(Luận văn thạc sĩ) inoculation of arbuscular mycorrhizal fungi to improve soybean growth under drought in the context of climate change

VIETNAM NATIONAL UNIVERSITY, HANOI VIETNAM JAPAN UNIVERSITY NGUYEN DUC TAM INOCULATION OF ARBUSCULAR MYCORRHIZAL FUNGI TO IMPROVE SOYBEAN GROWTH UNDER DROUHGT IN n THE CONTEXT OF CLIMATE CHANGE MASTER’S THESIS VIETNAM NATIONAL UNIVERSITY, HANOI VIETNAM JAPAN UNIVERSITY NGUYEN DUC TAM INOCULATION OF ARBUSCULAR MYCORRHIZAL FUNGI TO IMPROVE SOYBEAN GROWTH UNDER DROUHGT IN THE CONTEXT OF CLIMATE CHANGE n MAJOR: CLIMATE CHANGE AND DEVELOPMENT CODE: 8900201.02QTD RESEARCH SUPERVISOR: DR HOANG THI THU DUYEN DR DANG THANH TU Hanoi, 2021 PLEDGE I assure that this thesis is original and has not been published The use of results of other research and other documents must comply with regulations The citations and references to documents, books, research papers, and websites must be in the list of references of the thesis Author of the thesis Nguyen Duc Tam n TABLE OF CONTENT LIST OF TABLES i LIST OF FIGURES ii LIST OF ABBREVIATIONS iii ACKNOWLEDGMENT iv CHAPTER INTRODUCTION 1.1 Background and motivation of the study 1.2 The necessity of the research 1.2.1 Drought risk in Quang Nam province 1.2.2 Soybean in crop system in Quang Nam province 1.2.3 Mechanisms of soybean and arbuscular Mycorrhiza Fungi (AMF) symbiosis 1.2.4 Arbuscular Mycorrhiza Fungi (AMF) enhance phosphorus uptake by plants under drought 1.3 Research Framework: 1.4 The Research questions and hypotheses 10 1.5 Objectives and scope of the research 10 CHAPTER MATERIALS AND METHODOLOGIES 11 n 2.1 Methodology and experimental setup 11 2.1.1 Collecting data base and analysis 11 2.1.2 Fieldwork and soil sampling .11 2.1.3 Soil property measurement 11 2.1.4 Defining water holding capacity (WHC) 12 2.1.5 Experimental setup 12 2.2 Root, shoot biomass and root mycorrhiza inoculation .14 2.3 Microbial biomass phosphorus (MBP) .14 2.4 Microbial respiration 15 2.5 Statistical analysis 16 CHAPTER 3: RESULTS AND DISCUSSION 17 3.1 Characteristics of drought condition at the study site 17 3.2 AMF inoculation 20 3.3 Microbial Biomass Phosphorus (MBP) 21 3.4 Soil respiration 22 3.5 Root – Shoot characteristics .23 CHAPTER CONCLUSIONS AND RECOMMENDATIONS 26 4.1 Conclusions 26 4.2 Recommendations for future research 26 REFERENCES 28 n LIST OF TABLES Table Methodologies to analyze soil physic-chemical properties 11 n i LIST OF FIGURES Figure 1.1 Effects of drought on plant growth .2 Figure 1.2 Research framework .9 Figure 2.1 Determine Location of sampling area in Quang Nam province using Google Maps application .11 Figure 2.2 Soil was packed in total 16 transparent rhizoboxes 13 Figure 2.3 Microbial biomass P was measured in 96-wells microplates: a) the calibration was prepared for different concentrations of P and b) the microbial biomass was calculated based on the difference between fumigation and non-fumigation treatments .15 Figure 3.1 Average monthly sunshine hours (2000 – 2019) in Quang Nam .18 Figure 3.2 Average monthly temperature (2000 – 2019) in Quang Nam 18 Figure 3.3 Average monthly percipitation (2000 – 2019) in Quang Nam 19 Figure 3.4 Average monthly evaporation (2000 – 2019) in Quang Nam 20 Figure 3.5 The fungal structures 20 Figure 3.6 Microbial Biomass Phosphorus analysis results (μg P/ g soil) 21 n Figure 3.7 Soil respiration 23 Figure 3.8 The root nodules on the main roots .23 Figure 3.9 Average dry weight Root – Shoot (g) experimental box 24 Figure 3.10 Result Root – Shoot length (cm) .25 ii LIST OF ABBREVIATIONS C IPCC N Carbon Intergovernmental Panel on Climate Change Nitrogen TC Total carbon TN Total nitrogen WHC MBP Water holding capacity