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Etude Morphologique, Biochimique, Physiologique Et Biologique De Quelques Populations De Teucrium Polium L

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Etude Morphologique, Biochimique, Physiologique Et Biologique De Quelques Populations De Teucrium Polium L REPUBLIQUE ALGERIENNE DEMOCRATIQUE ET POPULAIRE MINISTERE DE L’ENSEIGNEMENT SUPERIEUR ET DE LA RECHERCHE SCIENTIFIQUE UNIVERSITE DES FRERES MENTOURI - CONSTANTINE 1 FACULTE DES SCIENCES DE LA NATURE ET DE LA VIE DEPARTEMENT DE BIOLOGIE ET ECOLOGIE VEGETALE N° d’ordre :…………………………. N° de Série : ………………………… THESE En vue de l’obtention du diplôme de DOCTORAT EN SCIENCES Filière : Biologie Végétale Spécialité : Production Végétale Thème Etude Morphologique, Biochimique, Physiologique et Biologique de quelques populations de Teucrium polium L. Capitatum dans l’Est algérien Présentée par : MALKI Samira Devant le jury : Président : Mr. D. KHELIFI Prof à Univ. Frères Mentouri - Constantine 1 Rapporteur : Mr. A. YAHIA Prof au Centre Univ. Abdelhafid Boussouf – Mila Examinateurs: Mr. H. GHERROUCHA Prof à Univ. Frères.Mentouri – Constantine 1 Mr. L. SAHAB Prof à Univ. Oum El Bouaghi Mr. A.A. OULDJAOUI M.C. Univ. Oum El Bouaghi Soutenue le : 26/01/ 2017 00 /00/000 Année : 2016/2017 Je dédie ce modeste travail à, La mémoire de ma chère mère, Mon cher frère omar, Mes chères sœurs soulef et wafia, et à tous ceux qui me sont chers. Samira Remerciements Je re mercie tout d’abord Allah qui m’ a donné la santé et le courage pour terminer cette thèse. Je tiens à exprimer ma reconnaissance à ma directeur de thèse Pr. A. YAHIA pour la confiance qu'elle m'a témoignée, et il a constamment porté un regard ouvert et constructif sur mes travaux. Je remercie Prof. D. KHELIFI Directeur du Laboratoire de Biochimie Génétique et Biotechnologies Végétales (Université Frères Mentouri, Constantine 1) qui m'a honoré en acceptant d'être président de ce jury, je tiens à le reme rcier pour sa contribution à la réalisation de l’étude biochimique . Je tiens à présenter ma profonde gratitude et mes sincères remerciements aux membres de Jury Prof. H. GHERROUCHA, Prof. L. SAHAB et Dr. A.A. OULDJAOUI et d’avoir accepté d’examiner cette thèse. Ma reconnaissance va également à Mr. Chekara Bouziani Mohamed qui m’a énormément aidé dans la partie biochimique et m’a fait bénéficier de son expérience dans ce domaine sans hésitation. Que toute personne ayant participé de près ou de loin dan s