Dissertations / Theses on the topic 'Parasitic plants Host plants'

Dodders (Cuscuta spp.) are parasitic plants that live by tapping into the vascular tissue of a host plant. Contents of the host phloem translocate readily into the parasite, and shared plasmodesmata have been documented between host cortical cells and dodder searching hyphae. Dodder is known to transmit viruses from one host to another, which is consistent with viral ability to traverse plasmodesmata (PD) with the aid of movement proteins (MPs). Plant endogenous mRNAs may also associate with specific proteins to pass through PD and traffic long distances in the phloem, a process that appears to play a role in coordination of development. We have evaluated the hypothesis that dodder is able to accumulate host phloem-mobile mRNAs by assaying lespedeza dodder (C. pentagona) for the presence of host transcripts. Reverse transcriptase PCR (RT-PCR) and tomato microarrays were used to probe RNA from dodder parasitizing tomato. Transcripts from four tomato genes were detected in dodder grown on tomato, but were not detected in control dodder grown on other hosts. Notable among these was LeGAI, a transcript previously shown to be phloem translocated. In addition, RT-PCR of RNA from dodder grown on pumpkin detected three mobile pumpkin mRNAs (CmNACP, CmSUTP1, and CmWRKYP). These results imply the existence of an extraordinary situation in which mobile mRNAs move from one plant into another, and raise questions about the role of this phenomenon in plant development and parasite pathogenicity.
Master of Science

Foraging theory predicts active responses by organisms upon encounter with a resource, as opposed to the passive responses of differential survivorship and growth. Stems of the parasitic plant Cuscuta subinclusa invest in resource acquisition (coil) relative to host quality in a way predicted by the marginal value theorem (MVT) in that: (1) stem coiling, the necessary antecedent and determinant of resource uptake, precedes exploitation of host materials; and (2) mean coiling on a host species is proportional to: (a) mean growth/haustorium, (b) mean biomass accumulation over the season, and (c) mean parasite growth/host individual. Coiling is correlated with growth/host individual for the 5 native host species examined, but not when a non-native species is added to the model, suggesting coiling response is a result of natural selection. Preliminary evidence indicates that coiling in C. subinclusa is induced by host bark chemicals. Resource-poor stems of C. subinclusa are more likely to coil, and coil more, than resource-rich stems, thus nutritional state of the parasite as well as host value affects foraging responses. Evidence from other experiments suggests that the costs of growth, or "search costs", may affect host acceptability. When water is readily available, transplanted C. subinclusa stems are less likely to coil on branches of Platanus racemosa. During the dry season, when cellular expansion is difficult, all p. racemosa branches were coiled upon. Large parasites are more likely to over-winter and set seed a second season, and parasites that start from over-wintered tissue are significantly larger at flowering than are those that have started from seed. Seed set is correlated with parasite size, thus linking foraging response and fitness of the plant. C. subinclusa's foraging response does not, however, predict population level patterns of host use. The principal determinant of host use by C. subinclusa is average proximity of a species to Malosma laurina. Parasite individuals infest many host species each season, but initially establish, set most seed, and over-winter only on M. laurina. Individual response of C. subinclusa contributes to the model of host use only after proximity to M. laurina is accounted for, suggesting that mechanisms maximizing exploitation of a host take effect after contact between host and parasite.

Une des contraintes biotiques majeures de la riziculture de bas-fond au nord du Togo est l’adventice parasite Rhamphicarpa fistulosa (Hochst.) Benth. Afin d’évaluer les options stratégiques pour contrôler efficacement R. fistulosa, notre étude a été conduite avec comme principaux objectifs : 1) l’identification de la répartition de R. fistulosa et l’étude des facteurs qui favorisent son infestation, 2) l’étude de l’impact des niveaux d’infestation sur la flore adventice du riz, et 3) la caractérisation des interactions hôte-parasite et de leurs impacts sur le développement de l’hôte et les performances reproductives du parasite. A l’issu des prospections de terrain, les plus fortes infestations ont été retrouvées sur des sols majoritairement à texture sableuse et très pauvres en matière organique. Une flore adventice très diversifiée a été retrouvée en présence de R. fistulosa, certaines adventices pouvant être parasitées et constituer éventuellement un réservoir pour accroître le stock semencier du parasite. En conditions contrôlées, R. fistulosa se développe et se reproduit aussi bien sur le riz que sur la tomate, alors que le soja retarde la reproduction du parasite. La germination des graines de R. fistulosa est maximale en présence de graines de plantes cultivées ou adventices. Les graines nouvellement récoltées ont une dormance de plus de six mois, mais leur germination a été stimulée par la présence de phytohormones. En conclusion, les meilleures stratégies de lutte doivent impliquer le contrôle par les semis tardifs, les rotations culturales avec les légumineuses comme le soja, l’arachide ou le niébé, et les cultures maraîchères
Main biotic constraints to rain-fed lowland rice cultivation in northern Togo include the parasitic weed Rhamphicarpa fistulosa (Hochst.) Benth. In order to evaluate efficient strategies to control R. fistulosa, our study was performed taking into account the following main objectives: 1) Identification of the distribution of R. fistulosa and study of the factors favouring its infestation, 2) Study of the impacts of infestation levels on rice weed communities, and 3) Characterization of the host-parasite interactions and their impacts on the host development and the parasite reproduction. Following fields investigations, highest infestations were observed on sandy soils with low levels of organic matter.In addition, weed flora found in the presence of R. fistulosa was much diversified and some weeds were found to been parasitized and could possibly constitute reservoirs to increase the seed stock of the parasite. In controlled experiments, R. fistulosa grown and reproduced better on rice and tomato crops, than on soybean which delayed the reproduction of the parasite. Germination of R. fistulosa seeds was stimulated in presence of cultivated crops or weeds. New collected seeds shown six months dormancy but their germination was stimulated in presence of phytohormones. In conclusion, the best strategies to control R. fistulosa must include delayed sowing, crop rotation with leguminous species (i.e. soybean, peanut or cowpea) and vegetable cropsKeywords: Rhamphicarpa fistulosa, rice, lowland, parasitism, host-parasite interaction, weeds

Kyoto University (京都大学)
0048
新制・課程博士
博士(人間・環境学)
甲第18605号
人博第701号
新制||人||167(附属図書館)
26||人博||701(吉田南総合図書館)
31505
京都大学大学院人間・環境学研究科相関環境学専攻
(主査)教授 加藤 眞, 教授 市岡 孝朗, 教授 瀬戸口 浩彰, 教授 宮本 嘉久, 教授 新宮 一成
学位規則第4条第1項該当

This study describes the effect of the root hemi-parasitic angiosperm Rhinanthus minor on the structure of the communities in which it lives and seeks to elucidate a mechanism through which the parasite acts to effect these changes in the community. Field manipulations reveal that R. minor suppressed the growth of grasses and legumes in a newly sown meadow whilst promoting the forbs within one growing season. In contrast the removal of R. minor from mature meadow plots did not influence their composition. After an additional growing season the parasite did not further influence the composition of the new meadows but removal did begin to benefit the biomass of mature plots. In isolation the parasite caused most damage to grasses whilst leaving legumes and forbs undamaged. Moreover, the parasite performed worst in terms of growth and photosynthesis when attached to the forbs. Consequently the parasite was able to moderate intra-specific competition between grasses and forbs. I thus hypothesised that forbs were able to prevent the parasite form abstracting resources where as grasses could not. Tracer experiments using isotopically e5N) labelled potassium nitrate confirmed this hypothesis showing that more of the resources taken up by the host were stolen by the parasite from grasses than from forbs. There was much variability in the translocation of resources from the legume studied. The reasons underlying the differential uptake of resources were highlighted using histological studies which showed that all of the forbs possessed successful resistance mechanisms to the parasite whilst no successful resistance was observed in the grasses or legumes. Two different resistance mechanisms were observed in the forbs; hypersensitive cell-death at the host-parasite interface and host lignification. I therefore propose that differential host resistance may underlie this parasite's community level effects as forbs possess a resistance capacity that other potential hosts do not.

Os nematoides que atualmente tem sido classificados como P. coffeae, demonstram ampla variabilidade morfológica, molecular e quanto à reação à diferentes hospedeiros. Por conseguinte, é essencial que se identifique corretamente as espécies e que se conheça a capacidade de parasitismo em alguns hospedeiros, para que se possa estabelecer medidas de controle e estimar eventuais riscos da entrada de um patógeno em uma área com hospedeiro suscetível. Neste sentido, a presente pesquisa propõe, numa primeira parte, uma medida de controle de P. jaehni (K5), que é muito agressivo à cafeeiro arábico, baseada no uso de plantas má hospedeiras. Na segunda parte deste trabalho é realizada uma caracterização da reação de diferentes espécies vegetais à quatro populações de Pratylenchus spp. Em virtude do exposto, este estudo objetivou avaliar a reação de diferentes cultivares de feijoeiro comum frente à P. jaehni (K5), visando seu possível uso no manejo de áreas cafeeiras infestadas e caracterizar a reação de diferentes espécies vegetais de importância econômica (café, porta-enxertos cítricos, banana e sorgo) frente à quatro populações de Pratylenchus spp. As populações inciais utilizadas nos experimentos variaram entre 180 e 200 nematóides. Em todos os experimentos, os nematóides foram extraídos das raízes pelo método de Coolen e D´Herde (1972) e, eventualmente, do substrato pelo método de Jenkins (1964). Foram realizados três ensaios: i) o primeiro com feijoeiro comum, onde todas as cultivares utilizadas foram resistentes à P. jaehni (K5), inclusive na réplica; ii) o segundo com porta-enxertos cítricos, no qual somente o limão-cravo foi hospedeiro de P. jaehni (K5); iii) o terceiro, no qual foi utilizado café, limão-cravo, banana e sorgo para o conhecimento das respectivas reações frente à 4 populações de Pratylenchus spp. Neste último experimento ocorreu uma reação hospedeira diferenciada para cada população. De acordo com os resultados, concluise que as cultivares de feijoeiro comum utilizadas apresentam potencial de uso em áreas cafeeiras infestadas por P. jaehni (K5), em consórcio ou em áreas de renovação de cafezal, e que as populações de Pratylenchus spp. são capazes de se reproduzir de forma diferenciada frente às espécies vegetais testadas.
Nematode populations that have been classified as P. coffeae show wide morphological, molecular and host range variability. Therefore, is essential the correct identification of the species and the knowledge about its parasitism capacity on some hosts, to ensure appropriate control measures and to estimate the entry risks of a pathogen in an area with susceptible host. In this sense, the present research report, firstly, propose a control measure of P. jaehni (K5), which is very aggressive to the arabic coffee, based on the use of poor host plants. In the second part, was carried out a host status evaluation of different plant species to four populations of Pratylenchus spp. In this context, were evaluated the reaction of different common bean cultivars to P. jaehni (K5), for its possible use in management of coffee areas infested with this nematode, and characterized the response of different economic important plant species (coffee, citrus rootstocks, banana and sorghum) to four populations of Pratylenchus spp (IB01P, IB02P, K5 e C1). The initial population used in experiments ranged between 180 and 200 nematodes. In all experiments, nematodes were extracted from roots by Coolen e D\'Herde´s method (1972) and eventually from the substrate by Jenkins´ method (1964). Three assays were conducted and results was as following: i) in the first one, with common bean, all tested cultivars were resistant to P. jaehni (K5), including the replica; ii) in the second one, with rootstocks, only rangpur lime was a good host of P. jaehni (K5); iii) in the third one, in which was used coffee, rangpur lime, banana and grain sorghum, was observed a differential host reaction for each nematode population evaluated. According to the present results, we suggest that common bean cultivars tested have great potential for use, or in intercropping or in crop rotation, in coffee areas infested by P. jaehni (K5) and populations of Pratylenchus spp. have different reproductive fitness in the plants species tested.

