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Arbuscular Mycorrhizal (Am) Fungal Inoculum Production Using in Vitro Technique for Revegetation of Degraded Sand Dunes

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Arbuscular Mycorrhizal (Am) Fungal Inoculum Production Using in Vitro Technique for Revegetation of Degraded Sand Dunes ARBUSCULAR MYCORRHIZAL (AM) FUNGAL INOCULUM PRODUCTION USING IN VITRO TECHNIQUE FOR REVEGETATION OF DEGRADED SAND DUNES A THESIS SUBMITTED TO S3j GOA UNIVERSITY FOR THE AWARD OF THE DEGREE OF DOCTOR OF PHILOSOPHY IN BOTANY FACULTY OF LIFE SCIENCES AND ENVIRONMENT BY KIM MARIA RODRIGUES 7 7 3> Research Guide ' PROF. B. F. RODRIGUES UGC-SAP DEPARTMENT OF BOTANY GOAj UA UNIVERSITYUIN1V JfcKSll \ Lf> ' t i t y f TALEIGAO GOA v. Z ^ INDIA May 2017 I Koh Joj'ax'f DECLARATION I hereby declare that the matter embodied in this thesis entitled, “Arbuscular mycorrhizal (AM) fungal inoculum production using in vitro technique for revegetation of degraded sand dunes” is the result of investigations carried out by me, under the supervision of Prof. B. F Rodrigues and it has not previously formed the basis for the award of any degree, diploma, associate ship, fellowship or the other such similar title. Goa University Ms. Kim Maria Rodrigues May 2017 (Candidate) CERTIFICATE This is to certify that the work incorporated in this thesis entitled, “Arbuscular mycorrhizal (AM) fungal inoculum production using in vitro technique for revegetation of degraded sand dunes” submitted by Ms. Kim Maria Rodrigues, constitutes her independent work and the same has not been previously submitted for the award of any other degree, diploma, associate ship, fellowship or the other such title. Goa University (Research Guide) May 2017 Department of Botany Goa University Acknowledgement am sincerely grateful to all the individuals who have kindly given me advice and support in various ways to the point I have reached for the submission of this thesis. As I got the opportunity to start working with ‘Arbuscular Mycorrhizal Fungi’ during my M. Sc. studies under the guidance of my research guide Prof. Bernard Felinov Rodrigues, which eventually led me to join for Ph. D. I would like to express my deep gratitude for his patient supervision. I further thank him for all his meticulous suggestions and helpful discussions that guided me in the right direction. 1 gratelully acknowledge the ‘Innovation in Science Pursuit for Inspired Research (INSPIRE) programme’, Department of Science and Technology (DST), Government of India, New Delhi for the fellowship awarded during the course of my Ph. D. I would also like to thank the Dean, Faculty of Life Sciences and Environment, and Department of Botany, Goa University for granting me the opportunity to cany out my research. 1 also would like to thank Prof. Vijaya Kerkar, Head of the Department of Botany, Goa University. In addition I would like to thank the teaching staff of the Botany Department for their help. In particular, I would like to thank my Vice Chancellors’ nominee Dr. Irene Furtado, Department of Microbiology, Goa University, for her scientific guidance and encouraging comments throughout my study period. My appreciation goes out to the non teaching staff at the Department of Botany, Goa University, for their constant help and cooperation. I owe a great deal of appreciation and gratitude to Dr. Gopal R. Mahajan, Scientist (Soil Sciences) Natural Resource Management Section, ICAR-CCARI, Ela, Old Goa, for plant nutrient analyses and the Director, ICAR-Central Coastal Agricultural Research Institute, Ela, Old Goa for extending help through these facilities. I add a note of thanks to the technical staff at Soil Sciences laboratory, Natural Resource Management Section, ICAR-CCARI, Ela, Old Goa for their help and cooperation. I would also like to thank the Assistant Director of Agriculture, Soil Testing Laboratory, Ela, Old Goa, Soil 1 esting Laboratory, Margao, Goa and Italab House (Goa) PVT. Ltd Industrial Testing and Analytical Laboratories, Margao, Goa for assisting with soil nutrient analysis. Special thanks to Professor B. R. Srinivasan, HOD Department of Chemistry, Goa University and Professor S. N. Dhuri, Department of Chemistry, Goa University for CUN analyses. 1 thank all the research students (past and present) at the Department of Botany and Goa University for their help. To all the members of the ‘Mycorrhiza lab’ for all those useful inputs, healthy discussions, friendship and mutual help; I have really appreciated the wonderful moments. Especial thanks go to my Family and close circle of Friends for their love, tremendous encouragement and support all the time. The root does anchor a plant, but it’s the mycorrhizae that become the main system to absorb water and nutrients from the soil. -Larry Simpson CONTENTS Page No, CHAPTER 1: Introduction................................................ 1-16 CHAPTER 2: Review of Literature............................................... 