Host Plant Tissues Research Articles

Piriformospora indica root colonization triggers local and systemic root responses and inhibits secondary colonization of distal roots.

Piriformospora indica is a basidiomycete fungus colonizing roots of a wide range of higher plants, including crop plants and the model plant Arabidopsis thaliana. Previous studies have shown that P . indica improves growth, and enhances systemic pathogen resistance in leaves of host plants. To investigate systemic effects within the root system, we established a hydroponic split-root cultivation system for Arabidopsis. Using quantitative real-time PCR, we show that initial P . indica colonization triggers a local, transient response of several defense-related transcripts, of which some were also induced in shoots and in distal, non-colonized roots of the same plant. Systemic effects on distal roots included the inhibition of secondary P . indica colonization. Faster and stronger induction of defense-related transcripts during secondary inoculation revealed that a P . indica pretreatment triggers root-wide priming of defense responses, which could cause the observed reduction of secondary colonization levels. Secondary P . indica colonization also induced defense responses in distant, already colonized parts of the root. Endophytic fungi therefore trigger a spatially specific response in directly colonized and in systemic root tissues of host plants.

Proteomic analysis of mycelium and secretome of different Botrytis cinerea wild-type strains

The necrotrophic fungus Botrytis cinerea is a very damaging phytopathogen of wide host range and environmental persistence. It is difficult to control because of its genetic versatility, expressed in the many phenotypical differences among isolates. The genomes of the B. cinerea B05.10 and T4 strains have been recently sequenced, becoming a model system for necrotrophic pathogens, and thus opening new alternatives for functional genomics analysis. In this work, the mycelium and secreted proteome of six wild-type strains with different host range, and grown in liquid minimal medium, have been analyzed by using complementary gel-based (1-DE and 2-DE) and gel-free/label-free (nUPLC-MS(E)) approaches. We found differences in the protein profiles among strains belonging to both the mycelium and the secretome. A total of 47 and 51 variable proteins were identified in the mycelium and the secretome, respectively. Some of them, such as malate dehydrogenase or peptidyl-prolyl cis-trans isomerase from the mycelium, and endopolygalacturonase, aspartic protease or cerato-platanin protein from the secretome have been reported as virulence factors, which are involved in host-tissue invasion, pathogenicity or fungal development. The necrotrophic fungus Botrytis cinerea is an important phytopathogen of wide host range and environmental persistence, causing substantial economic losses worldwide. In this work, the mycelium and secreted proteome of six B. cinerea wild-type strains with different host range have been analyzed by using complementary gel-based and gel-free/label-free approaches. Fungal genetic versatility was confirmed at the proteome level for both mycelium proteome and secreted proteins. A high number of hypothetical proteins with conserved domains related to toxin compounds or to unknown functions were identified, having qualitative differences among strains. The identification of hypothetical proteins suggests that the B. cinerea strains differ mostly in processes involved in adaptation to a particular environment or a growth condition, rather than in essential metabolic reactions. Proteomics can help in the identification of variable proteins related to the infection and colonization of host plant tissues, as well as of virulence and aggressiveness factors among different B. cinerea wild-type strains. This article is part of a Special Issue entitled: Trends in Microbial Proteomics.

The consequences of alternating diet on performance and food preferences of a specialist leaf beetle

The food quality of a given host plant tissue will influence the performance and may also affect the preference behavior of herbivorous animals. As nutrient contents and defense metabolite concentrations can vary significantly between different parts of a plant and change over time, herbivores are potentially confronted with diet differing in quality even when feeding on a single plant individual. Here we investigated to what extent feeding exclusively either on young or old, mature leaves of Brassica rapa or on a mixed diet of young and old leaves offered in alternating order affects the larval performance, food consumption, and the host preference behavior of adult mustard leaf beetles, Phaedon cochleariae. Analyzing different leaf quality traits, we found lower water contents, no changes in C:N ratio but more than threefold higher glucosinolate concentrations in young compared to old leaves. Individuals reared on mixed diet performed as well as animals reared on young leaves. Thus, compared to animals feeding exclusively on highly nutritious young leaves, diet-mixing individuals may balance the lower nutrient intake by a dilution of adverse secondary metabolites. Alternatively, they may integrate over the variation in their food, using a previously assimilated resource for growth at times of scarcity. Animals reared on old leaves grew less and had a prolonged larval developmental time, although they showed increased consumption indicating compensatory feeding. Additionally, we found that experience with a certain diet affected the preference behavior. Whereas individuals reared exclusively on young leaves preferred young over old leaves for feeding and oviposition, we did not find any preferences by animals reared exclusively on old leaves or by females reared on alternating diet. Thus, in contrast to positive feedbacks for animals reared on young leaves, an integrative growth of diet-mixing individuals potentially leads to a lack of feedback during development. Taken together, our results suggest that different diet regimes can lead to comparable performance of mustard leaf beetles but experienced feedbacks may differ and thus convey distinct diet preferences.

