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The Potential of the Fungicide Phosphite to Control Phytophthora Cinnamomi in Native Plant Communities Associated with Mining

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The Potential of the Fungicide Phosphite to Control Phytophthora Cinnamomi in Native Plant Communities Associated with Mining MINERALS AND ENERGY RESEARCH INSTITUTE OF WESTERN AUSTRALIA (MERIWA) REPORT NO. 209 THE POTENTIAL OF THE FUNGICIDE PHOSPHITE TO CONTROL PHYTOPHTHORA CINNAMOMI IN NATIVE PLANT COMMUNITIES ASSOCIATED WITH MINING Results of research carried out as MERIWA Project No. M280 at the School of Biological Sciences, Murdoch University by G Hardy, B Dell, I Colquhoun and J McComb December, 2000 Distributed by: MERIWA Mineral House 100 Plain Street EAST PERTH WA 6004 Crown copyright reserved To which all enquiries should be addressed ISBN 0 7309 8525 3 Contents Page Acknowledgments……………………………………………………………………………………1 Photographs…………………………………………………………………………………………..2 Executive Summary…………………………………………………………………………………. 4 Recommendations…………………………………………………………………………………… 7 Chapter 1 General introduction………………………………………………………………10 Chapter 2 Long-term effectiveness of phosphite to control Phytophthora cinnamomi……… 12 2.1 The long-term ability of phosphite to control Phytophthora cinnamomi in two native plant communities……………………………………………………. 13 2.2 The long-term ability of phosphite to control Phytophthora cinnamomi in five plant species native to Western Australia, in a glasshouse trial…………….. 34 2.3 The long-term prevention of death of jarrah forest plant species treated with phosphite and inoculated with Phytophthora cinnamomi………………….. 48 Chapter 3 Biology of phosphite-Phytophthora cinnamomi and host plant interactions……. 54 3.1 Sensitivity of Phytophthora cinnamomi isolates to phosphite in planta.………... 56 3.2 Sensitivity of Phytophthora cinnamomi isolates to phosphite in planta………… 61 3.3 The effect of phosphite on Phytophthora cinnamomi zoospore production in planta…………………………………………………………………………. 74 3.4 The efficacy of phosphite in reducing the infection and colonisation of the roots of a range of Western Australian native plant species by Phytophthora cinnamomi……………………………………………………………………….. 84 Chapter 4 Possible deleterious effects of phosphite………………………………………... 90 4.1 The effect of phosphite concentration on phytotoxicity in a range of plant species at Tiwest………………………………………………………………… 91 4.2 The effect of phosphite on seed production and seed viability………………….. 97 Chapter 5 Technical issue and operational trial…………………………………………….. 101 5.1.1 Facile high performance ion chromatographic analysis of ii phosphite and phosphate in plant samples……………………………... 102 5.1.2 The problems with phosphite analysis…………………………………. 108 5.2 Operational spray……………………………………………………………...… 110 Chapter 6 Associated studies……………………………………………………………….. 113 6.1 The effect of the fungicide phosphite on sexual reproduction of non-native and native plant species of the Eucalyptus marginata forest……………………. 114 6.2 The effect of phosphite on ectomycorrhiza……………………………………… 116 6.3 Phosphite distribution in E. marginata after foliar treatment with phosphite…... 117 Chapter 7 List of recommendations………………………………………………………… 118 References…………………………………………………………………………………………... 122 Appendix 1 Comparisons of phosphite concentrations in Eucalyptus calophylla tissues after spray, mist or soil drench applications with the fungicide phosphite……… 128 iii List of Tables Executive summary Table 1 Project objectives, the activities undertaken to meet the objectives and the outcomes of the project…………………………………………………..……... 4 Chapter 2.1 Table 2.1 Jarrah forest trial of phosphite application. Environmental conditions before inoculation and between inoculation and harvest……………………………..… 23 Table 2.2 Northern Sandplain trial of phosphite application. Environmental conditions before inoculation and between inoculation and harvest.……………………..… 27 Table 2.3 Effect of phosphite rate and time of application on visible phytotoxicity at 2 to 7 weeks and plant growth after >1.5 years after application………………... 29 Chapter 2.2 Table 2.4 Minimum and maximum temperatures (°C) between inoculation and harvest in Experiment 1..…………….………………..………………………………….. 39 Table 2.5 Average inhibition of Phytophthora cinnamomi growth in three plant species sprayed with 5 or 10 g phosphite/L and the average phosphite concentration detected in the stems of these plants (n=6)…………………….… 39 Table 2.6 The number of plants surviving after they were inoculated with Phytophthora cinnamomi at different time periods after phosphite was applied and the percent of live plants from which P. cinnamomi was recovered. There were 10 replicates in each treatment.……………………………………………………… 42 Chapter 2.3 Table 2.7 Schedule for the inoculation of Allocasuarina sp., Banksia grandis, Grevillea wilsonii and Adenanthos barbigera with Phytophthora cinnamomi isolate MP94-48………………………………………………………………… 49 Table 2.8 Death of Trymalium ledifolium after foliar application of 0, 5 or 10 g phosphite/L in November 1997.