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Ÿþa N N E X E FACULTE DES SCIENCES DE LA NATURE ET DE LA VIE DEPARTEMENT DE BIOLOGIE THÈSE Présentée par Mme DAHANE Née ROUISSAT Lineda En vue de l’obtention du DOCTORAT EN SCIENCES BIOLOGIQUES Spécialité : Biochimie végétale appliquée. Thème : Etude des effets nématicides et molluscicides des extraits de quelques plantes sahariennes. Soutenue le : 21 Décembre 2017 Devant le jury composé de : Mr HADJADJ- AOUL Seghir, Prof. Université Oran1 ABB Président Mr BELKHODJA Moulay, Prof. UniversitéOran1, ABB Examinateur Mme BENNACEUR Malika, Prof. Université Oran1, ABB Examinatrice Mr MEKHALDI Abdelkader Prof. Université de Mostaganem, Examinateur Mr. Marouf Abderrazak. Prof. Centre. Univ. Naama Directeur de thèse Mr. Cheriti Abdelkrim Prof. Université de Béchar. Co-directeur de thèse 2016-2017 RESUME Dans le présent travail, les parties aériennes de vingt et une plantes sahariennes (21) des différentes familles botaniques (Asteraceae ; Amaranthaceae ; Rhamnaceae ; Brassicaceae ; Plumbaginaceae ; Capparidaceae ; Caryophyllaceae ; Fabaceae ; Apocynaceae ; Solanaceae ; Verbenaceae et Euphorbiacaeae) ont été utilisées pour évaluer leurs extraits aqueux (par macération ou à reflux) et les extraits organiques (acétoniques et méthanoliques avec ces fractions : hexanique, éthérique, dichlorométanolique, chloroformique, butanoliques…) pour l’activité nématicide (vis-à-vis nématodes phytoparasites à kyste : Globodera sp. et Heterodera sp. et molluscicide (vis-à-vis aux mollusques d’eau douce transporteurs des parasites : Lymnaea acumunata et Bulinus truncatus ). Les résultats sont exprimées en LC50 (taux de mortalité est égale à 50% de la population testée) par l’analyse des probits. Après l’extraction et le criblage phytochimique des extraits, l’évaluation a été réalisée sous des conditions expérimentales convenables aux cycles de vie de chaque spécimen zoologique (Température 24°C avec l’humidité et l’aération). Nos résultats ont montré des valeurs en LC50 intéressantes au regard des durées d’exposition (de 24 h à 96 h) au dessous des valeurs en LC50 des témoins positifs (produits nématicide et molluscicide commercialisés). Les taux des rendements les plus importants sont aperçus aux macérats de Warionea saharae (40.61 %) et Moricandia arvensis (L.)DC. (34.49 %), tandis que pour les fractions organiques, la valeur maximale en taux de rendement est de 4%, a été enregistrée pour la fraction dichlorométhanique d’Asteriscus graveolens (Forsk.)Less subsp. graveolens. Le criblage phytochimique a mis en évidence la présence de saponosides, flavonides et tannins, chez toutes les plantes étudiées. Les résultats de l’activité nématicide des extraits aqueux des plantes d’étude ont montrés une très forte activité vis-à-vis Globodera sp. avec les décoctés des plantes : Haloxylon articulatum après 72h , LC50 = 16,354 mg/ml et le macérat de la même plante a divulgué une activité importante avec LC50 : 10,379 µg/ml mais après 96h d’exposition des kystes. En revanche pour les extraits organiques, les valeurs sont très inférieurs au témoin positif, autant que la fraction dichlorométhane de Lantana camara (LC50 :0,098 μg/mL) présente la valeur minimale après 96h