l’élaboration de ce travail, trouve ici l’expression de mes très vifs remerciements (Ali, Sorya, Hanane, Sonia, Souad, Asma, Imene). Résumé L’objet de notre travail a porté sur l’étude de Teucrium polium L. Capitatum une plante médicinale très répandue dans les steppes algériennes. Plusieurs volets d’étude ont été touchés tels que l’aspect morphologique, les constituants physio-biochimiques, et les effets biologiques . Les analyses morphologiques ont porté sur dix populations échantillonnées dans différents biotopes de l’Est algérien. Cette étude a montrée que les feuilles ont le plus faible nombre (cinq sur dix) de types de trichomes, suivies par les calices et les corolles avec huit types chacun . Ce qui confirme l’absence d’une variabilité intraspécifique chez les populations examinées . D’autre part, l’analyse des relations entre les caractères morphologiques et les facteurs du milieu d’origine, ont permis de montrer que le biotope intervient sur la longueur des trichomes. L’utilisation de la technique d ’électrophorèse monodimensionnelle sur gel polyacrylamide en présence d ’un détergent (SDS-PAGE) a permis de déterminer les profils des protéines totales des dix populations. Les coefficients de similitude des bandes polypeptidiques obtenus montrent des similitudes élevées entre les populations étudiées. Les analyses biochimiques sur les trois phénotypes de Teucrium polium L. Capitatum ont montré que les extraits des phénotypes mauves sont les plus riches en anthocyanines, en sucres solubles et en caroténoïdes par rapport aux phénotypes rose et blanc mettant ainsi l’importance de la couleur de la fleur. La combinaison de certaines méthodes d’analyses chimiques; spectrophotométrie (UV) et chromatographique (CLHP) ont pe rmis d’identifier 3 flavonoïdes dont un majoritaire la cyanidine et deux autres flavonoïdes; la quercétine et l’acide caféique. L’évaluation du pouvoir antioxydant qui a été réalisée en utilisant les méthodes du piégeage du radical libre DPPH et celle de l a réduction du fer FRAP a indiqué que l’extrait de la population de Biskra a montré une meilleure capacité réductrice par rapport à celui de Sétif et un fort piégeage des radicaux DPPH● . En ce qui concerne l’activité antibactérienne, il apparait que les ex traits des deux populations possèdent des propriétés antibactériennes acceptables. L’évaluation de l’effet pharmacologique de Teucrium polium L. Capitatum a mis en évidence une propriété diurétique intéressante. Mots clés: Teucrium