<p>Microorganisms, including two fungi, <i>Phytophthora cryptogea</i> and <i>Fusarium oxysporum</i> strain Fo-(IMI 386351), and one bacterium, <i>Pesudomonas</i> sp. strain MF30, were tested for their abilities to stimulate plant defence responses in tomato (<i>Lycopersicon esculentum</i> –L.) and to serve as effective biocontrol agents (<b>Bs</b>). The study included <i>in vivo</i> and <i>in vitro</i> characterization of biological attributes of the microorganisms, pertaining to their abilities to stimulate plant immunity against a fungal pathogen, <i>Fusarium oxysporum</i> f. sp. <i>lycopersici</i> (Fol), the causal agent of tomato wilt disease. Using <i>Lycopersicon esculentum</i> –L. as a model plant for examining some fundamental elements of the plant-microorganism interaction, the study reveals and clarifies some aspects of the close association and the complexity of such systems.</p><p>For each <b>B</b>, the results revealed a <b>B</b>-distinct plant-microorganism interaction, which included systemic induced resistance (SIR). A phylogenetic analyses of the partial sequences of two Fo-(IMI 386351) genes, a mitochondrial small subunit ribosomal DNA (mtSSU rDNA) and the nuclear translation elongation factor 1α (EF-1α), provided phylogenetic trees confirming that Fo-(IMI 386351) might be a member of Fol or of <i>F. oxysporum</i> f. sp. <i>melonis</i>, which have polyphyletic evolutionary origins. RFLP analysis (mtDNA), suggested that Fo-(IMI 386351) probably belongs to Fol. For routine and accurate differentiation between two morphologically indistinguishable <i>F. oxysporum formae speciales</i> strains, <i>F. oxysporum</i> f. sp. <i>lycopersici</i> and <i>F. oxysporum</i> f. sp. <i>radicis-lycopersici</i>, a molecular method (mtDNA RFLP analysis) was developed, and its usefulness for such differentiation was compared with that of two other methods: isozyme analysis and an osmotic method, revealed with high performance liquid chromatography (HPLC). The HPLC-spectra of Fo-(IMI 386351) had an extra peak for the two tested fractions, indicating that activation of the observed plant defence mechanism could have been at least partially the result of one of the products of the eliciting microbe. Preliminary results obtained by nuclear magnetic resonance spectrometry of those fractions suggest that the extra peak probably represents an oligosaccharide, which may have acted as a mobile signal and triggered the plant defence mechanisms.</p><p>We concluded that (1) our three tested microorganisms are able to stimulate plant defence mechanisms by triggering SIR (plant immunity), (2) the complexity and elaborateness of evolved plant-microbe interactions involving plant defence can, at least in some cases, be observed and studied in the laboratory, and (3) molecular tools can be a powerful means for identifying fungal strains and for clarifying their taxonomical relationships.</p>

The cabbage seedpod weevil, Ceutorhynchus obstrictus (Marsham) (Coleoptera: Curculionidae), is an important pest of brassicaceous oilseed crops, especially canola (Brassica napus L. and Brassica rapa L.) in North America and Europe. Application of foliar insecticide is the only method currently employed to control C. obstrictus populations; because this approach is environmentally unsustainable, alternatives including host plant resistance have been explored. White mustard, Sinapis alba L., is resistant to C. obstrictus and was chosen as a potential source of resistance for B. napus oilseed. Interspecific crosses of S. alba x B. napus have produced several lines that are resistant to C. obstrictus feeding and oviposition and yield fewer, lighter-weight weevil larvae that take longer to develop. I investigated potential mechanisms of this resistance, including assessing differences in visual and olfactory cues among resistant and susceptible genotypes, and antixenosis and antibiosis. Determining effects of visual cues associated with host plant resistance required investigation of weevil vision. Deployment strategies for resistant germplasm were assessed to evaluate incorporation of susceptible refugia to promote long-term durability of resistance traits. Results reported in Chapter 2 indicate that the C. obstrictus visual system is