Genotypic Variation For Tolerance To Low Temperature Stress In Rice Long Viet Ha Bachelor of Crop Science at Thai Nguyen University of Agriculture and Forestry, Vietnam in 2004 A thesis submitted for the degree of Master of Philosophy at The University of Queensland in 2015 School of Agriculture and Food Sciences Abstract Lack of irrigation water for flooded cultivation and low temperature during the reproductive phase are factors limiting rice production in the Riverina region of New South Wales, Australia The adoption of aerobic technique could be beneficial as it requires less irrigation water, but a major limitation of aerobic cultivation is that there is no insulation effect of flooded water in the field to protect young panicles from low air temperature, causing spikelet sterility Suitable varieties need to tolerate low temperature at the booting stage, grow well under aerobic condition and be adapted to other conditions such as different rates of nitrogen application Three controlled temperature glasshouse experiments were conducted at The University of Queensland, Brisbane, Australia from 2013-15 The objectives of this study were to identify genotypic variation for tolerance to low temperature stress in rice using a population developed from a cross between low temperature tolerant and susceptible parents, and to determine the genotypic consistency in spikelet sterility under different flood and aerobic conditions and levels of nitrogen application, when exposed to low temperature during the booting stage In all experiments two controlled temperature glasshouses were utilised The plants were grown in the warm glasshouse at 28oC/19oC except for 14 days when they were exposed to low temperature at 21oC/15oC at the booting stage Experiment phenotyped 101 F5 Kyeema//Kyeema/Norin PL8 (KKN) lines alongside the parent varieties, Kyeema and Norin PL8; and the recently released Australian low temperature tolerant variety Sherpa Experiment and consisted of 20 and 10 genotypes respectively, which were selected from Experiment In these experiments, there were also plants grown continuously in the warm glasshouse Experiment was grown under flooded conditions, while Experiment and were grown under aerobic and flooded conditions with Experiment also utilising two nitrogen application rates (0 and 150kg/ha, -N and +N treatments) The plants were exposed to low temperature at the late booting stage in Experiment and the early booting stage in Experiment and Spikelet sterility was used as the criterion for low temperature tolerance/susceptibility Most genotypes in this study showed a consistency in spikelet sterility under low temperature at the booting stage between experiments (r=0.83 between Experiment and 2, r=0.93 between Experiment and 3, and r=0.90 between Experiment and 3) under standard flood conditions Genotype’s response to low temperature under different growing conditions was also similar between flooded and aerobic conditions (r=0.90) in Experiment 2, flooded +N and aerobic +N (r=0.90) and flooded -N and flooded +N (r=0.97) in Experiment While there was no significant difference in mean spikelet sterility without low temperature exposure between aerobic and flooded conditions in both Experiment and 3, mean spikelet sterility under low temperature was significantly higher for aerobic conditions in Experiment (75.4% vs 51.7%) and Experiment (70.8% vs 56.8%) than those of flooded conditions The mean low water temperature in the flooded condition (15.3oC) was 1.4oC higher than the air temperature, and this may have caused soil temperature