Application of Biotechnology to Davidsonia Species - an Australian Native Plum Author Nand, Narendra Published 2008 Thesis Type Thesis (PhD Doctorate) School School of Biomolecular and Physical Sciences DOI https://doi.org/10.25904/1912/1940 Copyright Statement The author owns the copyright in this thesis, unless stated otherwise. Downloaded from http://hdl.handle.net/10072/365517 Griffith Research Online https://research-repository.griffith.edu.au Application of Biotechnology to Davidsonia species – an Australian Native Plum by Narendra Nand BSc GCEd. , MSc Hons (Biotech) School of Biomolecular and Physical Sciences Centre for Forestry and Horticultural Research Griffith University A thesis submitted in the fulfillment of the requirements of the degree of Doctor of Philosophy Griffith University September, 2007 Statement of Originality The material presented in this thesis is based on my own research and findings. The work has not previously been submitted for a degree or diploma in any university. To the best of my knowledge and belief, the thesis contains no material previously published or written by another person except where due reference is made in the thesis itself. ___________________ Narendra Nand ii Acknowledgements I thank all growers, producers and Bushfood industry groups who provided me with information for the bushfoods survey. I greatly appreciate the initial assistance provided by the editor of the Australian Bushfoods Magazine, Sammy Ringer for introducing me to the Bushfoods Indusrty Group. Ongoing assistance from Jan Sinclair of Mother Nature’s Bushtucker Nursery, Tumbulgum; Krish Kupsch of Burringbar; Martha Shepard and David Haviland of Galeru Bushfoods Plantation; Graham and Veronica of Witjuti Grub Nursery; John and Mary King of Rainforest Liqueurs; John Wrench and other members of Queensland Bushfoods Association for providing ongoing information and support. I would like to thank my laboratory colleagues, supervisors Dr Sarah E. Ashmore and Professor Roderick A. Drew and my friend Dr Cameron P. Peace for data analysis and for their assistance in writing and editing the thesis and research papers. Their continuous advice and encouragement enabled me to persevere and continue my part-time study, juggled between a full-time demanding job. A special thanks to my principal supervisor, Professor Rod Drew for his kindness, understanding and tolerance; the only source of inspiration during trying times. I greatly appreciate and thank my son Kaushik N. Nand and daughter Wanshika P. Nand for assisting me with the tricky computing skills and for sharing my work responsibilities. I also thank my mum for her support and encouragement throughout my studies and the health care she provided me during my recovery from surgery. Lastly, I would like to thank my wife, brother, sisters, other family and friends who appreciated and encouraged me to pursue my personal academic ambitions. Dedicated to my parents who inspired their nine children to attain the highest possible education. iii TABLE OF CONTENTS Statement of Originality ii Acknowledgements iii List of Abbreviations viii List of Tables ix List of Figures xii Abstract 1 1 Chapter 1: A survey of edible Australian native fruit species in the South- East Queensland and North-East New South Wales Coastal regions 5 1.1 Abstract 6 1.2 Introduction 7 1.2.1 Overview of Australian flora 7 1.2.2 The Australian bushfood industry 8 1.2.3 Application of biotechnology to Australian natives 11 1.3 Materials & Methods 17 1.3.1 The Survey 17 1.3.2 The questionnaire 18 1.4 Results 22 1.4.1 Questionnaire 22 1.4.2 The responses from the questionnaire 22 1.5 Discussion 29 1.5.1 Description of five main species 29 1.5.2 Questionnaire 32 1.5.3 Selection of species with high commercial potential 32 1.5.4 Propagation methods and constraints 34 1.5.5 Research and development 35 1.5.6 Commercialisation and constraints 36 1.6 Conclusions 37 Appendix to chapter 1 40 Appendix 1.1 41 iv Appendix 1.2 42 Appendix 1.3 43 2 Chapter 2: Micropropagation of two Australian native fruit species, Davidsonia pruriens (F. Muell.) and Davidsonia jerseyana (F. Muell. ex F.M. Bailey) G. Harden & J.B. Williams 44 2.1 Abstract 45 2.2 Introduction 46 2.3 Materials and methods 47 2.3.1 Plant material 47 2.3.2 Establishment of seedlings in vitro 47 2.3.3 In vitro shoot growth of juvenile tissue 48 2.3.4 In vitro shoot culture of adult tissues 48 2.3.5 In vitro root growth 49 2.3.6 Data analysis 50 2.4 Results 50 2.4.1 In vitro shoot development from juvenile tissues 50 2.4.2 In vitro shoot development from adult tissues 52 2.4.3 In vitro development 53 2.5 Discussion 57 2.6 Conclusions 61 3 Chapter 3: Organogenesis in Davidsonia species 62 3.1 Abstract 63 3.2 Introduction 64 3.3 Materials and methods 67 3.3.1 Plant material 67 3.3.2 Culture media 67 3.3.3 