d’exposition. Autrement, après 24h d’exposition aux fractions, la valeur minimale en LC50 : 0,166 μg/mL a été signalée pour la fraction d’acétate d’éthyle d’Euphorbia retusa. Tandis que pour Heterodera sp. , les fractions organiques les plus actives sont acétate d’éthyle d’ Anvillea garcini subsp. radiata (Coss. & Dur.) LC50=0.96 µg/ml après 48 heures. Les résultats obtenus de l’activité molluscicide montrent la richesse des extraits en métabolites responsables de la mortalité des mollusques d’eau douce (Bulinus et Lymnaea), le contrôle positif (LIMACIDE PRO) a révélé une valeur en LC50=36,148 mg/ml, alors qu’il existe des valeurs en LC50 très inférieurs pour le macérat de Launaea nudicaulis (L) Hook f. après seulement 24 heures d’exposition (LC50=8,178 mg/ml), ce qui signifie une très bonne activité molluscicide de cet extrait. L’activité nématicide des extraits aqueux a révélé un fort effet ovicide et larvicide de quelques plantes testés durant une période d’exposition (de 24h à 96 heures) in vitro, ces dernières ont été confirmées In vivo, avec quelques extraits aqueux les plus active vis-à-vis aux kystes d’Heterodera pour les cultures de blé. Mots clés : Plantes sahariennes, Macération, Reflux, Phytochimie, Nématicide, Molluscicide, Bulinus , Lymnaea, Globodera , Heterodera. المــــلــــــخــــص من خﻻل هذا العمل، قمنا باستخدام اﻷجزاء الهوائية لواحد و عشرين نبتة صحراوية )21(، من عائﻻت نباتية مختلفة )استراسيا، سرمقية، بذنجانية، لويزية، كباريدية، بﻻمباجينية، قرنفلية، بقولية، أسكلوبياداسية ( و ذلك من أجل تـــقــييــم المستخلصات المائية )عن طريق النقع أو الجزر( باﻹضافة إلى استعمال المستخلصات العضوية )اﻷستون، الميثانول، و أجزاؤها: الهكسان، اﻷثيري، ثنائي كلورو ميثان، الكلوروفورم، البيتانول( لنشاط مضاد الديدان الطفيلية الكيسية )النيماتسيد( Globoderaو Heterodera و مضاد للرخويات )ناقﻻت الطفيليات الموجودة في المياه العذبة: Lymnaea acuminataو Bulinus truncatus(. و قد تم التعبير عن النتائج LC50 )معدل وفيات 50% من العينات المختبرة(، من خﻻل تحليل اﻻحتمالية. بعد اﻻستخﻻص و الفحص الكيميائي للمستخلصات النباتية، أجرينا تقييما لنشاطها في ظل الظروف التجريبية المﻻئمة لدورة حياة كل عينة من الحيوانات المدروسة )درجة الحرارة المﻻئمة 24 °م و الرطوبة و التهوية(. و قد أظهرت النتائج قــيــما LC50 مهمة بعد استعمال المستخلصات لمدة 24 ساعة إلى 96 ساعة ، حيث أن هذه القيم أقل من قيمة LC50 للشواهد اﻻيجابية )المنتجات المسوقة موليسيسيدات و النيماتيسيدات(. سجل أعلى معدل للعوائد لمنقوع Warionea saharea)40.61%( و (Moricandia arvensis (34.49% ، بينما سجل أعلى معدل لعوائد المستخلصات العضوية لجزء ثـــنائي كلـــوروميثان لنبتــة Bubonium graveolens بقيمة 4% ، كما أظهر الفحص الكيميائي الـــــــنباتي وجـــــود الصابونين، الفﻻفونويدات و العفص عند أغلبية النباتات المدروسة. كما أظهرت نتائج جيدة للـــنشـــاط المضاد للديدان لمستخلصات المائية للنباتات المدروسة )بنسب كبيرة جدا ( ضــــــد Globoderaباستعمال منقوع النبتة Haloxylon articulatum بعد 72 ساعة كـــــانـــت قــــيمـــــتها LC50 =16.354mg/ml باﻹضافة إلى أصغر قيمة سجلت لنفس النبتة ولكن بــطريقـة الجزر ، سجلت LC50=10.379 mg/ml بعد 96 ساعة من التعرض للمستخلص ، خﻻفا لذلك سجلت أدنى قيمة LC50=0.098mg/ml و هو الحد اﻷدنى اﻷقل لنبتة Lantana camaraمن خﻻل مستخلص عضوي ثنائي الكلوروميثان . من ناحية أخرى، و بعد 24 ساعة من التـــــعـرض للمستخلصات العضوية، سجلت قيمة LC50=0.0166µg/ml لجزء اﻷسيتات