difference, resulting in the difference in spikelet sterility Even if grown under warm conditions, some genotypes showed high spikelet sterility (≥37%) in Experiment which was given high N application Experiment showed that N application increased tillering and spikelet number per plant, leading to increased spikelet sterility under low temperature condition Furthermore, spikelet sterility was found to be correlated with total spikelet number per plant under flooded low temperature +N among 13 genotypes tested in Experiment In this experiment, spikelet sterility was also significantly correlated with grain yield/plant but some genotypes which showed low spikelet sterility had low grain yield In this study, tolerant and susceptible genotypes were identified for flooded and aerobic conditions in controlled glasshouse experiments However, further research is required in the field in order to confirm the consistency in spikelet sterility of those genotypes Other future research needs such as identifying mechanisms for the genotypic variation in low temperature tolerance is also suggested Declaration by author This thesis is composed of my original work, and contains no material previously published or written by another person except where due reference has been made in the text I have clearly stated the contribution by others to jointly-authored works that I have included in my thesis I have clearly stated the contribution of others to my thesis as a whole, including statistical assistance, survey design, data analysis, significant technical procedures, professional editorial advice, and any other original research work used or reported in my thesis The content of my thesis is the result of work I have carried out since the commencement of my research higher degree candidature and does not include a substantial part of work that has been submitted to qualify for the award of any other degree or diploma in any university or other tertiary institution I have clearly stated which parts of my thesis, if any, have been submitted to qualify for another award I acknowledge that an electronic copy of my thesis must be lodged with the University Library and, subject to the policy and procedures of The University of Queensland, the thesis be made available for research and study in accordance with the Copyright Act 1968 unless a period of embargo has been approved by the Dean of the Graduate School I acknowledge that copyright of all material contained in my thesis resides with the copyright holder(s) of that material Where appropriate I have obtained copyright permission from the copyright holder to reproduce material in this thesis Long Viet Ha Publications during candidature No publications Publications included in this thesis No publications included Contributions by others to the thesis Significant contributions were made to the conception and design of the project as well as critical revision of the research data by Professor Shu Fukai, Dr Jaquie Mitchell and Dr Doug George Statement of parts of the thesis submitted to qualify for the award of another degree None Acknowledgements I sincerely thank my three supervisors from the School of Agriculture and Food Sciences, The University of Queensland, Professor Shu Fukai, Dr Jaquie Mitchell and Dr Doug George who offered continual encouragement, guidance and inspiration throughout my study I also sincerely thank professional English editor from the School of Agriculture and Food Sciences, The University of Queensland, Dr John M Schiller who offered encouragement and motivation, particularly for editing academic English