Les parasites sont une partie très importante des écosystèmes, néanmoins, dans certains cas tels que l’aquaculture ils causent des épidémies. Dans cette thèse j’ai étudié les interactions hôte-parasite chez les poissons coralliens, dans les milieux naturels et les milieux aquacoles. J’ai étudié l’arrangement des communautés de 13 espèces de monogènes de 34 espèces de poissons papillon dans l’Indo-Pacifique. Les résultats montrent qu’il existe un patron de distribution stable des espèces, ce qui, combiné avec les résultats de l’étude biogéographique, suggère que la spécificité parasitaire observée a pu se développer tout au long des épisodes biogéographiques passés. Notamment, seule une espèce, Chaetodon lunulatus, n’est jamais parasitée. J’ai étudié le mucus des poissons qui est décrit comme la première ligne de défense contre des agressions externes, pour analyser quels facteurs de C. lunulatus pourraient être reliés à l’absence des parasites. Le microbiome et le métabolome du mucus branchial des poissons papillons montrent une grande diversité. C. lunulatus présente une plus grande abondance de Fusobactéries qui est corrélée avec une surexpression de peptides pouvant dériver de la β-hémoglobine. La purification, la synthèse et l’évaluation des activités antiparasitaires des peptides sont en cours au laboratoire. J’ai aussi étudié l’utilisation des plantes médicinales comme alternative à des traitements chimiques dans l’aquaculture. J’ai testé l’activité antibactérienne et immunostimulatrice de plusieurs plantes et algues polynésiennes. L’algue rouge Asparagopsis taxiformis montre de fortes activités antibactériennes sur Vibrio harveyi et Tenacibaculum maritimum et induit une augmentation de l’expression de deux gènes liés à l’immunité chez Platax orbicularis
Fish parasites are an important part of ecosystems, however, in certain cases such as in aquaculture they can cause severe disease outbreaks. In this thesis I have studied host-parasite interactions in coral reef fishes, both in the natural and culture environments. I have studied the distribution of 13 dactylogyrid species from 34 butterflyfishes in the Indo West-Pacific. Composition of dactylogyrid communities was host specific and together with the biogeography results, where a turnover in the main Haliotrema species was observed, suggest that parasite specificity might result from host-parasite coevolution derived from past biogeographical episodes. Only one butterflyfish species, Chaetodon lunulatus, was never found parasitized by gill monogeneans. I have studied the butterflyfish mucus, which is the first barrier against pathogens, to investigate the C. lunulatus factors that might be related to the monogenean absence. Butterflyfish gill microbiome and metabolome revealed a high diversity. C. lunulatus presented a significantly higher abundance of Fusobacteria which was correlated to a higher expression of potentially derived β-hemoglobin peptides. Synthesis and evaluation of the peptide antiparasitic activities are being performed in the laboratory. I have also studied the use of medicinal plants as an alternative to chemotherapy in fish aquaculture. I investigated the antibacterial and immunostimulant activities of several local Polynesian plants and algae, and I found that Asparagopsis taxiformis displayed a potent antibacterial activity against Vibrio harveyi and Tenacibaculum maritmum and increased expression of two immune-related genes in Platax orbicularis

Les nématodes à galles (RKN), Meloidogyne spp. sont des petits vers parasites qui infectent les racines des plantes où ils induisent la formation de sites nourriciers. Les RKN sont des endoparasites à large gamme d'hôtes, englobant les principales espèces de plantes cultivées. Meloidogyne javanica, M. graminicola et M. incognita sont les principales espèces parasitant le riz (Oryza sativa). Le succès infectieux des RKN repose sur la production de protéines effecteurs de virulence, secrétées dans leurs glandes oesophagiennes et libérées dans les cellules de la plante hôte par leur stylet. La caractéristique principale des RKN est leur capacité à déréguler des cellules du parenchyme vasculaire pour induire la formation de cellules géantes multinucléées, à haute activité métabolique. Les processus moléculaires sous-jacents restent encore mal connus, alors que l’identification d’effecteurs de virulence et de leurs cibles végétales pourrait fournir de nouvelles perspectives pour le contrôle des RKN. Ainsi, les objectifs de cette étude étaient (1) d’évaluer le rôle de protéines de Meloidogyne sécrétées (MSP) au cours des interactions riz - RKN et (2) d'identifier des cibles des MSP parmi les principales protéines Hub d’Arabidopsis thaliana impliquées dans l'immunité des plantes, afin d'évaluer la fonction putative des MSP dans les cellules hôtes. Pour la première partie de notre étude, nous avons sélectionné trois MSP exprimées dans les glandes oesophagiennes et possiblement sécrétées. L’analyse de l’expression des gènes par RT-qPCR a montré que MSP2 est fortement exprimé dans les premiers stades du cycle du nématode, tandis que MSP18 et MSP19 sont activés au cours du parasitisme dans les racines du riz. Les essais de localisation subcellulaire dans les cellules d'oignon ont identifié le noyau (pour MSP2) et le cytoplasme (pour MSP7 et MSP18) comme compartiments cellulaires ciblés par les protéines du nématode. Des plants de riz (O. sativa Nipponbare) transgéniques ont été produits pour évaluer le rôle des MSP au cours des interactions riz-RKN. Des lignées de riz surexprimant MSP18 ont permis un taux de reproduction plus élevé de M. javanica et M. graminicola. Au contraire, des retards de développement et de reproduction de M. javanica ont été observés sur des lignées de riz exprimant des micro-RNAs capables de supprimer l’expression des gènes MSP2 ou MSP19. Ces données ont montré que MSP2, MSP18 et MSP19 peuvent être des gènes importants pour le parasitisme ou le développement du nématode. Les tests d'expression transitoire dans le tabac (Nicotiana benthamiana) ont montré que MSP18 peut interférer avec la mort cellulaire programmée déclenchée par INF1, ce qui suggère que MSP18 pourrait supprimer les voies de défense des plantes pour faciliter l’infection. Dans une deuxième partie de ce travail, des analyses systématiques en double-hybride chez la levure ont été menées pour vérifier les interactions protéine-protéine entre 6 MSP et 18 protéines Hub d’A. thaliana. Chez la levure, la protéine du nématode MSP400 interagit avec trois protéines Hub, l’Anaphase-Promoting-complex 8 (At-APC8) et les facteurs de transcription At-TCP14 et At-TCP15. L'interaction physique de MSP400 avec At-APC8, un régulateur clé du cycle cellulaire de la plante, a été confirmée in planta par complémentation bimoléculaire de fluorescence (BiFC). Ces résultats démontrent pour la première fois qu'un effecteur de nématode est capable d'interagir directement avec une protéine régulatrice du cycle cellulaire chez la plante, révélant un nouveau mécanisme utilisé par les RKN pour commander la machinerie du cycle de la cellule hôte et induire ainsi la formation du site d'alimentation. Les données obtenues dans cette étude élargissent considérablement notre connaissance des acteurs moléculaires qui contribuent à la pathogénicité des nématodes, mettant en évidence les différents mécanismes exploités par les RKN pour promouvoir la sensibilité des plantes
Root-knot nematodes (RKN), Meloidogyne spp. are small parasitic worms that infect plant roots where they induce the formation of highly specialized nutrient feeding sites. RKN are endoparasites with a wide host range encompassing major plant crops, impairing effective specific control. Meloidogyne javanica, M. graminicola, and M. incognita are the principal RKN species responsible for rice (Oryza sativa) production losses. Successful plant infection is likely achieved by nematode effector proteins produced in their esophageal gland cells and released into the host plant cells through their stylet. In particular, one of the striking features of RKN is their ability to deregulate vascular parenchyma cells to induce the formation of multinucleated giant cells with a high metabolic activity in the roots. The molecular processes underlying plant-RKN interactions still remain poorly understood. Identification of nematode virulence effectors and their plant targets may provide new insights for developing control strategies towards RKN. Thus, the aims of this study were to (1) assess the role of Meloidogyne secreted proteins (MSP) in rice – RKN interactions and (2) identify MSP targets among the major Arabidopsis thaliana Hub proteins involved in plant immunity, to assess the putative MSP function into host cells. For the first part of our study, we selected three Meloidogyne-genus specific proteins expressed in esophageal glands and predicted to be secreted. Gene expression analysis by RT-qPCR showed that MSP2 is highly expressed in the early stages of the nematode cycle, while MSP18 and MSP19 are up-regulated during parasitism in rice roots. Subcellular localization assays in onion cells identified the nucleus (for MSP2) and cytoplasm (for MSP7 and MSP18) as the main cellular compartments targeted by nematode proteins. Transgenic rice (O. sativa Nipponbare) plants expressing the MSP cDNAs or artificial micro-RNAs (amiRNAs) able to silence MSP genes were used to assess the role of MSPs during rice-RKN interactions. Homozygous transgenic lines were inoculated with pre-parasitic juveniles (J2) and (i) the number and developmental stage of nematodes present in roots after 21 days, (ii) the number of egg masses laid after 28 days and, (iii) the number of next-generation hatched J2 after 45 days were assessed. Rice lines overexpressing MSP18 allowed a higher reproduction rate of M. javanica and M. graminicola. On the contrary, impaired M. javanica development and reproduction was observed in rice lines expressing amiRNAs against MSP2 or MSP19 genes. These data showed that MSP2, MSP18, and MSP19 genes might be important genes for nematode parasitism or development. Transient expression assays in tobacco (Nicotiana benthamiana) revealed that MSP18 interfered with the INF1-triggered programmed cell death, suggesting that MSP18 could suppress the plant defense pathways to facilitate nematode parasitism. In the second part of this work, systematic yeast-two-hybrid paired assays were conducted to check for protein-protein interactions between 6 MSP and 18 A. thaliana Hub proteins. In yeast, the nematode MSP400 protein interacts with three Hub proteins, the Anaphase-Promoting-Complex 8 (At-APC8) and the transcription factors At-TCP14 and At-TCP15. Physical interaction of MSP400 with At-APC8, a key plant cell cycle regulator, was confirmed in planta by bimolecular fluorescence complementation (BiFC) assays. These results demonstrated for the first time that a plant parasitic nematode effector is able to directly interact with a cell cycle regulatory protein, revealing a novel mechanism utilized by RKN to control the host cell cycle machinery and thereby induce feeding site formation. The data obtained in this study significantly broaden our knowledge of the molecular players contributing to nematode pathogenicity, highlighting the different mechanisms exploited by RKN to promote plant susceptibility

Root-knot nematodes are serious economic pathogens of horticultural and crop plants in sub-tropical and tropical regions of the world including Australia. The nematodes invade the roots and are able to induce the redifferentiation of root cells into multinucleate feeding cells called 'giant cells'. These feeding cells form by repeated mitosis without cytokinesis and develop wall ingrowths typical of transfer cells. It is clear that the pronounced morphological and physiological changes associated with the formation of these specialised feeding cells are caused by altered gene expression in host cells. For this reason, the study of molecular responses induced in host cells by the pathogen will not only provide new information to the host-parasite relationship but could provide better options for control strategies. Differential display reverse transcription polymerase chain reaction (DDRT-PCR) was used as a tool to study changes in gene expression during the compatible interaction between tomatoes {Lycopersicon esculentum) and the root-knot nematode M javanica 25 days post-infection. A series of experiments were undertaken to develop this method, to improve sensitivity and reliability. Both short and long primers were used in the DD polymerase chain reaction (PCR). It was found that long primers in combination with a two-step PCR and the use of 33P as the radio-labelled nucleotide greatly improved the sensitivity and reproducibility of the technique. In addition, sequence-specific primers, which were originally designed for analysis of specific genes could be used as 5' arbitrary primers, further expanding the number of primers that could potentially be used in DD. Using a total of 16 primer combinations, 25 differentially expressed genes were identified, of which 12 were selected for further analysis. Although DD has several advantages over other methods for isolating differentially expressed genes, it is also prone to artefacts. An efficient methodology was developed that enabled false positives to be discarded after the cloning step via a reverse Northern blot analysis. This involves probing the clones with labelled cDNAs from healthy and giant cell enriched tissues to differentiate between true and false positives. Clones harbouring truly differentially displayed bands were sequenced and semi-quantitative RT-PCR was used to reconfirm their differential expression and to allow isolation of low abundance differential cDNAs. Use of the screening strategy described here led to the isolation of six cDNAs; three down-regulated (HA/HT1 and HAS) and three upregulated (IC1, IC9 and IA2) genes whose expression was as predicted by the display gels, whilst minimising the amount of RNA used. In order to determine the transcriptional activity of the genes more precisely, a relative real-time quantitative RT-PCR assay was developed that provides a quick, sensitive and reliable method of measuring and analysing expression patterns of genes. The starting materials from both healthy and giant cell enriched tissues were normalised to their relative amounts of actin in a TaqMan PCR assay. The expression pattern of the differentially expressed genes was then determined at various time points after infection (3, 15, 25 and 30 days) using SYBR Green I dye as the fluorescent dye, in TaqMan PCR. In healthy tissues, the expression pattern of the genes did not show any marked developmental regulation over time. However, significant changes in their transcriptional activity were observed in the giant cell enriched tissues particularly between 3 and 15 days after infection. Overall, the transcriptional regulation of the genes was up to 10-fold except for IC9 and HA/HT1, which showed a 69-fold increase and 28-fold decrease respectively, by 15 days post-infection. The real-time quantitative RT-PCR assay can be used to provide information on gene expression over the lifetime of the nematode plant association, and promises to be very widely applicable to study other systems. These results also demonstrate the sensitivity of DD to isolate genes that show subtle changes in gene expression. Of the six differentially expressed genes studied in detail, five were found to be of plant origin and one (IC1) was found to be of nematode origin. Database searches of these differentially expressed genes revealed that at the amino acid level, the deduced IA2 gene product shares 72% identity to cucumber basic protein (a plantacyanin). HA3 and IC1 showed weak similarity to Arabidopsis thaliana transcription factor MYB59 and C. elegans cosmid C respectively. IC1 could be a nematode mRNA contamination or it could be part of the stylet component that the nematode injects inside the plant cells. If this results stands and it is not a contaminant of tissue preparation, the result is novel and could be highly significant. Transcripts HT1 and HA both showed very strong homology to the previously identified Lycopersicon esculentum RB7 gene and interestingly in this study, the level of its mRNA was found to be down-regulated during infection relative to control tissues. The full-length sequence of IC9 was determined by 5' RACE and its cDNA showed 58% identity with the gibberellin 2- oxidase protein from Piswn sativum, indicating the involvement of gibberellic acid in giant cell or gall formation. Because most of the genes identified represent the 3' noncoding regions of the mRNA or they are members of gene families for which the function is unknown, their significance of regulation is difficult to interpret without any further characterisation. Nevertheless, the results obtained are new, and add to the pattern of genes with altered expression in feeding cells. The results presented in this thesis highlight the power of DD as a reliable, rapid and efficient method to systematically screen and identify differentially expressed genes in this system. The development of the SYBR Green PCR assay provides a novel approach to quantify native gene transcripts in giant cells compared to the use of a reporter gene assay. The genes isolated provide additional information on aspects of and function of nematode feeding cells. With the use of more primer combinations, many more genes transcriptionally regulated by the nematode can be revealed. This will lead to a better understanding of plant-nematode interactions and plant development, and may reveal ways to engineer sound and durable forms of synthetic resistance to the pathogen.