Involvement of two type 2C protein phosphatases BcPtc1 and BcPtc3 in the regulation of multiple stress tolerance and virulence of Botrytis cinerea

Type 2C Ser/Thr phosphatases (PP2Cs) are involved in various cellular processes in many eukaryotes, but little has been known about their functions in filamentous fungi. Botrytis cinerea contains four putative PP2C genes, named BcPTC1, -3, -5, and -6. Biological functions of these genes were analysed by gene deletion and complementation. While no phenotypes aberrant from the wild type were observed with mutants of BcPTC5 and BcPTC6, mutants of BcPTC1 and BcPTC3 had reduced hyphal growth, increased conidiation, and impaired sclerotium development. Additionally, BcPTC1 and BcPTC3 mutants exhibited increased sensitivity to osmotic and oxidative stresses, and to cell wall degrading enzymes. Both mutants exhibited dramatically decreased virulence on host plant tissues. All of the defects were restored by genetic complementation of the mutants with wild-type BcPTC1 and BcPTC3 respectively. Different from what is known in Saccharomyces cerevisiae, BcPtc3, but not BcPtc1, negatively regulates phosphorylation of BcSak1 (the homologue of S. cerevisiae Hog1) in B. cinerea, although both BcPTC1 and BcPTC3 were able to rescue the growth defects of a yeast PTC1 deletion mutant under various stress conditions. These results demonstrated that BcPtc1 and BcPtc3 play important roles in the regulation of multiple stress tolerance and virulence of B. cinerea.

Cell Death Control: The Interplay of Apoptosis and Autophagy in the Pathogenicity of Sclerotinia sclerotiorum

Programmed cell death is characterized by a cascade of tightly controlled events that culminate in the orchestrated death of the cell. In multicellular organisms autophagy and apoptosis are recognized as two principal means by which these genetically determined cell deaths occur. During plant-microbe interactions cell death programs can mediate both resistant and susceptible events. Via oxalic acid (OA), the necrotrophic phytopathogen Sclerotinia sclerotiorum hijacks host pathways and induces cell death in host plant tissue resulting in hallmark apoptotic features in a time and dose dependent manner. OA-deficient mutants are non-pathogenic and trigger a restricted cell death phenotype in the host that unexpectedly exhibits markers associated with the plant hypersensitive response including callose deposition and a pronounced oxidative burst, suggesting the plant can recognize and in this case respond, defensively. The details of this plant directed restrictive cell death associated with OA deficient mutants is the focus of this work. Using a combination of electron and fluorescence microscopy, chemical effectors and reverse genetics, we show that this restricted cell death is autophagic. Inhibition of autophagy rescued the non-pathogenic mutant phenotype. These findings indicate that autophagy is a defense response in this necrotrophic fungus/plant interaction and suggest a novel function associated with OA; namely, the suppression of autophagy. These data suggest that not all cell deaths are equivalent, and though programmed cell death occurs in both situations, the outcome is predicated on who is in control of the cell death machinery. Based on our data, we suggest that it is not cell death per se that dictates the outcome of certain plant-microbe interactions, but the manner by which cell death occurs that is crucial.

Enhanced Resistance toSpodoptera liturain Endophyte Infected Cauliflower Plants

Endophytic fungi, which live within host plant tissues without causing any visible symptom of disease, are important mediators of plant-herbivore interactions. These endophytes enhance resistance of host plant against insect herbivores mainly by productions of various alkaloid based defensive compounds in the plant tissue or through alterations of plant nutritional quality. Two endophytic fungi, i.e., Nigrospora sp. and Cladosporium sp., were isolated from Tinospora cordifolia (Thunb.) Miers, a traditional indian medicinal plant. Cauliflower (Brassica oleracea L.) plants were inoculated with these two endophytic fungi. The effect of endophyte infected and uninfected cauliflower plants were measured on the survival and development of Spodoptera litura (Fab.), a polyphagous pest. Endophyte infected cauliflower plants showed resistance to S. litura in the form of significant increase in larval and pupal mortality in both the fungi. Inhibitory effects of endophytic fungi also were observed on adult emergence, longevity, reproductive potential, as well as hatchability of eggs. Thus, it is concluded that antibiosis to S. litura could be imparted by artificial inoculation of endophytes and this could be used to develop alternative ecologically safe control strategies.

