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Saurashtra University Re – Accredited Grade ‘B’ by NAAC (CGPA 2.93) Saurashtra University Re – Accredited Grade ‘B’ by NAAC (CGPA 2.93) Maharshi, Anjisha R., 2012, “Studies on Phytohormone Producing Plant Pathogenic Fungi”, thesis PhD, Saurashtra University http://etheses.saurashtrauniversity.edu/id/eprint/575 Copyright and moral rights for this thesis are retained by the author A copy can be downloaded for personal non-commercial research or study, without prior permission or charge. This thesis cannot be reproduced or quoted extensively from without first obtaining permission in writing from the Author. The content must not be changed in any way or sold commercially in any format or medium without the formal permission of the Author When referring to this work, full bibliographic details including the author, title, awarding institution and date of the thesis must be given. Saurashtra University Theses Service http://etheses.saurashtrauniversity.edu [email protected] © The Author Acknowledgments This thesis arose in part out of years of research that has been done since I came to PPMB Lab. By that time, I have worked with a great number of people whose contribution in assorted ways to the research and the making of the thesis deserved special mention. It is a pleasure to convey my gratitude to them all in my humble acknowledgment. In the first place I would like to record my gratitude to Prof. Vrinda. S. Thaker for her supervision, advice, and guidance from the very early stage of this research as well as giving me extraordinary experiences throughout the work. Above all and the most needed, she provided me unflinching encouragement and support in various ways which made her a backbone of this research and so to this thesis. Her involvement with her originality has triggered and nourished my intellectual maturity that I will benefit from, for a long time to come. Her truly scientist intuition has made her as a constant oasis of ideas and passions in science, which exceptionally inspire and enrich my growth as a student, a researcher and a scientist want to be. I am indebted to her more than she knows. I gratefully thank to head of the Department of Biosciences Prof. S. P. Singh for providing me all the necessary laboratory facilities. I am also very indebted to other faculty members and nonteaching staff of Department of Biosciences for their help in direct or indirect manner. It is a pleasure to pay tribute also to the sample collaborators. I would like to thank to the Institute of Microbial Technology (IMTECH), Chandigarh, India, for providing me the fungal samples. I would also acknowledge to Vimal Research Society for Agro-biotech and Cosmic Powers (VIRSACO), Rajkot, Gujarat, India, for providing me the samples of essential oils. I convey special acknowledgement to Dr. Manish L. Vekaria (Managing Director, VIRSACO) and Dr. Virendra J. Patel (Director, VIRSACO) for endowing me with all the necessary laboratory facilities and instrument facilities at VIRSACO. I wish to acknowledge University Grant Commission (UGC), New Delhi, India for financial support as meritorious scholar, also thanks to Centre For advanced Studies in Plant Biotechnology & Genetic Engineering (CPBGE), state Government of Gujarat for providing lab facility. I convey special thanks to my rabbits who might have suffered a bit to accomplish my work; I also thank Jivanbhai for taking care of my rabbits. I thank Satyendarbhai, Govindbhai and Joshibhai for their help. I am also thankful to Saurabhbhai, Nitheshbhai and Purohitbhai for their all-time helping hand. Collective and individual acknowledgments are also owed to my colleagues Mr. Lalit Chariya whose present somehow perpetually refreshed, helpful, and memorable. I am also thankful to Ms. Kiran Chudasama for her company and the kind help. My special thanks go to my senior colleagues Dr. Snehal Bhagtaria, Dr. Vaishali Pawar, Dr. Rita Chudasama, Dr. Kunjal Bhatt, Dr. Madhavi Joshi, Dr. Rohan Pandya and Mr. Anil Nakum for their guidance and support. I am also grateful to all my junior colleagues especially Mr. Viral Mandaliya for all his skilful assistance and carefully reading my thesis script. I thank to Ms. Vibhuti Jhala and Ms. Dipika Kalariya for managing bioinformatics software for my work. I also thank to Bhavisha Sheth, Khyati Bhatt, Pooja Patel, Purvi Rakhasiya, Kausal Yadav and Smita Girnari for their assistant and help. It is a pleasure to express my gratitude wholeheartedly to my friends Jalpa and her family, Sarita, Shreya, Dr. Vivek, Ravi, Dhawal, Jahnavi and Dr. Rajan. They have helped me stay sane through these years. Their support, understanding and care helped me to stay focused on my work. I greatly value their friendship and I deeply appreciate their belief in me. I am also grateful to all the research scholar‘s of Department of Biosciences, who had helped me during my research period. Where would I be without my family? My parents deserve special mention for their inseparable support and prayers. It was not possible to complete this work without their support. My Father, R.B. Maharshi, in the first place is the person who put the fundament my