polium L. Capitatum, Lamiaceae, trichome, électrophorèse, CLHP, polyphenols, activité antioxydante, activité antibactérienne, effet diurétique. i ﻣﻠﺨﺺ ﺍﻟﻬﺪﻑ ﻣﻦ ﻫﺬﺍ ﺍﻟﺒﺤﺚ ﻫﻮ ﺩﺭﺍﺳﺔ ﻣﺮﻓﻮﻟﻮﺟﻴﺔ ،ﻛﻴﻤﻴﺎﺋﻴﺔ، ﻓﻴﺰﻳﻮﻟﻮﺟﻴﺔ ﻭ ﺑﻴﻮﻟﻮﺟﻴﺔ ﻟﻨﺒﺘﺔ ﻁﺒﻴﺔ ﻣﻌﺮﻭﻓﺔ ﻓﻲ ﺍﻟﺴﻬﻮﺏ ﺍﻟﺠﺰﺍﺋﺮﻳﺔ ﺑﺎﺳﻢ Teucrium polium L. Capitatum . ﺍﻟﺘﺤﺎﻟﻴﻞ ﺍﻟﻤﺮﻓﻮﻟﻮﺟﻴﺔ ﺗﻨﺎﻭﻟﺖ ﺑﺎﻟﺪﺭﺍﺳﺔ 10 ﻋﺸﺎﺋﺮ ﻣﻦ ﻣﻨﺎﻁﻖ ﻣﺨﺘﻠﻔﺔ ﻣﻦ ﺍﻟﺸﺮﻕ ﺍﻟﺠﺰﺍﺋﺮﻱ، ﺃﻭﺿﺤﺖ ﺍﻟﻨﺘﺎﺋﺞ ﺃﻥ ﺍﻷﻭﺭﺍﻕ ﺗﻤﺘﻠﻚ ﺃﻛﺒﺮ ﻋﺪﺩ ﻣﻦ ﺃﻧﻤﺎﻁ ﺍﻷﻭﺑﺎﺭ (5 ﻣﻦ 10 ﺃﻧﻤﺎﻁ ) ، ﻳﻠﻴﻬﺎ ﺍﻟﺴ ﻼﺒ ﺕ ﻭ ﺍﻟﺒﺘﻼﺕ ﺏ : 8 ﺃﻧﻤﺎﻁ ﻟﻜﻞ ﻋﻀﻮ، ﺍﻷﻣﺮ ﺍﻟﺬﻱ ﻳﺒﻴﻦ ﻏﻴﺎﺏ ﺍﻟﺘﻨﻮﻉ ﺗﺤﺖ ﺍﻟﻨﻮﻋﻲ ﺑﻴﻦ ﺍﻟﻌﺸﺎﺋﺮ ﺍﻟﻤﺪﺭﻭﺳﺔ ، ﻭﻣﻦ ﺟﻬﺔ ﺃﺧﺮﻯ، ﺗﺤﺎﻟﻴﻞ ﺍﻻﺭﺗﺒﺎﻁﺎﺕ ﺑﻴﻦ ﺍﻟﺨﺼﺎﺋﺺ ﺍﻟﻤﺮﻓﻮﻟﻮﺟﻴﺔ ﻭ ﺍﻟﻌﻮﺍﻣﻞ ﺍﻟﺒﻴﺌﻴﺔ ﺳﻤﺤﺖ ﺑﺘﺄﺛﻴﺮ ﻭﺳﻂ ﺍﻟﻤﻌﻴﺸﺔ ﻋﻠﻰ ﺃﻁﻮﺍﻝ ﺍﻷﻭﺑﺎﺭ . ﺍﺳﺘﺨﺪﺍﻡ ﺗﻘﻨﻴﺔ ﺍﻟﻔﺼﻞ ﺍﻟﻜﻬﺮﺑﺎﺋﻲ ﺃﺣﺎﺩﻳﺔ ﺍﻷﺑﻌﺎﺩ ﺑﻬﻼﻡ ﻣﺘﻌﺪﺩ ﺍﻷﻛﺮﻳﻼﻣﻴﺪ ﻓﻲ ﻭﺟﻮ ﺩ SDS-PAGE ، ﺳﻤﺤﺖ ﺑﺘﺤﺪﻳﺪ ﻣﻼﻣﺢ ﺍﻟﺒ ﺮﻭﺗﻴﻨﺎﺕ ﺍﻟﻜﻠﻴﺔ ﻟﻠﻌﺸﺎﺋﺮ ﺍﻟﻤﺪﺭﻭﺳﺔ ، ﺣﻴﺚ ﺟﺎءﺕ ﻧﺴﺒﺔ ﺍﻟﺘﺠﺎﻧﺲ ﻋﺎﻟﻴﺔ ﺑﻴﻦ ﻭﺣﺪﺍﺕ ﻣﺘﻌﺪﺩﺍﺕ ﺍﻟﺒﻴﺘﻴﺪ . ﺃﻣﺎ ﺍﻟﺘﺤﺎﻟﻴﻞ ﺍﻟﻔﻴﺰﻳﻮ ﻛﻴﻤﻴﺎﺋﻴﺔ ﻟﻸﻧﻤﺎﻁ ﺍﻟﻤﻈﻬﺮﻳﺔ ﺍﻟﺜﻼﺛﺔ ﻟﻨﺒﺘﺔ : Teucrium polium L. Capitatum ﺑﻴﻨﺖ ﺃﻥ ﻣﺴﺘﺨﻠﺼﺎﺕ ﺍﻟﻨﻤﻂ ﺍﻟﻤﻈﻬﺮﻱ ﺍﻟﺒﻨﻔﺴﺠﻲ ﻫﻲ ﺍﻷﻛﺜﺮ ﻭﻓﺮﺓ ﻣﻦ ﺍﻷﻧﺘﻮﺳﻴﺎﻧﻴﻦ ، ﺍﻟﺴﻜﺮﻳﺎﺕ ﺍﻟﻤﻨﺤﻠﺔ ﻭ ﺍﻟﻜﺎﺭﻭﺗﻴﻨﻮﻳﺪﺍﺕ ﻣﻘﺎﺭﻧﺔ ﻣﻊ ﺍﻟﻨﻤﻄﻴﻦ ﺍﻟﻤﻈﻬﺮﻳﻦ ﺍﻟﻮﺭﺩﻱ ﻭ ﺍﻷﺑﻴﺾ. ﺍﻟ ﺘﺪﺍﺧﻞ ﺑﻴﻦ ﻁﺮ ﻕ ﺍﻟﺘﺤﻠﻴﻞ ﺍﻟﻜﻴﻤﻴﺎﺋﻲ : ﺟﻬﺎﺯ ﺍﻟﻄﻴﻒ ﺍﻟﻀﻮﺋﻲ (UV-visible) ﻭ ﺍﻟﻜﺮﻭﻣﺎﺗﻮﻏﺮﺍﻓﻴﺎ ﺳﺎﺋﻠﺔ ﻋﺎﻟﻴﺔ ﺍﻷﺩﺍء(CLHP) ، ﻋﺪﺎﺳ ﻓﻲ ﺗﺤﺪﻳﺪ 3 ﻓﻼ ﻓﻮﻧﻮﻳﺪﺍﺕ ﻬﺎﺒﻏﻠﺃ cyanidine ، ﻭﺍﻟ ﻔﻼ ﻓﻮﻧﻮﻳﺪﺍﺕ ﺍﻟﻤﺘﺒﻘﻴﺔ ﻋﺒﺎﺭﺓ ﻋﻦ : quercétine ﻭ acide caféique . ﻓﻴﻤﺎ ﻳﺨﺺ ﺗﻘﺪﻳﺮ ﺍ ﻟﻔﻌﺎﻟﻴﺔ ﺍﻟﻤﻀﺎﺩﺓ ﻟﻸﻛﺴﺪﺓ ﺑﻄﺮﻳﻘﺘﻲ: ﻣﺤﺎﺻﺮﺓ ﺍﻟﺠﺬﺭ ﺍﻟﺤﺮ DPPH ﻭ ﺇﺭﺟﺎﻉ ﺍﻟﺤﺪﻳﺪ ( FRAP ) ﺃﻅﻬﺮﺕ ﺗﻔﻮﻕ ﻟﻤﺴﺘﺨﻠﺺ ﺍﻻﻳﺜﺎﻧﻮﻟﻲ ﻟﻨﺒﺎﺕ ﻣﻨﻄﻘﺔ ﺑﺴﻜﺮﺓ ﻋﻠﻰ ﻧﻈﻴﺮﻩ ﻣﻦ ﻣﻨﻄﻘﺔ ﺳﻄﻴﻒ . ﺃﻣﺎ ﺍﻟﻨﺸﺎﻁ ﺍﻟﻤﻀﺎﺩ ﻟﻠﺒﻜﺘﻴﺮﻳﺎ ، ﺃﺛﺒﺖ ﺃﻥ ﻛﻼ ﺍﻟﻤﺴﺘﺨﻠﺼﻴﻦ ﻟﻤﻨﻄﻘﺘﻲ ﺑﺴﻜﺮﺓ ﻭ ﺳﻄﻴﻒ ﻳﻤﻠﻜ ﺎﻥ ﺧﺼﺎﺋﺺ ﻣﻀﺎﺩﺓ ﻟﻠﺒﻜﺘﻴﺮﻳﺎ ﻣﻘﺒﻮﻟﺔ . ﻭ ﺃﺧﻴﺮﺍ ﺗﻢ ﺗﺜﻤﻴﻦ ﺍﻟﺪﻭﺭ ﺍﻟﺼﻴﺪﻻﻧﻲ ﻟﻠﻨﺒﺘﺔ ﻣﻦ ﺧﻼﻝ ﺇﻅﻬﺎﺭ ﺍﺩﺭﺍﺭﻫﺎ ﻟﻠﺒﻮﻝ . ﺍﻟﻜﻠﻤﺎﺕ ﺍﻟﻤﻔﺘﺎﺣﻴﺔ: Teucrium polium L. Capitatum ، ﺍﻟﻌﺎﺋﻠﺔ ﺍﻟﺸﻔﻮﻳﺔ ، ﺍﻷﻭﺑﺎﺭ ، ﺍﻟﻔﺼﻞ ﺍﻟﻜﻬﺮﺑﺎﺋﻲ، CLHP، ﺍﻟﻮﻅﻴﻔﺔ ﺍﻟﻤﻀﺎﺩﺓ ﻟ ﻸﻛﺴﺪﺓ، ﺍﻟﻮﻅﻴﻔﺔ ﺍﻟﻤﻀﺎﺩﺓ ﻟﻠﺒﻜﺘﻴﺮﻳﺎ ، ﻓﻌﻞ ﺍﺩﺭﺍﺭ ﺍﻟﺒﻮﻝ. ii ABSTRACT The purpose of our work has focused on the study of Teucrium polium L. Capitatum , which is a widespread medicinal plant in algerian steppes. Several study topics were investigated such as the morphology, physio-biochemical constituents and biological effects. The morphological analysis consisted of ten populations sampled at different habitats in eastern Algeria. This study revealed that the leaves have the lowest number (five out of ten) of types of trichomes, followed by the calyx and corolla with eight types for each. This confirms the absence of intraspecific variability within the study populations. Furthermore, the