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Biologie, Ecologie Et Conservation Animales ETUDE ECO-BIOLOGIQUES DES UROPLATE

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Biologie, Ecologie Et Conservation Animales ETUDE ECO-BIOLOGIQUES DES UROPLATE UNIVERSITE D’ANTANANARIVO FACULTE DES SCIENCES DEPARTEMENT DE BIOLOGIE ANIMALE DEPARTEMENT DE BIOLOGIE ANIMALE Latimeria chalumnae MEMOIRE POUR L’OBTENTION DU Diplôme d’Etudes Approfondies (D.E.A.) Formation Doctorale : Sciences de la vie Option : Biologie, Ecologie et Conservation Animales ETUDE ECO-BIOLOGIQUES DES UROPLATES ET IMPLICATION DES ANALYSES PHYLOGENETIQUES DANS LEUR SYSTEMATIQUE Présenté par : Melle RATSOAVINA Mihaja Fanomezana Devant le JURY composé de : Président : Madame RAMILIJAONA RAVOAHANGIMALALA Olga, Professeur Titulaire Rapporteurs : Madame RAZAFIMAHATRATRA Emilienne, Maître de Conférences Examinateurs: Madame RAZAFINDRAIBE Hanta, Maître de conférences Monsieur EDWARD LOUIS, Doctor of Veterinary Medicine, PhD Soutenu publiquement le 27 Février 2009 ________________________________________________________________Remerciements REMERCIEMENTS Cette étude a été réalisée dans le cadre de l’accord de collaboration entre le Département de Biologie Animale, Faculté des Sciences, Université d’Antananarivo et Henry Doorly Zoo Omaha. Nebraska. USA, dans le cadre du projet Madagascar Biogeography Project (MBP). Je tiens à exprimer ma profonde reconnaissance à: - Madame RAMILIJAONA RAVOAHANGIMALALA Olga, Professeur Titulaire, Responsable de la formation doctorale, d'avoir accepté de présider ce mémoire et consacré son précieux temps pour la commission de lecture. - Madame RAZAFIMAHATRATRA Emilienne, Maître de conférences, encadreur et rapporteur qui a accepté de diriger et de corriger ce mémoire. Je la remercie pour ses nombreux conseils. - Madame RAZAFINDRAIBE Hanta, Maître de conférences, qui a accepté de faire partie de la commission de lecture, de juger ce travail et de figurer parmi les membres du jury. - Monsieur Edward LOUIS, Doctor of Veterinary Medecine, PhD d’être parmi les membres du jury et qui m’a permis d’avoir intégrer l’équipe de HDZ et de m’avoir inculqué les a priori des travaux de recherche. « Thank you for all the opportunities ». Je remercie également : - Dr Achille RASELIMANANA, membre de la commission de lecture ; - Dr Rick BRENNEMAN Vice- Principal Investigateur de MBP ; - Tous les enseignants et les personnels techniques et administratifs du Département de Biologie Animale et de la Faculté des Sciences ; i ________________________________________________________________Remerciements - Tous les techniciens de laboratoire au Center for Conservation and Research (CCR) ; - Le Henry Doorly Zoo, tous les guides, Hubbard Family Foundation, Dr Lee Simmons et Dr Doug Amstrong; - L’équipe Pinus et tous mes amis qui ont investi leur temps et leur efforts pour m’aider à réaliser la présente étude. Mes chaleureux remerciements s’adressent également à mes sœurs, mes frères, et toute la famille pour leur soutien moral et leurs encouragements permanents. Pour terminer, je remercie très vivement toutes les personnes qui, de près ou de loin, ont contribué à la réalisation de ce mémoire. ii _____________________________________________________________________Résumé Résumé Ce travail porte sur une étude éco-biologique et phylogénétique des uroplates dans les forêts humides tropicales de l’Est et du Sud-Est de Madagascar. Les espèces Uroplatus fimbriatus, U. lineatus, U. phantasticus, U. sikorae sikorae et U. sikorae sameiti ont été inventoriées dans la RNI de Betampona (Septembre 2005) et le PN de Ranomafana (Janvier et Mars 2004). 75 individus ont été capturés dans les deux sites. La taille, le poids et le sexe de chaque animal ont été notés. Toutes les mesures concernant la taille montrent des valeurs supérieures à celles trouvées par les chercheurs précédents sauf pour U. s. sameiti qui est de 104,7 mm. Le type de support, la distribution verticale et la période d’activité sont aussi bien collectés pour chaque espèce. La plupart des individus trouvés préfèrent la branche comme perchoir. Mais U. phantasticus est rencontrée aussi bien que sur des feuilles, que des fines branches et des lianes en raison de sa petite taille. Les espèces de grande taille sont souvent trouvées sur les troncs d’arbre et les branches. A Betampona, les individus sont trouvés de 0,5 m à 6 m du sol. A Ranomafana, les animaux évoluent à partir de 0,5 m jusqu’à 8 m du sol. Une large répartition verticale est notée chez U. s. sikorae de 0,5 à 8 m. Une extraction de l’ADN total à partir du tissu caudal, une amplification par PCR de l’ADN cible et son séquençage ont été effectués pour un total de 93 individus d’uroplates et 2 Phelsuma comme groupe externe. 