Coevolutionary interactions change across landscapes, leading to the formation of geographic selection mosaics. Analyses of coevolutionary dynamics have so far focused on interactions between two trophic levels. The tritrophic interactions between the pierid butterfly Anthocharis cardamines (the orange-tip), its brassicaceous host-plants Cardamine pratensis (lady's smock) and Alliaria petiolata (garlic mustard), and primary parasitoids Phryxe vulgaris (Tachinidae) and Cotesia saltator (Braconidae), are known to vary across mainland Britain and Continental Europe. In Britain, northern A. cardamines populations tend to utilize C. pratensis and southern ones A. petiolata; the primary parsitoid in this country is P. vulgaris. In Sweden, the butterfly does not show a strong preference for any brassicaceous hostplant, and the primary parasitoid is C. saltator. In this thesis, I investigate likely selection pressures operating on these interactions in a single study site on the Wirral peninsula in northern England. Host use affects the emergence timing and dispersal of A. cardamines males; specimens utilizing C. pratensis emerge earlier than those utilizing A. petiolata, and are also smaller. Since small size is linked with depressed dispersal, utilization of C. pratensis results in an "emerge early and wait" mate seeking strategy; conversely, utilization of A. petiolata is associated with an "emerge late and rove" strategy. These alternative strategies are likely adapted to the varying density of locally emerging females across the landscape, with high density populations tending to be associated with C. pratensis and low density ones with A. petiolata. In dense populations, late emerging males will be at a disadvantage since females mate only once; in the study population, their predicted fitness always declines to < 1 in late season. This is coupled with a 'stay-or-go' response, in which a proportion of late emerging males immediately emigrate to a low density continuum outside the study area, where early emergence is less critical for fitness. Such a response could help maintain sink populations by averting Allee effects (decreased population growth due to low mate encounter rates). Late instar A. cardamines larvae are heavily parasitized by P. vulgaris. This could select for early vacation of the host-plant at small larval size, whereas the size-fecundity relationship in females should select for prolonged growth to larger size. Mathematical models indicate that high rates of parasitization are sufficient to overturn the fecundity benefit of large size, but fall short of maintaining strong stabilizing selection for an optimal winglength. The tendency of some larvae to move off their host-plant before the final instar resting phase is probably a direct evolutionary response to parasitism risk. A. cardamines larvae are pre-dispersal seed-predators; an early flowering ecotype of C. pratensis has likely been selected to avoid egg-laying A. cardamines females. In turn, the butterfly appears to be invading this host-plant's phenological space, with selection favouring small, early emerging females which oviposit on it. This cautions against interpreting the recent advance in A. cardamines' phenology solely in terms of a response to climate change. Spatio-temporal variation in the intensity of these effects likely contributes to the ongoing coevolutionary dynamics within this tritrophic system.

Une invasion biologique réussie se décompose en trois étapes : l'introduction d'une population allochtone sur une nouvelle aire de répartition géographique, l'établissement de populations viables et leur prolifération exponentielle dans l'aire d'introduction. En 2002, Shea et Chesson ont énoncé l'hypothèse d'Opportunité de Niche dans le but d'expliquer la réussite de l'établissement et de la prolifération des espèces envahissantes. Selon ces auteurs, les espèces exogènes introduites sur une nouvelle aire géographique peuvent y profiter d'une part de la diminution de la pression exercée par leurs ennemis naturels et d'autre part d'une augmentation de la disponibilité de leurs ressources. Drosophila suzukii est une de ces espèces dites envahissantes. Originaire d'Asie, sa présence est aujourd'hui largement recensée en Amérique du Nord et dans tout l'ouest de l'Europe. Depuis son introduction en 2008, en Europe notamment, l'invasion de D. suzukii est particulièrement rapide et les dégâts engendrés dans les cultures par cette espèce qui se développe sur les fruits en cours de maturité sont très préoccupants. Au cours de nos travaux, nous avons mis en évidence certains caractères chez cette espèce comme étant des éléments facilitant le succès de son invasion. D. suzukii présente tout d'abord une grande résistance face à ses principaux ennemis naturels, les parasitoïdes. Nous avons ensuite montré l'existence d'un lien entre sa capacité à mettre en échec les parasitoïdes larvaires et son importante charge hémocytaire. Par ailleurs, grâce à un échantillonnage très large, nous avons illustré la grande polyphagie de cette espèce. Ce caractère lui assure potentiellement une disponibilité en ressources tout au long de l'année. Enfin, à l'aide d'une étude portant sur les relations tritrophiques, nous avons montré que la qualité nutritive des plantes hôtes utilisées par D. suzukii peut avoir un impact sur le niveau de sa résistance aux entomophages. D. suzukii parvient en effet à augmenter sa résistance à plusieurs espèces de parasitoïdes lorsqu'elle réalise son développement sur un substrat riche en alcaloïde. Nos derniers résultats suggèrent qu'elle serait même capable de réaliser une forme d'automédication. Les différentes études réalisées au cours de cette thèse semblent confirmer que D. suzukii profite d'une opportunité de niche en Europe tempérée
Successful biological invasion is divided into three stages: the introduction of a non-native population on a new geographical area, the establishment of viable populations and their exponential growth in the area of introduction. In 2002, Shea and Chesson proposed the Opportunity Niche Hypothesis in order to explain the success of the establishment and the spread of invasive species. According to these authors, alien species introduced into a new range can jointly benefit from a decrease of the pressure applied by natural enemies and from an increase in the availability of resources. Drosophila suzukii has been reported to be an invasive species. Native of Asia, it has been now widely observed in North America and throughout Western Europe. Since its introduction in 2008, especially in Europe, the invasion of D. suzukii is particularly rapid and, as it grows on ripe fruits, a lot of concerns have been raised on the damage it causes on crops. In our work, we have highlighted some characters of this species as elements that facilitate the success of its invasion. D. suzukii is highly resistant against parasitoids, known as its major natural enemies. We have then showed the existence of a link between the ability to resist to larval parasitoids and a high hemocyte load. Furthermore, thanks to a very large sample, we have illustrated the great polyphagia of this species. This property provides available resources throughout the year. Finally, with a study of tritrophic interactions, we have shown that the nutritional quality of host plants used by D. suzukii can have an impact on the level of resistance to entomophages. Indeed, D. suzukii increases its resistance to several species of parasitoids when it growths on an alkaloid-rich substrate. Our recent results suggest that it would even be able to achieve a form of self-medication. The different studies performed in this thesis seem to confirm that D. suzukii makes the most of niche opportunity to efficiently invade temperate Europe

Doutoramento em Biologia
O género Botryosphaeria é bem conhecido devido ás espécies que causam doenças numa grande diversidade de plantas. A taxonomia deste género e seus géneros anamórficos, assim como das espécies tem sido algo confusa. Neste trabalho foram caracterizadas espécies de Botryosphaeria associadas a Quercus spp. e outros hospedeiros lenhosos, com base em características morfológicas e sequências nucleotídicas de um ou vários genes. A utilização desta abordagem permitiu a clarificação da taxonomia de algumas espécies já descritas, bem como a identificação e descrição de novos taxa. O género Dothiorella foi reintroduzido para acomodar os anamorfos de Botryosphaeria que possuem conídios castanhos e 1-septados. Esta decisão baseou-se no estudo das características morfológicas da espécie tipo do género bem como em análises filogenéticas. A análise filogenética de sequências nucleotídicas de vários genes confirmou que os géneros Diplodia e Lasiodiplodia representam taxa distintos. Demonstrou-se que as estrias longitudinais em conídios não têm valor taxonómico para distinguir entre os dois géneros. O género Lasiodiplodia distingue-se do género Diplodia pela presença de paráfises. Os métodos baseados em PCR, ARDRA, MSP-PCR e rep-PCR foram utilizados para discriminar entre espécies do género Botryosphaeria e espécies anamórficas relacionadas. O método ARDRA permitiu a diferenciação de 10 espécies, enquanto os métodos MSP-PCR e rep-PCR permitiram a identificação de 24 espécies. Estes métodos representam procedimentos simples e rápidos que podem ser utilizados para a identificação rotineira de isolados de Botryosphaeria ao nível da espécie. Os métodos MSP-PCR e rep- PCR também se revelaram úteis para estudar a variabilidade intraespecífica. Foi realizada uma análise sistemática molecular da família Botryosphaeriaceae recorrendo a sequências nucleotídicas de diversos genes. Esta análise mostrou que o género Botryosphaeria sensu lato é parafilético, sendo composto por várias linhagens filogenéticas. Estas linhagens representam géneros distintos que são caracterizados essencialmente com base na morfologia das formas anamórficas. O género Melanops foi reintroduzido para acomodar uma espécie anteriormente classificada no género Botryosphaeria. Não foi possível definir qual a posição da família Botryosphaeriaceae na classificação dos Ascomicetos. De acordo com os dados disponíveis prevê-se que esta família deverá ser incluída numa nova ordem distinta das ordens Pleosporales e Dothideales.
The genus Botryosphaeria is well known by the species that cause disease in a wide range of plant hosts. The taxonomy of Botryosphaeria and associated anamorph genera as well as of the species has been confused in the past. In this work, Botryosphaeria species occurring on Quercus spp. and other woody hosts were characterised on the basis of morphology and nucleotide sequence data from single or multiple genes. Using this approach it was possible to clarify taxonomic uncertainties related to some known species and also to identify and describe new species. Through a study of the type species of Dothiorella this genus was reinstated to accommodate anamorphs of Botryosphaeria with brown, one-septate conidia. This was further supported by phylogenetic analyses. Phylogenetic analyses of combined multi-gene sequence data supported the genera Diplodia and Lasiodiplodia as distinct. It was shown that conidial striations are not a taxonomically meaningful character to separate both genera, and that Lasiodiplodia is distinguished from Diplodia by the presence of paraphyses. The PCR-based methods ARDRA, MSP-PCR and rep-PCR were used to discriminate between species of the genus Botryosphaeria as well as associated anamorphic species. The ARDRA procedure allowed for the differentiation of 10 species while MSP-PCR and rep-PCR allowed a clear identification of 24 species. These methods provide simple and rapid procedures that can be used for routine identification of Botryosphaeria isolates at the species level. MSP-PCR and rep-PCR also proved useful to study intraspecific variability. A molecular systematic analysis of the family Botryosphaeriaceae was performed using multi-gene sequence data. The genus Botryosphaeria sensu lato was shown to be paraphyletic, and composed of different phylogenetic lineages. These represent distinct genera which are characterised essentially on the basis of morphological features of the anamorphs. The genus Melanops was reinstated to accommodate a species previously included in Botryosphaeria. The position of the Botryosphaeriaceae within the higher classification of Ascomycetes could not be indubitably established. Given the current data it is envisaged that this family should be included in a new order distinct from the Pleosporales and Dothideales.
FCT - SFRH/BD/10389/2002
FSE
POCTI/AGR/44348/2002
POCTI/AGR/56140/2004

The obligately outbreeding root hemiparasite Striga hermonthica (Orobanchaceae)is a serious threat to subsistence agriculture in sub-Saharan Africa. Resistance to this parasite in its crop hosts, such as rice, sorghum and maize, is not common, and the evolution of host adaptations that are able to overcome new sources of resistance is an ever-present risk. Research into host adaptation in S. hermonthica has generally sought to correlate the genetic relationship between Striga individuals with host identity; however, such approaches must be supported by lab-based evidence of host adaptation, otherwise ad hoc field sampling may result in the confounding of host identity with isolation-by-distance. Additionally, genetic variation used to reconstruct relationships is unlikely to provide an insight into relationships at functional loci underlying host adaptation. In this thesis, I use a range of new approaches to investigate several different aspects of parasite adaptation in the S. hermonthica-Sorghum bicolor pathosystem. Host adaptations, or pre-adaptations, are commonly revealed using tests for differential virulence between Striga populations and host genotypes; that is, by demonstrating population-level genotype-by-genotype interactions. Evidence for such interactions was found between three West African populations of S. hermonthica and five sorghum cultivars. These interactions were shown to be strongest at the parasite post-attachment life stage, and to depend on the parasite virulence metric used. Environmental influences on host-parasite interactions were strong and variable between years and sites. Candidate genetic loci for virulence, responding to selection in a micro-evolutionary fashion, were identified by Fst differentiation-based approaches ('outlier analyses') that aim to uncover associations between particular loci and environmental drivers, such as host identity. An AFLP outlier analysis was used on Striga plants parasitising nine sorghum cultivars in a field trial in Burkina Faso. Significant locus-specific differentiation was detected at 14 out of 1275 loci. However, predicted allele frequencies at these loci did not correlate with a field measure of Striga virulence across host-selected sub-populations. Simulation results suggested that the estimated levels of Fst at outlying loci could mean that alleles underlying host adaptation exist at intermediate frequencies in populations. A three-generation pedigree, created from a cross between S. hermonthica individuals from an East African and a West African population, enabled further insights into the genetics of adaptation. Individuals from a pseudo-backcrossed F1 (BCF1) generation, grown on two different sorghum hosts and in axenic culture, indicated significant host-related segregation distortion. Analyses of virulence in the BCF1 also provided strong evidence for epistasis, and for an effect of maternal identity. A second outlier analysis of host adaptation, investigating the East African population used in the pedigree, indicated some correspondence between outlier loci and loci found to be differentially segregating between different hosts in the BCF1 generation, and demonstrated the differing genomic extents of these phenomena. The results accumulated across these experiments provide evidence for a complex, polygenic basis to virulence in S. hermonthica.