Microbial Biomass Phosphorus AMF Arbuscular Mycorrhiza Fungi n iii ACKNOWLEDGMENT To complete this thesis, I would like to thank the lecturers and staff of the Program on Climate Change and Development, Vietnam Japan University, Hanoi National University and the lecturers of Experimental center, Faculty of Forestry of the Vietnam National University of Forestry for their supports My research cannot be successful without their advices and collaboration First and foremost, I would like to express my gratitude and sincere thanks to Dr Hoang Thi Thu Duyen, who directly supervised my research implementation, for her enthusiastic instruction and dedication to orient my research topic, sampling soil, conducting lab experiment, data processing, and analysis My deepest and most heartful thanks to my sub supervisor - Dr Dang Thanh Tu, my advisors - Dr Kotera Akihiko, Dr Nguyen Van Quang, MS Hoa for their dedication and valuable advice to the thesis In addition, the author also highly appreciates the financial support of the VNU n Project (QG.20.63), without this support, the implementation of the thesis would not be possible Finally, I would like to dedicate this thesis to my parents, my wife, my children and my friends as a gesture of thanks for their support and putting their trust in me In the process of studying, researching and implementing the topic, I have also received a lot of valuable attention, suggestions and support from teachers, colleagues, friends from Vietnam Japan University iv CHAPTER INTRODUCTION 1.1 Background and motivation of the study Climate change (CC) is a natural process however anthropogenic activities have fosterred this process and worsened its impacts (IPCC, 2012) The increase in CO2 concentration in the atmosphere over the past centuries has caused global warming and led to a series of unpredictable weather phenomena Prolonged drought that becomes more severe in high-risk areas is one of the consequences of climate change (IPCC, 2019) Global temperature is expected to increase by 1.5 to 2°C between 2081 and 2100 (Collins et al., 2013) Each increase in the air temperature results in an increase in the air humidity by 7% (Bui et al., 2019) Therefore, rain becomes more concentrated during the year and the dry season is longer In arid sensitive regions such as the Mediterranean, Northeast Asia, West Asia, parts of South America, and much of Africa (IPCC, 2019), global warming tend to intensify During the years 1999 to 2018, the Climate Risk Indicators (CRI) (Eckstein et al., 2019) showed that Vietnam ranked 6th out of the 10 countries most affected by extreme weather events especially n an increase in the level of drought The severity of drought is threatening agricultural production in these regions Along with global warming, natural drought is one of the global challenges to agriculture to meet the food demand for the growing world population Droughts occur when the effective amount of water in the soil drops while the air humidity is low, causing the water to continuously evaporate or releasing to the air Many researches have demonstrated that droughts inhibit the expansion and development of plant cells (Shao et al., 2008), reducing dry and fresh biomass accumulation in plants (Farooq et al., 2009) Especially when a drought occurs during the development of cereal crab triggering the decrease of grain yield (Kamara et al., 2003; Monneveux et al., 2006) In general, drought seriously affects the growth of plants (Fig.1) Quang Nam is located in the South Central region of Vietnam with its diverse topography and harsh climate, strongly influenced by drought According to the report by the People's Committee of Quang