apparently trichromatic and incorporates receptors with response maxima near 350, 450, and 550 nm. Modelling indicated that UV light alone reduced weevil responses but the interaction of yellow and UV light increased responses at a threshold reflectance level of UV. Results reported in Chapter 3 indicated that differences in yellow and UV reflectance among host plant flowers influence host selection in C. obstrictus. Results described in Chapter 4 determine differential attraction to the odours of S. alba and B. napus and among resistant and susceptible accessions. Inferences of the identities of glucosinolates found in varying amounts among susceptible and resistant genotypes suggested that 2-phenylethyl glucosinolate influenced attractiveness. Results described in Chapter 5 indicate differences in adult feeding and oviposition preferences among resistant and susceptible genotypes. Oocyte development, larval biomass and larval development time varied among weevils feeding on resistant and susceptible genotypes. Based on results of Chapter 4, 1-methoxy-3-indolylmethyl glucosinolate was implicated as contributing to antixenosis and antibiosis resistance. Results reported in Chapter 6 describe effects of mixed plots of resistant and susceptible genotypes on weevil spatial distribution and oviposition. These results are consistent with associational resistance and attributed to reduced apparency of susceptible plants in mixtures and antixenosis resistance associated with resistant germplasm. / Plant Science

Microorganisms, including two fungi, Phytophthora cryptogea and Fusarium oxysporum strain Fo-(IMI 386351), and one bacterium, Pesudomonas sp. strain MF30, were tested for their abilities to stimulate plant defence responses in tomato (Lycopersicon esculentum –L.) and to serve as effective biocontrol agents (Bs). The study included in vivo and in vitro characterization of biological attributes of the microorganisms, pertaining to their abilities to stimulate plant immunity against a fungal pathogen, Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of tomato wilt disease. Using Lycopersicon esculentum –L. as a model plant for examining some fundamental elements of the plant-microorganism interaction, the study reveals and clarifies some aspects of the close association and the complexity of such systems. For each B, the results revealed a B-distinct plant-microorganism interaction, which included systemic induced resistance (SIR). A phylogenetic analyses of the partial sequences of two Fo-(IMI 386351) genes, a mitochondrial small subunit ribosomal DNA (mtSSU rDNA) and the nuclear translation elongation factor 1α (EF-1α), provided phylogenetic trees confirming that Fo-(IMI 386351) might be a member of Fol or of F. oxysporum f. sp. melonis, which have polyphyletic evolutionary origins. RFLP analysis (mtDNA), suggested that Fo-(IMI 386351) probably belongs to Fol. For routine and accurate differentiation between two morphologically indistinguishable F. oxysporum formae speciales strains, F. oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis-lycopersici, a molecular method (mtDNA RFLP analysis) was developed, and its usefulness for such differentiation was compared with that of two other methods: isozyme analysis and an osmotic method, revealed with high performance liquid chromatography (HPLC). The HPLC-spectra of Fo-(IMI 386351) had an extra peak for the two tested fractions, indicating that activation of the observed plant defence mechanism could have been at least partially the result of one of the products of the eliciting microbe. Preliminary results obtained by nuclear magnetic resonance spectrometry of those fractions suggest that the extra peak probably represents an oligosaccharide, which may have acted as a mobile signal and triggered the plant defence mechanisms. We concluded that (1) our three tested microorganisms are able to stimulate plant defence mechanisms by triggering SIR (plant immunity), (2) the complexity and elaborateness of evolved plant-microbe interactions involving plant defence can, at least in some cases, be observed and studied in the laboratory, and (3) molecular tools can be a powerful means for identifying fungal strains and for clarifying their taxonomical relationships.

