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Characterising the Growth Response and Pathogenicity of Phytophthora Agathidicida in Soils from Contrasting Land-Uses

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Characterising the Growth Response and Pathogenicity of Phytophthora Agathidicida in Soils from Contrasting Land-Uses Lincoln University Digital Thesis Copyright Statement The digital copy of this thesis is protected by the Copyright Act 1994 (New Zealand). This thesis may be consulted by you, provided you comply with the provisions of the Act and the following conditions of use: you will use the copy only for the purposes of research or private study you will recognise the author's right to be identified as the author of the thesis and due acknowledgement will be made to the author where appropriate you will obtain the author's permission before publishing any material from the thesis. Characterising the growth response and pathogenicity of Phytophthora agathidicida in soils from contrasting land-uses A thesis submitted in partial fulfilment of the requirements for the Degree of Master of Science at Lincoln University by Kai Lewis Lincoln University 2018 Abstract of a thesis submitted in partial fulfilment of the requirements for the Degree of Master of Science. Characterising the growth response and pathogenicity of Phytophthora agathidicida in soils from contrasting land-uses by Kai Lewis The genus Phytophthora (Oomycetes, Peronosporales, Pythiaceae) is responsible for several forest declines worldwide (i.e. jarrah dieback in Australia (P. cinnamomi) and sudden oak death in California and Europe (P. ramorum)). The recently described pathogen, P. agathidicida, is the causal agent of dieback in remnant stands of New Zealand kauri (Agathis australis), and poses a significant threat to the long-term survival of this iconic species. However, what is least understood are how key physicochemical parameters (e.g. soil pH and soil organic matter) influence growth and pathogenicity of P. agathidicida. This study examined the effects of three contrasting land-uses (kauri forest, grazed pasture, short-rotation pine plantation (Pinus radiata) on the growth and pathogenicity of P. agathidicida in soils sampled from Waipoua Forest, Northland, New Zealand. This was investigated usin: 1) Growth response assay, 2) Pathogenicity trait study with blue lupin (Lupinus angustifolius), and 3) Pasture and pine alternative host infection study. Experiment 1 found that significantly greater sporangia (p < 0.001) and oospore (p <0.01) counts occurred within pasture and pine soils compared to kauri soils, suggesting that they favour P. agathidicida in the early stages of establishment. Additionally, significant increases in oospores (p <0.01) over time in the pine soils potentially suggest their enhanced capacity to act as pathogen reservoirs. Furthermore, two new Phytophthora spp. to New Zealand (P. pini and P. gregata) were identified in this study. Experiment 2 identified non-significant land-use effects on pathogenicity traits (e.g. lesion presence, lesion length etc.) of P. agathidicida inoculation of blue lupin. Experiment 3 also confirmed the potential for pasture and pine to act as reservoirs for P. agathidicida. Overall, the findings of this study revealed that contrasting land-use affects the growth of P. agathidicida in soil, and further detailed study of the activity, distribution and pathogenicity of P. agathidicida in fragmented landscapes is warranted. Keywords: Phytophthora agathidicida, Waipoua Forest, Kauri (Agathis australis), land-use impact, growth response, pathogenicity, alternative hosts ii Table of Contents Characterising the growth response and pathogenicity of Phytophthora agathidicida in soils from contrasting land-uses ..................................................................................... i Table of Contents ............................................................................................................... iii List of Tables ..................................................................................................................... vii List of Figures ..................................................................................................................... ix Acknowledgements .......................................................................................................... xiv Chapter 1 : Literature review ........................................................................................... 1 1.1 The history and current conservation status of New Zealand Kauri (Agathis australis) ...................................................................................................................... 1 The role of Phytophthora agathidicida in kauri dieback ................................ 2 1.2 History of the genus Phytophthora.............................................................................. 3 1.3 The pathology of Phytophthora agathidicida and kauri dieback ................................ 4 Discovery, identification and distribution ....................................................... 4 Symptoms ........................................................................................................ 7 Life cycle ......................................................................................................... 8 1.4 Effect of abiotic and biotic factors on Phytophthora ................................................ 11 Abiotic factors ............................................................................................... 11 Biotic factors .................................................................................................. 13 1.5 Managing the effects of Phytophthora spp. .............................................................. 16 Detection tools ............................................................................................... 16 Preventing pathogen-host contact .................................................................. 17 Host resistance ............................................................................................... 18 Fungicide usage ............................................................................................. 19 1.6 Summary and project aims ........................................................................................ 20 Summary ........................................................................................................ 20 Project aims ................................................................................................... 20 iii Chapter 2 : Characterising the growth and disease expression of Phytophthora agathidicida in soils of contrasting land-uses .................................................................. 22 2.1 Introduction ............................................................................................................... 22 2.2 Research aim ............................................................................................................. 23 2.3 Materials and Methods .............................................................................................. 24 Study area ...................................................................................................... 24 Site history ..................................................................................................... 24 Sampling period ............................................................................................. 25 Sample collection and processing .................................................................. 26 Selective medium .......................................................................................... 27 Growth response assay .................................................................................. 27 Spore counts .................................................................................................. 30 Phytophthora baiting ..................................................................................... 32 Soil chemical/physical analyses .................................................................... 33 Statistical analyses ......................................................................................... 34 2.4 Results ....................................................................................................................... 35 Soil baiting ..................................................................................................... 35 Analysis of physicochemical data between land-uses ................................... 37 Spore count data analysis .............................................................................. 40 Influence of environmental variables on spore counts .................................. 46 2.5 Discussion.................................................................................................................. 47 Phytophthora populations associated with land-use soils ............................. 47 Kauri Dieback Survey ................................................................................... 48 Physicochemical variable analysis ................................................................ 48 Count variables .............................................................................................. 50 Environmental drivers of Phytophthora spore numbers ................................ 54 2.6 Conclusion ................................................................................................................. 56 Chapter 3 : Impact of contrasting land-uses on Phytophthora agathidicida pathogenicity using blue lupin (Lupinus angustifolius) as a model plant system ........ 57 3.1 Introduction ..............................................................................................................
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