversified composition. The ECM is involved in determining tissue texture, screening cells from the outside medium, development, and innate immunity, all of which are essential features in the biology of multicellular eukaryotes. This review addresses the origin and evolution of the ECM, with a focus on multicellular marine algae. We show that in these lineages the expansion of extracellular matrix played a major role in the acquisition of complex multicellularity through its capacity to connect, position, shield, and defend the cells. Multiple innovations were necessary during these evolutionary processes, leading to striking convergences in the structures and functions of the ECMs of algae, animals, and plants

BASES BIOCHIMIQUES ET CELLULAIRES DES INTERACTIONS PLANTE-PATHOGENE DANS LE SYSTEME CHONDRUS-CRISPUS-ACROHAETE OPERCULATA

PARIS-BIUSJ-Physique recherche (751052113) / SudocCentre Technique Livre Ens. Sup. (774682301) / SudocSudocFranceF

Développement d'enzymes recombinants issus des bactéries marines P. carrageenovora et SW5 pour la production d'oligo-fucoïdanes et d'oligo-ë-carraghenane

Ce travail visait à caractériser et produire deux enzymes originaux hydrolysant d'une part un polysaccharide pariétal d'algue rouge (le g-carraghénane), et d'autre part un polysaccharide d'algue brune (le fucoïdane). Ces deux endo-hydrolases extracellulaires sont produites par deux bactéries marines saprophytes, P. carrageenovora, et SW5. Suite à la purification des deux enzymes sauvages, leur[s] gène[s] ont été clonés puis séquencés. L'activité recombinante obtenue par surexpression dans E. coli a validé les séquences obtenues. L'analyse de ces dernières montre l'architecture modulaire des deux protéines. Cependant, le domaine catalytique de la g-carraghénase n'a pas pu être identifié. Il s'agit donc d'un membre orphelin des glycoside hydrolases, distinct des carraghénases préalablement caractérisées. A l'inverse, les similitudes partagées par la séquence de la fucoïdanase avec deux autres fucoïdanases, a permis de définir une nouvelle famille de glycoside hydrolases.This work aimed to characterize and produce two new biocatalysts which hydrolyze two polysaccharides extracted from the cell wall of red algae (g-carrageenan) and brown algae (fucoidan). These extracellular endo-hydrolases are produced by two saprophytic marine bacteria, Pseudoalteromonas carrageenovora, (y-Proteobacteria), and SW5 (Bacteroidetes). Following the purification ofwild-type proteins, their genes were cloned and sequenced. The recombinant activity obtained by overexpression in E. coli confirmed that the cloned sequences coded for corresponding enzymes. Sequence analysis showed that the enzymes have a modular structure. The catalytic domain of the g-carrageenase was net identified. This enzyme is therefore different from previously described glycoside hydrolases, and aise distinct from previously known carrageenases. The fucoidanase sequence shares similarity with two other bacterial putative fucoïdanase and these three enzymes define a new glycosidase family.COMPIEGNE-BU (601592101) / SudocSudocFranceF

Caractérisation biochimique et moléculaire des haloperoxydases dépendantes du vanadate chez l'algue brune Laminaria digitata

PARIS-BIUSJ-Thèses (751052125) / SudocSudocFranceF

Effets d'oligosaccharides sur l'embryogenèse de microspores de brocoli : (mode d'action et transduction du signal)

L'induction embryogène dans des cultures de microspores isolées est un processus dépendant d'un stress, qui peut être déclenché par un choc thermique, une privation de saccharose ou de nitrogène ou par l'utilisation d'inhibiteurs de la polymérisation des microtubules. Connus pour mimer des stress biotiques, les oligosaccharides ont été testés en tant que source de composés inducteurs de l'embryogenèse de microspores de Brassica napus var. italica. Parmi les huit séries d'oligosaccharides étudiés et leurs polymères respectifs (à savoir des pectines, des alginates, des fucanes, des laminarines, des agars et des ?-, ?-, ?-carraghénanes) seuls les oligo-carraghénanes présentaient des effets significatifs sur l'induction des microspores. Ajoutés en combinaison avec un choc thermique, ils augmentaient très fortement les rendements finaux en embryons dérivés de microspores. Ainsi, un doublement du rendement était observé dans le cas du traitement le plus efficace, c'est-à-dire en présence d'oligo-?-carraghénanes. Le mode d'action des oligo-carraghénanes a été étudié en suivant le devenir d'oligosaccharides fluorescents. Les oligo-carraghénanes peuvent entrer dans les cellules ; toutefois, l'utilisation d'oligo-carraghénanes biotinylés immobilisés sur des billes de streptavidine a révélé que les molécules exerçaient probablement leurs effets au niveau extracellulaire. Les réponses ont été comparées avec celles obtenues lors de traitements avec des peptides contenant la séquence RGD (site de reconnaissance universel des molécules d'adhésion des cellules animales) sur des cultures de microspores. Les traitements à l'aide de peptides RGD présentaient des résultats similaires sur l'induction de l'androgenèse et semblaient entrer en compétition avec les sites de liaisons des oligo-carraghénanes. Des études pharmacologiques ont permis de montrer que la signalisation des oligo-carraghénanes et des peptides RGD présentent des voies de transduction communes, impliquant des protéines kinases et des MAP kinases, éléments distincts de la voie de transduction du choc thermique. Ces données ont permis de proposer un modèle hypothétique de l'induction de l'embryogenèse par modification de la polarité des microspores. Le développement propre des grains de pollen nécessite une adhésion polarisée pour établir de possibles connections au niveau cortical...RENNES1-BU Sciences Philo (352382102) / SudocSudocFranceF

Oligoguluronates Elicit an Oxidative Burst in the Brown Algal Kelp Laminaria digitata

Oligomeric degradation products of alginate elicited a respiratory and oxidative burst in the sporophytes of the kelp Laminaria digitata. The generation of activated oxygen species (AOS), O(2)(−), and H(2)O(2) was detected at the single cell level, using nitroblue tetrazolium precipitation and a redox-sensitive fluorescent probe, respectively. The oxidative burst involved diphenyleneiodonium-sensitive AOS-generating machinery and its amplitude depended on the type of tissue. After a first elicitation plants were desensitized for about 3 h. The activity of alginate oligosaccharides was dose dependent, saturating around 40 μm. It was also structure-dependent, with homopolymeric blocks of α-1,4-l-guluronic acid, i.e. the functional analogs of oligogalacturonic blocks in pectins, being the most active signals. The perception of oligoguluronate signals resulted in a strong efflux of potassium. Pharmacological dissection of the early events preceding the emission of AOS indicated that the transduction chain of oligoguluronate signals in L. digitata is likely to feature protein kinases, phospholipase A(2), as well as K(+), Ca(2+), and anion channels