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Parkinsonia Aculeata Investigating the cause of dieback in the invasive plant, Parkinsonia aculeata BY TRACEY VIVIEN STEINRUCKEN A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy at Western Sydney University in 2017 This page has been intentionally left blank “Watch with glittering eyes the whole world around you because the greatest secrets are always hidden in the most unlikely places. Those who don't believe in magic will never find it” -- Roald Dahl This page has been intentionally left blank Acknowledgements I would like to thank my advisors Rieks van Klinken (CSIRO Health & Biosecurity), Andrew Bissett (CISRO Oceans & Atmosphere) and Jeff Powell (Hawkesbury Institute for the Enivronment, Western Sydney University) for their excellent mentoring, patient communication across borders and constant support. This research project was supported by Meat and Livestock Australia via a technical assistance grant (B.STU.0271). My PhD was supported by the Australian Government via an Australian Postgraduate Award and Western Sydney University via a top-up stipend. The Hawkesbury Institute for the Environment also supported my work with an annual research allocation and conference attendance funding. Thanks to Patricia Hellier, David Harland, Ian Anderson and Lisa Davison at HIE for administrative support. Thank-you to Kelli Pukallus (Biosecurity Queensland), Andrew White (CSIRO), Eva Pôtet (Agro Campus Oest, Paris), Marcus Klein (HIE at WSU), Donald Gardiner (CSIRO), Shamsul Hoque (CSIRO), Ryan O’Dell (DAFF) and Dylan Smith (UC Berkeley) for field and technical support in various chapters throughout this thesis. Huge thanks to my CSIRO Biosecurity team: Gio Fichera, Ryan Zonneveld, Brad Brown, Andrew White and Jeff Makinson for technical support in Chapter 3. Also to the rest of the ESP staff from CSIRO Health & Biosecurity, CSIRO Agriculture & Food and CSIRO Land and Water for many morning teas, encouragement and advice, career inspiration and long-lasting friendships. Thanks to Giles Hardy, Sonia Aghighi and Briony Williams from CPSM at Murdoch University for their work on Chapter 4. Thanks also to the Australian-American Fulbright Commission, the various Fulbright and IIE chapters in the USA, Prof Matteo Garbelotto, lab manager Doug Schmidt, and colleagues Laura Sims, Tina Popenuck and Mike Johnson from the Forest Pathology and Mycology Laboratory at UC Berkeley. Their support made Chapters 5 and 6 possible. Thank-you Louise Morin, Donald Gardiner, Kylie Ireland, Gavin Hunter and Luke Barrett (CSIRO) for internal reviews. Thanks to my incredible parents, Avril and Viv, for giving me the tools and confidence to challenge everything and for always believing in me. And to my brothers, Grant and Warren, who are my unceasing inspiration and have always offered support and understanding. Finally, to Bastian, for your sacrifices, patience, encouragement, neck rubs, our fieldwork adventures, your understanding and your love: you are the reason I can look back on this experience with an abundance of fond memories. Jag älskar dig. This page has been intentionally left blank 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. Signed: ___ __ Tracey Vivien Steinrucken This page has been intentionally left blank TABLE OF CONTENTS List of Figures ................................................................................................................................ v List of Tables ............................................................................................................................... vii Abbreviations .............................................................................................................................. viii Thesis Abstract ............................................................................................................................. ix CHAPTER 1: Microbe-plant interactions, their influence on invasion ecology, and the use of dieback in the biocontrol of invasive plants: a review ......................................................... 1 1.1 Introduction ........................................................................................................................... 1 1.2 Plant-microbe interactions ..................................................................................................... 4 1.3 The roles of endophyte and other microorganisms in plant invasion .................................... 6 1.4 Plant dieback ......................................................................................................................... 8 1.4.1 Pathogen-related dieback of trees .................................................................................. 10 1.5 Dieback in Weeds of National Significance (WONS) ........................................................ 13 1.5.1 Parkinsonia aculeata ..................................................................................................... 14 1.5.2 The potential for using dieback in the biological control of parkinsonia ....................... 22 1.6 Summary ............................................................................................................................. 23 1.7 Thesis outline ...................................................................................................................... 24 1.8 Publications ......................................................................................................................... 26 CHAPTER 2: Endophyte community composition is associated with dieback occurrence in an invasive tree ........................................................................................................................ 30 2.1 Abstract ............................................................................................................................... 30 2.2 Introduction ......................................................................................................................... 31 2.3 Materials and methods ........................................................................................................ 36 2.3.1 Sampling and sample preparation .................................................................................. 36 2.3.2 Nucleic acid analysis ..................................................................................................... 40 2.3.3 Statistical data analysis .................................................................................................. 41 2.4 Results ................................................................................................................................. 42 2.4.1 Sampling effort .............................................................................................................. 42 2.4.2 Plant part and disease status effects ............................................................................... 42 2.4.3 Indicator species analysis .............................................................................................. 46 2.5 Discussion ........................................................................................................................... 46 2.6 Conclusion........................................................................................................................... 51 CHAPTER 3: Triggering dieback in an invasive plant: endophyte diversity and pathogenicity ... 52 3.1 Abstract ............................................................................................................................... 52 3.2 Introduction ......................................................................................................................... 53 3.3 Materials and method .......................................................................................................... 57 3.3.1 Sampling, identification and analysis of the fungal endophyte community .................. 57 i 3.3.2 Glass house pathogenicity trial ...................................................................................... 59 3.4 Results ................................................................................................................................. 64 3.4.1 Fungal endophytes in healthy and dieback-affected Parkinsonia aculeata ................... 64 3.4.2 Pathogenicity testing of isolates with a water stress interaction .................................... 69 3.5 Discussion ........................................................................................................................... 73 CHAPTER 4: First report of oomycetes associated with the invasive tree Parkinsonia aculeata (family: Fabaceae) .............................................................................................................. 78 4.1 Abstract ............................................................................................................................... 78 4.2 Introduction ......................................................................................................................... 79 4.3 Materials and Methods ........................................................................................................ 82 4.3.1 Field survey and sampling ............................................................................................
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