………………………………………………… 50 Table 2.9 The effect of foliar application of 0, 5 or 10 g phosphite/L on the survival of plants which have been inoculated with Phytophthora cinnamomi………... 51 Table 2.10 The recovery of Phytophthora cinnamomi from buried colonised Banksia stem pieces………………………………………………………………………. 52 Chapter 3.2 Table 3.1 Mean phosphite and phosphate concentrations in the stems of glasshouse grown plants which had been sprayed with phosphite 3 months previously……. 66 Table 3.2 The minimum and maximum temperatures (°C) and rainfall (mm) during the periods inoculated plants were present on the minesite……………………... 67 Table 3.3 The average percent inhibition of five Phytophthora cinnamomi isolates inoculated into Eucalyptus marginata seedlings which had been sprayed with 5 or 10 g phosphite/L………………………………………………………. 69 Table 3.4 Phosphite and phosphate concentrations in the stems of Eucalyptus marginata seedlings sprayed with 0, 5 or 10 g phosphite/L (n=5)……………... 70 Chapter 3.3 Table 3.5 Numbers of sporangia and the percentage of dehisced sporangia observed on the stems of glasshouse grown Eucalyptus marginata and Banksia grandis seedlings which had been inoculated with Phytophthora cinnamomi, sprayed with phosphite and flooded and sampled after 14 days………………… 79 Table 3.6 Phosphite concentration in the stem of Eucalyptus marginata seedlings sprayed with 0, 5 or 10 g phosphite/L (n=4)……………………………………. 80 Chapter 3.4 Table 3.7 The effect of foliar applied phosphite on the infection and colonisation by Phytophthora cinnamomi zoospores of the roots of a range of native plant iv species grown aeroponically. Also, the phosphite levels in the roots of phosphite treated Pattersonia occidentalis……………………………………… 87 Table 3.8 Minimum and maximum temperatures for the time between inoculation and harvest ……………………………………………………………..…………….. 88 Chapter 4.1 Table 4.1 The average percent of plant affected by foliar application of phosphite at Tiwest (Cataby). Plants were rated 2 weeks and 8½ months after phosphite was applied (n=7)……………………………………. …………………………. 95 Chapter 4.2 Table 4.2 Germination of Trymalium ledifolium seeds which had been collected from phosphite treated plants (number of plants=2)……………………..……...…….. 99 Chapter 5.1.1 Table 5.1 Comparison of the extraction efficiency of 500µg/g phosphite dosed plant samples by water and succinic acid.…………………………………………….. 104 Table 5.2 The effect of time on the efficiency of extraction of phosphite and phosphate…. 105 Table 5.3 Analysis of the stability of the phosphite/phosphate ration during water and succinic acid extractions……………………………………………………. 105 Chapter 6.1 Table 6.1 Maximum duration in weeks of a significant effect (p<0.05) on pollen fertility or seed germination by sprays of (a) 2.5, (b) 5 or (c) 10 g phosphite/L… 114 Chapter 6.3 Table 6.2 Phosphite concentration (µg/g dry wt) detected in various tissue types of E. marginata seedlings 7 days after foliar treatment with 5 or 10 g phosphite/L……………………………………………………………………… 117 Appendix 1 Table 1 Published data on the use of phosphite on plants and their subsequent concentration of phosphite in plant tissue………………………………………. 129 Table 2 Rating system for foliar phytotoxicity symptoms of phosphite in Eucalyptus calophylla plants treated in Experiment 2……………………………………….. 131 Table 3 Mean phosphite concentrations in mature roots, stems, mature leaves and y oung fully expanded leaves in Eucalyptus calophylla 7 days after being sprayed (0, 0.25, 0.5, 1%) or misted (10, 20, 40%) with phosphite (Experiment 1 results only)……………………………………………………… 134 v List of Figures Chapter 2.1 Figure 2.1a The effect of foliar application of phosphite (0, 5, 10 or 20 g/L) applied in spring (n=7) on the average growth rate of Phytophthora cinnamomi in the stems of plant species from the Eucalyptus marginata forest of Western Australia. Points with different letters indicate a significant (p<0.05) difference in the growth rate of P. cinnamomi within each plant species and time of inoculation……………………..…………………………………… 19 Figure 2.1b The effect of foliar application of phosphite (0, 5, 10 or 20 g/L) applied in spring (n=7) on the average growth rate of Phytophthora cinnamomi in the stems of plant species from the Eucalyptus marginata forest of Western Australia. Points with different letters indicate a significant (p<0.05) difference in the growth rate of P. cinnamomi within each plant species and time of inoculation……………….…..…………………………...………… 21 Figure 2.2 The average phosphite concentration (ug g-1 dry wt ± SE) in the stems of0 plant species from the Eucalyptus marginata forest which were treated with phosphite in (a) spring or (b) autumn. 0 g phosphite/L (n=2), 5, 10 and 20 g phosphite/L (n=6)………………………………………………………….. 21 Figure 2.3 The effect of foliar application of phosphite (0, 5 or 10 g/L) applied in (a) spring
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