In tomato (Solanum lycopersicum), race-specific resistance against the fungal wilt pathogen Verticillium dahliae race 1 (Vd1) is established in the stem. However, the molecular factors and mechanisms leading to this resistance response are still unknown. In this study, Craigella resistant (CR) and susceptible (CS) tomato plants were successfully infected with Vd1 and this was verified by fungal quantification and symptom score assays. Previous microarray results showed interesting patterns of defence gene expression that correlated with biological phenomena. Plant defence genes code for proteins that are responsible for or associated with the plant resistance response. Through RT-PCR, this thesis set out to confirm these microarray observations and also to generate expression data for genes in which sensitivity was an issue in the microarray. The standard RT-PCR data confirmed a number of the microarray results, but some conflicts remained. From the defence genes investigated, there was agreement between the microarray data and the RT-PCR data for pre-mRNA processing factor 8, class IV chitinase, cyclin-dependent kinase inhibitor and IMP dehydrogenase/GMP reductase. Partial agreement was observed for genes coding for ethylene response factor 2, phenylalanine ammonia lyase and P6 protein. However, there was total disagreement for 14-3-3, beta-glucanase, P1a, RNA-binding protein, calcium-binding protein and S-Adenosyl-L-methionine: hydroxide adenosyltransferase. Real-time RT-PCR was attempted to clarify the remaining issues but further discrepancies arose, particularly in the Ve resistance genes. To resolve these discrepancies, two approaches were designed: (1) one based on the use of a universal internal control and (2) another based on restriction enzyme digestion. In general, the results were more consistent with standard RT-PCR. Overall, this study showed that standardization of a system involving vascular pathogens, leading to reproducible analysis, was possible but only with proper controls and additional validation. Standard RT-PCR appeared to offer a more accurate picture of the expression of defence genes in the tomato-Verticillium pathosystem. The defence gene expression results confirmed in this study remain as potential insights into the molecular mechanisms for Verticillium resistance in tomato plants.

No description available.

Abstract Grazing by reindeer (Rangifer tarandus L.) affects vegetation and soil microbial processes in tundra ecosystems. It is considered that grazing can induce two alternative vegetation states that differ in plant species composition and the rate of nutrient cycling. I hypothesised that these alternative vegetation states differ in ecosystem responses to climate warming. I tested the hypothesis using a factorial warming and fertilisation experiment on long-term lightly grazed (LG) and heavily grazed (HG) tundra. The reindeer grazing induced vegetation shift from dwarf shrubs to graminoids increased microbial activities for SOM decomposition. The grazer-induced shifts in vegetation and microbial activities in combination with the fertilisation via urine and faeces had important consequences on soil N availability and soil C quality that determined the ecosystem-level consequences of climate warming. Due to higher soil N availability, warming increased plant productivity (GEP) on HG but not on LG tundra, where N limitation prevented the warming-increased plant production. The varying effects of warming on GEP at different grazing intensities determined the effects of warming on ecosystem net C sink, which was unaffected by warming on HG but decreased on LG tundra. Reindeer grazing reduced the soil C quality, as soils under LG stored a higher proportion of carbohydrates vulnerable to microbial decomposition than soils under HG. According to laboratory soil incubations, the grazer-induced reduction