La compréhension du processus d’adaptation des populations de parasites à leur plante-hôte est une question fondamentale en écologie évolutive. C’est également un enjeu majeur de recherche finalisée qui a des retombées pour la protection des cultures. L’oomycète Plasmopara viticola, agent causal du mildiou de la vigne, attaque les espèces du genre Vitis. Dans un contexte où l’enjeu principal des programmes d’amélioration est la durabilité des résistances, des connaissances nouvelles sur l’écologie et l’évolution de l'interaction entre le parasite et son hôte sont nécessaires afin d’évaluer le potentiel du mildiou à surmonter ces résistances. Dans ma thèse, je me suis intéressée au rôle de la plante-hôte comme facteur d’évolution des populations de mildiou, en posant cette question à différentes échelles évolutives : (i) dans le bassin d’origine du pathogène (Amérique du Nord), j’ai cherché à évaluer le degré de spécialisation du parasite sur sa gamme d’hôtes sauvages et cultivés; (ii) en Europe, où le mildiou de la vigne a été introduit récemment, j’ai étudié l’évolution des populations de mildiou soumis à la pression de sélection des résistances des nouvelles variétés de vigne. Pour comprendre la spécialisation plante-hôte dans ce pathosystème où plusieurs espèces cryptiques ont été identifiées, nous avons réalisé des tests d’inoculations croisées entre espèces hôtes (Vitis spp.) et agent pathogène (P. viticola). Les données phénotypiques et morphologiques apportent les preuves d’une spécialisation plante-hôte au sein des populations de P. viticola : les espèces A et D de mildiou sont spécialisées sur leur plante-hôte, tandis que le processus de spécialisation est en cours pour les espèces B et C. Même si aucune différenciation génétique n’a été montrée au sein de l’espèce C, il existe deux groupes distincts au sein de l’espèce B. Les isolats du compartiment cultivé sont en moyenne plus agressifs que les isolats issus des vignes sauvages, indiquant une adaptation des isolats cultivés sur leur plante hôte. A partir d’un large échantillonnage, nous avons étudié la distribution des espèces de mildiou sur leurs plantes-hôtes sauvages et cultivées. Ce travail a permis d’identifier une nouvelle espèce cryptique et a confirmé la spécialisation plante-hôte. En Europe, nos résultats montrent que le déploiement limité de variétés à résistantes partielles a conduit à des modifications des populations de mildiou: apparition d’isolats virulents (i.e. contournant un QTL majeur de résistance), et augmentation de l’agressivité sur Vitis vinifera. Dans le but de comprendre les mécanismes à l’origine de la spécialisation et du contournement des résistances, nous nous sommes intéressés au répertoire d’effecteurs du parasite. Une centaine d’effecteurs candidats ont été identifiés en utilisant les données disponibles sur le génome de P. viticola. L’analyse du polymorphisme de 32 candidats sur une sélection d’isolats montre que trois d’entre eux évoluent sous sélection positive. Ces résultats soulignent l’importance de la plante-hôte comme facteur de diversification des populations de l’agent pathogène et révèlent que le mildiou s’adapte rapidement aux résistances de la vigne. Il est désormais nécessaire de mieux appréhender le déploiement des résistances de la vigne afin qu’elles puissent être durables
Understanding the process of adaptation of parasite populations to their host-plant is a key issue in evolutionary ecology. It is also a major subject in applied research that has implications for crop protection. The oomycete Plasmopara viticola, the causal agent of downy mildew, attacks the species of the Vitis genus. In a context where the main concern of the breeding programs is the durability of resistance, new knowledge about the ecology and evolution of the interaction between parasite and host is needed in order to evaluate the potential of downy mildew to overcome the resistance. In my thesis, I addressed the role of the host-plant as an evolutionary factor for downy mildew populations, by asking this question at two different evolutionary scales: (i) in the pathogen region of origin (North America) I assessed the degree of specialization of the parasite on its wild and cultivated host range (ii) in Europe, where downy mildew has been introduced recently, I studied the evolution of downy mildew populations subject to the selection pressure imposed by resistant grapevine varieties. To understand the host-plant specialization in this pathosystem, where several cryptic species have been identified, we performed cross inoculations between different host (Vitis spp.) and pathogen (P. viticola) species. Morphological and phenotypic data provide evidence of host-plant specialization in P. viticola populations: downy mildew species A and D are specialized on their host-plant, while the specialization process is ongoing for species B and C. Although no genetic differentiation has been shown inside species C, there are two distinct groups within species B. Isolates from the cultivated compartment are on average more aggressive than isolates from wild vines, indicating an adaptation of isolates growing on cultivated host-plants. Finally, a large-scale study of the distribution of downy mildew species on both their wild and cultivated host-plants resulted in the identification of a new cryptic species and confirmed the host-plant specialization. In Europe, our results show that the limited deployment of resistant varieties has led to changes in downy mildew populations: emergence of virulent isolates (i.e. breakdown of a major QTL for resistance), and increased aggressiveness on Vitis vinifera. In order to understand the mechanisms at the origin of specialization and resistance breakdown, we examined the parasite’s effector repertoire. Over one hundred effector candidates were identified using available data on the P. viticola genome. The polymorphism of 32 candidate genes revealed that three of them evolve under positive selection. Our results reveal the strong ability of downy mildew to adapt to its host plant and to plant resistance. They should be taken into account when devising strategies for the deployment of grapevine resistances in order to guarantee their durability

Endoparasitoids of herbivorous lepidopterans have intimate relationships with their host species as well as the plant on which their host develops. Characteristics of both hosts and plants can affect parasitoid success in tri-trophic systems and thus, drive diversification. Genetic differentiation was estimated for Cotesia congregata (Say) collected from two distinct host-plant complexes, Manduca sexta L. on tobacco (Nicotiana tabacum L.) and Ceratomia catalpae, (Haworth) on catalpa (Catalpa bignonioides Walker), in the mid-Atlantic region of North America using seven microsatellite loci and the mitochondrial COI locus. Microsatellite allele frequencies were differentiated based on host-plant complex, and COI haplotypes from individuals on the same host-plant were identical despite geographic distances between catalpa sites of up to 830 km and distances between tobacco sites of up to 294 km. Results indicate genetic differentiation of subpopulations of C. congregata based on host-plant complex and not geographic distance, and were designated as host races. Cotesia congregata is a gregarious parasitoid, meaning that many individuals develop in a single host larva. Superparasitism, or repeated egg-laying events in the same host larva, is likely to occur in gregarious species. Brood size was not a good predictor of superparasitism in C. congregata, but within-brood male allele diversity indicates either superparasitism or multiple mating by female wasps.

Cuscuta pentagona is an obligate parasitic plant that hinders production of crops throughout the world. Parasitic plants have unique morphological and physiological features, the most prominent being the haustorium, a specialized organ that functions to connect them with their host's vascular system. The Cuscuta haustorium is remarkable in that it enables mRNA movement to occur between hosts and parasite, but little is known about the mechanisms regulating cross-species mRNA transfer or its biological significance to the parasite. These questions were addressed with genomics approaches that used high throughput sequencing to assess the presence of host mRNAs in the parasite as well as parasite mRNAs in the host. For the main experiment Cuscuta was grown on stems of Arabidopsis thaliana and tomato (Solanum lycopersicon) hosts because the completely sequenced genomes of these plants facilitates identification of host and parasite transcripts in mixed mRNA samples. Tissues sequenced included the Cuscuta stem alone, the region of Cuscuta-host attachment, and the host stem adjacent to the attachment site. The sequences generated from each tissue were mapped to host reference genes to distinguish host sequences, and the remaining sequences were used in a de novo assembly of a Cuscuta transcriptome. This analysis revealed that thousands of different Arabidopsis transcripts, representing nearly half of the expressed transcriptome of Arabidopsis, were represented in the attached Cuscuta. RNA movement was also found to be bidirectional, with a substantial proportion of expressed Cuscuta transcripts found in host tissue. The mechanism underlying the exchange remains unknown, as well as the function of mobile RNAs in either the parasite or host. An approach was developed to assay potential translation of host mRNAs by detecting them in the Cuscuta translatome as revealed by sequencing polysomal RNA and ribosome-protected RNA. This work highlights RNA trafficking as a potentially important new form of interaction between hosts and Cuscuta.
Ph. D.

The obligate stem parasite Cuscuta pentagona is able to take up host plant mRNA through a specialized organ known as the haustorium. Direct cell-to-cell symplastic connections between two different organisms are rare, and the translocation mechanisms and fate of these RNAs in the parasite is not understood. To characterize this phenomenon, mobile Arabidopsis and tomato mRNAs were identified from microarray and transcriptome sequencing projects and quantified in the host-parasite system. Mobile RNAs were quantified using real time (qRT)-PCR and were found to vary substantially in their rate of uptake and distribution in the parasite. Transcripts of tomato Gibberellic Acid Insensitive (SlGAI) and Cathepsin D Protease Inhibitor (SlPI) can be traced over 30-cm of parasite stem. SlPI was abundant in the C. pentagona stem, but the number of copies decreased substantially within the first eight hours post detachment. Additional studies of mobile RNAs from Arabidopsis, Translationally Controlled Tumor Protein (AtTCTP), Auxin Response Factor (AtARF) and a Salt-inducible Zinc Finger Protein (AtSZFP) supported the idea that mRNA molecules differ in their mechanisms of uptake and mobility between host and parasite. Known phloem-mobile RNAs (SlGAI and AtTCTP) have uptake patterns that differ from each other as well as from other RNAs that are not reported to be phloem mobile (SlPI and AtSZF1). The function of RNAs in plants extend beyond protein translation to include post transcriptional gene silencing or long distance signaling, and mobile RNA in C. pentagona systems offers novel insights into this aspect of plant biology. Studies of cell-to-cell trafficking of RNAs and other macromolecules would be facilitated by the ability to manipulate individual cells. To this end, work was initiated to explore alternative approaches to understanding single cell biology using laser-mediated approaches. Optoperforation, or the use of multiphoton processes to form quasi-free electron plasmas to initiate transient pore formation in plasma membranes, has been demonstrated, but not in cells of an intact plant. This work details a protocol for optoperforation of Arabidopsis epidermal cells to allow for uptake of external dye-labeled dextrans and retention for up to 72 hours, and has the potential for transformation and molecular tagging applications.
Ph. D.

Prior exposure to plants cues can enhance assortative mating in insects. We hypothesized that, as previously reported for females, males of Cotesia congregata would display inherent responses to plant cues that could be modified by postemergence experience and further, that males originating from two different host-plant complexes (HPCs) would display different behavioral responses to these HPCs. In no-choice contact assays with a non-host plant, searching responses of males and females increased sharply at Day 2 and remained stable through Day 4. In no-choice assays with potential host plants, males searched longer on catalpa than tobacco; responses were not modified by postemergence experience. In choice assays with both HPCs, naïve males did not display orientation preferences; however, males experienced with their natal plant preferred their natal HPC. Results indicate that postemergence experience on the natal host plant induces an orientation preference for the natal HPC and thus, can facilitate assortative mating.