of my thesis I would like to thank Mr Christopher Proud for his technical support - without him this study would not have been conducting well Thank you to Mr Thinh Tran who provided invaluable assistance in the analysis of the experiments during the study I acknowledge Ms Zuziana Susanti who was willing to help and share knowledge during the conduct of the experiments Finally, thank you to my wife, Anh Vu and my daughter, Anh Ha for their love and encouragement, especially during the final stages of the study when our beautiful daughter Chi was born Keywords Rice, low temperature tolerance, glasshouse, flooded condition, aerobic condition, low temperature during the booting stage and nitrogen application Australian and New Zealand Standard Research Classifications (ANZSRC) ANZSRC code: 070302, Agronomy, 80% ANZSRC code: 070305 Crop and Pasture Improvement, 20% Fields of Research (FoR) Classification FoR code: 0703, Crop Pasture and Production, 80% FoR code: 0701, Agriculture, Land and Farm Management, 20% TABLE OF CONTENTS CHAPTER INTRODUCTION 16 1.1 GENERAL INTRODUCTION 16 CHAPTER LITERATURE REVIEW 18 2.1 INTRODUCTION 18 2.2 RICE GROWTH AND DEVELOPMENT 19 2.2.1 VEGETATIVE PHASE 20 2.2.1.1 Seed germination 20 2.2.1.2 Seedling emergence 20 2.2.1.3 Tillering 21 2.2.1.4 Stem elongation 21 2.2.2 REPRODUCTIVE PHASE .21 2.2.2.1 Panicle initiation to heading stage 22 2.2.2.2 Flowering 23 2.2.2.3 Grain filling and ripening 24 2.3 RICE YIELD AND THE EFFECT OF ENVIRONMENTAL CONDITIONS .24 2.4 THE EFFECT OF LOW TEMPERATURE AT DIFFERENT GROWTH STAGES 26 2.4.1 Vegetative phase 26 2.4.1.1 Germination stage 26 2.4.2 Reproductive phase 27 2.4.2.1 Booting stage 27 2.4.2.2 Mechanisms of low temperature induced male sterility 29 Anther respiration 30 Cytological abnormalities 30 Carbohydrate metabolism 31 Enzyme activity 31 2.4.2.3 Flowering stage 32 2.7 AEROBIC RICE 32 2.5 NITROGEN EFFECT ON RICE GROWTH 33 2.6 CONCLUSIONS OF LITERATURE REVIEW AND FOCUS OF THIS STUDY 34 CHAPTER EXPERIMENT GENOTYPIC VARIATION IN SPIKELET STERILITY WITHIN A POPULATION EXPOSED TO LOW TEMPERATURE AT THE BOOTING STAGE 35 3.1 INTRODUCTION 35 3.2 MATERIALS AND METHODS 36 3.2.1 Genetic materials 36 3.2.2 Cultural details 36 3.2.3 Data collections and analysis 37 3.3 RESULTS .38 3.3.1 Air temperatures 38 3.3.2 Heading date 39 3.3.3 Spikelet Sterility 40 3.3.4 Other characters 43 3.4 DISCUSSION 44 3.5 CONCLUSIONS 46 CHAPTER EXPERIMENT THE EFFECTES OF LOW TEMPERATURE AT THE BOOTING STAGE AND AEROBIC CONDITIONS ON SPIKELET STERILITY IN 20 SELLECTED GENOTYPES AND VARIETIES 48 4.1 INTRODUCTION 48 4.2 MATERIALS AND METHODS 49 4.2.1 Genetic materials 49 4.2.2 Cultural details 49 4.2.3 Water and temperature treatments 50 4.2.4 Measurements 50 4.2.5 Data Analysis 51 4.3 RESULTS .51 4.3.1 Air and water temperatures 51 4.3.2 Time of heading 51 4.3.3 Plant height 53 4.3.4 Spikelet Sterility 55 4.3.5 Spikelet number per panicle 57 4.4 DISCUSSION 59 4.5 CONCLUSIONS 60 CHAPTER EXPERIMENT GENOTIPIC VARIATION FOR SPIKELET STERILITY BETWEEN FLOODED AND AEROBIC CONDITIONS UNDER TWO DIFFERENT LEVEL OF NITROGEN FERTILIZATION EXPOSED TO LOW TEMPERATURE AT THE BOOTING STAGE 62 5.1 INTRODUCTION 62 5.2 MATERIALS AND METHODS 63 5.2.1 Genetic materials 63 10 Fageria, N K & Baligar, V C 2001a Lowland rice response to nitrogen fertilization Communications in Soil Science and Plant Analysis, 32, 1405-1429 Farrell, T C., Williams, R L & Fukai, S 2001b The cost of low temperature to the NSW rice industry In: Proceeding of The 10th Australian Agronomy Conference, Australian Society of Agronomy, www.regional.org.au/au/asa/2001/1/d/farrell.htm