Seed health testing of selected forest tree seeds and their control using sodium hypochlorite and capsicum extract soap

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY JERZYNE ANNE ABRERA CORPUZ SEED HEALTH TESTING OF SELECTED FOREST TREE SEEDS AND THEIR CONTROL USING SODIUM HYPOCHLORITE AND CAPSICUM EXTRACT SOAP SOLUTION BACHELOR THESIS Study Mode: Major: Faculty: Batch: Type: Full-time Environmental Science and Management Advanced Education Program 2015 – 2018 On- The Job Training (OJT) Thai Nguyen, 25/09/2018 DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Environmental Science and Management Student Name Jerzyne Anne A Corpuz Student ID DTN1454290095 Thesis Title Seed Health Testing of Selected Forest Tree Seeds and Their Control Using Sodium Hypochlorite and Capsicum Extract Soap Supervisor(s) Marcelina V Pacho (M.A.) &Ly Thi Duong (M.A.) Supervisor’s signature (s) Abstract: Seed must be genetically and physically pure, has high vigor and high germination But what if seeds are contaminated by seed borne pathogens Seed borne pathogens can cause infection that leads to reduction of germination plant growth /height That is the reduction of crop productivity both terms in quality and quantity Seed health testing is a procedure by which one can determine whether the seeds are healthy or has pathogen infection or microbial contamination Sodium Hypochlorite (50 ml from 15 % NaOCl with 50 ml distilled water) and Capsicum Extract Solution (CESS, extract from 25 pcs Capsicum: L Distilled water) was applied to treat the fungus infected/ contaminated seed/ separately per test species The result of this study showed that Sodium Hypochlorite (NaOCl) and Capsicum Extract Soap Solution (CESS) is effective in reducing the fungal infection on the forest tree seeds However in terms of the Molave seeds there is no significant effect on reducing the fungal infection.There is a possibility that the level of concentration of the two treatments need to be increased But still the seed borne pathogen of Molave seed was slightly reduce some fungi species based on the results i Keywords: Number of Pages Date of Submission Seed health testing, fungal infection, Sodium Hypochlorite, Capsicum Extract soap solution, seed borne pathogens 86 September 15 , 2018 ii ACKNOWLEDGMENT “The LORD is my strength and my shield; my heart trusts in him, and he helps me.” — Psalm 28:7 I would like to thanks of gratitude to my Adviser Marcelina Pacho who gave me the golden opportunity to this wonderful project on the topic (Seed health Testing of Selected Forestry Seed and their Control Using Sodium Hypochlorite and Capsicum Extract Soap Solution) of Ecosystem Research Development Bureau (ERDB) as well as Mrs Ly Thi Duong which also help me to improve my research topic and I came to know about so many new things I am really thankful to them Secondly I would also like to thank my parents “Daddy” Virgilio and Ledivina and loving brothers “Kuyas” Jefferson and Jay Mark, to my Aunt Evangelina I wouldn’t be able to achieve this success without your financial support, love and guidance To all friends, Especially Hannah Ayson, Nicole de Castro, Alexis Alcantara, Mark Glodo, Angelika Aragones, Ghia Sarmiento, Colleene Lanzador, Mishel Ranada, Kenneth Kua and ERDB family for the ideas, support and modivation To the Advance Education Program Thai Nguyen University of Agriculture and Forestry for providing the opportunity of study abroad and grateful to the Ecosystem Research Development Bureau (ERDB) for accepting me to conduct my thesis/ OJT and whose made all the laboratory experiment possible without further expenses.To batch, K46-A AEP, for sharing knowledge and crazy memories along the journey in Vietnam iii Dear God from the bottom of my heart I want to thank you for being with me all the way Jerzyne Anne A Corpuz iv TABLE OF CONTENTS List of Figures List of Tables List of Abbreviations CHAPTER I INTRODUCTION 1.1.Rationale 1.2.Objectives 1.2.1 General 1.2.2 Specific 1.3.Content of Work 1.4.Profile of the Cooperating Agency 1.4.1 Vision 1.4.2 Mission 1.4.3 ERDB Main Office Function 1.4.4 ERDB Research Centers Function 1.4.5 ERDB’S Research, Development and Extension Program 1.4.6 Special Projects 10 1.4.7 Adviser 11 1.5.Description of the Activity 11 1.6.Timeline 11 CHAPTER II: REVIEW OF RELATED LITERATURE 13 2.1 Seedborne Pathognes 13 2.2 Seed Treatments 14 2.3.Methodology 17 2.4.Seed Borne Fungi of Some Forest and Their Control 19 v 2.5.Impact of Seed Mycoflora and Forest Seed on See Quality anf Their Management 20 CHAPTER III STATUS OF THE CONSIDERED ISSUE AT THE TRAINING AGENCY 21 3.1 Title 