learning character, showing me the joy of intellectual pursuit ever since I was a child. Words fail me to express my gratitude towards my Mother, Bhagyawanti Maharshi is the one who sincerely raised me with her caring and gently love. She always gives me strength, courage and managed my time. I am thankful to Ashish Maharshi and Avdhendra Maharshi for being supportive and caring siblings. I also thankful to my cousin Manoj Prabhakar for his kind support at each and every step of my life. Finally, I would like to thank everybody who was important to the successful realization of thesis, as well as expressing my apology that I could not mention personally one by one. Anjisha Maharshi Index No. Chapter Page No. 1 General Introduction 1-13 Isolation, Growth and Identification of 2 Plant Pathogenic Fungi 14-33 3 Phytohormone Production by Plant Pathogenic Fungi 3A Indole Acetic Acid Production 34-51 Abscisic Acid and Gibberellic Acid 3B Production 52-62 4 Biocontrol Approach Biological approach by Plant extract 4A and Essential Oils 63-77 Immunological Approach Against Cell 4B Wall Degrading Enzymes 78-90 5 Summary 91-102 References 103-173 Fungi form a large and heterogeneous eukaryotic group of living organisms characterized by their lack of photosynthetic pigment and their chitinous cell wall. It has been estimated that the fungal kingdom contains more than 1.5 million species, but only around 100,000 have so far been described, with yeast, mold, and mushroom being the most familiar (Hawksworth, 1991). Although the majority of fungal species are saprophytes, a number of them are parasitics, in order to complete their biological cycle in animals or plants. With around 15,000 of fungi, the majority was belonging to the Ascomycetes and Basidiomycetes, causing disease in plants (Reddy, 2001). Fungal diseases are, in nature, more the exception than the rule. Thus, only a limited number of fungal species are able to penetrate and invade host tissues, avoiding recognition and plant defense responses, in order to obtain nutrients from them, causing disease and sometimes host death. In agriculture, annual crop losses due to pre- and post harvest fungal diseases exceed 200 billion euros, and in the United Stated alone, over $600 million are annually spent on fungicides (Gonzalez-Fernandez et al., 2010). Fungal pathogens have complicated life cycles, with both asexual and sexual reproduction and stages involving the formation of different infective, vegetative, and reproductive structures (Glawe, 2008). Current taxonomic identification of filamentous fungi is based on micro- and macro-morphological characteristics, such as cultural morphologies including colony and color characteristics on specific culture media, the size, shape, and development of sexual and asexual spores and spore-forming structures, and/or physiological characteristics such as the ability to utilize various compounds as nitrogen and carbon sources. The advent of molecular methods has supplemented traditional taxonomic methods with DNA-based tools with which to examine phylogenetics and systematics of fungi (Bowman et al., 1992; Bruns et al., 1992; Guerra-García et al., 2008). Accurate identification and early detection of pathogens is a crucial step in health care, agriculture and environmental monitoring including the fight against bioterrorism (Schaad and Frederick, 2002; Ivnitski et al., 2003; 1║ P a g e Schaad et al., 2003) plant disease management. The failure to adequately identify and detect plant pathogens using conventional, culture based morphological techniques has led to the development of nucleic acid based molecular approaches (López et al., 2003a; Lievens et al., 2005). Molecular diagnostic began to develop a real momentum after the introduction of polymerase chain reaction (PCR) in the mid 1980s and the first PCR based detection of a pathogen in diseased plants was published in the beginning of 1990s (Rasmussen and Wulff, 1991). In the last two decades, DNA marker technologies have been revolutionized the plant pathogen genomic analysis and have been extensively employed in many fields of molecular plant pathology (Bridge et al., 2003). Molecular markers offer also the possibility of faster and accurate identification and early detection of plant pathogen (Schaad et al., 2003; Michailides et al., 2005; Ma and Michailides, 2007). On the other hand, with the advances of molecular biology many complex questions could be answered such as the sources of inoculum, changes in their population‟s structures and the population dynamics of the disease they cause. Currently, numerous diagnostics laboratories are adopting molecular methods for plant pathogen detection (Lopez et al., 2003b; Schaad et al., 2003; Hernandez-Delgado, 2009). Among others important applications of these technologies which are increasingly developed as resourceful tools for quickly and sometimes cheaply assessing diverse aspects of plant pathogen genomes. These include genetic variation characterization, genome fingerprinting, gene mapping and tagging, genome evolution analysis, population genetic diversity, taxonomy and phylogeny of plant pathogen taxa.
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