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Agricultural University of Athens Department of Crop Science Laboratory of Pesticide Science AGRICULTURAL UNIVERSITY OF ATHENS DEPARTMENT OF CROP SCIENCE LABORATORY OF PESTICIDE SCIENCE EVALUATION OF BIOACTIVITY OF PHYTOTOXINS FROM PATHOGENIC FUNGI OF OROBANCHE SP SUH CHRISTOPHER Ph.D THESIS Athens March 2011 Ph.D THESIS EVALUATION OF BIOACTIVITY OF PHYTOTOXINS FROM PATHOGENIC FUNGI OF OROBANCHE SP SUH CHRISTOPHER Supervisor: Prof. Maria Chrysayi Three member supervisory committee Prof. Maria Chrysayi Prof. Moschos Polissiou Asst. Prof Anastasios Markoglou Seven member examining committee Prof. Maria Chrysayi – Agricultural University of Athens Prof. Moschos Polissiou- Agricultural University of Athens Asst. Prof Anastasios Markoglou - Agricultural University of Athens Prof. Panagiotis Efthimiadis - Agricultural University of Athens Prof. Konstantinou Violeta- Agricultural University of Athens Prof. George Vassiliou - Demokritos University of Thrace Dr. Nikolaos Fokialakis- National and Kapodistrian University of Athens Athens March 2011 Η παρούσα διατριβή εκπονήθηκε με υποτροφία από το Ίδρυμα Κρατικών Υποτροφιών (ΙΚΥ) «Η έγκριση της διδακτορικής διατριβής από τη Γενική Συνέλευση του Τμήματος Επιστήμης Φυτικής Παραγωγής του Γεωπονικού Πανεπιστημίου Αθηνών, δεν υποδηλώνει αποδοχή των γνωμών του συγγραφέα» Νόμος 5343/32, άρθρο. 202§2 APPRECIATION It is with pleasure that I show appreciation to all those who have contributed in one way or the other to see me through this uphill task. First and foremost, I am grateful to God almighty for his love and protection throughout my studies. I am especially grateful to my supervisor, Prof. Maria Chrysayi for her relentless efforts to see that this work is done. In fact her motherly role in encouraging me gave me the courage not to give up even when the road was rough. Prof, I am really grateful. Thanks also go to the members of my supervising committee Prof. Polissiou Moschos and Asst. Prof Anastasios Markoglou for their useful contributions in shaping this thesis. To Dr. Nikolas Fokialakis, I lack words to express my appreciation to him for his valuable contributions to this work. I am also grateful to members of the examining panel for their useful contributions. I cannot forget to give thanks to the government of Greece through the state scholarship foundation, IKY for granting me a scholarship to undertake this research. I wish to express my sincere appreciation to Dr. Kostas Aliferis, Prof. Konstantinou Violeta and Ms. Georgia Paziotou for their scientific contributions during my research. I am grateful to all the staff and students of the laboratory of Pesticide science, Agricultural university of Athens for their various assistances to me during my research. It is worthwhile also thanking the staff and students (especially Job Tchoumtchoua and Zéphyrin Djiogue) of the laboratory of Pharmacognosy of the faculty of pharmacy, Kapodistrian University of Athens for their scientific contribution. I am also grateful to Dr. Malandraki Anastasios for assisting me in the computer aspects of this work. When I couldn`t really support my family economically during my studies, the following stood in for me: H.E. Mr. Angelos Papadopoulos, the Hon. Consul General of Cameroon in Greece; Aliki Kiritsi; Maria Kiritsi and Konstantina Kiritsi. I say a big thank you to you all. To all my Cameroonian friends in Greece, who encouraged me during the difficult periods, I say God bless you all. Thanks also go to my sister Winifred Sirri and the husband George Mbaku for taking over my responsibilities of caring for my mother. To my beloved mother, I am highly indebted to you for your prayers and encouragement. I am also thanking all my siblings and relations who depend on me, for their understanding. Finally I wish to express my gratitude to my dear wife, Lillian Suh and my children Suh Chrysayis and Suh Theokleai for understanding with me even when I deprived them of the love I owe them because of my studies. DEDICATION To GOD ALMIGHTY for giving me the perseverance to carry this work to the end in spite of all the hurdles. 7 TABLE OF CONTENTS LIST OF ABBREVIATIONS ................................................................................................. 11 Περίληψη Διδακτορικής Διατριβής του ............................................................................. 12 ΕΙΣΑΓΩΓΗ ............................................................................................................................. 12 1. Γενικά περί Οροβάγχης....................................................................................................... 12 2. Βιολογία της Orobanche crenata ...................................................................................... 12 3. Μέθοδοι αντιμετώπισης οροβάγχης ................................................................................... 