Are endophytes an important link between airborne spores and allergen exposure?

Grasses represent one of the Earth’s most common plant groups, and natural and cultivated habitats dominated by grasses cover about 40% of the land surface. In turn, each grass species hosts multiple fungal species which can behave as endophytes. An analysis of the endophytic taxa identified in surveys conducted in 14 grass species showed that some of the most frequent taxa on each grass were also present across several host grasses. These taxa were Alternaria, Epicoccum, Cladosporium, Fusarium, and a few others. A similar analysis of airborne fungi surveyed at 41 different locations throughout the world showed that some of the most geographically widespread, and most locally frequent airborne fungi belonged to the same genera that are dominant endophytes in grasses (i.e. Cladosporium, Alternaria, Fusarium, etc.). Therefore, airborne spores of genera that are ubiquitous in grasses are common worldwide and attain high atmospheric concentrations. In addition, spores of the above mentioned fungi are also important respiratory allergens. Direct observation indicates that saprobic colonization and sporulation of non-systemic grass endophytes could become unrestrained when their host plant tissue dies. Subsequently, when appropriate environmental conditions favour sporulation on grass host surfaces, the natural cycle for airborne conidia initiates, and large numbers of these conidia disperse as inoculum for new endophytic infections. Therefore, the cycle of endophytism may be an important link between climate, plant biology and public health.

Developmental pathway from leaves to galls induced by a sap-feeding insect on Schinus polygamus (Cav.) Cabrera (Anacardiaceae)

Galling sap-feeding insects are presumed to cause only minor changes in host plant tissues, because they usually do not require development of nutritive tissues for their own use. This premise was examined through comparison of the histometry, cytometry and anatomical development of non-galled leaves and galls of Calophya duvauae (Scott) (Hemiptera: Calophyidae) on Schinus polygamus (Cav.) Cabrera (Anacardiaceae). Cell fates changed from non-galled leaves to galls during the course of tissue differentiation. C. duvauae caused changes in dermal, ground, and vascular systems of the leaves of S. polygamus. Its feeding activity induced the homogenization of the parenchyma, and the neoformation of vascular bundles and trichomes. The histometric and cytometric data revealed compensatory effects of hyperplasia and cell hypertrophy in the epidermis, with hyperplasia predominating in the adaxial epidermis. There was a balance between these processes in the other tissues. Thus, we found major differences between the developmental pathways of non-galled leaves and galls. These changes were associated with phenotypic alterations related to shelter and appropriate microenvironmental conditions for the gall inducer. The nondifferentiation of a typical nutritive tissue in this case was compared to other non-phylogenetically related arthropod gall systems, and is suggested to result from convergence associated with the piercing feeding apparatus of the corresponding gall-inducer.

Detection of potential suberinase-encoding genes in Streptomyces scabiei strains and other actinobacteria

Streptomyces scabiei causes common scab, an economically important disease of potato tubers. Some authors have previously suggested that S. scabiei penetration into host plant tissue is facilitated by secretion of esterase enzymes degrading suberin, a lipidic biopolymer of the potato periderm. In the present study, S. scabiei EF-35 showed high esterase activity in suberin-containing media. This strain also exhibited esterase activity in the presence of other biopolymers, such as lignin, cutin, or xylan, but at a much lower level. In an attempt to identify the esterases involved in suberin degradation, translated open reading frames of S. scabiei 87-22 were examined for the presence of protein sequences corresponding to extracellular esterases of S. scabiei FL1 and of the fungus Coprinopsis cinerea VTT D-041011, which have previously been shown to be produced in the presence of suberin. Two putative extracellular suberinase genes, estA and sub1, were identified. The presence of these genes in several actinobacteria was investigated by Southern blot hybridization, and both genes were found in most common-scab-inducing strains. Moreover, reverse transcription - polymerase chain reaction performed with S. scabiei EF-35 showed that estA was expressed in the presence of various biopolymers, including suberin, whereas the sub1 gene appeared to be specifically expressed in the presence of suberin and cutin.