In host-parasitoid systems, there is a continuous coevolutionary arms race where each species imposes a strong selection pressure on the other. The host needs to develop defence strategies in order to escape parasitism and the parasitoid must evolve counter-defence strategies in order to overcome the host’s immune defence and successfully reproduce. This makes host-parasitoid systems excellent model systems for understanding evolutionary processes underlying host race formation and speciation. In order to gain a better understanding of the complexity of host-parasitoid interactions several aspects must be considered, such as search behaviour and host selection in the parasitoid, the development of immune responses in the host and counter-defence strategies in the parasitoid. In this thesis, I investigate interactions and coevolution in a natural host-parasitoid system, consisting of five species of Galerucella leaf beetles and three species of Asecodes parasitoids, by combining behavioural ecology with chemical ecology and immunology. In the studies performed, I found that pheromone production and responses in the beetles are connected to the phylogenetic relatedness between the Galerucella species (Paper I). I found no evidence that Asecodes exploits the adult pheromone to locate host larvae, but observed an ability in the parasitoids to distinguish a better host from a less suitable one based on larval odors (Paper II). The studies also revealed large differences in immune competence between the Galerucella species, which were linked to differences in hemocyte composition in the beetle larvae (Paper III, IV). Further, the results suggest that parasitism success in polyphagous Asecodes is strongly affected by former host species of the parasitoid (Paper IV). In conclusion, the results of this thesis suggest an on-going evolution in both parasitoid virulence and host immune responses in the Asecodes-Galerucella system.

At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Manuscript. Paper 4: Manuscript.

Resums pendents
Les comunitats ecològiques són entitats dinàmiques l’estructura i composició de les quals està conformada per processos ecològics i per variabilitat ambiental. Degut a que les espècies interaccionen entre elles, els canvis en l’estructura i la composició de les comunitats poden alterar les xarxes d’interacció i el funcionament dels ecosistemes. En aquesta tesi estudio com un procés ecològic (la colonització d’un hàbitat per una nova espècie) i un component important de la variabilitat espacial (la grandària de l’hàbitat) afecta a les comunitats de pol·linitzadors, d’abelles i vespes, així com a les seves interaccions mutualistes i antagonistes. També investigo si aquests canvis afecten l’estructura de la xarxa d’interaccions i les conseqüències potencials sobre la funció ecosistèmica (pol·linització, reproducció de les plantes, i parasitisme).Els canvis en la composició de les comunitats de plantes són cada vegada més freqüents en el context actual de les invasions biològiques. Tot i així, les invasions de plantes normalment representen escenaris extrems de dominància ecològica i els seus efectes han sigut estudiats relativament bé. Contràriament, els efectes de la colonització dels hàbitats per espècies autòctones (per exemple, com a resultat dels canvis en la distribució geogràfica mediats pel canvi climàtic) han rebut molta menys atenció.Al Capítol 1 analitzem els efectes de l’arribada d’una nova espècie de planta autòctona a un matollar, incloent els canvis en la composició de la comunitat de pol·linitzadors, els patrons d’interacció i l’estructura de la xarxa planta-pol·linitzador, y les conseqüències per a la pol·linització i per a l’èxit reproductiu de les espècies de plantes residents. Per a això vam manipular experimentalment taques de matollar introduint una planta autòctona (planta colonitzadora d’ara endavant), ajustant el número de flors introduïdes a la quantitat de recursos florals localment disponibles. La planta colonitzadora va atraure borinots, facilitant les interaccions a les plantes residents. Les comunitats colonitzades van exhibir una generalització de la xarxa més gran, un aniuament més gran, i modificacions en la composició dels mòduls de la xarxa. Tots aquests canvis van resultar en una deposició de pol·len heteroespecífic (però no conespecífic) més gran, i va tenir efectes contrastats en l’èxit reproductiu de les plantes residents (neutral, positiu, o negatiu depenent de l’espècie). El nostre estudi mostra que les interaccions planta-pol·linitzador es reorganitzen ràpidament com a resposta a noves situacions (inclús quan la nova planta no és excessivament dominant), amb conseqüències funcionals importants en la pol·linització i en la reproducció de les plantes. La relació espècies-àrea (increment en la riquesa d’espècies amb l’àrea) és una de les lleis de l’Ecologia més antigues i fonamentals i, recentment, ha sigut estesa a la riquesa d’interaccions (relació enllaços-àrea) i a l’estructura de la xarxa (relació xarxa-àrea). Tot i així, tot i que les relacions espècies-àrea i enllaços-àrea han rebut un ampli suport d’estudis empírics, la relació entre l’estructura de la xarxa i l’àrea roman en gran part inexplorada. Entendre aquesta relació és important perquè la fragmentació antropogènica dels hàbitats està tenint impactes profunds en la configuració dels hàbitats i està reduint la grandària dels mateixos.Al Capítol 2 testem vàries hipòtesis relacionades amb les relacions espècies-àrea, enllaços-àrea, i xarxa-àrea utilitzant comunitats d’abelles i vespes nidificants en cavitats preestablertes així com la fauna associada als seus nius (hostes i parasitoides d’ara endavant, respectivament) com a sistema d’estudi. Per a obtenir nius d’abelles i vespes vam instal·lar estacions de niadors en taques d’hàbitat obert (favorable) –envoltats de bosc dens (hàbitat desfavorable)- de tres categories de grandària (petites, ~0.1 ha; mitjanes, ~5ha; i grans, hàbitat obert continu). Els nius obtinguts en cada taca van ser disseccionats i els seus contingut van ser analitzats anotant la identitat i l’abundància de cada espècie d’hoste i parasitoid. La riquesa d’hostes, parasitoides, i interaccions va incrementar de les taques petites a les mitjanes, però no de les taques mitjanes a les grans. Els enllaços per espècie van incrementar amb la grandària de la taca, però la taxa de parasitisme no va seguir cap tendència relacionada amb la grandària de la taca. L’especialització de la xarxa va disminuir de les taques petites a les mitjanes, però les taques grans van mostrar una especialització similar a la de les taques petites i mitjanes. La modularitat va disminuir de les taques petites a les mitjanes, però no va haver-hi diferències entre les taques mitjanes i les grans. El número de mòduls va incrementar amb la grandària de la taca. Els nostres resultats estan, majoritàriament, parcialment d’acord amb les relacions espècies-àrea i enllaços-àrea. L’absència de diferències entre les taques mitjanes i grans indica l’existència d’un llindar per damunt del qual la riquesa d’espècies i d’interaccions ja no està limitada per la grandària de l’hàbitat, amb implicacions importants per a la conservació. D’altra banda, els nostres resultats estan majoritàriament en desacord amb les prediccions de la relació xarxa-àrea. Suggerim que l’empobriment de les comunitats a les taques petites resulta en una limitació en l’elecció dels hostes per part dels parasitoides generalistes, donant lloc a nivells alts d’especialització ecològica i de modularitat.S’espera que els canvis en la composició de plantes afectin a les comunitats planta-pol·linitzador de múltiples maneres depenent del context floral, de l’abundància relativa de la planta colonitzadora, del seu grau de generalització, i del seu síndrome floral. La facilitació o la competència pels pol·linitzadors poden resultar en canvis predictibles en l’estructura de la xarxa i en la pol·linització, però s’espera que els efectes sobre la reproducció de les plantes siguin altament contingents a cada espècie. S’espera que la grandària de l’hàbitat afecti a les espècies i a les interaccions seguint les relacions espècies-àrea i enllaços-área. Unint les evidències empíriques amb les prediccions teòriques, desenvolupem un marc de referència per a la relació xarxa-àrea. Les respostes de les xarxes mutualistes i antagonistes als canvis són predictibles. Tot i així, les conseqüències sobre la funció ecosistèmica segueixen sent altament contingents.
Las comunidades ecológicas son entidades dinámicas cuya estructura y composición está conformada por procesos ecológicos y por variabilidad ambiental. Debido a que las especies interaccionan entre ellas, los cambios en la estructura y composición de las comunidades pueden alterar las redes de interacción y el funcionamiento de los ecosistemas. En esta tesis estudio cómo un proceso ecológico (la colonización de un hábitat por una nueva especie) y un componente importante de la variabilidad espacial (el tamaño del hábitat) afecta a las comunidades de polinizadores, de abejas y avispas, así como a sus interacciones mutualistas y antagonistas. También investigo si estos cambios afectan a la estructura de la red de interacciones y las consecuencias potenciales sobre la función ecosistémica (polinización, reproducción de las plantas, y parasitismo).Los cambios en la composición de las comunidades de plantas son cada vez más frecuentes en el contexto actual de las invasiones biológicas. Sin embargo, las invasiones de plantas normalmente representan escenarios extremos de dominancia ecológica y sus efectos han sido estudiados relativamente bien. Por el contrario, los efectos de la colonización de los hábitats por especies autóctonas (por ejemplo, como resultado de los cambios en la distribución geográfica mediados por el cambio climático) han recibido mucha menos atención.En el Capítulo 1 analizamos los efectos de la llegada de una nueva especie de planta autóctona en un matorral, incluyendo los cambios en la composición de la comunidad de polinizadores, los patrones de interacción y la estructura de la red planta-polinizador, y las consecuencias para la polinización y para el éxito reproductivo de las especies de plantas residentes. Para ello manipulamos experimentalmente parches de matorral introduciendo una planta autóctona (planta colonizadora de ahora en adelante), ajustando el número de flores introducidas a la cantidad de recursos florales localmente disponibles. La planta colonizadora atrajo a abejorros, facilitando las interacciones a las plantas residentes. Las comunidades colonizadas exhibieron una mayor generalización de la red, un mayor anidamiento, y modificaciones en la composición de los módulos de la red. Todos estos cambios resultaron en una mayor deposición de polen heteroespecífico (pero no conespecífico) y tuvo efectos contrastados en el éxito reproductivo de las plantas residentes (neutral, positivo, o negativo dependiendo de la especie). Nuestro estudio muestra que las interacciones planta-polinizador se reorganizan rápidamente en respuesta a nuevas situaciones (incluso cuando la nueva planta no es excesivamente dominante), con consecuencias funcionales importantes en la polinización y en la reproducción de las plantas.La relación especies-área (incremento de la riqueza de especies con el área) es una de las leyes de la Ecología más antiguas y fundamentales y, recientemente, ha sido extendida a la riqueza de interacciones (relación enlaces-área) y a la estructura de la red (relación red-área). Sin embargo, a pesar que las relaciones especies- y enlaces- área han recibido un soporte amplio de estudios empíricos, la relación entre la estructura de la red y el área permanece en gran parte inexplorada. Entender esta relación es importante porque la fragmentación antropogénica de los hábitats está teniendo impactos profundos en la configuración de los hábitats y está reduciendo el tamaño de los mismos.En el Capítulo 2 testamos varias hipótesis relacionadas con las relaciones especies-área, enlaces-área y red-área utilizando comunidades de abejas y avispas nidificantes en cavidades preestablecidas así como la fauna asociada a sus nidos (huéspedes y parasitoides de ahora en adelante, respectivamente) como sistema de estudio. Para obtener nidos de abejas y avispas instalamos estaciones de nidales en parches de hábitat abierto (favorable) –rodeados por bosque denso (hábitat desfavorable)- de tres categorías de tamaño (pequeños, ~0.1 ha; medianos, ~5 ha; y grandes, hábitat abierto continuo). Los nidos obtenidos en cada parche fueron diseccionados y sus contenidos fueron analizados anotando la identidad y la abundancia de cada especie de huésped y parasitoide. La riqueza de huéspedes, parasitoides, e interacciones incrementó de los parches pequeños a los medianos, pero no de los parches medianos a los grandes. Los enlaces por especie incrementaron con el tamaño del parche, pero la tasa de parasitismo no siguió ninguna tendencia relacionada con el tamaño del hábitat. La especialización de la red disminuyó de los parches pequeños a los medianos, pero los parches grandes mostraron una especialización similar a la de los parches pequeños y medianos. La modularidad disminuyó de los parches pequeños a los medianos, pero no hubo diferencias entre los parches medianos y los grandes. El número de módulos incrementó con el tamaño del parche. Nuestros resultados están, en su mayor parte, parcialmente de acuerdo con las relaciones especies-área y enlaces-área. La ausencia de diferencias entre los parches medianos y grandes indica la existencia de un umbral por encima del cual la riqueza de especies y de interacciones ya no está limitada por el tamaño del hábitat, con implicaciones importantes para la conservación. Por otro lado, nuestros resultados están mayoritariamente en desacuerdo con las predicciones de la relación red-área. Sugerimos que el empobrecimiento de las comunidades en los parches pequeños resulta en una limitación en la elección de los huéspedes por parte de los parasitoides generalistas, dando lugar a altos niveles de especialización ecológica y de modularidad.Se espera que los cambios en la composición de plantas afecten a las comunidades planta-polinizador de múltiples maneras dependiendo del contexto floral, de la abundancia relativa de la planta colonizadora, de su grado de generalización, y de su síndrome floral. La facilitación o la competencia por los polinizadores pueden resultar en cambios predecibles en la estructura de la red y en la polinización, pero se espera que los efectos sobre la reproducción de las plantas sean altamente contingentes en cada especie. Se espera que el tamaño del hábitat afecte a las especies y a las interacciones siguiendo las relaciones especies-área y enlaces-área. Uniendo las evidencias empíricas con las predicciones teóricas, desarrollamos un marco de referencia para la relación red-área. Las respuestas de las redes mutualistas y antagonistas a los cambios son predecibles. Sin embargo, las consecuencias sobre la función ecosistémica siguen siendo altamente contingentes.
Ecological communities are dynamic entities whose structure and composition are shaped by ecological processes and environmental variability. Because species interact with one another, changes in community structure and composition are likely to alter interaction networks and ecosystem functioning. In this thesis I study how an ecological process (the colonization of a habitat by a new species) and an important component of spatial variability (habitat area) affect pollinator and bee-wasp communities and their mutualistic and antagonistic interactions. I also investigate whether these changes affect interaction network structure and the potential consequences on ecosystem function (pollination, plant reproduction, and parasitism). Changes in plant community composition are becoming increasingly frequent in the current context of pervasive biological invasions. However, plant invasions usually represent extreme scenarios of ecological dominance and their effects have been relatively well studied. By contrast, the effects of habitat colonization by native species (for example, as a result of changes in geographical distribution mediated by climate change) have received much less attention.In Chapter 1 we analyze the effects of the arrival of a new native plant into a scrubland community, including changes in pollinator community composition, plant–pollinator interaction patterns and network structure, and the consequences for pollination and reproductive success of resident plant species. To do so, we experimentally manipulated patches of scrubland by introducing a native plant (henceforth colonizing plant), adjusting the number of flowers introduced to the amount of floral resources locally available. The colonizing plant attracted bumblebees, facilitating interactions to resident plants. Colonized communities exhibited higher network generalization, nestedness, and modifications in the composition of network modules. All these changes resulted in higher heterospecific (but not conspecific) pollen deposition and had contrasting effects on the reproductive success of resident plants (neutral, positive, or negative depending on the species). Our study shows that plant-pollinator interactions are rapidly rearranged in response to novel situations (even when the new plant is not overly dominant), with important functional consequences on pollination and plant reproduction.The species-area relationship (increasing species richness with area) is one of the oldest and most fundamental laws in Ecology and, in recent years, has extended to interaction richness (link-area relationship) and network structure (network-area relationship). However, although the species- and link-area relationships have received ample support from empirical studies, the relationship between network structure and area remains largely unexplored. Understanding this relationship is important because anthropogenic habitat fragmentation is having a profound impact on habitat configuration and effectively reducing the size of natural habitats.In Chapter 2 we test several hypothesis related to the species-area, link-area and network-area relationships using cavity-nesting bee-wasp communities and their nest associates (henceforth hosts and parasitoids, respectively), as a study system. To obtain bee-wasp nests we set up nest-trapping stations in patches of (favorable) open habitat -surrounded by dense forest (unfavorable habitat)- of three size categories (small, ~0.1 ha; medium, ~5 ha; and large, continuous open habitat). The nests obtained in each patch were dissected and their contents analyzed by noting the identity and abundance of each host and parasitoid species. Host, parasitoid, and interaction richness increase from small to medium patches, but not from medium to large patches. Links per species increased with patch size, but parasitism rate did not follow any patch-size-mediated trend. Network specialization decreased from small to medium patches, but large patches showed similar specialization to that of small and medium patches. Modularity decreased from small to medium patches but there were no differences between medium and large patches. The number of modules increased with patch size. Our results are mostly in partial agreement with the species-area and the link-area relationships. The lack of differences between medium and large patches indicates the existence of a threshold above which local species/link richness are no longer limited by habitat size, with important implications for conservation. On the other hand, our results are mostly in disagreement with the predictions of the network-area relationship. We suggest that community impoverishment in small patches results in limited host choice by generalist parasitoids, leading to high levels of ecological specialization and modularity. Changes in plant composition are expected to affect plant-pollinator communities in multiple ways depending on the flower context, the relative abundance of the colonizing plant, its degree of generalization, and its pollination syndrome. Facilitation or competition for pollinators may result in predictable responses in network structure and pollination, but the effects on plant reproduction are expected to be highly contingent on each species. Habitat size is expected to affect species and interactions following the species-area and the link-area relationships. Merging empirical evidence with theoretical predictions, we developed a framework for the network-area relationship. Mutualistic and antagonistic network responses to changes are predictable. However, consequences on ecosystem function remain highly contingent.

