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Distribution of Soil Nematodes Associated with Grapevine Plant in Central Horticultural Centre (CHC), Kirtipur, Kathmandu

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Distribution of Soil Nematodes Associated with Grapevine Plant in Central Horticultural Centre (CHC), Kirtipur, Kathmandu Distribution of Soil Nematodes associated with Grapevine plant in Central Horticultural Centre (CHC), Kirtipur, Kathmandu ANU DESHAR TU Registration No: 5-2-0037-0278-2011 TU Examination Roll No: 308 Batch: 2072 A thesis submitted in partial fulfillment of the requirements for the award of the degree of Master of Science in Zoology with special paper Parasitology. Submitted to Central Department of Zoology Institute of Science and Technology Tribhuvan University Kirtipur, Kathmandu Nepal September, 2019 i DECLARATION I here by declare that the work presented in this thesis entitled “Distribution of Soil Nematodes Associated with Grape Vine Plant in Central Horticultural Centre (CHC), Kirtipur, Kathmandu.” has been done by myself and has not been submitted elsewhere for the award of any degree. All sources of the information have been specifically acknowledged by references to the author (s) or institution (s). Anu Deshar Date……………….. ii RECOMMENDATION This is to recommend that the thesis entitled “Distribution of Soil Nematodes Associated with Grape Vine Plant in Central Horticultural Centre (CHC), Kirtipur, Kathmandu.” has been carried out by Ms. Anu Deshar for the partial fulfillment of Master‟s Degree of Science in Zoology with special paper Parasitology. This is her original work and has been carried out under my supervision. To the best of my knowledge this thesis work has not been submitted for any other degree in any other institutions. Date………….. ……………….. Supervisor Prof. Dr. Mahendra Maharjan Central Department of Zoology Tribhuvan University Kirtipur, Kathmandu, Nepal iii LETTER OF APPROVAL On the recommendations of supervisor “Dr. Mahendra Maharjan” this thesis submitted by Anu Deshar entitled “Distribution of Soil Nematodes Associated with Grape Vine Plant in Central Horticultural Centre (CHC), Kirtipur, Kathmandu.” is approved for the examination and submitted to the Tribhuvan University in partial fulfillment of the requirements for Master‟s Degree of Science in Zoology with special paper Parasitology. Date: ……………… ………………… Head of Department Prof. Dr. Tej Bahadur Thapa Central Department of Zoology Tribhuvan University Kirtipur, Kathmandu, Nepal iv CERTIFICATE OF ACCEPTANCE This thesis work submitted by Ms. Anu Deshar entitled “Distribution of Soil Nematodes Associated with Grape Vine Plant in Central Horticultural Centre (CHC), Kirtipur, Kathmandu.” has been accepted as a partial fulfillment for the requirements of Master's Degree of Science in zoology with special paper Parasitology. EVALUATION COMMITTEE ………………………….. ….............................. Supervisor Head of Department Prof. Dr. Mahendra Maharjan Prof. Dr. Tej Bahadur Thapa Central Department of Zoology Central Department of Zoology …………………….. …………………. External examiner Internal examiner Prof. Dr. Arvinda K. Keshari Pitamber Dhakal Patan Multiple Campus Lecturer Central Department of Zoology Date of Examination : 5th September, 2019 v ACKNOWLEDGEMENTS I extend my first and foremost gratitude to my supervisor, Prof. Dr. Mahendra Maharjan, for his continuous encouragement, support and valuable suggestions from the project formulation phase to completion of the thesis. I would like to thank Late Mr. Ashok Bahadur Bam for his valuable support and encouragement to do my thesis on soil nematodes. I am eminently thankful to Prof. Dr. Tej Bahadur Thapa, Head of Central Department of Zoology for his kind support, suggestions and encouragements. I am deeply indebted to officials of Central Horticulture Centre, Kirtipur for co-operation during field. I would like to thank Bashu Dev Subedi, Department Head of CHC for providing me permission for field. I am heartily thankful to Ms. Rabina Dhakal for helping me in field and my family for continuous encouragement. Anu Deshar Examination Roll No. 308 Batch: 2072 vi CONTENTS DECLARATION ...................................................................................................................... ii RECOMMENDATION .......................................................................................................... iii LETTER OF APPROVAL ......................................................................................................iv CERTIFICATE OF ACCEPTANCE ...................................................................................... v ACKNOWLEDGEMENTS .....................................................................................................vi LIST OF PHOTOGRAPHS ..................................................................................................... x ABBREVATION ..................................................................................................................... xi ABSTRACT............................................................................................................................. xii 1. INTRODUCTION ................................................................................................................ 1 1.1 General Introduction .............................................................................................. 1 1.2. Medical Importance: ............................................................................................ 1 1.3 Disease in Grapevine ............................................................................................. 1 1.4 Nematodes in grape vine ........................................................................................ 2 1.5 Common plant parasitic nematodes ........................................................................ 2 1.2 Objectives .............................................................................................................. 4 1.2.1 General objective ................................................................................................ 4 1.2.2 Specific objectives ........................................................................................... 4 1.3 Rationale of study .................................................................................................. 4 2. LITERATURE REVIEW .................................................................................................... 5 3. MATERIALS AND METHODS ........................................................................................ 9 3.1 Study Area ............................................................................................................. 9 3.2 Study Period .......................................................................................................... 9 3.3 Sampling ................................................................................................................ 9 3.4 Physical Requirements ......................................................................................... 10 3.5 Chemical Required ............................................................................................... 11 3.6 Preparation of chemicals ...................................................................................... 11 3.6.1 TAF solution ................................................................................................. 11 3.6.2 Glycerol-alcohol ............................................................................................ 11 3.7 Processing of soil samples .................................................................................... 11 3.8 Killing and fixation of nematodes ........................................................................ 13 3.9 Dehydration of nematodes .................................................................................... 13 3.12 Morphometric analysis for species identification ................................................ 14 vii 3.13 Callibration of microscope ................................................................................. 15 4. RESULT ............................................................................................................................. 16 4.1 Taxonomic treatment of plant parasitic nematodes of grape vine .......................... 16 4.2 Distribution and density of plant parasitic nematodes. .......................................... 20 4.3Distribution and density of predator and free living nematodes ............................. 20 5. DISCUSSION ..................................................................................................................... 23 6. CONCLUSION AND RECOMMENDATIONS ............................................................. 27 REFERENCES ....................................................................................................................... 28 viii LIST OF TABLE S.N. Title of table Page Table 1: Morphometric Data for Species Detection 27 Table 2: Distribution and Density of Parasitic nematode 31 Table 3: Distribution and Density of Predator and Free living nematode 32 ix LIST OF PHOTOGRAPHS S.N. Title of Photographs Pages 1. Map of Central Horticultural Centre 10 2. Collection of soil sample 11 3. Processing of soil Sample by Seiving method 13 4. Baermann funnel method of soil nematode extraction 14 5. Dehydration of nematode in dessicator 14 6. Preparation of slides 15 7. Observation of nematode under microscope 15 8. Tylenchorhynchus sp. 18 9. Xiphinema sp. 19 10. Anterior part of Xiphinema sp. 19 11. Posterior part of Xiphinema sp. 19 12. Anterior part of Longidorus sp. 20 13. Posterior part of Longidorus sp. 20 14. Anterior part of Mononchus sp. 22 15. Posterior part of Mononchus sp. 22 16. Rhabditis sp. 22 x ABBREVATION Cacl2 -
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