ARISTOTLE UNIVERSITY OF THESSALONIKI FACULTY OF SCIENCES, SCHOOL OF BIOLOGY DEPARTMENT OF BOTANY SHARAF MUSA AL-TARDEH B. Sc. Biological Sciences M.Sc. Food Quality Management MORPHOLOGICAL AND ANATOMICAL ADAPTATIONS OF THE PERENNIAL GEOPHYTE URGINEA MARITIMA (L.) BAKER (LILIACEAE) TO THE MEDITERRANEAN CLIMATE DOCTORAL DISSERTATION Thessaloniki 2008 SHARAF MUSA AL-TARDEH B. Sc. Biological Sciences M.Sc. Food Quality Management MORPHOLOGICAL AND ANATOMICAL ADAPTATIONS OF THE PERENNIAL GEOPHYTE URGINEA MARITIMA (L.) BAKER (LILIACEAE) TO THE MEDITERRANEAN CLIMATE DOCTORAL DISSERTATION Submitted to the School of Biology, Faculty of Sciences, Aristotle University of Thessaloniki Presented for public criticism on the 28th of November 2008 MORPHOLOGICAL AND ANATOMICAL ADAPTATIONS OF THE PERENNIAL GEOPHYTE URGINEA MARITIMA (L.) BAKER (LILIACEAE) TO THE MEDITERRANEAN CLIMATE Figures on the cover page: (A) TEM micrograph of nectary of Urginea maritima showing the nectary epithelium cell with nectar in crystallized form at the old stage; (B) Photo of inflorescence stalk; (C) SEM micrograph of anther; (D) LM micrograph of leaf with parallel veins and veinules; (E) TEM micrograph of leaf with sieve pore between the sieve element; (F) LM micrograph of bulb scale revealing cells stained red with Schiff’s reagent; (G) LM micrograph revealing cells stained brown-black with Sudan Black B; (H) LM micrograph showing idioblastic cortical cells with bundles of raphides and mucilage stained with TBO; (I) LM micrograph of root showing the 10-arch vascular cylinder; (J) LM micrograph of root showing series of idioblastic cells with bundles of raphides. To my family To my lovely wife “WOROD” To those who defend their rights SHARAF ELABORATION OF THE DOCTORAL DISSERTATION Place Aristotle University of Thessaloniki, Faculty of Sciences, School of Biology Department of Botany Time February 2004 – November 2008. Examining committe • Stylianos Delivopoulos, Professor, Department of Botany, School of Biology, A.U.Th. Supervisor • Thomas Sawidis, Associate Professor, Department of Botany, School of Biology, A.U.Th. Cosupervisor • Barbara-Evelin Diannelidis, Assistant Professor, Department of Botany, School of Biology, A.U.Th. Cosupervisor • Bosabalidis Artemios, Professor, Department of Botany, School of Biology, A.U.Th. Examiner • Eleftheriou Eleftherios, Professor, Department of Botany, School of Biology, A.U.Th. Examiner • Moustakas Michael, Associate Professor, Department of Botany, School of Biology, A.U.Th. Examiner • Pantis John, Associate Professor, Department of Ecology, School of Biology, A.U.Th. Examiner i © Sharaf Musa Al-Tardeh © AUTh. Morphological and anatomical adaptations of the perennial geophyte Urginea maritima (L.) Baker (Liliaceae) to the mediterranean climate. ISBN Approval of the present Doctoral Dissertation by the Department of Botany, School of Biology, Aristotle University of Thessaloniki does not imply the acceptance of the opinions of the author (N. 5343/1932, law 202, paragraph 2). ii Sharaf Al-Tardeh Table of contents TABLE OF CONTENTS ELABORATION OF THE DOCTORAL DISSERTATION……………………....... i TABLE OF CONTENTS…………………………………………………………...... iii PROLEGOMENA ……...