This thesis examines a broad spectrum of physiological and silvicultural features of the highly valued woody angiosperm hemi-parasite Santalurn album L. (Indian sandalwood) in relation to its culture in plantations in northern Western Australia. Topics covered include allometry of host and Santalum when grown as single plant pairings in both field and pot culture, nutritional interactions between Santalum and beneficial and non-beneficial hosts, deleterious influences of parasitism on plantation productivity and heartwood induction in young trees. In Western Australia sandalwood is grown in the nursery for 8 months before establishment in the field and during this time a pot host is introduced. Survival of Santalurn after field establishment and its subsequent growth were significantly affected by the time of introduction of the pot host, Alternanthera nana. Increasing the period of the Santalum : Alternanthera association in the nursery to 109 days prior to field establishment markedly increased early growth of Salztalum plantations. Introduction at 134 days prior to field establishment was detrimental to the parasite as the Alternanthera was too vigorous for the small Santalum seedlings. Santalurn plants had a lower root : shoot ratio lower when cultured with Alternanthera in the nursery prior to field establishment compared with seedlings grown without Alternanthera. Alterrzantlzera survival in the field was high when it had been grown with Santalum for 12 weeks or more in the nursery prior to field establishment. After 1 1 weeks in the field a strong negative linear relationship was shown between Santalunz root : shoot ratio and Alternarzthera dry weight, and a positive linear relationship between Salztalum DW and Alternanthera DW. In Western Australia Santalu~n is established in the field with an intermediate host which nourishes the parasite for 3-5 years before Santalum becomes dependent on its long-term host and the intermediate host dies. The relationship between Santalum and several species tested as intermediate hosts was examined by pairing Santalum seedlings with intermediate host seedlings in 25 litre pots over a 10 month period. Growth of Santalum in pot culture with three N2-fixing woody intermediate hosts (Sesbania forrnosa, Acacia traclzycarpa and A. ampliceps), the woody non N2-fixing Eucalyptus camaldulensis or without a host varied considerably between host treatments. Santalum growth was greater and root : shoot ratio lower for seedlings grown with N2-fixing hosts compared with seedlings grown with E. carnaldulensis or with no host. The root : shoot ratio of unattached Santalum increased exponentially over time, whereas for all other treatments it remained relatively constant. An assessment of the value of the hosts, termed host use efficiency, was computed as Santalum shoot DW / host shoot DW. The host use efficiency of A. trachycalpa was greater than that of the other hosts. The xylem sap of hosts and Sarztalum, and ethanolic extracts of endophytic tissue of haustoria of Santalzkm were analysed for amino acids, organic acids and sugars to determine which solutes were available in the host and which were extracted by the Santalum haustoria from different hosts. There were similarities between Santalum and legume hosts in concentration and composition of xylem sap amino acids, and in the amino acid spectra of the corresponding Santalum endophytic tissue, whereas there were low N levels in xylem sap of E. camaldulensis and dissimilarities between its amino acid composition and that of Santalum. This indicated substantial direct intake of xylem N by Santalum from legume hosts but little N from the xylem sap of E. canzaldulensis. There were high concentrations of asparagine, glutamate, aspartate and y-amino glutamate in the xylem sap of the legume hosts, while in the non-legume the most common amino acids were glutamate, aspartate, glutamine and arginine. Proline, the predominant amino acid in the xylem sap of Santalum acurninatum growing in natural vegetation (Tennakoon et al. 1997) was not detected or present in very low concentrations in Santalurn album under these conditions. in the non-legume. Xylem sap of hosts contained variable amounts of sugars (sucrose, glucose and fructose) and organic acids (fumaric, citric and malic acid), whereas that of the parasitic Santalum was dominated by fructose and malic acid. Dissimilarities in the proportional amounts of xylem-borne sugars and organic acids were particularly evident for the E. camaldulensis : Santalum partnership. Diurnal profiles of photosynthesis and transpiration of Santalum were closely similar to those for corresponding hosts, whereas the midday leaf water potential of Santalum was consistently more negative than that of corresponding hosts. Net photosynthesis and water use efficiency was lower, but transpiration rates were similar to that of corresponding hosts. Nitrogen concentrations of foliage of Santalum were higher than their hosts, and higher when on legume hosts than on E. camaldulensis, or without a host. Nitrogen concentrations of Santalum foliage was strongly correlated with net photosynthesis and water use efficiency of Santalum. 813C values of shoot dry matter of Santalum were poorly correlated with instantaneous water use efficiency of Santalum. Tissue water relations of Santalum were similar to that of water-stress tolerant species. S. formosa proved the best host followed by Acacia ampliceps and A. traclzycarpa based on dry matter gains of Santalum. Estimates of heterotrophic gain of C of Santalum when grown in association with the legume hosts over a nine week period indicate 57.9% of C was derived from A. ampliceps, 45.5% from A. trachycarpa and 34.6% fiom S. fomosa. Abundance of haustorial attachments on roots of hosts was poorly correlated to Santalum shoot DW. Root nodules of legume hosts were parasitised by a small proportion of Santalum haustoria. Sodium and phosphorus concentrations of foliage of Santalum were generally higher than that of corresponding hosts. Net gains of calcium, potassium, phosphorus and sodium in Santalum was greatest when grown in association with hosts richest in the corresponding element. Net losses or only small gains of calcium, potassium, phosphorus and sodium were recorded when Santalum was grown with E. camaldulensis or without a host suggesting that Santalum has limited ability for uptake of those minerals through its own root system. To understand the effect of hosts on the productivity of a Santalum plantation a young plantation of Santalum with three host species Cathormion umbellatum, Sesbania formosa and Acacia anuera was selected to study the relationship between host quality and distance of hosts from Santalunz on Santalum health. The selected plantation showed marked decline in health and vigour of both Santalum and hosts between years 3 and 5. Parameters of the host plants were assessed to select the best predictor of Santalunz crown health. The height and diameter growth increment of Santalum between years 3 and 5 was strongly correlated to Santalum crown health. Santaluin crown health and growth increased as host quality increased, and the distance of host fiom Santalum decreased. An index, which combined host quality and the distance of the host from that of Santalum, was a better predictor of Santalum crown health than host distance or quality alone. The age at which heartwood is initiated in Santalum album under plantation conditions in Western Australia in unknown, but in natural stands in India it occurs between 10-13 years of age (Rai 1990). A field experiment was conducted to determine the efficacy of stem injections of paraquat andlor ethrel in initiating heartwood formation in five year old Santalum trees in a plantation. Trees injected with paraquat alone had a significantly greater extension of induced heartwood, both radially and vertically, than those trees injected with ethrel alone or distilled water. Eight months after treatment with paraquat or ethrel or a combination of these chemicals induced heartwood was formed, which had high lipid, and low starch and polysaccharide concentrations compared to the sapwood. Induced heartwood from both chemical treatments and their combinations contained total volatile oil and santalol oil (alpha and beta santalol) concentrations that were equal to or greater than that of naturally formed heartwood and greater than that of sapwood. Moisture content, and concentrations of K and Mg, and in some treatments Ca of induced heartwood were significantly lower than that of sapwood. The thesis concludes with a synthesis of the findings and suggestions for future research, with special reference to mid-rotation aspects of Santaltrm plantation silviculture.

