The Effects Of Bacterial And Jasmonic Acid

THE EFFECTS OF BACTERIAL AND JASMONIC ACID TREATMENTS ON INSECTS OF CANOLA by Katherine Marie Bergen
A thesis submitted to the Faculty of Graduate Studies of The University of Manitoba
in partial fulfilment of the requirements for the degree of
Master of Science
Department of Entomology University of Manitoba Winnipeg, MB
Copyright © July 2008 by Katherine Marie Bergen

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THE UNIVERSITY OF MANITOBA FACULTY OF GRADUATE STUDIES
COPYRIGHT PERMISSION
The Effects of Bacterial and Jasmonic Acid Treatments on Insects of Canola
BY
Katherine Marie Bergen
A Thesis/Practicum submitted to the Faculty of Graduate Studies of The University of Manitoba in partial fulfillment of the requirement of the degree Of
MASTER OF SCIENCE
Katherine Marie Bergen © 2008
Permission has been granted to the University of Manitoba Libraries to lend a copy of this thesis/practicum, to Library and Archives Canada (LAC) to lend a copy of this thesis/practicum, and to LAC's agent (UMI/ProQuest) to microfilm, sell copies and to publish an abstract of this
thesis/practicum. This reproduction or copy of this thesis has been made available by authority of the copyright owner solely for the purpose of private study and research, and may only be reproduced and copied as permitted by copyright laws or with express written authorization from the copyright owner.

ACKNOWLEDGEMENTS
I would like to sincerely thank everyone that was involved with my project, especially Dr. Holliday, Dr. Fernando and Dr. Lamb for their guidance and advice. I would also like to thank Lisa Babey, Paula Parks and Alvin Iverson for their assistance in the field and laboratory, and Alicia Leroux, Heather Collins, Rajesh Ramarafhnam, Yu Chen, Dr. Genyi Li, Abdel el Hadrami, Zhixia Niu, Lars Andreassen, Lome Adam and Ian Brown for everything they have helped with over the past few years. Funding for this project was provided by the Canola Council of Canada and the Agronomic Research and Development Initiative of the Province of Manitoba.
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ABSTRACT
Two strains of plant growth-promoting rhizobacteria, Pseudomonas chlororaphis (PA23) and Bacillus amyloliquefaciens (BS6), can control some fungal diseases of canola through production of bacterial metabolites and through induced systemic resistance, which is initiated by the signalling molecule jasmonic acid. Direct application of jasmonic acid activates defence-related compounds and influences insect herbivory in canola. Field and laboratory studies investigated the effects of the two bacteria and of jasmonic acid on insects of canola. In the field there were no consistently significant effects of treatment on insects sampled by beat cloth or sweep net, level of flea beetle injury, canola yield or quality. In the laboratory, jasmonic acid significantly increased oviposition and decreased larval feeding in diamondback moth {Plutella xylostella) and slowed development and reduced reproduction in turnip aphid (Lipaphis erysimi). The effects of jasmonic acid on canola were systemic. Analysis of leaf tissue showed significant effects of treatment on defence-related compounds.
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TABLE OF CONTENTS

ACKNOWLEDGEMENTS

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ABSTRACT

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LIST OF TABLES

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LIST OF FIGURES

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1. INTRODUCTION

1

2. LITERATURE REVIEW

3

2.1 Canola.

3

2.2 Insects and pathogens on canola

3

2.3 Induced plant defences

6

2.3.1 Systemic acquired resistance

6

2.3.2 Induced systemic resistance

8

2.3.3Wound-induced defences

10

2.3.4 Differential induction with type of damage

11

2.3.5 Activation of defence responses

12

2.3.6 Cross-talk between responses to pathogens and herbivores

15

2.4 Potential for use in agriculture

17

2.4.1 Potential for disease control by PGPR

17

2.4.2 Potential for insect control by PGPR

19

2.4.3 Potential for insect control by jasmonic acid

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2.5 Research objectives

23

3. FIELD STUDY OF THE EFFECTS OF PSEUDOMONAS CHLORORAPHIS

STRAIN PA23, BACILLUS AMYLOLIQUEFACIENS STRAIN BS6 AND JASMONIC

ACID ON INSECTS OF CANOLA

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Abstract

27

3.1 Introduction

28

3.2 Methods

31

3.2.1 Treatments

32

3.2.2 Flea Beetle injury

33

3.2.3 Beat cloth and sweep net samples

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3.2.4 Cabbage maggot sampling

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3.2.5 Yield and seed quality

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3.3 Results.

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3.3.1 Flea Beetle injury

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3.3.2 Beat cloth and sweep net samples

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3.3.3 Cabbage maggot sampling

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3.3.4 Yield and seed quality