Farrell, T C., Fox, K M., Williams, R L & Fukai, S 2006a Genotypic variation for cold tolerance during 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Plant Science, 168, 527-534 93 APPENDIX Appendix Genotypes exposed to low temperature at 120C in Experiment No Pop Rep Temperature SS (%) Average 216 12°C for days 62.5 62.5 221 12°C for days 89.5 89.5 9225 12°C for days 89.6 12°C for days 91.9 12°C for days 90.2 12°C for days 55.2 12°C for days 90.5 12°C for days 59.8 12°C for days 54.9 12°C for days 73.1 12°C for days 54.9 12°C for days 85.6 12°C for days 87.5 230 231 90.7 78.6 62.6 10 216 10 222 10 223 12°C for days 75.0 75.0 10 229 12°C for days 75.3 75.3 10 10 232 12°C for days 35.9 12°C for days 80.5 11 11 220 12°C for days 91.6 91.6 12 11 227 12°C for days 92.9 92.9 14 11 231 12°C for days 46.9 46.9 15 12 218 12°C for days 100.0 100.0 16 12 225 12°C for days 75.5 75.5 17 12 228 12°C for days 72.5 72.5 18 12 229 12°C for days 86.8 86.8 19 12 230 12°C for days 77.3 77.3 12°C for days 64.5 12°C for day 97.0 12°C for days 34.2 20 12 231 54.9 86.5 58.2 65.2 21 13 227 12°C for days 89.4 89.4 22 13 231 12°C for days 88.1 88.1 23 14 219 12°C for days 95.6 96.2 94 12°C for days 96.8 24 14 221 12°C for days 90.8 90.8 25 14 222 12°C for days 97.3 97.3 26 14 227 12°C for days 58.2 58.2 27 15 222 12°C for days 85.1 85.1 28 15 228 12°C for days 92.2 12°C for days 97.8 29 15 229 12°C for days 90.3 30 15 230 15 231 32 15 232 12°C for days Norin 94.7 92.4 92.8 12°C for days 12°C for days 33 90.3 89.9 31 95 76.3 50.6 62.5 63.4 62.5 29.1 12°C for days 63.3 49.5 95 47.3 Appendix The average number of days to heading, percentage of spikelet sterility, auricle distance at the time of transfer, auricle distance development rate and spikelet number per panicle of 96 genotypes and varieties in Experiment Genotype Days to Heading sterility (%) 215 95.0 42.7 Auricle distance at the time of transfer (cm) -1.2 91.0 53.8 217 93.7 57.9 218 95.3 219 Auricle distance development rate (cm/day) Spikelet number/panicle 0.4 180.0 1.9 0.4 104.0 0.6 0.4 63.0 44.1 0.7 0.2 73.3 95.3 48.8 1.3 0.5 153.7 220 95.0 23.9 0.7 0.6 119.3 221 90.3 29.7 0.9 0.6 112.0 222 92.0 64.6 0.9 0.4 120.3 223 97.0 14.5 0.7 0.3 69.3 224 99.0 21.6 2.5 0.3 49.0 225 75.7 40.4 2.2 0.5 79.3 226 94.7 86.2 1.3 0.5 129.3 227 83.0 29.2 1.0 0.6 203.0 228 94.3 36.5 0.5 0.4 120.3 10 216 86.3 23.3 1.6 0.7 157.3 10 217 88.0 40.9 3.2 0.2 89.0 10 218 91.5 31.7 2.3 0.6 187.5 99.0 66.9 1.7 0.4 103.0 10 220 97.3 72.1 -0.3 0.5 165.3 10 221 101.5 34.1 1.2 0.3 76.5 10 222 82.3 23.5 0.9 0.5 89.7 10 223 89.0 41.0 0.7 0.5 112.0 10 224 98.7 32.8 0.7 0.5 116.3 10 227 84.0 35.2 0.7 0.5 159.3 10 228 93.7 14.8 1.0 0.4 99.0 10 229 83.7 47.1 0.7 0.4 98.7 10 230 83.7 17.8 -0.4 0.5 170.7 10 231 88.7 50.8 1.3 0.6 130.0 10 232 79.3 46.1 1.5 0.7 102.0 11 215 92.0 29.1 0.8 0.6 155.0 11 216 98.3 24.1 1.6 0.6 179.0 11 217 99.3 24.6 1.2 0.4 113.3 11 218 94.0 72.2 3.2 0.4 98.0 86.0 50.6 0.3 0.5 93.0 100.7 35.6 1.5 0.6 164.0 216 10 219 11 220 11 222 Spikelet 96 97.3 59.8 0.6 0.5 136.7 101.7 29.2 2.7 0.4 102.0 11 226 96.7 37.3 0.1 0.4 96.3 11 227 81.0 28.3 0.4 0.6 135.7 11 228 84.5 35.4 1.1 0.4 89.0 11 229 100.5 32.9 3.7 0.2 121.0 11 230 85.3 53.6 -0.1 0.5 99.0 11 231 83.0 79.5 0.3 0.7 141.7 11 232 89.0 27.5 1.8 0.7 139.7 12 215 93.0 34.1 -0.2 0.6 155.0 12 216 96.0 21.6 1.3 0.4 112.7 12 218 84.0 12.7 1.8 0.6 100.3 12 219 100.0 62.4 0.5 0.4 93.3 12 221 96.7 67.5 0.3 0.5 169.0 91.0 77.8 0.7 0.6 175.7 12 223 100.3 28.9 0.2 0.6 204.0 12 224 95.3 46.3 -0.3 0.5 120.7 87.5 55.3 1.3 0.4 107.5 12 226 83.0 60.2 1.0 0.6 204.0 12 227 