Development and evaluation of a system for the study of mineral nutrition of sacred lotus (Nelumbo nucifera). David. J. Hicks B.Hort.Sci. (Hons.) University of Western Sydney, Hawkesbury A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy Centre for Horticulture and Plant Sciences University of Western Sydney, Hawkesbury. Australia July 2005 Acknowledgements In the capacity of supervisor(s) I would like to thank Dr Tony Haigh (Principal supervisor) for advice and guidance in the areas of experimental design, results interpretation, thesis construction, editing, and patience with a student who in hindsight must have been a frustrating experience. Similarly, Dr Vong Q. Nguyen (Co-supervisor) provided a perspective of the thesis within the Asian vegetable scenario as it is in Australia, helping with employment opportunities in similarly oriented work, patience with my many deadline far misses and his infectious enthusiasm. On the technical front I would like to note the input of Emma Borchet, Aaron Simmons, Leonie Noakes and Colonel James Sheehy for greasing the wheel with assistance in various trial technical setups and material acquisitions. Further, the staff in Horticulture at UWSH Richmond, too many to identify individually, require recognition for the many small but necessary activities which help to gel the whole, especially CHAPS for funding the tissue analysis. Initially tissue tests were conducted by staff at CSIRO and Agrifoods Technology. Secondary testing was performed by the team at the Waite campus of Adelaide University. At the industry level, the Rural Industries Research and Development Corporation (RIRDC) deserve recognition for initial funding of the ‘Agronomic and physiological studies of Nelumbo nucifera for export to Asia’ project. Kim and Vicky Jones warrant a mention for allowing the frequent inspection of their fledgling lotus farm and the provision of ‘Green Jade’ rhizomes for planting at Richmond. Thanks to Peter McLaughlin and assorted folk, at the now defunct NORADA, for an unwavering belief in my abilities, encouragement and motivational support. On the personal side the most thanks are reserved for my family, especially mum and dad for emotional, financial and domestic support including delivery of food preparations, lawn mowing, dishwashing and vacuuming of my seedy hovel. Thanks to brothers Steve and Col for encouragement and motivation. Respect to friends, Dave Gearin, Dean Francis, Matt Barnett, Steve Anderson, Jo Campbell, Matt Carroll, Liz Reichardt, Peter Lister, Jacqui O’Brien and Martin Ayers for enduring support and tolerance. This thesis is dedicated to the lamb of Dog, the Rt. Rev. Dr Jello Fruits, and my lotus flower Imogen. Statement of Authentication The work presented in this thesis is, to the best of my knowledge and belief, original except as acknowledged in the text. I hereby declare that I have not submitted this material, either in full or in part, for a degree at this or any other institution. ………………………………………… David. J. Hicks. Table of Contents Volume I Development and Evaluation of a System for the Study of Mineral Nutrition of Sacred Lotus (Nelumbo nucifera). Abbreviations vi Abstract vii Chapter 1: General Introduction 1 1.1 Background 1 1.1.1 Asian Vegetables 1 1.1.2 Australian Position 2 1.2 Lotus: Old Crop, New Opportunities 3 1.2.1 Lotus Descriptors 3 1.2.2 Uses of Lotus 6 1.2.3 Commercial Potential 7 1.2.4 Crop production: Techniques and Feasibility 8 1.2.5 Constraints to Production 10 1.3 Solution Culture 14 1.3.1 Definitions and History 14 1.3.2 Techniques 15 1.3.3 Media 17 1.3.4 Solutions 17 1.3.5 Uses 19 1.3.6 Rationale for Choice of System for Lotus 20 1.4 Scope of Research 22 1.4.1 Development of a System for Replicated Trials for Lotus 22 1.4.2 Examine the Effects on Growth and Plant Organ Major Nutrient Concentration 23 Chapter 2 Seeds 26 2.1 Introduction 26 2.2 Seed Generation 27 2.2.1 Experiment 1 Observation of the Generation of Lotus (Nelumbo nucifera) Seeds. 27 2.2.3 Materials and Methods 27 2.2.3 Results and Discussion 28 2.3 Seed Germination 29 2.3.1 Experiment 2 Effect of Temperature on the Germination of Lotus (Nelumbo nucifera) Seed.29 2.3.1.2 Materials and Methods 29 2.3.1.3 Results and Discussion 30 2.3.2 Experiment 3 Estimation of the Number of i Seeds to be Sown for the Production of Uniform Lotus (Nelumbo nucifera) Seedlings. 31 2.3.2.2 Materials and Methods 31 2.3.2.3 Results and Discussion 32 2.4 Discussion 34 Chapter 3 Plant Culture 37 3.1 Introduction 37 3.2 Container Trials 38 3.2.1 Experiment 4 Observation Trial on the Effects of N, P and K on Growth and Tissue Composition of Lotus (Nelumbo nucifera) Cultivated in 1 m3 Containers. 