植物内生菌促进宿主氮吸收与代谢研究进展

Endophytes are microbes that reside within the living tissues of host plants without substantively harming them.They are ubiquitous in most plant species,latently residing in or actively colonizing the tissues.Infection by endophytes can improve the level of host plant nitrogen assimilation and metabolism.The mechanisms mediated by endophytes are complex;when symbiotic with plants,the ability to fix and solubilize mineral nutrients otherwise unavailable to plants allows plant-associated bacteria to act as biofertilizers.Endophytes are able to improve the level of host plant nitrogen assimilation and metabolism by promoting the growth and development of roots and by the fixation of nitrogen(diazotrophy).Nitrogen is often a limiting factor for plant growth because atmospheric nitrogen exists as dinitrogen(N2),a form that is inaccessible to all organisms except a few specially adapted prokaryotes.For all other organisms(including plants),nitrogen needs to be in the form of either ammonia or nitrate before it can be used.Diazotrophic bacteria possess nitrogenase,an O2-sensitive enzyme that catalyzes the reduction of atmospheric nitrogen to ammonia.Higher plants cannot carry out this process in the absence of these bacteria.Endophytes can also produce multiple ectoenzymes,such as laccase and proteolytic enzymes.These ectoenzymes facilitate the hydrolyzing and mineralizing of some polypeptides and soluble proteins for plant use.The function of ectoenzymes suggests that endophytes can help the host plants to use organic nitrogen as well as atmospheric.In addition,endophytes also clearly enhance the activities of key enzymes in the nitrogen assimilation and metabolism pathways of their hosts.In plants,glutamine synthetase(GS) and nitrate reductase(NR) are the enzymes that are mainly responsible for the assimilation of ammonium,and enhanced activity in these two key enzymes greatly improves plant metabolism.Furthermore,endophytes are also able to promote the production of phytohormones such as ethylene,auxins,indole-3-acetic acid(IAA),cytokinins,abscisic acid or combinations thereof,as well as the production of vitamins such as zeatin,vitamin B1,and vitamin B6.Both phytohormones and vitamins are important in nitrogen metabolism.Interestingly,endophyte infection results in higher NH+4 concentrations in the plant tissue,but NO-3 concentrations are decreased.Tiller production,chlorophyll content and photosynthetic rate are also affected by endophytes;infected plants produce more tillers and have greater chlorophyll content than uninfected ones,and photosynthetic rates are also increased.In this thesis,we will give a brief summary of the aspects that deserve special attention in researching the role of endophytes in enhancing plant nitrogen assimilation and metabolism.We will also attempt to outline the mechanisms of endophyte-affected plant nitrogen assimilation and metabolism to highlight future research prospects.

Review: Plant Binary Vectors of Ti Plasmid in <i>Agrobacterium tumefaciens</i> with a Broad Host-Range Replicon of pRK2, pRi, pSa or pVS1

This review chronicles the development of the plant binary vectors of Ti plasmid in Agrobacterium tumefaciens during the last 30 years. A binary vector strategy was designed in 1983 to separate the T-DNA region in a small plasmid from the virulence genes in avirulent T-DNA-less Ti plasmid. The small plant vectors with the T-DNA region have been simply now called binary Ti vectors. A binary Ti vector consist of a broad host-range replicon for propagation in A. tumeraciens, an antibiotic resistance gene for bacterial selection and the T-DNA region that would be transferred to the plant genome via the bacterial virulence machinery. The T-DNA region delimited by the right and left border sequences contains an antibiotic resistance gene for plant selection, reporter gene, and/or any genes of interest. The ColEI replicon was also added to the plasmid backbone to enhance the propagation in Escherichia coli. A general trend in the binary vector development has been to increase the plasmid stability during a long co-cultivation period of A. tumefaciens with the target host plant tissues. A second trend is to understand the molecular mechanism of broad host-range replication, and to use it to reduce the size of plasmid for ease in cloning and for higher plasmid yield in E. coli. The broad host-range replicon of VS1 was shown to be a choice of replicon over those of pRK2, pRi and pSA because of the superior stability and of small well-defined replicon. Newly developed plant binary vectors pLSU has the small size of plasmid backbone (4566 bp) consisting of VS1 replicon (2654 bp), ColE1 replicon (715 bp), a bacterial kanamycin (999 bp) or tetracycline resistance gene, and the T-DNA region (152 bp).