………………………………………………………......... vii LIST OF ORIGINAL ARTICLES……….………....................................................... ix LIST OF ABBREVIATIONS AND SYMBOLS………………………………..…… x SUMMARY...………………………………………………………..…………......... xii ΠΕΡΙΛHΨΗ…………………………………………………………………………... xvi CHAPTER I: INTRODUCTION…………...……………………………………… 1 1.1 Mediterranean climate……………………………………………………………. 1 1.1.1 Introduction…………………...…………………………………………….… 1 1.1.2 Spatial patterns of temperature in the Mediterranean climate…………….…... 1 1.1.3 Spatial patterns of precipitation in the Mediterranean climate………………... 2 1.2 Geophytes……………………………………………………………………….... 3 1.2.1 Introduction…………………………………………………………………… 3 1.2.2 Taxonomy and origin of geophytes…………………………………………... 3 1.2.3 Diversity of geophytes………………………………………………………... 3 1. 2.3.1 Bulbous plants………………………………………………………......... 4 1.2.3.2 Tuberous plants…………………………………………………………… 4 1.2.4 The body of geophytes……………………………………………………...... 5 1. 2.4.1 Storage organs and tissues…………….…………………………………. 5 1.2.4.2 Aerial organs and tissues………..………………………………………… 6 1. 2.4.3 Roots……………………………………………………………………… 6 1.2.4.4 Tunics…………………………………………..…………………………. 7 1.2.5 Periodicity and dormancy of flower bulbs…………………………………….. 7 1.2.5.1 Definitions…………………………………………………………………. 7 1.2.5.2 Factors that affect bulb dormancy…………………………………………. 8 1.2.6 The flowering process……………………………………………………......... 9 1.2.7 Types of hysteranthous geophytes…………………………………………….. 9 1.2.8 Factors affecting bulb growth and development……………………………… 10 1.2.8.1 The size of the bulb……………………………………………………....... 10 1.2.8.2 Temperature………………………………………………………………... 10 1.2.8.3 Light……………………………………………………………………….. 11 1.2.8.4 Soil features……………………………………………………………....... 11 1.2.9 World production of geophytes……………………………………………..... 12 1.3 Chlorophyll contents and fluorescence…………………………………………... 13 1.4 Nectaries and nectar…………………………………………………………….... 17 1.4.1 Introduction…………………………………………………………………… 17 1.4.2 Definitions of nectary…………………………………………………………. 17 1.4.3 Basic types of floral nectaries.…..…………………………………………….. 18 ii Sharaf Al-Tardeh Table of contents 1.4.4 Floral and extrafloral nectaries………………………………………………... 20 1.4.5 Gynopleural (septal) nectaries……………………………………………….... 20 1.4.5.1 Distribution of the gynopleural nectaries………………………………….. 20 1 4.5.2 Structure of the gynopleural nectaries…………………………………....... 21 1.4.5.3 Nectar secretion in the gynopleural nectaries…………………………….... 22 1.4.5.4 Fate of the gynopleural nectaries…………………………………………... 22 1.4.6 Nectar production and presentation…………………………………………… 23 1.4.6.1 Nectar secretion mechanism…………………………….............................. 23 1. 4.6.2 Nectar presentation………………………………………………………... 24 1.4.6.3 Fate of nectar and nectarines………………………………………………. 26 1.5 The summer flowering geophyte: Urginea maritima (L.) Baker (Liliaceae)…...... 27 1.5.1 Urginea maritima overview…………………………………………………... 27 1.5.2 Morphology…………………………………………………………………… 27 1.5.3 Growth, development and flowering……………………………………….… 28 1.5.4 Nectary of U. maritima……………………………………………………….. 29 1.5.5 Pollination in U. maritima as a model of hysteranthous geophytes………....... 29 1.5.6 Bulb production and/or propagation………………………………………….. 