100.3 30.9 0.3 0.5 104.0 12 228 92.7 22.9 0.9 0.5 137.7 12 229 86.7 52.4 0.9 0.5 131.0 12 231 86.0 30.7 1.3 0.6 108.7 12 232 85.7 56.3 0.6 0.7 156.7 13 215 94.3 53.5 1.7 0.5 126.3 13 216 99.0 18.0 0.6 0.5 178.3 13 217 97.7 83.0 0.9 0.3 98.7 13 218 87.7 73.0 0.9 0.2 93.7 13 220 92.7 44.6 0.6 0.5 121.7 13 221 100.0 65.9 1.2 0.6 151.0 95.7 46.1 0.4 0.5 91.7 13 223 97.0 71.3 0.4 0.3 88.0 13 225 83.0 31.2 1.8 0.4 76.0 13 227 84.0 47.1 1.4 0.5 121.3 13 228 98.5 30.5 -0.2 0.5 145.5 13 230 96.0 19.1 1.1 0.2 95.3 13 231 84.3 49.1 0.8 0.6 118.7 13 232 90.7 51.2 0.0 0.5 131.0 14 215 89.3 38.2 1.3 0.6 198.7 14 216 97.0 13.9 -0.1 0.5 149.3 14 217 87.0 61.1 0.2 0.4 106.0 14 218 89.7 46.4 1.6 0.3 125.3 11 223 11 224 12 222 12 225 13 222 97 82.3 43.3 1.1 0.5 107.0 95.0 34.2 0.4 0.5 160.3 14 221 89.3 24.2 0.4 0.5 97.3 14 222 79.5 58.9 1.2 0.7 96.5 14 223 100.5 63.5 1.6 0.4 82.5 14 225 93.5 30.6 0.8 0.4 131.0 14 226 99.0 18.8 0.6 0.3 98.0 14 227 83.5 38.1 1.3 0.4 84.0 14 228 97.3 49.6 1.0 0.4 71.3 14 230 100.0 53.7 1.1 0.5 155.0 15 215 96.0 64.1 1.0 0.5 156.7 15 218 98.3 55.6 0.4 0.4 75.3 15 221 86.3 32.5 0.1 0.8 135.3 15 222 90.0 31.2 1.6 0.6 107.7 91.0 20.4 1.1 0.5 150.3 15 224 96.3 76.0 1.0 0.4 120.3 15 227 97.5 66.5 0.9 0.3 106.0 88.7 35.9 1.3 0.4 92.0 15 232 82.0 36.0 -0.2 0.7 104.0 Kyeema 100.7 30.6 0.7 0.5 194.3 Norin 73.0 47.3 1.4 0.7 97.7 Sherpa 90.3 36.7 0.3 0.4 116.7 Minimum least significant difference 11.3 28.3 -0.5 0.23 67.0 Average least significant difference 11.8 32.6 0.9 0.24 70.7 Maximum least significant difference 17.0 49.35 4.0 0.35 116.8 14 219 14 220 15 223 15 229 98 Appendix Air and water temperatures in the warm and low temperature glasshouses in Experiment 99 Appendix Air temperatures in the warm and low temperature glasshouses in Experiment 100 Appendix Correlation coefficient matrix of spikelet number/panicle and spikelet sterility/main stem of 10 genotypes and varieties under warm and low temperature conditions (flooded -N and +N and aerobic +N) in Experiment (Spi/p: Spikelet number/panicle; %SS/ms: % spikelet sterility/main stem; Fl: Flooded; A: Aerobic) Spi/p (Fl warm +N) Spit/p (A warm +N) Spi/p (Fl cold -N) Spi/p (Fl cold +N) Spi/p (A cold +N) %SS/ms (Fl warm -N) %SS/ms (Fl warm +N) %SS/ms (A warm +N) %SS/ms (Fl cold -N) %SS/ms (Fl cold +N) %SS/ms (A cold +N) Spi/p Spi/p Spit/p Spi/p Spi/p Spi/p %SS/ms (Fl warm -N) (Fl warm +N) (A warm +N) (Fl cold -N) (Fl cold +N) (A cold +N) (Fl warm -N) %SS/ms (Fl warm +N) %SS/ms %SS/ms %SS/ms (A warm +N) (Fl cold -N) (Fl cold +N) 0.67** 0.71** 0.55* 0.80** 0.45ns 0.58* 0.58* 0.77** 0.45ns 0.40ns 0.52ns 0.44ns 0.85** 0.35ns 0.34ns -0.12ns 0.18ns -0.03ns -0.09ns -0.23ns -0.06ns -0.21ns 0.14ns -0.05ns -0.23ns 0.06ns 0.21ns 0.16ns 0.03ns 0.14ns -0.10ns -0.18ns 0.09ns 0.14ns 0.08ns 0.89** 0.11ns 0.37ns 0.52ns 0.23ns 0.50ns 0.42ns 0.04ns 0.42ns 0.20ns 0.07ns 0.34ns 0.46ns 0.18ns 0.49ns 0.40ns -0.05ns 0.51ns 0.27ns 0.96** 0.06ns 0.42ns 0.33ns 0.16ns 0.50ns 0.27ns 0.09ns 0.63* 0.43ns 0.91** 101 0.90** ... were to identify genotypic variation for tolerance to low temperature stress in rice using a population developed from a cross between low temperature tolerant and susceptible parents, and to determine... were to identify genotypic variation for tolerance to low temperature stress in rice using a population developed from a cross between low temperature tolerant and susceptible parents, and to determine... temperature, including prolonged periods of low temperature, is one of the main production constraints in temperate rice producing countries in general, and in Australia in particular For instance,