اﻻثيل لـــنـــبتــة Euphorbia retusa المضاد ل Globodera. بينما ل Heterodera، فان جزء اﻷسيتات اﻻثيل المؤثر لنبتة LC50=0.96µg/ml ( Anvillea radiata( قــــضـى عليها بعد 48 ساعة . أظهرت النتائج المتحصل عليها لنشاط المضاد للرخويات )Molluscicide( ثرائـــه بالمستخلصات اﻷيضية المسؤولة عن وفيات رخويات المياه العذبة ) Bulinusو Lymnaea( حيث كشفت العينة اﻻيجابية )LIMACIDE PRO( المسؤولة عن وفيات قواقع المياه العذبة ) Bulinusو Lymnaea( على قيمة LC50=36.148mg/ml ، بينما توجد قيمة أقل منها بكثير لمنقوع نبتة LC50=8.178 mg/ml( Launaea nudicaulis(، و هذا يدل على النشاط الجيد لهذا المستخلص . أظهر النشاط المضاد للديدان الطفيلية الكيسية )nématicide( للمستخلصات المائية نشاطا مثبطا للبيوض و اليرقات، اختبرناها خﻻل فترة التعرض 24 ساعة إلى 96 ساعة في المختبر . و تأكدت هذه النتائـــج بمكان نـــموها الطبيعي بإضافة بعض المستخلصات المائية اﻷكثر نشاطا بالنسبة ل Heterodera في مزارع تجريبىة للقمح. الكلمات المفتاحية : النباتات الصحراوية، النقاعة، الجزر، النشاط المضاد للديدان الطفيلية الكــــيسية ،Bulinus ، Lymnaea، Globodera )النـــــيماتيسيد(، مضاد للرخويات )الموليسيسيد(، Heterodera Abstract In this work, We yoused aerial parts of twenty-one Saharan plants (21) from the different botanical families (Asteraceae, Amaranthaceae , Rhamnaceae, Brassicaceae, Plumbaginaceae, Capparidaceae, Caryophyllaceae, Fabaceae, Apocynaceae, Solanaceae, Verbenaceae and Euphorbiacaeae) To evaluate their aqueous extracts (by maceration or at reflux) and the organic extracts (acetonic and methanolic with these fractions: hexanic, etheric, dichlorometanolic, chloroformic, butanolic, etc.) for nematicidal activity (against nematodes phytoparasites with cyst: Globodera sp., and Heterodera spp.), and molluscicidal activity (opposite to freshwater molluscs carrying parasites: Lymnaea acumunata and Bulinus truncatus) The results are expressed in LC50 (mortality rate is equal to 50% of the population Tested) by probit analysis. After extracting and phytochemical screening of extracts, Evaluation was carried out under experimental conditions suitable for the life cycles of each zoological specimen (temperature 24 °C with humidity and aeration). Our results showed interesting LC50 values with respect to exposure times (24 h to 96 h) below the LC50 values of the positive controls (nematicide and molluscicide products marketed). The highest rates of yield are seen in the macerates of Warionea saharae (40.61%) and Moricandia arvensis (L.)DC. (34.49%), while for organic fractions, the maximum value in terms of yield is 4%, was recorded for fraction Dichloromethane of Asteriscus graveolens (Forsk.) Less subsp. graveolens. The phytochemical screening revealed the presence of saponosides, flavonides and tannins in all the plants studied. The results of the nematicidal activity of the aqueous extracts of the study plants showed a very high activity against Globodera sp. With the plant decocts: Haloxylon articulatum after 72h, LC50 = 16,354 μg / ml and the macerate of the same plant disclosed a significant activity with LC50: 10,379 μg / ml but after 96h of exposure of the cysts. For organic extracts, the values are much lower than the positive control, as much as Lantana camara dichloromethane fraction
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