22 3.2 Time and Place 22 3.4 Statement of the Problem 22 3.5 Hypothesis 23 3.6 Scope and Limitation 23 3.7 Materials 23 3.7.1 Four selected forestry seeds species 24 3.7.2 Petri Plates 24 3.7.3 Autoclave 24 3.7.4 Compound Microscope 24 3.7.5 Hot air oven 24 3.7.6 Distilled water 24 3.7.7 Filter paper 24 3.7.8 Forceps 25 3.7.9 De- natured Alcohol and Alcohol lamp 25 3.7.10 Glass slides and Glass cover 25 3.8 Methods 25 3.9 Research Approach 26 3.10 Research Design 27 3.11 Data Gathering 27 3.12 Statistical Analysis 28 3.13 Results 28 vi 3.13.1 Seedborne Pathogens 28 3.13.2 Fungal Infection 29 3.13.3 Bacteria 31 3.13.4 Aspegillus flavus 33 3.13.5 Verticillium sp 35 3.13.6 Fusarium spp 37 3.13.7 Asperillus niger 39 3.13.8 Chaetonium sp 41 3.13.9 Germination Rate 43 3.14 Discussion 45 4.1 Discussion 49 4.2 Lesson Learned 51 CHAPTER V: CONCLUSION 53 References 54 APPENDICES 58 vii LIST OF FIGURES Figure 1: Research Approach 26 Figure 2: Research Design 27 LIST OF TABLES Table 1: Timeline of Working Days of On-The-Job Training 12 Table 2: List of Seedborne Pathogens/Contaminants 28 Table 3: Percentage of infection of Fungi on four slected forestry Seeds and Effect of Pre-treatment 29 Table 4: Percentage of Bacteria of Four Selected Forestry Seeds with Pretreatment Using NaOCl and CESS 31 Table 5: Percentage of As flavus of four selected Forestry Seeds and effect of Pre-treatment Using NaOCl andd CESS 33 Table 6: Percentage of Verticilium sp on Four Seleceted Forestry Seeds and Effect of Pre-treatment Using NaOCl and CESS 35 Table 7: Percentage of Fusarium ssp on Four Selected Forestry Seeds andEffect of Pre-treatment Using NaOCl and CESS 37 Table 8: Percentage of As niger on Four Selected Forestry Seeds and Effect of Pre-treatment Using NaOCl and CESS 39 Table 9: Percentage of Chaetonium sp on Four Selected Forestry Seeds and Effect of Pre-treatment Using NaOCl and CESS 41 Table 10: Percentage of Germiantion Rate of Four Selected Forestry Seeds as Affected by NaOCl and CESS 43 Apppendix E: Letter that ws used in Internship 64 Apppendix F: Certificate of Completion 65 Appendix E: Results 1.) Lanete Table E- 1: Lanete Seed s with Control Treatment ControlTreatment Replication Infected Germination Bacterium As Fusarium Verticillium flavus 5 0 7 2 6 2 6 5 10 8 2 Table E- 2: Lanete Seeds with Sodium Hypochlorite (NaOCl) Sodium Hypochlorite As Fusarium flavus Replication Infected Germination Bacterium Verticillium 0 9 1 5 0 0 0 7 0 7 0 7 0 8 0 10 0 66 Table E- 3: Lanete Seeds with Capsicum Extract Soap Solution (CESS) Capsicum Replication Infected Germination Bacterium 2 As Fusarium Verticillium 0 0 0 0 5 0 0 0 10 0 0 10 5 0 flavus 2.) Molave Table E- 4: Molave Seeds with Control Treatment Control Treatment Replicate Infecte Germinatio Bacteriu As.flavu s d n m s 10 10 2 10 10 10 10 As Chaetoniu Verticilliu m m 4 10 10 10 10 10 10 10 10 10 10 10 10 3 10 10 10 2 nige r 67 Table E- 5: Molave Seeds with NaOCl Sodium Hypochlorite Replicate Infecte Germinat Bacteriu As.flavu As Chaetoniu Verticilliu s d e m s niger m m 10 10 2 10 10 10 10 0 10 10 0 0 10 10 0 0 10 10 2 10 10 10 10 0 10 10 0 10 10 10 0 Chaetoniu Verticilliu m m Table E- 6: Molave Seeds with CESS Capsicum As Replicate Infecte Germinat Bacteriu As.flavu s d e m s 10 10 10 10 0 10 10 2 10 10 0 10 10 0 10 10 0 10 10 0 10 10 0 2 10 10 0 10 10 10 nige r 68 Ipil-ipil Table E- 7: Ipil-ipil Seeds with Control Treatment ControlTreatment Replicates Infected Germinate Bacterium fusarium Chartonium As.niger As flavus 10 10 1 10 0 6 0 10 4 2 10 6 3 0 10 2 10 10 3 Table E- 8: Ipil-ipil Seeds with NaOCl Sodium Hypochlorite Replicates Infected Germinate Bacterium Fusarium Chartonium As.niger As flavus 0 0 0 2 2 0 3 3 0 3 0 5 1 0 2 0 0 0 0 0 0 10 1 0 69 Table E- 9: Ipil-ipil with CESS Capsicum Replicates Infected Germinate Bacterium Fusarium Chartonium As.niger As flavus 4 1 4 0 3 3 0 5 1 0 0 0 6 1 5 2 1 0 3 0 10 2 1 0 3.) Taluto Table E- 10: Taluto Seeds with Control Treatment ControlTreatment Replicates Infected Germinate Bacterium Fusarium 3 3 2 5 5 2 6 4 4 3 10 4 4 70 Table E- 11: Taluto Seeds with NaOCl Sodium Hypochlorite Replicates Infected Germinate Bacterium Fusarium 1 10 1 1 1 5 5 10 0 10 0 10 2 3 10 2 10 0 Table E- 12: Taluto Seeds with CESS Capsicum Replicates Infected Germinate Bacterium Fusarium 0 1 3 3 4 0 0 0 0 0 0 10 2 71 Appendix F: ANOVA Table Using Turkey’s Studentized Range (HSD (Multiple Comparison) 1.) Lanete Seeds Table E- 1: ANOVA Table of Lanete Seeds of Fungal Infection Sum of Source Df Mean Square F value p-value Squares Model 29 14.34666667 0.494712644 2.29 0.0003 Treatment 7.08666667 3.54333333 16.38