12 4. Σκοπός της παρούσης έρευνας: .......................................................................................... 13 5.ΥΛΙΚΑ ΚΑΙ ΜΕΘΟΔΟΙ: .................................................................................................... 13 Υλικά ...................................................................................................................................... 13 Μέθοδοι .................................................................................................................................. 14 Καλλιέργεια μυκήτων και οροβάγχης – θρεπτικά υλικά .................................................. 14 Δοκιμές παθογένεσης ............................................................................................................ 14 Βιοδοκιμές φυτοτοξικότητας ............................................................................................... 15 Δοκιμές παθογένεσης ............................................................................................................ 15 Εκχύλιση και χημικός προσδιορισμός των μεταβολιτών .................................................. 16 Βιοδοκιμές φυτοτοξικότητας ............................................................................................... 16 SUMMARY ............................................................................................................................ 18 CHAPTER 1. .......................................................................................................................... 19 GENERAL INTRODUCTION ........................................................................................... 19 1.1. DESCRIPTION OF OROBANCHE SPECIES ................................................................ 19 1.2 ORIGIN & DISTRIBUTION OF OROBANCHE ............................................................. 23 1.3 BIOLOGY OF OROBANCHE CRENATA. ...................................................................... 24 1.4 ECONOMIC ASPECTS OF OROBANCHE INFESTATION ........................................ 26 1.5 In vitro GROWTH OF OROBANCHE SPECIES ........................................................... 27 1.6 GERMINATION STIMULATION AND ATTACHMENT ............................................. 29 1.7. CONTROL METHODS OF OROBANCHE CRENATA AND THE CONSTRAINTS. ...................................................................................................................................................... 31 1.7 .a. Preventive control. .......................................................................................... 32 1.7. b. Biological control............................................................................................ 32 1.7. b.1. Insect as control agents. ............................................................................ 33 1.7. b.2. Breeding for resistant crop varieties. ........................................................ 34 1.7.b.3. Allelopathy. ................................................................................................ 35 1.7.b.4. Microbial agents (in general). ................................................................... 36 1.7.b.5. Microbial agents (Orobanche species). ...................................................... 39 1.7.c. Cultural control method. ................................................................................... 39 1.7. d. Chemical methods. ........................................................................................... 46 1.8 FUNGI IN BIOLOGICAL CONTROL .............................................................................. 47 1.9 FUSARIUM IN OROBANCHE CONTROL ..................................................................... 50 1.10 FUSARIUM PHYTOTOXINS .......................................................................................... 51 8 1.11. MODE OF ACTION OF PHYTOTOXINS .................................................................... 54 9. Scirpenol derivatives. .......................................................................................................... 69 1.12 TESTING CHEMICALS ON OROBANCHE ................................................................. 76 1.12. a. Seed germination test. ................................................................................... 76 1.12.b. Leaf puncture test. ........................................................................................... 76 1.12.c. In vitro test. ..................................................................................................... 76 1.12.d. field test. ......................................................................................................... 77 1.13 IDENTIFYING FUSARIUM METABOLITES .............................................................
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