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Thrips Obscuratus in New Zealand Vineyards: It's Biology and Effects On 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. Thrips obscuratus in New Zealand vineyards: its biology and effects on Botrytis cinerea infection A thesis submitted in partial fulfillment of the requirements for the Degree of Doctor of Philosophy At Lincoln University By K. Schmidt Lincoln University 2007 Scientific publications of this thesis Chapter 2 Section 1 K. Schmidt, D.A.J. Teulon, M. V. Jaspers (2006). Phenology of the New Zealand flower thrips (Thrips obscuratus) in two vineyards. New Zealand Plant Protection 59, 323-329. Chapter 6 Submitted to 'Molecular Ecology Notes' Katrin Schmidt, Shelley L. Ball, Steve D. Wratten, Marlene V. Jaspers: Molecular identification of T. obscuratus (Thysanoptera) using DNA barcoding. Abstract of a thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy Thrips obscuratus in New Zealand vineyards: its biology and effects on Botrytis cinerea infection. By Katrin Schmidt This research project investigated aspects of Thrips obscuratus biology and its interaction with grapevines and botrytis bunch rot in two New Zealand vineyards. Flight activity, phenology and short-distance movement of T. obscuratus were assessed in the field and the influences of the fungus and the insect on B. cinerea incidence and severity, skin scarring and phenolics and grape yield were investigated in greenhouse and field experiments. The numbers of airborne adult thrips were monitored at Canterbury House Vineyard (now Waipara Hills Vineyard), Waipara, and at Neudorf Vineyard, Nelson, from spring to autumn in the 2003/04 and 2004/05 season. Thrips numbers were highest during the grapevine flowering period, being mostly T. obscuratus (90%), and higher at Neudorf Vineyard than at Canterbury House Vineyard. In the latter vineyard, thrips numbers per inflorescence at full bloom were higher in the later flowering Sauvignon Blanc than in Riesling. T. obscuratus numbers at different life-cycle stages were also monitored in the two vineyards, from two weeks before full bloom amongst Sauvignon Blanc and Riesling varieties. Flowers from grapevines in both regions were stained to show thrips eggs, which were commonly observed in the pedicels of flowers. Only small numbers of larvae were found in the traps underneath the vine canopy and no emerging adults were caught in the emergence traps placed on the ground, indicating that little reproduction had occurred in the vineyards. At Canterbury House Vineyard in spring 2005, T. obscuratus movement from three flowering inter-row plant species, buckwheat (Fagopyrum escu[entum), phacelia (PhaceUa tanacetifoUa) and alyssum (Lobu[aria maritima), to grape flowers was investigated to determine whether such floral resources could be alternative hosts of T. obscuratus, or act as trap plants. Thrips on these plants were marked with a foliar spray of 2000 ppm rubidium chloride once during full bloom of the grapevines. Numbers of marked T. obscuratus adults collected from grape flowers on adjacent vines were very low for all inter-row species, indicating little movement between them. An investigation into the potential role of T. obscuratus in vectoring B. cinerea conidia to grapevine flowers showed that 20 thrips exposed to sporulating B. cinerea deposited 800 viable conidia per inflorescence and caused 88% flower rot. Scanning electron microscopy of T. obscuratus revealed that many conidia were distributed over the cuticle of the insect, most being trapped between the cilia on the wings or attached to the numerous setae of the body. The behaviour of T. obscuratus on grape flowers, observed in the laboratory by continuous video recording over 10 minute periods, showed that T. obscuratus spent most time walking and feeding on nectar, pollen and stigma tissues. The grooming behaviour between feeding and walking events and the feeding positions were considered sufficient for it to deposit B. cinerea conidia on the naturally susceptible and wounded areas of the flowers. Greenhouse experiments conducted in 2004/05 and 2005/06 used Mullins' plants of the white varieties Riesling and Sauvignon Blanc in both seasons and Pinot Noir in the first season only. Silk bags enclosed 10 T. obscuratus collected from the field and conidia of the pathogenic B. cinerea marker strain 879-1 nit1 around each inflorescence for 7 days. In 2004/05, Pinot Noir berries had much lower incidence of B. cinerea infection than did the white varieties, whose overall incidence increased from fruit set to harvest in