This thesis examines a broad spectrum of physiological and silvicultural features of the highly valued woody angiosperm hemi-parasite Santalurn album L. (Indian sandalwood) in relation to its culture in plantations in northern Western Australia. Topics covered include allometry of host and Santalum when grown as single plant pairings in both field and pot culture, nutritional interactions between Santalum and beneficial and non-beneficial hosts, deleterious influences of parasitism on plantation productivity and heartwood induction in young trees. In Western Australia sandalwood is grown in the nursery for 8 months before establishment in the field and during this time a pot host is introduced. Survival of Santalurn after field establishment and its subsequent growth were significantly affected by the time of introduction of the pot host, Alternanthera nana. Increasing the period of the Santalum : Alternanthera association in the nursery to 109 days prior to field establishment markedly increased early growth of Salztalum plantations. Introduction at 134 days prior to field establishment was detrimental to the parasite as the Alternanthera was too vigorous for the small Santalum seedlings. Santalurn plants had a lower root : shoot ratio lower when cultured with Alternanthera in the nursery prior to field establishment compared with seedlings grown without Alternanthera. Alterrzantlzera survival in the field was high when it had been grown with Santalum for 12 weeks or more in the nursery prior to field establishment. After 1 1 weeks in the field a strong negative linear relationship was shown between Santalunz root : shoot ratio and Alternarzthera dry weight, and a positive linear relationship between Salztalum DW and Alternanthera DW. In Western Australia Santalu~n is established in the field with an intermediate host which nourishes the parasite for 3-5 years before Santalum becomes dependent on its long-term host and the intermediate host dies. The relationship between Santalum and several species tested as intermediate hosts was examined by pairing Santalum seedlings with intermediate host seedlings in 25 litre pots over a 10 month period. Growth of Santalum in pot culture with three N2-fixing woody intermediate hosts (Sesbania forrnosa, Acacia traclzycarpa and A. ampliceps), the woody non N2-fixing Eucalyptus camaldulensis or without a host varied considerably between host treatments. Santalum growth was greater and root : shoot ratio lower for seedlings grown with N2-fixing hosts compared with seedlings grown with E. carnaldulensis or with no host. The root : shoot ratio of unattached Santalum increased exponentially over time, whereas for all other treatments it remained relatively constant. An assessment of the value of the hosts, termed host use efficiency, was computed as Santalum shoot DW / host shoot DW. The host use efficiency of A. trachycalpa was greater than that of the other hosts. The xylem sap of hosts and Sarztalum, and ethanolic extracts of endophytic tissue of haustoria of Santalzkm were analysed for amino acids, organic acids and sugars to determine which solutes were available in the host and which were extracted by the Santalum haustoria from different hosts. There were similarities between Santalum and legume hosts in concentration and composition of xylem sap amino acids, and in the amino acid spectra of the corresponding Santalum endophytic tissue, whereas there were low N levels in xylem sap of E. camaldulensis and dissimilarities between its amino acid composition and that of Santalum. This indicated substantial direct intake of xylem N by Santalum from legume hosts but little N from the xylem sap of E. canzaldulensis. There were high concentrations of asparagine, glutamate, aspartate and y-amino glutamate in the xylem sap of the legume hosts, while in the non-legume the most common amino acids were glutamate, aspartate, glutamine and arginine. Proline, the predominant amino acid in the xylem sap of Santalum acurninatum growing in natural vegetation (Tennakoon et al. 1997) was not detected or present in very low concentrations in Santalurn album under these conditions. in the non-legume. Xylem sap of hosts contained variable amounts of sugars (sucrose, glucose and fructose) and organic acids (fumaric, citric and malic acid), whereas that of the parasitic Santalum was dominated by fructose and malic acid. Dissimilarities in the proportional amounts of xylem-borne sugars and organic acids were particularly evident for the E. camaldulensis : Santalum partnership. Diurnal profiles of photosynthesis and transpiration of Santalum were closely similar to those for corresponding hosts, whereas the midday leaf water potential of Santalum was consistently more negative than that of corresponding hosts. Net photosynthesis and water use efficiency was lower, but transpiration rates were similar to that of corresponding hosts. Nitrogen concentrations of foliage of Santalum were higher than their hosts, and higher when on legume hosts than on E. camaldulensis, or without a host. Nitrogen concentrations of Santalum foliage was strongly correlated with net photosynthesis and water use efficiency of Santalum. 813C values of shoot dry matter of Santalum were poorly correlated with instantaneous water use efficiency of Santalum. Tissue water relations of Santalum were similar to that of water-stress tolerant species. S. formosa proved the best host followed by Acacia ampliceps and A. traclzycarpa based on dry matter gains of Santalum. Estimates of heterotrophic gain of C of Santalum when grown in association with the legume hosts over a nine week period indicate 57.9% of C was derived from A. ampliceps, 45.5% from A. trachycarpa and 34.6% fiom S. fomosa. Abundance of haustorial attachments on roots of hosts was poorly correlated to Santalum shoot DW. Root nodules of legume hosts were parasitised by a small proportion of Santalum haustoria. Sodium and phosphorus concentrations of foliage of Santalum were generally higher than that of corresponding hosts. Net gains of calcium, potassium, phosphorus and sodium in Santalum was greatest when grown in association with hosts richest in the corresponding element. Net losses or only small gains of calcium, potassium, phosphorus and sodium were recorded when Santalum was grown with E. camaldulensis or without a host suggesting that Santalum has limited ability for uptake of those minerals through its own root system. To understand the effect of hosts on the productivity of a Santalum plantation a young plantation of Santalum with three host species Cathormion umbellatum, Sesbania formosa and Acacia anuera was selected to study the relationship between host quality and distance of hosts from Santalunz on Santalum health. The selected plantation showed marked decline in health and vigour of both Santalum and hosts between years 3 and 5. Parameters of the host plants were assessed to select the best predictor of Santalunz crown health. The height and diameter growth increment of Santalum between years 3 and 5 was strongly correlated to Santalum crown health. Santaluin crown health and growth increased as host quality increased, and the distance of host fiom Santalum decreased. An index, which combined host quality and the distance of the host from that of Santalum, was a better predictor of Santalum crown health than host distance or quality alone. The age at which heartwood is initiated in Santalum album under plantation conditions in Western Australia in unknown, but in natural stands in India it occurs between 10-13 years of age (Rai 1990). A field experiment was conducted to determine the efficacy of stem injections of paraquat andlor ethrel in initiating heartwood formation in five year old Santalum trees in a plantation. Trees injected with paraquat alone had a significantly greater extension of induced heartwood, both radially and vertically, than those trees injected with ethrel alone or distilled water. Eight months after treatment with paraquat or ethrel or a combination of these chemicals induced heartwood was formed, which had high lipid, and low starch and polysaccharide concentrations compared to the sapwood. Induced heartwood from both chemical treatments and their combinations contained total volatile oil and santalol oil (alpha and beta santalol) concentrations that were equal to or greater than that of naturally formed heartwood and greater than that of sapwood. Moisture content, and concentrations of K and Mg, and in some treatments Ca of induced heartwood were significantly lower than that of sapwood. The thesis concludes with a synthesis of the findings and suggestions for future research, with special reference to mid-rotation aspects of Santaltrm plantation silviculture.

Les plantes, en tant que producteurs primaires, peuvent être consommées/attaquées par de nombreux organismes, notamment des virus, des pathogènes, des insectes herbivores ou encore des nématodes phytophages. D'une part, les plantes sont une ressource alimentaire principale pour ces organismes ; d'autre part, les plantes peuvent se défendre contre ces consommateurs en mobilisant des défenses constitutives et/ou induites. En cas de défenses induites suite à l’attaque d’un ravageur, des interactions indirectes médiées par la plante peuvent apparaitre entre ce ravageur et d’autres ravageurs partageant le même hôte i.e. la présence d’un ravageur va provoquer des modifications au niveau de la plante qui peuvent à leur tour impacter les autres ravageurs présents sur la même plante. Tout d’abord, les effets sublétaux de la bêta-cyperméthrine, un pesticide de type pyréthrinoïdes, ont été évalués sur les interactions interspécifiques entre le puceron spécialiste Aphis glycines et le puceron généraliste Aulacorthum solani sur soja. La concentration sublétale (LC5) de bêta-cyperméthrine accélère la compétition interspécifique entre les deux espèces de pucerons. La sensibilité des plants de soja aux pucerons induite par une pré-infestation d’une autre espèce de puceron pourrait ainsi être considérablement réduite par une exposition préalable à la bêta-cyperméthrine sublétaux. Dans un deuxième temps, nous nous sommes intéressés aux facteurs biotiques susceptibles de moduler les interactions indirectes. Dans ce cadre, divers ravageurs, à savoir le puceron, la chenille, le phytopathogène et le nématode, ont été introduits et suivis sur des plants de tomate, Solanum lycopersicum. La performance des ravageurs arthropodes, (e.g. le nombre d'individus de pucerons et le taux de développement des larves de lépidoptères) était corrélée au nombre d’interactions indirectes (i.e. au nombre d’espèces de ravageurs partageant le même hôte), que les ravageurs attaquent les plants de tomate séquentiellement ou simultanément. En effet, lorsque les ravageurs étaient mis en présence d’un plant de tomate de manière simultanée, la performance des insectes était positivement corrélée au nombre d’interactions indirectes. A l’inverse, lorsque les ravageurs étaient mis en présence de plants de tomate de façon séquentielle, la performance des insectes ravageurs était négativement corrélée à l'abondance des interactions indirectes. Enfin, aucune corrélation n'a pas été détectée pour les pathogènes, champignons et nématodes. D’autre part, nous avons pu mettre en évidence que la présence de nématodes et d’oïdium réduisait à court-terme i.e. à l’échelle d’une génération l’abondance de pucerons sur un plant de tomate attaqués par ces trois ravageurs. Nous avons voulu tester cette interaction indirecte sur du long-terme i.e. à l’échelle d’un suivi de la dynamique de populations des pucerons. De plus, nous avons ajouté à cette expérimentation des parasitoïdes du puceron afin de voir l’impact de ces interactions indirectes sur le troisième niveau trophique. La population de pucerons impliquée dans les interactions indirectes testées a été réduite à court et à long terme parce-que la survie des nymphes diminuait. De plus, le nombre de pucerons ailés a augmenté. Enfin, le taux de parasitisme de l’ennemi naturel Aphidius ervi. Ainsi, ce travail démontre la présence d’interactions indirectes fortes entre les différents ravageurs partageant une même plante hôte et ces interactions sont susceptibles d’être modulées par de nombreux facteurs comme le nombre ou encore la diversité des ravageurs. De plus, elles sont susceptibles de modifier le service de biocontrôle rendu par les auxiliaires des cultures ainsi que le rendement de la culture. Il est donc nécessaire de comprendre et prendre en compte ces interactions indirectes dans le cadre d’attaques multiples pour adopter les stratégies IPM les plus adéquates
In the agroecosystem, plant crops are the primary trophic producer that can be consumed/parasitized by a plethora of organisms, including viruses, bacteria, fungi, nematodes and many herbivore insects. On one hand, plants support these organisms as food resources; on the other hand, plants defend themselves against those consumers by employing constitutive and induced defenses. Both features might be involved in plant-mediated indirect interactions when multiple pests share the same host plant. It means that the presence of one pest might have an indirect effect on another plant through the modification of some plant host features (nutritional quality or defense). During my Ph.D. study, I assessed the modulating factors (both abiotic and biotic factors) that might affect the plant-mediated indirect interactions, and also investigated the influences of such indirect interactions at long term and on a tri-trophic system, and finally analyzed the underlying phytochemical mechanisms by mechabolomics analyses. Firstly, sublethal effects of beta-cypermethrin, a pyrethroid pesticide as the abiotic factor, were assessed on the interspecific interactions between the specialist aphid Aphis glycines, and the generalist aphid Aulacorthum solani on soybeans. Sublethal concentration (LC5) of beta-cypermethrin accelerated the interspecific competition between the two species of aphids. Induced susceptibility of soybean plants to one species of aphids caused by the other species would be significantly impaired by the exposure to sublethal beta-cypermethrin.Secondly, various pest species abundance and feeding strategies, i.e. the piercing-sucking aphid, the chewing caterpillar, the plant fungus and the root-knot nematode, were introduced on tomato plants, Solanum lycopersicum to evaluate the plant-mediated indirect interactions among these pests. In the results, insect performances, i.e. the number of aphid individuals and development rate of caterpillar larva, were correlated to the abundance of indirect interactions whether pests attacked tomato plants sequentially or simultaneously. When pests attacked tomato plants at the same time, insect performances were positively correlated to the abundance of indirect interactions. Nevertheless, when pests infested tomato plants in sequence, the insect performances were negatively correlated to the abundance of indirect interactions. However, such a correlation was not detected with the pathogenic organisms (fungi and nematodes). Thirdly, the population dynamic of aphid M. euphorbiae was also monitored under a tri-trophic system. We found that the aphid population, involved in the above-belowground interactions, slowed down in both the short term and long term via reduced survival of nymph aphids and facilitating induction of winged aphids. Furthermore, parasitism efficiency of the natural enemy Aphidius ervi to aphids and tomato yields were also reduced by such above-belowground interactions. Lastly, following an untargeted metabolomic approach, we detected many significant chemical variations between non-infested and infested plants, disregarding the pest infestation. Most of them are probably induced by the presence of nematodes, the only belowground pest. On the opposite, we did not observe many chemical differences in roots metabolome when we compared the different types of infestation, meaning that the presence of nematodes probably lead the metabolomic signal in infested roots, disregarding the presence of other pests aboveground. All the studies may contribute to a comprehensive understanding of the diverse interactions among plants, pesticides, various pests, and natural enemies, and optimizing the integrated pest management in the agroecosystem

Plant feeding insects comprise about 25% of all animal species on earth and play an important role in all ecosystems. Although we understand that their association with plants is a key-factor driving the diversification in this group, we still have large gaps in our knowledge of the underlying processes of this relationship. Female choice of host plant is an important event in the insect life-cycle, as it is a major determinant of the larval food plant. In this Thesis I studied different aspects of insect host plant choice and used butterflies from the family Nymphalidae as my study system. I found that butterflies have a well developed olfactory system and that they use odors when searching for food or host plants. However, the information obtained from the odor of host plants does not seem to be sufficient for the studied species to make a distinction between plants of different qualities. Interestingly, even when in full contact with the leaf they do not make optimal decisions. I show for example that a sub-optimal female choice may be mitigated by larval ability to cope with unfavorable situations. Moreover, species that utilize a broader set of host plants may not be very well adapted to all the hosts they use, but at the same time they may survive in areas where there is only a subset of the plants available. Lastly, differences in the evolution of life-history traits between species can account for differences in how each species realizes its lifestyle. Thus, by incorporating findings on mechanisms of host plant choice with the ecological and evolutionary context of a species, our ability to explain the dynamics of host plant choice and insect-plant interactions can be improved.