Callus induction on specific explants 69 3.3.4 Organogenesis 70 3.3.5 Organogenesis – optimisation of shoot induction 71 3.3.6 Organogenesis – optimisation of duration of exposure of best explants on shoot induction medium 72 3.4 Results 73 3.4.1 Callus induction 73 3.4.2 Organogenesis 77 3.4.3 Organogenesis – optimisation of shoot induction in various explants 79 v 3.4.4 Organogenesis - optimisation of duration of exposure of best explants on shoot induction medium 83 3.5 Discussion 87 3.6 Conclusions 92 Appendix to chapter 3 93 Appendix 3.1 94 Appendix 3.2 95 Appendix 3.3 99 Appendix 3.4 100 4 Chapter 4: Improved micropropagation protocol for Davidsonia pruriens and Davidsonia jerseyana 101 4.1 Abstract 102 4.2 Introduction 103 4.3 Materials and methods 105 4.3.1 Plant material 105 4.3.2 Culture media 106 4.3.3 Recording and analysis of data 106 4.3.4 Effect of GA3 treatment on various explants 107 4.3.5 Effect of residual GA3 on root response 107 4.3.6 Effect of various concentrations of G A3 in combination with optimum shoot growth medium 108 4.3.7 Effect of 32.2 µM IBA on root response 108 4.3.8 Effect of 1.0 µM G A3 concentration in with SIM 108 4.4 Results 109 109 4.4.1 Effect of GA3 treatment on various explants 109 4.4.2 Effect of lower range of GA3 concentration on nodal explants 110 4.4.3 Effect of residual GA3 on root response 111 4.4.4 Effect of various concentrations of GA3 in combination with optimum shoot growth medium 113 4.4.5 Effect of 32.2 µM IBA on root response 115 4.4.6 Effect of 1.0 µM G A3 concentration in combination with SIM 116 4.5 Discussion 119 4.6 Conclusions 123 vi 5 Chapter 5: Genetic Analysis of Davidsonia species with Randomly Amplified DNA Fingerprinting (RAF) markers 124 5.1 Abstract 125 5.2 Introduction 126 5.3 Methods and materials 128 5.3.1 Plant material 128 5.3.2 Genomic DNA extraction and purification 129 5.3.3 Selection of primers 130 5.3.4 DNA amplification by PCR 130 5.3.5 Data analysis 131 5.4 Results 131 5.5 Discussion 141 5.6 Conclusions 146 6 Chapter 6: Conclusions 148 Bibliography 158 Appendix 1 189 Appendix 2 200 vii List of Abbreviations ANBIC – Australian Native Bushfood Industry Committee ANFIL – Australian Native Food industry Limited CSIRO – Commonwealth Scientific and Industrial Research Organisation DIA – Davidsonia Industry Association DPI – Department of Primary Industries NSW – New South Wales QBA – Queensland Bushfoods Association QDPI – Queensland Department of Primary Industries Qld. – Queensland RIRDC – Rural Industries Research and Development Corporation SGAP – Society for the Growers of Australian Plants Scientific Abbreviations: BA - N6-benzyladendine 2,4-D - 2,4-dichlorophenoxyacetic acid GM - growth medium IBA – Indole Butyric Acid js - juvenile seedling jm - juvenile micropropagated mf - mature field mg - mature green house mm - mature micropropagated MS - Murashige and Skoog NAA - 1-naphthalene acetic acid PGR - plant growth regulator SIM - shoot induction medium TDZ - thidiazuron viii LIST OF TABLES Chapter 1 Table 1.1 Australian native fruit species selected by other authors in their reports 21 Table 1.2 Number and percentage of growers and sellers of plant or plant products of Australian native fruit species in south east Queensland and north east New South Wales 23 Table 1.3 Type of native plant products of top 15 species being currently sold 23 Table 1.4 Selection of five native fruit species of high commercial potential, listed by the number and percentage of growers and or sellers 24 Table 1.5.1 The source of seeds or fruits and plants obtained by the number and percentage of growers 24 Table 1.5.2 Methods of propagation for top 15 species used by the number of growers for the species selected by the growers 25 Table 1.6 Knowledge of type and level of research being currently undertaken on some Australian native edible species, listed by the growers 26 Table 1.7 Research and development support from organizations and institutions obtained by growers, processors and product sellers of bushfood industry 26 Table 1.8 Number of respondents indicating the type of problems in the commercialisation of Australian native fruit species 28 Table 1.9 Additional general comments/ suggestions provided by some respondents 29 Table 1.10 The major constraints or problems faced by the number of growers in the propagation of Australian native fruit species 41 Table 1.11 The number of processors having problems in processing of their Australian native fruits 41 Table 1.12 Number of respondents indicating the difficulty in finding market for their products 41 ix Table 1.13 The most common native species and their products being developed for commercial purposes and their estimated volume and value 43 Chapter 2 Table 2.1 Effect of various cytokinin concentrations on the growth response of axillary shoots of D.