in soil C quality mitigated the responses of soil microbial activity to prolonged warming. Warming caused a stronger decrease in concentrations of phenolics, an important means of plant defence against biotic and abiotic stresses, in Empetrum nigrum ssp. hermaphroditum under HG than LG. Grazing history by reindeer, with the associated vegetation shift from dwarf shrubs to graminoids, can significantly alter the ecosystem-level consequences of climate warming. Overall, this thesis highlights that the effects of reindeer grazing on soil properties, soil N availability and C quality, are important determinants of the ecosystem responses to climate warming. Therefore, future research on climate warming should take into account herbivores and aim towards a more holistic approach that includes both aboveground and belowground components of the ecosystem. / Tiivistelmä Tundralla porolaidunnus vaikuttaa kasvillisuuteen ja maaperän mikrobien toimintaan. Porolaidunnus voi aikaansaada kaksi vaihtoehtoista kasvillisuuden tilaa, jotka eroavat toisistaan paitsi kasvilajiston myös ravinnekierron suhteen. Esitin hypoteesin, jonka mukaan ilmaston lämpenemisen vaikutukset eroavat tundratyypeillä, jotka edustavat vaihtoehtoisia kasvillisuustiloja. Testasin hypoteesia faktoriaalisen lämmitys- ja lannoituskokeen avulla tundra-alueilla, joilla poron laidunnuspaine on ollut pitkäaikaisesti joko kevyttä tai voimakasta. Poron aiheuttama kasvillisuusmuutos varpuvaltaisesta heinävaltaiseksi lisäsi maaperän mikrobien hajotusaktiivisuutta. Poron aiheuttamat erot kasvillisuudessa ja mikrobiaktiivisuuksissa yhdessä virtsan ja papanoiden lannoittavan vaikutuksen kanssa muuttivat maaperän typen saatavuutta sekä hiilen laatua. Erot maaperän ominaisuuksissa puolestaan ohjasivat ilmaston lämpenemisen ekosysteemitason vaikutuksia. Lämmitys kasvatti ekosysteemituotantoa ravinteikkaalla voimakkaasti laidunnetulla tundralla mutta ei kevyesti laidunnetulla tundralla, joka oli typpirajoitteinen. Lämmityksen erilaiset vaikutukset ekosysteemituotantoon eri laidunpaineissa määrittivät lämmityksen vaikutuksen ekosysteemin hiilinieluun, joka pysyi muuttumattomana voimakkaasti laidunnetulla tundralla mutta pieneni kevyesti laidunnetulla tundralla. Porolaidunnus alensi maaperän hiilen laatua, ja kevyesti laidunnetulla tundralla maaperässä oli enemmän mikrobien hajotukselle alttiita hiilihydraatteja kuin voimakkaasti laidunnetulla tundralla. Laboratoriossa suoritetun inkubaatiokokeen perusteella maaperän hiilen alhaisempi laatu lievensi mikrobien hajotusaktiivisuuden vastetta pitkäaikaiseen lämmitykseen. Lämmitys vähensi pohjanvariksenmarjan fenoliyhdisteiden, jotka ovat tärkeä osa kasvien puolustusta bioottisia ja abioottisia stressitekijöitä vastaan, pitoisuuksia enemmän voimakkaasti kuin kevyesti laidunnetulla tundralla. Poron laidunnushistoria yhdessä kasvillisuusmuutoksen kanssa voi merkittävästi vaikuttaa ilmaston lämpenemisen ekosysteemitason seurauksiin. Tämän väitöstyön tulokset osoittavat, että poron vaikutukset maaperän typpeen ja hiileen määrittävät lämpenemisen vaikutukset ekosysteemissä. Tulevaisuudessa ilmastonmuutostutkimuksen pitäisikin kokonaisvaltaisemmin huomioida sekä herbivorian että ekosysteemin eri osien merkitys.

Chez les mammifères, le glutamate est un neuromédiateur bien connu au niveau du système nerveux central et plus récemment un rôle immunomodulateur lui a été reconnu. Le glutamate est le ligand de récepteurs ionotopiques (iGluRs) qui sont des récepteurs-canaux perméables à divers cations dont le calcium (non-selective cation channels, NSCC). Chez Arabidopsis thaliana, une famille de 20 gènes de iGluRs homologues des iGluRs de mammifères a été identifiée et leur implication dans divers processus biologiques est suggérée. Dans ce travail où nous utilisons des suspensions de cellules de tabac (Nicotiana tabacum var Xanthi), divers arguments suggèrent