17-60 CHAPTER 3: To identify the dominant AM fungal species from the sand dune ecosystem (Objective 1)............................... 61-79 3.1: Introduction................................................................................... 61-63 3.2: Materials and M ethods................................................................ 63-68 3.3: R esults............................................................................................ 69-73 3.4: D iscussion..................................................................................... 73-78 3.5: Conclusion..................................................................................... 78-79 CHAPTER 4: To prepare pure culture inoculum using trap and pot cultures (Objective 2 ) .......................................................... 80- 84 4.1: Introduction................................................................................... 80-81 4.2: Materials and M ethods................................................................ 81- 82 4.3: R esults............................................................................................ 83 4.4: D iscussion..................................................................................... 83-84 CHAPTER 5: To prepare and standardize the protocol for in vitro culture technique for dominant AM fungal species 85-98 (Objective 3 ) ....................................................................................... 5.1: Introduction.................................................................................. 85- 86 5.2: Materials and M ethods............................................................... 86- 89 5.3. Results........................................................................... 89-90 ^•4* Discussion........................ 91-97 5.5: Conclusion.................................................................... 9g CHAPTER 6: To develop viable inoculum using suitable carrier for re-inoculation (Objective 4 ) ............................................ 99-115 6.1: Introduction...................................................................................... 99-102 6.2: Materials and Methods................................................................... 102-107 6.3: R esults............................................................................................... 108-110 6.4: D iscussion........................................................................................ 111-114 6.5: Conclusion......................................................................................... 115 CHAPTER 7: To maximize the shelf life of the in vitro prepared inoculum (Objective 5 ) ....................................................... 116-124 7.1: Introduction...................................................................................... 116-117 7.2: Materials and M ethods................................................................... 118-121 7.3: R esults................................................................................................. 121 7.4: D iscussion........................................................................................ 122-123 7. 5 : Conclusion....................................................................................... 123-124 CHAPTER 8: To study the effect of in vitro produced carrier based bio-inocula on selected plant species suitable for U(,5CU 125-140 revegetation of the sand dunes (Objective 6 )................................ 125-127 8.1: Introduction............... 127-133 8.2: Materials and Methods 8.3: R esults....... 133-136 8.4: D iscussion................ 136-139 8.5: Conclusion................ 140 CHAPTER 9: Summary 141-144 R eferences....................... 145-218 Synopsis............................ 219-241 Appendix 242-245 LIST OF TABLES Table No. Title After Page No. 2.1 Consensus classification of the Glomeromycota by Redecker et al. (2013) ................................................... 23 2.2 Arbuscular mycorrhizal (AM) species cultivated on Root Organ Culture (R O C ).............................................................................................. 46 3.1 Physico-chemical properties of sand from selected study sites............... 69 3.2 AM association in sand dune vegetation at Betalbatim (S I)................... 69 3.3 AM association in sand dune vegetation at Colva (S2)........................... 69 3.4 AM association in sand dune vegetation at Benaulim (S3)..................... 70 3.5 AM association in sand dune vegetation at Varca (S4)........................... 70 3.6 AM association in sand dune vegetation at Sinquerim (S5).................... 70 3.7 AM association in sand dune vegetation at Candolim (S6)..................... 70 3.8 AM association in sand dune vegetation at Vagator (S7)....................... 70 3.9 AM association in sand dune vegetation at Moijim (S8).......................
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