Host‐plant quality adaptively affects the diapause threshold: evidence from leaf beetles in willow plantations

1. Voltinism of herbivorous insects can vary depending on environmental conditions. The leaf beetle Phratora vulgatissima L. is univoltine in Sweden but will sometimes initiate a second generation in short‐rotation coppice (SRC) willow plantations.2. The study investigated whether increased voltinism by P. vulgatissima in plantations can be explained by (i) rapid life‐cycle development allowing two generations, or (ii) postponed diapause induction on coppiced willows.3. In the field, no difference was found in the phenology or development of first‐generation broods between plantations (S. viminalis) and natural willow habitats (S. cinerea). However, the induction of diapause occurred 1–2 weeks later in SRC willow plantations.4. Laboratory experiments indicated no genetic difference in the critical day‐length for diapause induction between beetles originating from plantations and natural habitats. Development time was unaffected by host‐plant quality but critical day‐length was prolonged by almost an hour when the beetles were reared on a non‐preferred willow species (S. phylicifolia). When reared on new leaves from re‐sprouting shoots of recently coppiced willow plants, diapause incidence was significantly less than when the beetles were reared on mature leaves from uncoppiced plants.5. The study suggests that P. vulgatissima has a plastic diapause threshold influenced by host‐plant quality. The use of host‐plant quality as a diapause‐inducing stimulus is likely to be adaptive in cases where food resources are unpredictable, such as when new host‐plant tissue is produced after a disturbance. SRC willows may allow two beetle generations due to longer growing seasons of coppiced plants that grow vigorously.

Overexpression of a novel biotrophy-specific Colletotrichum truncatum effector, CtNUDIX, in hemibiotrophic fungal phytopathogens causes incompatibility with their host plants.

The hemibiotrophic fungus Colletotrichum truncatum causes anthracnose disease on lentils and a few other grain legumes. It shows initial symptomless intracellular growth, where colonized host cells remain viable (biotrophy), and then switches to necrotrophic growth, killing the colonized host plant tissues. Here, we report a novel effector gene, CtNUDIX, from C. truncatum that is exclusively expressed during the late biotrophic phase (before the switch to necrotrophy) and elicits a hypersensitive response (HR)-like cell death in tobacco leaves transiently expressing the effector. CtNUDIX homologs, which contain a signal peptide and a Nudix hydrolase domain, may be unique to hemibiotrophic fungal and fungus-like plant pathogens. CtNUDIX lacking a signal peptide or a Nudix motif failed to induce cell death in tobacco. Expression of CtNUDIX:eGFP in tobacco suggested that the fusion protein might act on the host cell plasma membrane. Overexpression of CtNUDIX in C. truncatum and the rice blast pathogen, Magnaporthe oryzae, resulted in incompatibility with the hosts lentil and barley, respectively, by causing an HR-like response in infected host cells associated with the biotrophic invasive hyphae. These results suggest that C. truncatum and possibly M. oryzae elicit cell death to signal the transition from biotrophy to necrotrophy.

Quantitative Real-Time PCR Analysis of ‘Candidatus Phytoplasma mali’ Without External Standard Curves

Apple proliferation caused by ‘Candidatus Phytoplasma mali’ is an economically important disease of apple (Malus ´ domestica). The availability of a simple quantitative approach to assess pathogen load in infected host plants would certainly contribute to a better understanding of pathogenesis and epidemiology. This study proposes a quantification approach not requiring the analysis of external standard curves. It is based on the simultaneous detection of the 16S rRNA gene of the pathogen and the 1-aminocyclopropane-1-carboxylate oxidase gene of the host plant in a single-tube reaction using TaqMan chemistry. The quantity of the phytoplasma relative to its host plant is determined as the difference between CT values of the two target genes (ΔCT). To assess the agreement of the relative quantification approach with a standard curve-based method, a dataset of 450 DNA samples from infected apple trees was reanalysed. Comparison of the ΔCT-based relative quantities with the corresponding absolute values revealed high degrees of agreement between the relative and absolute quantification methods. The ΔCT procedure can thus be considered adequate for quantification of the phytoplasma in infected host plant tissue. In addition, this approach ensures improved methodological standardisation and increased analysis throughput.