analysis of relationships between morphological characteristics and environmental factors of their habitat showed that the latter influences the length of trichomes. The use of one-dimensional electrophoresis technique on polyacrylamide gel in the presence of detergent (SDS-PAGE) allowed to determine the total protein profiles of the ten study populations. The similarity coefficients of polypeptide bands indicated high similarity between populations. Biochemical analyzes of the three phenotypes of T. polium L. Capitatum showed that extracts of purple phenotypes were richest in anthocyanins, soluble sugars and carotenoids compared with pink and white phenotypes enlightening there by the importance of flower color. The combination of certain chemical analysis methods, spectrophotometry (UV-visible) and chromatographic (HPLC) have identified three flavonoids including mostly the cyanidin and two other flavonoids, quercetin and caffeic acid. The evaluation of the antioxidant potency, that was conducted using the method of trapping the free radical DPPH and that of iron reduction FRAP, revealed that the extract of Biskra population showed higher reducing capacity compared to that of Sétif, with a strong trapping of DPPH● radicals. As regards to the antibacterial activity, it appears that extracts from two populations have acceptable antibacterial properties. The pharmacological effect of T. polium L. Capitatum showed an interesting diuretic property. Keywords: Teucrium polium L. Capitatum , Lamiaceae, trichomes, electrophoresis, HPLC, polyphenols, antioxidant activity, antibacterial activity, diuretic effect. iii Table des matières Résumés ……………………………………………………………………………………… i Table des matières …………………………………………………………………………. .. iv Liste des tableaux …...……………………………………………………………………… viii Liste des planches …………………………………………………………………………… ix Liste des figures ………………………………………………………………………… .…. x Liste des abréviations ………..……………………………………………………...….... .... xii Liste des annexes …………………………………………………………………………… . xiii Introduction générale ……………………………………………………………………… 1 Chapitre 1 : Recherche bibliographique I. Etude ethnobotanique de la plante Teucrium polium L. Capitatum …………………. 3 Introduction …………………………………………………………………………………. 3 1. Caractères morphologiques de Teucrium polium ssp. Capitatum ………………………... 3 2. Taxonomie et systématique………………………………………………………………. 4 3. Habitat et répartition géographique ………………………………………………………
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