38 3.2.1.2 Materials and Methods 38 3.2.1.2 Results and Discussion 39 3.2.2 Experiment 5 Observation of the Growth of Lotus (Nelumbo nucifera) in Containers of Differing Dimensions. 42 3.2.2.1 Materials and Methods 42 3.2.2.2 Results and Discussion 43 3.3 Solution Quality 44 3.3.1 Experiment 6 Effect of Solution Electrical Conductivity on Growth of Lotus (Nelumbo nucifera). 44 3.3.1.1 Materials and Methods 44 3.3.1.2 Results and Discussion 45 3.3.2 Experiment 7 Effect of Solution pH on Growth of Lotus (Nelumbo nucifera). 51 3.3.2.1 Materials and Methods 51 3.3.2.2 Results and Discussion 52 3.3.3 Experiment 8 Observation of Growth of Lotus (Nelumbo nucifera) from Cuttings. 57 3.3.3.1 Materials and Methods 57 3.3.3.2 Results and Discussion 57 3.3.4 Experiment 9 Observation on the effect of Algaecides on the Growth of Lotus (Nelumbo nucifera) and Control of Solution Algae. 59 3.3.4.1 Materials and Methods 59 3.3.4.2 Results and Discussion 59 3.4 Discussion 61 ii Chapter 4 Analysing the Interaction between Nitrogen and Lotus 64 4.1 Introduction 64 4.1.1 Experiment 10 Effect of Nitrogen on Growth and Tissue Nutrient Concentration of Lotus (Nelumbo nucifera) 67 4.2 Materials and Methods 67 4.2.1 Plant Culture 67 4.2.2 Experimental Design 69 4.3 Results 72 4.3.1 Observations of Nitrogen Supply on Visual Growth Expression. 72 4.3.2 Nitrogen Supply Effect on Growth Parameters. 74 4.3.3 Effect of Nitrogen Supply on Organ Nutrient Concentration. 79 4.3.4 Analysis of Major Nutrient Concentration as a Function of Nitrogen Concentration. 84 4.3.5 Analysis of Growth as a Function of Organ Nitrogen Concentration. 90 4.3.6 Analysis of Total Dry Mass as a Function of Nitrogen Affected Growth Parameters. 96 4.3.7 Analysis of Growth Parameters Affected by Nitrogen Concentration, as a Function of Nitrogen Concentration Affected Organ Nutrient Concentration. 98 4.4 Discussion 103 Chapter 5 Analysing the Interaction between Phosphorous and Lotus 116 5.1 Introduction 116 5.1.1 Experiment 11 Effect of Phosphorous on Growth and Tissue Nutrient Concentration of Lotus (Nelumbo nucifera) 119 5.2 Materials and Methods 119 5.3 Results 120 5.3.1 Observations of Phosphorous Supply on Visual Growth Expression. 120 5.3.2 Phosphorous Supply Effect on Growth Parameters. 121 5.3.3 Effect of Phosphorous Supply on Organ Nutrient Concentration. 126 5.3.4 Analysis of Major Nutrient Concentration as a Function of Phosphorous Concentration. 131 5.3.5 Analysis of Growth as a Function of Organ Phosphorous Concentration. 138 5.3.6 Analysis of Total Dry Mass as a Function of Phosphorous Affected Growth Parameters. 145 iii 5.3.7 Analysis of Growth Parameters Affected by Phosphorous Concentration, as a Function of Phosphorous Concentration Affected Organ Nutrient Concentration. 147 5.4 Discussion 152 Chapter 6 Analysing the Interaction between Potassium and Lotus 157 6.1 Introduction 157 6.1.1 Experiment 12 Effect of Potassium on Growth and Tissue Nutrient Concentration of Lotus (Nelumbo nucifera) 159 6.2 Materials and Methods 159 6.3 Results 160 6.3.1 Observations of Potassium Supply on Visual Growth Expression. 160 6.3.2 Potassium Supply Effect on Growth Parameters. 161 6.3.3 Effect of Potassium Supply on Organ Nutrient Concentration. 166 6.3.4 Analysis of Major Nutrient Concentration as a Function of Potassium Concentration. 171 6.3.5 Analysis of Growth as a Function of Organ Potassium Concentration. 177 6.3.6 Analysis of Total Dry Mass as a Function of Potassium Affected Growth Parameters. 181 6.3.7 Analysis of Growth Parameters Affected by Potassium Concentration, as a Function of Potassium Concentration Affected Organ Nutrient Concentration. 182 6.4 Discussion 185 Chapter 7 Analysing the Interaction between Calcium and Lotus 189 7.1 Introduction 189 7.1.1 Experiment 13 Effect of Calcium on Growth and Tissue Nutrient Concentration of Lotus (Nelumbo nucifera) 192 7.2 Materials and Methods 192 7.3 Results 193 7.3.1 Observations of Calcium Supply on Visual Growth Expression. 193 7.3.2 Calcium Supply Effect on Growth Parameters.194 7.3.3 Effect of Calcium Supply on Organ Nutrient Concentration. 198 7.3.4 Analysis of Major Nutrient Concentration as a Function of Calcium Concentration. 203 iv 7.3.5 Analysis of Growth as a Function of Organ Calcium Concentration. 209 7.3.6 Analysis of Total Dry Mass as a Function of Calcium Affected Growth Parameters. 214 7.3.7 Analysis of Growth Parameters Affected by Calcium Concentration, as a Function of Calcium Concentration Affected Organ Nutrient Concentration.