FgVelB globally regulates sexual reproduction, mycotoxin production and pathogenicity in the cereal pathogen Fusarium graminearum

The velvet genes are conserved in ascomycetous fungi and function as global regulators of differentiation and secondary metabolism. Here, we characterized one of the velvet genes, designated FgVelB, in the plant-pathogenic fungus Fusarium graminearum, which causes fusarium head blight in cereals and produces mycotoxins within plants. FgVelB-deleted (ΔFgVelB) strains produced fewer aerial mycelia with less pigmentation than those of the wild-type (WT) during vegetative growth. Under sexual development conditions, the ΔFgVelB strains produced no fruiting bodies but retained male fertility, and conidiation was threefold higher compared with the WT strain. Production of trichothecene and zearalenone was dramatically reduced compared with the WT strain. In addition, the ΔFgVelB strains were incapable of colonizing host plant tissues. Transcript analyses revealed that FgVelB was highly expressed during the sexual development stage, and may be regulated by a mitogen-activated protein kinase cascade. Microarray analysis showed that FgVelB affects regulatory pathways mediated by the mating-type loci and a G-protein alpha subunit, as well as primary and secondary metabolism. These results suggest that FgVelB has diverse biological functions, probably by acting as a member of a possible velvet protein complex, although identification of the FgVelB-FgVeA complex and the determination of its roles require further investigation.

Molecular Identification of Synanthedonini Members (Lepidoptera: Sesiidae) using Cytochrome Oxidase I

Abstract Many North American sesiid moths within Synanthodonini have been studied extensively because their feeding activity can cause detrimental economic and esthetic impacts to many commercially important ornamental and native plant species. Recent discoveries of nonnative clearwing moth pest introductions [e.g., Synanthedon myopaeformis (Borkh.)], reinforce the need for reliable and accurate molecular diagnostic tools that can be used by nontaxonomic experts, particularly when juvenile life stages are recovered from infested host-plant tissues. Cytochrome oxidase I (cox I) previously has been used to successfully identify species and resolve species complexes. In this study, the cox I phylogeny inferred from sequences generated from 21 species of sesiid moths classified within Synanthedonini confirms the close evolutionary relationship between sesiid species. As other authors have suggested in previous works, we observed that Synanthedon rileyana H. Edwards appears atypical for the genus, as it paired with Carmenta bassiformis (Walker) one node removed from, but not sister to, a large well-supported Synanthedon-rich clade. Sannina uroceriformis Walker and Podosesia Möschler were observed nested deeply within the aforementioned well-supported clade (posterior probability [PP] of clade = 100) comprised of all Synanthedon species sampled, except S. rileyana. Placement of these two taxa conflicts with results from previous morphological studies. These placements were immune from repeated attempts to delete perceived nearby long branches within the data set. Despite these few conflicts and overall low statistical support for most interspecific and higher relationships, our data suggest that all species examined possess unique genetic signatures that lend themselves to accurate identification of all life history stages of these clearwing pests.

A preliminary histological investigation of gall induction in an unconventional galling system

In an unusual case involving a candidate biological control agent, the histologically complex stem galls of the weevil, Rhinusa pilosa (Coleoptera: Curculionidae) on yellow toadflax (Linaria vulgaris), are rapidly induced during oviposition and reach full size by larval hatch. To investigate gall induction, the oviposition behavior of R. pilosa was described. We experimentally disrupted ovipositing weevils at three key points in the oviposition sequence and compared host-plant tissue responses post disruption, to what occurs during normal gall induction using histological methods. De novo xylem production, intercellular spaces in the cortex, and hyperplasia and hypertrophy of the procambium and pith parenchyma surrounding the egg were some of the tissue- and cellular-level modifications observed only 3–5 days after normal oviposition. Normal gall development was not observed after any of the oviposition disruption treatments, although some of the cellular and tissue responses resembled those found after undisrupted oviposition. Feeding by the female during oviposition canal formation induced wound meristem and callus tissue formation, but no other modifications consistent with gall formation. When females were disrupted about 20 s into oviposition, a homogenously dense substance was observed, which was suspected to be ovipositional fluid. There was minor stem swelling 10 days later and histologically, periclinal cell divisions, de novo xylem, and pith cells with numerous stained plastids were observed as in normal gall development, thus suggesting that ovipositional fluid plays a role in gall induction.