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3.4 Discussion,

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4. LABORATORY STUDIES ON THE EFFECTS OF PSEUDOMONAS CHLORORAPHIS STRAIN PA23, BACILLUS AMYLOLIQUEFACIENS STRAIN AND JASMONIC ACID ON THE DIAMONDBACK MOTH (PLUTELLA XYLOSTELLA) AND THE TURNIP APHID (LIPAPHIS ERYSIMI) Abstract
4.1 Introduction 4.2 Methods
4.2.1 Plants 4.2.2 Treatments
4.2.2.1 Spray preparation 4.2.2.2 Spray application 4.2.2.3 PA23+pathogen inoculation 4.3 Studies with Plutella xylostella 4.3.1 Oviposition preference 4.3.2 Larval feeding and growth rate 4.3.3 Test of systemic influences 4.3.4 Changes in plant chemistry 4.3.5 Peroxidase activity 4.3.6 Phenol concentration 4.3.7 Glucosinolates 4.4 Studies with Lipaphis erysimi 4.5.1 Development and reproduction 4.5 Statistical analyses 4.6 Results 4.6.1 Effects of methanol on bacterial growth 4.6.2 Studies with Plutella xylostella 4.6.2.1 Oviposition preference 4.6.2.2 Larval feeding and growth rate 4.6.2.3 Test of systemic influences 4.6.2.4 Changes in plant chemistry 4.6.2.5 Peroxidase activity 4.6.2.6 Phenol concentration 4.6.2.7 Glucosinolates 4.6.3 Studies with Lipaphis erysimi 4.6.3.1 Development and reproduction 4.7 Discussion 4.7.1 Implications of insect responses for use of PGPR

BS6
59 60 61 65 65 66 66 67 68 69 70 71 72 73 74 74 75 77 77 79 81 81 81 81 82 83 83 84 84 85 87 87 88 102

5. GENERAL DISCUSSION

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LITERATURE CITED

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LIST OF TABLES

Table 2.1 Examples of plant growth-promoting rhizobacteria-induced systemic

resistance

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Table 3.1. Mean number of insects (± SEM) from beat cloth samples collected 26 July

and 9 August 2006 (N=5, df=3,12)

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Table 3.2. Mean number of insects (± SEM) from sweep net samples collected 26 July

and 9 August 2006 (N=5 df=3,12)

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Table 3.3. Results from repeated measures analyses for the effect of treatment on the

overall numbers of insects collected from both sampling dates (between subjects) and the

effect of treatment on patterns of temporal change (within subjects) for beat cloth

samples (df=3,12)

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Table 3.4. Results from repeated measures analyses for the effect of treatment on the

overall numbers of insects collected from both sampling dates (between subjects) and the

effect of treatment on patterns of temporal change (within subjects) for sweep net

samples (df=3,12).

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Table 3.5. Species composition of emerged insects in each treatment

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Table 4.1. Levels of peroxidase in relation to treatments and the presence or absence of

insects

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Table 4.2. Analysis of variance for levels of peroxidase

114

Table 4.3. Phenol activity in relation to treatments and the presence or absence of

insects

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Table 4.4. Analysis of variance for phenol activity

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Table 4.5. Ratio of peak area (Mean ±SEM) of the glucosinolate glucobrassicin relative

to the area of the known standard (sinigrin) in relation to treatments and the presence or

absence of insects (N=4)

116

Table 4.6. Analysis of variance for glucobrassicin

116

Table 4.7. Ratio of peak area (Mean ±SEM) of the glucosinolate neoglucobrassicin

relative to the area of the known standard (sinigrin) in relation to treatments and the

presence or absence of insects (N=4)

117

Table 4.8. Analysis of variance for neoglucobrassicin

117

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Table 4.9. Effects of treatment on aphid development time, reproduction and intrinsic rate

of increase (N=23)

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LIST OF FIGURES

Figure 3.1. Field layout (randomized complete block design) used for 15 May field and 5

June fields

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Figure 3.2. Mean (±SEM) flea beetle injury ratings (N=5) for 10 plant-samples at

cotyledon stage prior to application of treatments (8 June, pre-treatment) and one week

following treatment (15 June, post-treatment)

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Figure 3.3. Mean (±SEM) flea beetle injury ratings (N=5) for 10 plant-samples at true

leaf stage prior to application of treatments (8 June, pre-treatment) and one week

following treatment (15 June, post-treatment)

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Figure 3.4. Mean (±SEM) number of root maggot larvae per plant sampled on 27 July

and 11 August 2006 (N=5)

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Figure 3.5. Mean (±SEM) root maggot damage rating (N=5) for samples collected on 27

July, 11 August and 6 September 2006. Root damage was rated on a scale of 0-4

according to Dosdall et al. (1994)

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Figure 3.6. Mean (±SEM) seed weight of canola for each treatment harvested 31 August

2006 (N=5)

51

Figure 3.7. Mean (±SEM) glucosinolate content (umoles/g) of canola seeds harvested 31

August 2006 (N=5)

52

Figure 3.8. Percent (%) protein and oil (dry weight) of canola seeds harvested 31 August

2006 (N=5)

53

Figure 4.1. Mean (±SEM) number of eggs laid per plant for each treatment (N=29) 104

Figure 4.2. Mean (±SEM) number of eggs laid on bottom, petiole or top of leaves 1-2, 3 -

4 and 5-6 for each treatment (N=29)

105

Figure 4.3. Frequency distribution of Plutella xylostella head capsule widths

106

Figure 4.4. Mean (±SEM) leaf area consumed by larvae in each treatment (N=31) 107

Figure 4.5. Mean (±SEM) relative growth rate of larvae in each treatment (N=29) 108

Figure 4.6. Mean (±SEM) number of larval feeding initiation sites per leaf in each

treatment (N=31)

109

Figure 4.7. Mean (±SEM) biomass conversion efficiency (mg/cm ) for larva in each

treatment (N=31)

110

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