que ces iGluR sont fonctionnels dans le tabac : influx de calcium et élévation rapide et transitoire de la concentration en calcium cytosolic libre en réponse à l’addition de glutamate, inhibition de ces effets par 4 antagonistes de iGluRs animaux (compétitifs ou non compétitifs), désensibilisation, et pH dépendance des effets. Pour la première fois chez les plantes nous montrons que le glutamate induit la production de NO très vraisemblablement via l’activation de iGluRs. De plus, nous démontrons que ce(s) iGluRs sont impliqués dans le mode d’action, via les flux de calcium, de la cryptogéine une protéine de 10 kDa de Phytophthora cryptogea, éliciteur des réactions de défense chez le tabac. Néanmoins, à ce niveau, les iGluRs ne sont pas impliqués dans la plupart des événements calcium-dépendants induits par la cryptogéine dont l’activation des MAPKs et de canaux anioniques, la production de H2O2 (activation de la NADPH-oxydase) et la réponse hypersensible. En revanche, ils sont tout ou partiellement responsables de la production de NO décrite pour la première fois par le passé en réponse à la cryptogéine. Ces résultats suggèrent que différents types de canaux calciques activés par divers médiateurs, génèrent, via le calcium, des messages spécifiques décodés par des protéines associées à chacun de ces types de canaux et impliquées dans des réponses biologiques différentes. Dans le mode d’action de la cryptogéine, nous démontrons que l’activation des iGluRs est possible grâce à l’exocytose de glutamate dans l’apoplaste, induite par la cryptogéine. Ainsi, ce travail est la première démonstration du rôle de iGluRs potentiels dans la défense chez les plantes et de leur implication dans la production de NO. Nos résultats sont un argument supplémentaire à la conservation des mécanismes de la défense dans le monde vivant et posent le problème du rôle du glutamate dans la signalisation chez les plantes. / Glutamate is recognized as the primary excitatory neurotransmitter in the mammalian central nervous system (CNS) but recent studies have shown that glutamate has an important additional immunomodulator role. Glutamate is the ligand of ionotropic glutamate receptors (iGluRs), which are non-selective cation channels (NSCC), permeable to calcium. In plants, animal iGluR homologs were found that were involved in many developmental processes. Here we demonstrate the involvement of putative iGluRs in calcium signalling in response to cryptogein which is a 10 kDa protein secreted by the oomycete Phytophthora cryptogea and is an elicitor of defence in tobacco. Using transformed tobacco cell suspensions expressing aequorin in the cytosol or in the nucleus, our results have shown that glutamate induces a strong and transient [Ca2+]cyt elevation without [Ca2+]nuc changes. Glutamate-induced [Ca2+]cyt elevation was a result of calcium influx from the extracellular medium and was inhibited by different GluR inhibitors. This data suggest the presence of functional calcium channels of GluRs-type in tobacco. Nevertheless, glutamate does not induce some of the calcium-dependent characteristic events of the defence pathways, which are H2O2 production, MAPK activation and hypersensitive response, but promoted NO production. Further, Ca2+ influx,[Ca2+]cyt elevation and NO production induced by cryptogein were shown to be partially inhibited by the glutamate receptor inhibitors, suggesting that cryptogein treatment could activate a calcium channel of the GluR-type leading to plant defense signalling through NO production. We have also demonstrated that cryptogein induces an efflux of glutamate in the apoplast by the process of exocytosis thus activating the GluRs in tobacco. This is the first demonstration for a potential GluR(s) involvement in plant defense signalling, furthermore by mechanisms that showed homology with glutamate effect on neuronal cells.