30 1.5.7 Horticultural usage……………………………………………………………. 32 1.5.8 Biochemical principles and their action………………………………………. 32 1.5.8.1 Toxic principle……………………………………………………………... 32 1.5.8.2 Defense mechanisms………………………………………………………. 33 1.5.8.3 Pharmaceutical advantages of U. maritima……………………………….. 34 1.6 Aim of the thesis…………………………………………………………………. 35 CHAPTER II: MATERIALS AND METHODS ……………………………….... 37 2.1 Materials………………………………………………………………………….. 37 2.1.1 Plant samples………………………………………………….……………..... 37 2.1.2 Apparatuses……….…………………………………………...……………… 38 2.1.3 Experimental design…………………………………………………………... 40 2.2 Methods…………………………………………………………………………… 42 2.2.1 Water content………………………………………………………………….. 42 2.2.2 Fixation………………………………………………………………………... 43 2.2.2.1 Solution preparations………………………………………………………. 43 2.2.2.2 Procedure…………………………………………………………………... 44 2.2.3 Sectioning……………………………………………………………………... 45 2.2.4 Staining techniques…………………………………………………………… 46 2.2.4.1 Staining with toluidine blue O (0.1%)…………………………………….. 46 2.2.4.2 Staining with Sudan Black B (1%)………………………………………... 46 2.2.4.3 Staining with Periodic acid-Schiff’s reaction (PAS reaction)…………….. 47 2. 2.4.4 Staining with uranyl acetate and lead citrate…………………………….. 48 2. 2.5 Scanning Electron Microscopy (SEM)……………………………………….. 48 iii Sharaf Al-Tardeh Table of contents 2.2.6 Maceration…………………………………………………………………….. 49 2.2.6.1 Maceration of soft material………………………………………………... 49 2.6.2 Maceration of hard material…………………………………...…………….. 50 2.2.7 Leaf chlorophyll assay………………………………………………………... 50 2.2.8 Chlorophyll a fluorescence…………….……………………………………… 50 2.2.9 Morphometry…………………………………………………………………. 52 2. 2.10 Mathematical analysis of morphological and anatomical features of the plant tissues………………………………………………………………….... 52 2.3 Statistical analysis………………………………………………………………... 52 CHAPTER III: RESULTS ………………..……………………………………….. 53 3.1 ANATOMICAL STUDIES ON THE ADVENTITIOUS ROOTS……………... 53 3.1.1 Root morphology ………………………………………………………..……. 53 3.1.2 Root anatomy………………………………………………………………….. 53 3.1.3 Root histochemistry…………………………………………………………… 56 3.1.4 Root morphometry……………………………………………………………. 56 3.2 Biomass and reserve allocation patterns within the bulb………………………… 58 3.2.1 Bulb morphology……………………………………………………………… 58 3.2.2 Bulb anatomy…………………………………………………………………. 58 3.2.3 Bulb histochemistry…………………………………………………………… 60 3.2.4 Bulb morphometry and reserves allocation patterns………………………….. 61 3.2.5 Biomass allocation patterns and water status…………………………………. 62 3.3 MORPHO-ANATOMICAL FEATURES OF THE LEAVES…………………. 65 3.3.1 Leaf anatomy………………………………………………………………….. 65 3.3.2 Leaf morphology and morphometry…………………………………………... 67 3.3.3 Leaf histochemistry…………………………………………………………..... 69 3.3.4 Leaf chlorophyll assay……………………………………………………….... 69 3.3.5 Chlorophyll a fluorescence……………………………………………………. 70 3.4 MORPHO-ANATOMY OF THE INFLORESCENCE…………………………. 73 3.4.1 Flower morphology……………………………………………………………. 73 3.4.2 Anatomy and development of the gynopleural (septal) nectary………………. 74 3.4.2.1 Young stage nectary……………………………………………………….. 74 3.4.2.2 Nectary at the intermediate stage…………………………………………... 75 3.4.2.3 Nectary at old stage………………………………………………………...