Nam province (2010), prolonged drought caused losses of 3.841 out of 4.500 hectares of summer-autumn rice crop In addition, there REFERENCES n A.B.M Firoz, A Nauditt, M Fink, L Ribbe, 2018 Quantifying human impacts on hydrological drought using a combined modelling approach in a tropical river basin in central Vietnam Hydrology and Earth System Sciences 22, pp 547-565 Abdelmoneim T.S., Moussa, T.A.A., Almaghrabi O.A., Alzahrani, H.S., Abdelbagi, I., 2014 Increasing Plant Tolerance to Drought Stress by Inoculation with Arbuscular Mycorrhizal Fungi Life Sci J 2014; 11(1): 10-17] (ISSN: 1097-8135) Abrams, M.D.; Hock, W.K., 2006 Annual growth rings and the impact of Benlate 50 DF fungicide on citrus trees in seasonally dry 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respectively with ethanol 70%, 45o with the open side like a door facing hydropeoxit 10% and rinsed again down in a greenhouse chamber at 20-22 times with distilled water oC Harvest Roots were pale, whereas fungal structures remained black 34 n 35 Appendix A: The results of drought indices analysis Table B.1: Results Microbial biomass Phosphorus Microbial biomass P (μg P/ g soil) Rep Rep Rep drought conditions 60% 0,064 0,057 0,046 drought conditions Control 20% 0,024 0,023 0,023 drought conditions 60% 0,073 0,078 0,069 drought conditions AMF 20% 0,032 0,032 0,037 AVE 0,055±0.0053 0,023±0.0003 0,073±0.0026 0,033±0.0015 Table B.2: Phosphorus Microbial Biomass data table when comparing different drought conditions in ANOVA analysis control control AMF AMF AMF n control 60% 0,064 20% 0,024 0,057 0,046 0,073 0,078 0,069 0,023 0,023 0,032 0,032 0,037 Table B.3: MBP results after statistical analysis by ANOVA Test of Homogeneity of Variances Levene Statistic df1 df2 Sig VAR 1,289 ,320 60% VAR 9,846 ,035 20% N Mean Descriptives Std Std 95% Deviatio Error Confidence Minimu m Maximu m 36 n VA R 60% VA R 20% 1.00 ,0557 ,00907 2.00 ,0733 ,00451 Total ,0645 ,01161 1.00 ,0233 ,00058 2.00 ,0337 ,00289 Total ,0285 ,00596 Interval for Mean Lower Upper Bound Bound ,0052 ,0331 ,0782 ,0026 ,0621 ,0845 ,0047 ,0523 ,0767 ,0003 ,0219 ,0248 ,0016 ,0265 ,0408 ,0024 ,0222 ,0348 ,05 ,06 ,07 ,08 ,05 ,08 ,02 ,02 ,03 ,04 ,02 ,04 ANOVA VAR 20% Mean Square ,000 ,000 ,000 ,000 df n VAR 60% Sum of Squares Between ,000 Groups Within ,000 Groups Total ,001 Between ,000 Groups Within ,000 Groups Total ,000 F 9,120 Sig ,039 36,962 ,004 Table B.4: Phosphorus microbial biomass data table for comparison between AMF and control in ANOVA analysis Control AMF 60% 0,064 0,073 60% 60% 20% 20% 20% 0,057 0,046 0,024 0,023 0,023 0,078 0,069 0,032 0,032 0,037 37 Table B.5: MBP results after statistical analysis by ANOVA Test of Homogeneity of Variances Levene Statistic df1 df2 Sig VAR 5,380 ,081 Control VAR ,352 ,585 AMF VAR 1.00 Contro l 2.00 Tota l 1.00 2.00 Tota l Minimu m ,05 Maximu m ,06 ,02 ,02 ,02 ,06 ,07 ,08 ,03 ,04 ,03 ,08 n VAR AMF N Mea n ,055 ,023 ,039 ,073 ,033 ,053 Descriptives 95% Confidence Interval for Mean Std Deviatio Std Lower Upper n Error Bound Bound ,00907 ,0052 ,0331 ,0782 ,00058 ,0003 ,0219 ,0248 ,01862 ,0076 ,0200 ,0590 ,00451 ,0026 ,0621 ,0845 ,00289 ,0016 ,0265 ,0408 ,02199 ,0089 ,0304 ,0766 ANOVA Sum of Squares VAR Between ,002 Control Groups Within ,000 Groups Total ,002 VAR Between ,002 AMF Groups Within ,000 Groups Total ,002 Mean Square ,002 df F 37,940 Sig ,004 ,000 ,002 164,663 ,000 ,000 38 Table B.5: Data sheet Root – Shoot mass Control Drought 60% Control Drought (20%) AMF Drought 60% AMF Drought (20%) S2 Root Shoot (g) (g) 0,38 0,76 S7 Root Shoot 0,21 0,38 S10 Root Shoot 0,62 1,21 S22 Root Shoot 0,27 0,54 S3 Root Shoot (g) (g) 0,37 0,79 S8 Root Shoot 0,21 0,41 S11 Root Shoot 