Does exogenic food benefit both partners in an ant-plant mutualism? The case of Cecropia obtusa and its guest Azteca plant-ants

In the mutualisms involving the myrmecophyte Cecropia obtusa and Azteca ovaticeps or A. alfari, both predatory, the ants defend their host trees from enemies and provide them with nutrients (myrmecotrophy). A. ovaticeps provisioned with prey and then 15N-enriched food produced more individuals than did control colonies (not artificially provisioned). This was not true for A. alfari colonies, possibly due to differences in the degree of maturity of the colonies for the chosen range of host tree sizes (less than 3m in height). Myrmecotrophy was demonstrated for both Azteca species as provisioning the ants with 15N-enriched food translated into higher δ15N values in host plant tissues, indicating that nitrogen passed from the food to the plant. Thus, the predatory activity of their guest ants benefits the Cecropia trees not only because the ants protect them from defoliators since most prey are phytophagous insects but also because the plant absorbs nutrients.

Aliphatic Esters as Targets of Esterase Activity in the Parsnip Webworm (Depressaria pastinacella)

As a specialist on the reproductive structures of Pastinaca sativa and species in the related genus Heracleum, the parsnip webworm (Depressaria pastinacella) routinely encounters a distinctive suite of phytochemicals in hostplant tissues. Little is known, however, about the detoxification mechanisms upon which this species relies to metabolize these compounds. In this study, larval guts containing hostplant tissues were homogenized, and metabolism was determined by incubating reactions with and without NADPH and analyzing for substrate disappearance and product appearance by gas chromatography-mass spectrometry. Using this approach, we found indications of carboxylesterase activity, in the form of appropriate alcohol metabolites for three aliphatic esters in hostplant tissues-octyl acetate, octyl butyrate, and hexyl butyrate. Involvement of webworm esterases in hostplant detoxification subsequently was confirmed with metabolism assays with pure compounds. This study is the first to implicate esterases in lepidopteran larval midgut metabolism of aliphatic esters, ubiquitous constituents of flowers and fruits. In addition, this method confirmed that webworms detoxify furanocoumarins and myristicin in their hostplants via cytochrome P450-mediated metabolism, and demonstrated that these enzymes also metabolize the coumarin osthol and the fatty acid derivative palmitolactone.

Methyl isonicotinate induces increased walking and take‐off behaviour in western flower thrips, Frankliniella occidentalis

AbstractTimely detection of western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), is important to help prevent economic damage to crops. Coloured traps are commonly used for thrips monitoring and their efficacy can be markedly improved with the addition of semiochemicals such as methyl isonicotinate (MI). However, the behavioural response of thrips to these compounds is poorly understood. In leaf/petal disc laboratory experiments the effect of MI on adult female WFT was examined with respect to walking and take‐off behaviour. Clean air or air with MI was blown over the leaf/petal discs at an angle of 30° via a glass pipette connected to a wash bottle containing a cotton dental roll with or without a defined amount of MI. At airspeeds of 0.6–1.2 m s−1 and at doses of 1, 10, 50, 100, and 1000 μl (equivalent to indicative concentrations of 1.7, 4.5, 7.3, 8.8, and 14.9 ng MI l−1 air blown over the leaf discs), WFT spent a greater proportion of time walking on chrysanthemum leaf discs in the presence of MI than the non‐attractive thrips compound β‐caryophyllene or clean air. Western flower thrips showed a dose‐dependent take‐off response to the thrips attractant MI, but not to β‐caryophyllene (virtually no take‐off). At 100 μl of MI (8.8 ng l−1 air), WFT spent a greater proportion of time walking on host plant tissue than in clean air, depending on plant disc substrates. Furthermore, WFT took off more often from plant discs exposed to an air current containing MI than from those exposed to clean air. Western flower thrips exposed to MI took off at a similar rate from chrysanthemum leaves, chrysanthemum flower petals, and sweet pepper leaves, but much less readily from bean leaves. The results show that MI stimulates walking and take‐off behaviour for adult female WFT under these experimental conditions and these responses are dose‐dependent and mediated by the host plant. These results are placed in the context of previous research on the behavioural response of thrips to semiochemicals and their implications for thrips pest management are discussed.