893 paires de bases de l’ADN mitochondrial ont été alignées pour les gènes de la sous-unité 4 de la deshydrogénase-NADH (ND4), l’ARNtHis, l’ARNtSer et l’ARNtLeu . Le logiciel PAUP version 4.0b5. est utilisé pour obtenir des phylogrammes par les méthodes Neighbor-joining et Maximum parcimonie. Le diagramme en réseau a été construit à l’aide du programme Network. La fiabilité de chaque analyse est donnée par un test de Bootstrap avec 1000 répétitions. La monophylie au sein du genre Uroplatus est confirmée par l’analyse. Les individus collectés dans chaque site forment chacun un clade et même étant géographiquement proches, la distance génétique entre les espèces est grande et implique la notation en type. Les espèces déjà décrites sont confirmées par l’analyse. Mais, elle met aussi en cause le statut des taxa puisque la distance génétique entre deux espèces différentes est inférieure à celle entre deux types ou sous-espèces. Même si la description de nouvelle forme dépasse le cadre du présent travail, Il encourage de faire des recherches plus étendues pour avancer dans la connaissance de l’histoire naturelle ainsi que la phylogénie et la conservation du groupe. Mots clés : Uroplatus, Ecologie, ADN mitochondrial, PCR, Séquençage, Phylogénie, Ranomafana, Betampona, Madagascar. iii _____________________________________________________________________Abstract Abstract This work reports the ecobiology and the phylogenetic study of the genus Uroplatus in East and South-East Rainforest of Madagascar. Uroplatus fimbriatus, U. lineatus, U. phantasticus, U. sikorae sikorae and U. sikorae sameiti are assessed in Natural Reserve Betampona (September 2005) and Ranomafana National Park (January and March 2004). 75 individuals were captured in the two sites. Body length, Weight and sex identification are accomplished for each specimen. Body length recorded go over data from previous study except for U. s. sameiti which is 104.5 mm. Support type, vertical distribution and time of activity are defined; branches are the most used by Uroplatus. But U. phantasticus is observed on leaves, little branches and lianas for its small size. Larger species such as U. fimbriatus are found on the tree trunk and larger branches. In Betampona, individuals are found between 0.5 to 6 m above ground. In Ranomafana, leaftail geckos are seen up to 8 m. U. s. sikorae is found to have the largest vertical distribution; 0.5 to 8 m. A molecular genetics study was conducted for the Uroplatus sampled from the study sites with tissue from the tail used as genetic material. DNA extraction, PCR amplification and sequence data were generated for 93 leaf tailed gecko individuals and 2 specimens of Day geckos (genus Phelsuma) were utilized as outgroup taxa. 893 base pairs from the mitochondrial DNA are aligned, studied fragment are: ND4, tRNA His, tRNA Ser and tRNA Leu. Neighbor-joining and Maximum parsimony trees are obtained by PAUP version 4.0b5 program. Star tree are given by Network program to illustrate the speciation among the population. Bootstrap, probability statistic, was conducted to show the analysis liability with 1000 replications. The monophyly of the genus Uroplatus was confirmed by the present study. Each sample from a specific site is gathered in one clade; even geographically close, huge genetic divergence is found out, thus the nomination as Type. The following study allows confirming described species; also it puts in doubt the state of each species, because genetic distance between two different species is less than found in two subspecies or types. Even describing a new species is beyond this survey, it is recommended to conduct wide and further researches to obtain knowledge on natural history and progress on phylogeny to set better conservation strategy for Uroplatus group. Keywords: Uroplatus, Ecology, mitochondrial DNA, PCR, Sequencing, Phylogeny, Ranomafana, Betampona, Madagascar. iv ____________________________________________________________________Sommaire SOMMAIRE INTRODUCTION ......................................................................................................................... 1 CHAPITRE I : PRESENTATION DU GENRE UROPLATUS ................................................ 3 I.1- Classification du genre Uroplatus .............................................................................. 3 I.2- Historique du genre Uroplatus ................................................................................... 4 I.3- Morphologie ............................................................................................................... 4 I.4- Biologie ...................................................................................................................... 6 I.5- Ethologie ..................................................................................................................... 6 I.6- Distribution géographique .......................................................................................... 7 I.7- Menaces .....................................................................................................................
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