both seasons. Presence of T. obscuratus increased infection of grape inflorescences of both white varieties in both seasons but not of Pinot Noir. The presence of T. obscuratus and B. cinerea at flowering also caused significantly lower bunch weights in some treatments, lower numbers of berries per bunch and higher incidences and severity of B. cinerea, with Riesling being most susceptible. ii Field experiments conducted at Canterbury House Vineyard and Neudorf Vineyard in 2004/05 and 2005/06 with the same treatments demonstrated similar effects on infection incidence, severity and bunch sizes. In 2005/06, assessment of grape bunch scarring by T. obscuratus and B. cinerea in greenhouse and field experiments showed increased scarring for some B. cinerea treatments and more consistent increases with T. obscuratus treatments in both varieties. Analysis of total phenolics in berry skins of Riesling reflected the trends in scarring, being higher in the B. cinerea and T. obscuratus treatments than in the control. When one application of the insecticide pyrethrum was made in the field at full bloom, there were fewer B. cinerea infections higher yields and lower percentages of scars than when flowers were untreated. An identification method was developed for T. obscuratus by DNA sequencing of the 'Folmer' fragment of the mitochondrial gene COL DNA sequences of larvae and adults were established that could allow them to be distinguished from other thrips, which could formerly be done only for the adults using traditional morphological identification methods. This study has provided information about the interaction between T. obscuratus and B. cinerea infection pathways in grapevines under laboratory, greenhouse and field conditions, which may improve the developments of control strategies in the future. Keywords: Botrytis cinerea, Thrips obscuratus, grapevine, phenology, population dynamics, rubidium chloride, small-scale video analysis, scarring, total phenolics, DNA barcoding iii Table of Contents Abstract Table of contents iv List of Tables ix List of Figures xiii List of Plates xvi CHAPTER 1 General Introduction 1 1.1. The grapevine 1 1. 1. 1. History and taxa 1 1.1.2. Grapevine growth stages 3 1.2. Grapevine pests 5 1.2.1. Mites 5 1.2.2. Mealybug 5 1.2.3. Phyloxera 5 1.2.4. Leafroller 6 1.3. Thrips 7 1.3.1. Thrips obscuratus taxonomy 7 1.3.2. Distribution 8 1.3.3. Life cycle 8 1.3.4. Ecology of Thrips obscuratus 10 1.3.5. Damage and economic importance 12 1.3.6. Control practices 14 1.4. Grapevine diseases 15 1.4.1. Downy mildew 15 1.4.2. Powdery mildew 15 ~4.3. Bunch rot 15 1.5. Botrytis cinerea 16 1.5.1. Taxonomy 16 1.5.2. Morphology 17 1.5.3. Disease cycle 18 1.5.4. Symptoms on the grapevine 19 1.5.5. Economic importance 21 1.5.6. Control practices 21 1.6. Interactions between T. obscuratus and B. cinerea on grapevine 23 1.7. Aims and objectives of this study 25 1.7.1. Thesis structure 26 1.8. References 29 CHAPTER 2 The biology of Thrips obscuratus in two vineyards in the South Island of New Zealand 34 2.1. Chapter introduction 34 iv SECTION 1 Seasonal flight activity and infestation levels of the New Zealand Flower Thrips (Thrips obscuratus) in two South Island vineyards 35 2.2. Introduction 35 2.3. Materials and methods 36 2.3.1. Flower sampling 37 2.3.2. Sampling with water traps 37 2.3.3. Data analysis 38 2.4. Results 39 2.4.1. Flower sampling 39 2.4.2. Sampling with water traps 39 2.5. Discussion 43 2.6. References 45 SECTION 2 Population dynamics of Thrips obscuratus in vineyards 47 2.7. Introduction 47 2.8. Materials and methods 47 2.8.1. Monitoring of different life stages in the vineyard 47 2.8.2. Data analysis 50 2.9. Results 50 2.9.1. Monitoring of different life stages in the vineyard 50 2.10. Discussion 53 2.11. References 56 SECTION 3 Movement of Thrips obscuratus between flowering inter-row plants and grapevines 58 2.12. Introduction 58 2.13. Materials and methods 60 2.13.1. Study site 60 2.13.2. Application of rubidium chloride and collection of insect and plant material 60 2.13.3. Sample preparation and rubidium analysis 62 2.13.4. Data analysis 63 2.14. Results 63 2.14.1. Background concentrations of rubidium 63 2.14.2. Labelled insects and plants 63 2.15. Discussion 64 2.16. References 67 CHAPTER 3 Interactions between Thrips obscuratus and Botrytis cinerea in two South Island vineyards 71 3.1. Chapter introduction 71 v SECTION 1 T. obscuratus and B. cinerea infestation of grape flowers in greenhouse experiments 74 3.2. Introduction 74 3.3. Materials and methods 75 3.3.1.
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