At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 4: Manuscript.

In many developing countries, farmers and pastoralists still rely on their indigenous knowledge, practices and locally available plants to control nematode parasitic infections, both in livestock and humans. The overall aim of my thesis was to undertake bioassay-guided phyto-chemical study of extracts and their constituents from Ethiopian anti-parasitic plants used by healers to control gastrointestinal nematode parasites in livestock to validate their ethno-medicinal use and to characterise and identify their active ingredients. As a first experiment (Chapter Three), four types of crude extracts (water, 70% methyl-alcohol, absolute methanol and acetone) of four indigenous Ethiopian medicinal plants (Adenia species, Cissus ruspolii, Ipomoea eriocarpa and Euphorbia thymifolia) were screened against Teladorsagia circumcincta egg hatching in vitro, not only as a first step to validate the traditional healers claim but also to choose the most promising plant extract(s) for further phyto-chemical studies. The egg hatching inhibition (EHI) test results revealed that the anti-parasitic properties of these plants depended on plant species, dose, and solvent polarity. The water extracts of both C. ruspolii and Adenia sp. exhibited largest, up to 100% EHI but also larval migration inhibition activities, and were selected for further studies. The second experiment (Chapter Four) assessed the nature of active constituents in these extracts by physico-chemical methods. It was observed that the major constituents of both plant extracts responsible for the EHI activities are likely highly polar, water-soluble, small and moderately heat-labile molecules. The third and fourth experiments (Chapters Five and Six) consisted of separating Cissus ruspolii and Adenia sp. water extracts into discrete fractions by gel-permeation chromatography, EHI tests of Bio-Gel P-2 fractions followed by thin layer chromatography (TLC) profiling of these fractions to detect separated spots (in day light, under UV-light or after staining with various staining reagents) and also to see how elution patterns of separated spots affected by column parameters. The EHI tests on the fractions obtained revealed that the active constituents of C. ruspolii and Adenia sp. water crude extracts were eluted into few fractions based on their molecular sizes. The TLC profilings of these fractions identified spot patterns of active and inactive fractions, which allowed pooling of active constituents based on their EHI and TLC profiling into three pools for each plant. The fifth experiment (Chapter Seven) was to isolate and purify compounds from these pools using various preparative planar and column chromatographic methods. Sequential applications of column chromatography followed by preparative thin layer chromatography isolated and purified five active compounds from C. ruspolii and two active compounds from Adenia sp. The sixth experiment (Chapter Eight) was to characterize and propose/elucidate structures of compounds from the active fractions using chromatographic, analytical and spectroscopic methods. In this regard, the structures of two oleanane type triterpenoid saponins isolated from one of active fractions of Adenia sp. were proposed based on their mass spectrometry (MS) and nuclear magnetic resonance (NMR) data with support of compounds property, TLC and literature. Similar outcomes for C. ruspolii were not achieved due to lack of sufficient sample to run 13C-nuclear magnetic resonance spectroscopy and distortionless enhancement by polarization transfer (DEPT), contamination of some purified compounds with ill-characterised substance from the preparative TLC matrix and in some cases mass spectrometry (MS) and nuclear magnetic resonance (NMR) data did not support each other. The last experiment (Chapter Nine) was to assess anthelmintic efficacy and safety of C. ruspolii and Adenia sp. crude water extracts in Heligmosomoides bakeri infected mice. This in vivo test revealed that both plant extracts exhibited significant reduction in worm burdens and worm egg excretion, with moderate effects on haematology and organ weights at tolerated dosages. In conclusion, both in vitro and in vivo data revealed that Adenia sp. and C. ruspolii have anthelmintic properties, thus validating traditional healer claims and supporting ethno-medicinal use. The bioassay-guided phytochemical study resulted in the isolation of a number of active compounds from these plants, for some of which a structure has been proposed.

Parasitic weeds of the family Orobanchaceae attach to the roots of host plants via haustoria capable of drawing nutrients from host vascular tissue. Species in this family span the spectrum of host nutrient dependency, allowing comparisons that provide insight into parasite adaptation. A key aspect of this is the relationship between parasite metabolism and the metabolite profile of its host. To what extent does the metabolite profile of the parasite depend on that of the host? Do parasites that differ in host-dependency also differ in their metabolism or do they use common metabolic strategies? These questions were addressed using comparative profiling of primary metabolites to gain insight into carbon and nitrogen assimilation by the obligate holoparasite Phelipanche aegyptiaca and the facultative hemiparasite Triphysaria versicolor. First, metabolite profiles of these parasites and their hosts were compared during the key life stages before and after haustorial attachment. Second, the impact of specific variations in host metabolism was analyzed for P. aegyptiaca growing on Arabidopsis thaliana hosts that had mutations in amino acid metabolism but otherwise identical genetic backgrounds. Comparison of P. aegyptiaca and T. versicolor metabolite profiles identified substantial differences in the stages spanning the transition from pre-haustorial development through post-haustorial feeding. Each parasite species is distinct from the other and from their hosts. For parasites growing on host lines that differ in amino acid content, the size of P. aegyptiaca tubercles decreased when grown on the aap6 mutant line, which has decreased levels of asparagine in the phloem sap compared to the wild type. However, altered amino acid levels in other lines did not impact P. aegyptiaca growth, indicating that this parasite has ability to compensate for variation in host metabolic composition. This research highlights the importance of aspartate and asparagine to early post-attachment metabolism in both P. aegyptiaca and T. versicolor and through host deficiencies possibly associated with decreased growth in P. aegyptiaca. Overall, this work provides insights both into the metabolism of parasitic plants and lays the foundation for the development of new metabolism-based control strategies.
Ph. D.
Parasitic weeds of the plant family Orobanchaceae attach to the roots of host plants via haustoria. Parasite haustoria embed into the host plant and are capable of drawing nutrients from host vascular tissue. Species in this family span the spectrum of the extent to which a parasitic plant may depend on its host for nutrients. This allows comparisons that provide insight into the ways in which parasites adapt. A key aspect of this is the relationship between the metabolite profile of the parasite and the metabolite profile of the host. To what extent does the metabolite profile of the parasite depend on that of the host? Do parasites that differ in host-dependency also differ in their metabolism or do they use common metabolic strategies? These questions were addressed using comparative profiling of primary metabolites to gain insight into carbon and nitrogen assimilation by the obligate parasite Phelipanche aegyptiaca (which cannot perform photosynthesis) and the facultative parasite Triphysaria versicolor (which can perform photosynthesis). First, metabolite profiles of these parasites and their hosts were compared during the key life stages before and after haustorial attachment. Second, the impact of specific variations in host metabolism was analyzed for P. aegyptiaca growing on Arabidopsis thaliana hosts. These hosts had mutations in enzymes related to amino acid metabolism but otherwise identical genetic backgrounds. Comparison of P. aegyptiaca and T. versicolor metabolite profiles identified substantial differences in the stages spanning the transition from pre-haustorial development through post-haustorial feeding. Each parasite species is distinct from the other and from their hosts. For parasites growing on host lines that differ in amino acid content, the size of P. aegyptiaca tubercles decreased when grown on the aap6 mutant line, which has decreased levels of asparagine in the phloem sap compared to the wild type. However, altered amino acid levels in other lines did not impact P. aegyptiaca growth, indicating that this parasite has ability to compensate for variation in host metabolic composition. Overall, this work provides insights both into the metabolism of parasitic plants and lays the foundation for the development of new metabolism-based control strategies.

Interactions between host plants and aphids under water stress conditions were examined. Two controlled environment room studies, a Veld experiment and phenological modelling analysis were used to investigate three different plant-aphid relationships. The aim of these studies was to determine the results of water shortage on plant and pest populations, which are likely to result under climate change scenarios. The central hypothesis was that mild drought stress would lead to greater aphid populations through a beneficial increase of small nitrogenous molecules in the phloem sap providing improved diet quality — the Plant Stress Hypothesis (White, 1969). Myzus persicae (the peach–potato aphid) is a broad-range feeder with a preference for senescent foliage. Contrary to expectations, on young cabbage plants, Brassica oleracea var capitata, it was found to increase faster and to a greater extent on well-watered hosts. Aphids were also found to prevent osmoregulation in droughted plants. Despite this, the aphid infestation led to a drought-like response in all plants irrespective of watering regime, causing significantly reduced growth. Metopolophium dirhodum (the rose–grain aphid) overwinters as eggs on Rosa spp. but most of its life-cycle is spent on grasses and cereal crops. As with M. persicae, aphid infestation caused drought-like symptoms even in well-watered plants. The mechanism of reduction in plant health is most likely through aphid damage to the efficiency of Photosystem a within 7 days of infestation. On young barley plants, Hordeum vulgare, the rose–grain aphid was found to increase faster and to greater extent on droughted hosts. In contrast to M. persicae, the study strongly supported the Plant Stress Hypothesis, Finding that even mild water stress of approximately −0.3 MPa could lead to conditions favourable to rapid aphid population increase. The difference in the response of different species of aphids may be associated more with the levels of soluble nitrogen in their diet than specifically host plant water status. Drepanosiphum platanoidis is an aphid specific to sycamore, Acer pseudoplatanus and spends its entire life-cycle on that species. In the Veld, the total number of aphids supported by a single tree was found to correlate most strongly with soluble leaf nitrogen. In addition, the number of aphids on individual leaves was found to correlate strongly and negatively with stomatal conductance and leaf water potential. Finally, analysis of historical aphid suction-trap data was carried out. UK Rothamsted suction-traps are 12.2 m tall and sample air at 0.75 m3 s-1, with daily aphid sampling between April and November. The traps are designed to catch the largest proportion of aphids in the total insect sample and to reduce the effect of surrounding land-use on the sample. The area represented by a trap may be as much as 700 km2 (Harrington and Woiwod, 2007). The dataset is the longest and most geographically extensive for aphid populations, comprising 46 sites across Europe as of 2011. In this analysis, aphid and meteorological data from the Edinburgh East Craigs site were used. This also enabled comparison with D. platanoidis numbers collected in the Veld, which showed a strongly significant positive relationship between the two. Since the data collected in suction-traps are correlated with numbers in the Veld, aphid population analysis can be improved with use of these longer datasets, without reliance on shorter-term Veld surveys. The various population characteristics examined were found to be strongly correlated with each other, but were unconnected to data from previous years. The date of first catch was found to be significantly advancing over time, with strong influences from daily minimum temperatures and vapour pressure deficits of the air within the year. Higher temperatures with lower humidity tend to bring forward the date of first catch. Lower tends to depress the population in the summer. The overall hypothesis that mild drought would benefit aphid populations cannot be accepted, but in two cases of the experiment sections of the work there were strong relationships between aphid numbers and both leaf temperature and soil moisture. The meteorological data analysis indicates that climatic changes towards a hotter, drier climate could substantially alter population dynamics. Harrington, R. and Woiwod, I. (2007). Foresight from hindsight: The Rothamsted Insect Survey. Outlooks on Pest Management, 18, 9–14. White, T. (1969). An index to measure weather-induced stress of trees associated with outbreaks of psyllids in Australia. Ecology, 50, 905–909.

3 pp.
Originally published: 2003
Dwarf mistletoes are parasitic flowering plants that grow within host plants for about two years before producing characteristic yellow to orange or green to brown leafless aerial shoots on the outside of infected host tissue. They occur only on conifers in the pine family in Arizona and are usually host specific. This article gives information about the disease cycle, the symptoms and prevention and control methods for dwarf mistletoes.

Previous studies have shown that: (1) plant defensive compounds may have medicinal properties; and (2) defensive compounds present in aposematic insects are often sequestered from their host-plant(s). This study addresses whether aposematic insects can be used as guides to detect plants containing medicinally-active compounds. First, ten tropical medicinally-active plants and ten non-active plants, selected using previous ICBG bioassay results, were observed regularly to determine their insect populations. Aposematic insects were found more frequently on active than non-active plants ( X2=8.167, P=0.01). Second, three aposematic insects feeding on Tithonia diversifolia were examined chemically to determine the fate of the plant's pharmaceutically-active compounds. They were not found to sequester or excrete these compounds. Therefore, using aposematic insects could increase the likelihood of finding plants with medicinally-active compounds; however, these insects may not necessarily utilize these compounds for defensive purposes. The underlying basis for this significant association between aposematic insects and medicinally-active plants requires further investigation.