0,64 1,22 S23 Root Shoot 0,31 0,53 S4 Root Shoot (g) (g) 0,34 0,7 S1 Root Shoot 0,22 0,53 S15 Root Shoot 0,65 1,17 S17 Root Shoot 0,27 0,66 AVE root AVE shoot 0,36±0.012 0,75±0.026 0,21±0.003 0,44±0.045 0,64±0.008 1,2±0.015 0,28±0.013 0,57±0.041 Table B.5: Data sheet Root – Shoot length S3 Root Shoot (cm) (cm) 23 34,5 S7 Root Shoot 22 24 S11 Root Shoot 24 45 S22 Root Shoot 19 31 n Control Drought 60% Control Drought 20% AMF Drough 60% AMF Drough 20% S2 Root Shoot (cm) (cm) 23 30 S6 Root Shoot 22 27 S10 Root Shoot 24 46 S21 Root Shoot 22 28 S4 Root Shoot (cm) (cm) AVE root AVE shoot 23 30 7,66±0 10,5±1.5 S1 Root Shoot 21,5 24,5 7,28±0.166 8,38±0.928 S15 Root Shoot 25 39 8,11±0.333 14,44±2.186 S23 Root Shoot 22 30 7±1 9,89±0.882 39 Appendix C: Learning outcome Results of the Other outcomes of the Master’s Master’s thesis Program Learning Outcomes (PLOs) of MCCD Soil Map analysi s thesis samplin g, field practice skills Labora tory skills PLO1: Accumulating and mastering the basic knowledge on principles of Marxism Leninism, Political Theory and Ideology of Ho Chi Minh; and general knowledge about administration and management PLO2: Mastering fundamental, n interdisciplinary the knowledge and methodologies to assess and address actual problems (fate and features) related to CC x mitigation, adaptation for x sustainable development at global, national and local levels PLO3: Understanding and developing systematic thinking; necessary knowledge on science, technology, innovation and governance related to CC response for development; identifying, analyzing, x x x x assessing and forecasting the issues related to CC and CCR; predicting the developing trend of CC science 40 PLO4: Applying knowledge to solve the problems in CC and CCR: planning and approaching the works in field of CC; proposing the initiatives as well as the researches on CC; solutions on implementing science, x the technology, mechanism, policy and finance for CCR and development PLO5: Having skills of cooperation with personal, agencies, organizations domestically and internationally to solve the CC issues, communication in works, projects x x x on CC; and organizing, managing and administrating advanced career development PLO6: Accumulating soft skills to self- n directed and adapt to competitive working environment such as English proficiency (at level 4/6 according to English competencies Framework for Vietnam), Japanese communication skills; having skills on time x x x x and x x x x management; using the basic computer skills proficiently; working and researching independently; having skills of research and development; and using technologies creatively in academic and professional fields PLO7: Dynamic, enthusiastic, and confident, persistent, risk-taking management 41 Having PLO8: social/community’s responsibility and professional morality, especially for the scientific research results; being able to adapt to multicultural environment, ensure the harmony between the stakeholders, CCR and development; x x making expert decisions on climate change x x having good social morality, assist the vulnerable people to climate change; compliance with the law; discipline at work and positive lifestyle; having good attitude to their career in climate change response for sustainable development PLO9: Having responsibility for researching, creating new knowledge, and offering new ideas on climate change n response in different complex situations; adapting and guiding other people and x x response; managing research, having high responsibility in learning in order to develop professional knowledge, and creating new ideas in new process; and having good lifelong learning capacity 42