Les racines de la plupart des arbres forment des symbioses ectomycorhiziennes avec les champignons mutualistes du sol. Le basidiomycète L. bicolor (Maire) P.D. Orton secrète de petites protéines effectrices (MiSSP) afin d'établir les structures symbiotiques. Toutefois, les protéines de l'hôte ciblées par les MiSSPs ne sont pas connues. Dans notre étude, nous démontrons, à l'aide du système double hybride chez la levure (Y2H), que la protéine MiSSP7 interagit avec les co-récepteurs de l'acide jasmonique (AJ) JAZ5 et JAZ6 de P. trichocarpa. Cette interaction entraine un blocage de la voie de signalisation de l'AJ et favorise le développement symbiotique. Des transformants de L. bicolor, dont l'expression de MiSSP7 est fortement réduite, ne sont plus capables de mycorhizer les racines du peuplier. Une variation transgénique de la transcription de PtJAZ6 ou l'inhibition de la voie de signalisation de l'AJ complémente ce phénotype. Nous avons également montré que la protéine PtJAZ6 interagit avec une protéine de type 14-3-3 et un facteur de transcription de type MYC, formant un complexe de régulation. Deux autres protéines effectrices, MiSSP8 et MiSSP17, sont sécrétées et essentielles au développement symbiotique. Les résultats des analyses Y2H suggèrent que MiSSP8 et MiSSP17 pourraient aider au contournement des réactions de défense de la plante-hôte. Au cours du développement symbiotique, le champignon est le siège d'une reprogrammation génétique importante. Les facteurs de transcription (TFs) sont les principaux acteurs de ces changements génétiques. Nous avons donc étudié les TFs de L. bicolor afin d'obtenir un inventaire complet des TFs régulés par la mycorhization / Roots of most trees form symbiosis with mutualistic soil-borne fungi. The ectomycorrhizal basidiomycete L. bicolor (Maire) P.D. Orton relies on mycorrhizal-induced small secreted proteins (MiSSP) to establish symbiotic tissues in the host-plant. The host proteins targeted by these fungal effectors are yet unknown. In the present study, we used the binary yeast two-hybrid (Y2H) system to determine direct interactions between MiSSP7 and the plant proteins in the L. bicolor-P. trichocarpa ectomycorrhizae. We showed that MiSSP7 interact with the jasmonic acid (JA) co-receptors JAZ5 and JAZ6 of P. trichocarpa, blocking JA signaling and promoting mutualism. L. bicolor transformants with severely reduced expression of MiSSP7 did not enter into symbiosis with poplar roots, a phenotype that could be complemented by transgenically varying the transcription of PtJAZ6 or through inhibiting JA signalling. Additional Y2H assays showed that PtJAZ6 protein form a regulatory complex involving 14-3-3 protein(s) and MYC transcriptional factors. Two others L. bicolor effector-like proteins, MiSSP8 and MiSSP17, are secreted and are essential for the symbiosis development. Y2H assays suggested that these MiSSPs interact with plant proteins involved in plant defence signalling pathways. During symbiosis development, L. bicolor experiences important genetic reprogramming required for root colonization. Transcription factors (TFs) are key players of these genetic changes. Here, we developed high throughput analysis of TFs in L. bicolor to obtain a comprehensive inventory of significantly regulated transcription factors in ECM

Almost all conifer ovules produce a liquid secretion as part of reproduction. This secretion, termed an ovular secretion, is produced during ovule receptivity and is involved in pollen capture and transport. Historically, examinations of the ovular secretion have focused on how they are part of pollination mechanisms. As a result, the chemical composition of the ovular secretion has not been examined systematically. Investigations into the constituents of the ovular secretion were limited to analyses for simple water soluble compounds such as sugars, minerals, amino acids and organic acids. More recently, the protein component of the secretion has been investigated using mass spectrometry-based proteomics. Proteins involved in processes such as carbohydrate modification, proteolysis, and defence have been identified in conifer ovular secretions. This biochemical complexity suggests a broader view of the function of the ovular secretion is warranted. However, protein identifications only provide putative information on function. Functional characterization of these proteins is needed in order to fully understand how they contribute to ovular secretion function. The research outlined in this dissertation describes the first functional characterizations of proteins found in conifer ovular secretions. Three proteins - invertase, chitinase, and thaumatin-like protein - were characterized in the ovular secretions of Douglas-fir (Pseudotsuga menziesii) and hybrid yew (Taxus × media). The Douglas-fir ovular secretion is capable of converting sucrose to glucose and fructose, confirming that invertases present in the secretion are functional. The invertase activity was maximal at pH 4.0. Activity was 77% of maximal at pH 4.5, the physiological pH. This indicates that post-secretory hydrolysis of sucrose occurs in situ in the Douglas-fir ovular secretion. Invertases in the ovular secretion are likely involved in controlling the movement of carbohydrates to developing pollen and could facilitate pollen selection. Chitinases present in the Douglas-fir ovular secretion are functional at physiological conditions. All three modes of chitinolytic activity, i.e. endochitinase, chitobiosidase and β-N-acetylglucosaminidase, were detected at physiological pH. β-N-acetylglucosaminidase activity was 80 % of maximal at physiological pH. Chitinases are pathogenesis-related proteins capable of hydrolysing chitin in fungal cell walls. These results suggest the ovular secretion is capable of defending the ovule against infection by phytopathogens. Thaumatin-like protein was immunolocalized to the cell wall and amyloplasts in Douglas-fir and yew nucellar tissue in a pattern consistent with a defensive role. It was also localized to the cell wall of fungal spores and germinating hyphae that were present in the micropyle of a yew ovule. These results provide additional evidence for an antifungal role for the ovular secretion. Functioning enzymes involved in pollen-ovule interactions and ovule defence are present in the conifer ovular secretion. The ovular secretion has functions beyond pollen capture. A revised functional model for the conifer ovular secretion is proposed. / Graduate / 2021-08-17

New Zealand’s isolation and periods of marine transgression have limited its biota to an extent which can be considered depauperate, even by island standards. Endemic vertebrates are rare and prominent invertebrate families, such as the renowned forest defoliators of the Lymantriidae, are absent. The proven vulnerability of the flora to introduced vertebrates reaffirms a belief in the invasiveness of islands and fuels the contingency plans aimed at averting similar devastation from further alien invertebrate defoliators. Nothofagus is a dominant element of the climax forests of New Zealand and the larger landmasses bordering the South Pacific Ocean. Assessments of the resistance to defoliation of continental and New Zealand species of Nothofagus, and a range of other forest genera endemic to New Zealand, was undertaken using bioassays of naïve polyphagous defoliators. The bioassays were undertaken in Europe, utilising gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae) and the fall webworm, Hyphantria cunea (Lepidoptera: Arctiidae) as defoliators, fed foliage plants growing in European arboreta. In New Zealand, bioassays utilised Australian painted apple moth, Teia anartoides (Lepidoptera: Lymantriidae) and tree species from local arboreta, gardens and natural populations. Larval growth rate was the primary parameter recorded to assess plant resistance. The relevance of growth rate was investigated by comparison with other recorded parameters and resistance to a surrogate pathogen, in the form of commercially available bio-insecticide. Larval growth rate was positively correlated with survivorship, potential fecundity, mating success and resistance to disease. The growth rate of larvae fed Nothofagus was positively correlated to the species-specific leaf nitrogen content. The results of the bioassays showed that despite the accepted paradigms, New Zealand’s flora was largely resistant to exotic defoliators. As an explanation of this apparent anomaly, the Island Resource Allocation (IRA) hypothesis was developed and posits that ‘the palatability iv of a plant to invertebrate herbivores is proportional to the geographic range of the plant’. The basis for the IRA hypothesis proposes a redefinition of the fundamental ecological principle of the species: area relationship. Islands, or similarly geographically constrained ecosystems, which support lower biodiversity, have impoverished trophic levels and consequently have weaker top-down regulation of herbivores by natural enemies. The IRA hypothesis argues that island ecosystem stability is achieved through the bottom-up process of plant defence. The IRA hypothesis was tested intra-specifically using bioassays using painted apple moth in which larvae were offered foliage of specimens from naturally discontinuous populations of Nothofagus truncata. The results supported the hypothesis in that the smallest populations of N. truncata exhibited the greatest resistance to the defoliator. The IRA hypothesis and a demonstrated mechanism for a differential resistance in Nothofagus species could resolve a number of enduring debates in ecology. Habitat area appears to explain the relative strengths of top-down and bottom-up regulation of herbivores. It also predicts the strengths of reciprocal evolution within the geographic mosaic of co-evolution and highlights the influence of biodiversity in invasive ecology. It may also help to resolve the contentious and extremely relevant debate of the role of biodiversity in ecosystem function.