Search

Organisms of the third trophic level can indirectly interact with plants. However, whether parasitoids of herbivores have a positive effect on plant fitness has been controversial. In addition to possible effects on plant fitness, parasitoid‐mitigated herbivory can modify plant physiological responses and thereby alter the plant‐mediated indirect interactions between different herbivore species. These types of indirect multitrophic interactions remain largely unexplored. Thus, to understand the full effect of the third trophic level on plants, it is necessary to consider the context of the community of interacting species, both herbivores and their enemies. Here, we investigated whether parasitoids of leaf‐feeding caterpillars affect plant fitness (seed quantity and quality) and the consequences for seed‐dwelling insects at the second and third trophic levels through plant‐mediated effects. To test this, we exposed lima bean plants (Phaseolus lunatus), under controlled field conditions, to caterpillars (Spodoptera latifascia) that were unparasitized or parasitized by the parasitoid species Cotesia marginiventris. Later in the season, we measured seed production and infestation by seed beetles and their parasitoids. We found that parasitoids significantly reduced the leaf damage inflicted by the caterpillars, such that the plants suffered no loss in seed production. Yet, parasitoids had no effect on the emergence of seed beetles (Zabrotes subfasciatus and Acanthoscelides obtectus), which was equally reduced in plants attacked by unparasitized and by parasitized caterpillars. Seeds from undamaged plants were significantly more attacked by Z. subfasciatus beetles. Parasitism rates of seed beetle larvae were similar for all treatments. Although parasitized caterpillars did not damage the plants enough to reduce seed production (unlike unparasitized caterpillars), the damage they inflicted induced resistance against other herbivores. Taken together, these results show how parasitoids can indirectly enhance plant fitness in the context of the local multitrophic ecological networks. These findings have significant implications for natural and agricultural systems since they reveal that the indirect interaction between plants and parasitoids can be beneficial in communities with multiple herbivore species.

PREMISE Plant responses to herbivores and their elicitors include changes in traits associated with phenology, defense, and reproduction. Induced responses by chewing herbivores are known to be hormonally mediated by the jasmonate pathway and can cascade and affect late-season seed predators and pollinators. Moreover, herbivore-induced plant responses can be transmitted to the next generation. Whether herbivore-induced transgenerational effects also apply to phenological traits is less well understood. METHODS Here, we explored responses of wild lima bean plants (Phaseolus lunatus) to herbivory and jasmonate treatment and possible transgenerational effects of herbivore-induced early flowering. In a controlled field experiment, we exposed lima bean plants to herbivory by leaf beetles or methyl jasmonate sprays (MJ). We then compared plant development, phenology, reproductive fitness and seed traits among these treatments and undamaged, untreated control plants. RESULTS We found that MJ and leaf herbivory induced similar responses, with treated plants growing less, flowering earlier, and producing fewer seeds than undamaged plants. However, seed size, phenolics and cyanogenic glycosides concentrations did not differ among treatments. Seed germination rates and flowering time of the offspring were similar among maternal treatments. CONCLUSIONS Overall, the results confirm that responses of lima bean to herbivory by leaf beetles are mediated by jasmonate; however, effects on phenological traits are not transmitted to the next generation. We discuss why transgenerational effects of herbivory might be restricted to traits that directly target herbivores.

Immune infiltration is typically quantified using cellular density, not accounting for cellular clustering. Tumor-associated macrophages (TAM) activate oncogenic signaling through paracrine interactions with tumor cells, which may be better reflected by local cellular clustering than global density metrics. Using multiplex immunohistochemistry and digital pathologic analysis we quantified cellular density and cellular clustering for myeloid cell markers in 129 regions of interest from 55 samples from 35 patients with metastatic ccRCC. CD68+ cells were found to be clustered with tumor cells and dispersed from stromal cells, while CD163+ and CD206+ cells were found to be clustered with stromal cells and dispersed from tumor cells. CD68+ density was not associated with OS, while high tumor/CD68+ cell clustering was associated with significantly worse OS. These novel findings would not have been identified if immune infiltrate was assessed using cellular density alone, highlighting the importance of including spatial analysis in studies of immune cell infiltration of tumors.SIGNIFICANCEIncreased clustering of CD68+ TAMs and tumor cells was associated with worse overall survival for patients with metastatic ccRCC. This effect would not have been identified if immune infiltrate was assessed using cell density alone, highlighting the importance of including spatial analysis in studies of immune cell infiltration of tumors.

It is increasingly evident that plants interact with their outside world through the production of volatile organic compounds,1-5 but whether the volatiles have evolved to serve in plant defense is still a topic of considerable debate.3,6-8 Unharmed leaves constitutively release small amounts of volatiles, but when the leaves are damaged by herbivorous arthropods, they emit substantially more volatiles. These herbivore-induced plant volatiles (HIPVs) attract parasitoids and predators that kill insect herbivores,9-12 and this can benefit the plants.13,14 As yet, however, there is no tangible evolutionary evidence that this tritrophic interplay contributes to the selection forces that have shaped the volatile emissions of plants.2,3,5-8,15 With this in mind, we investigated the evolutionary changes in volatile emissions in invasive common ragwort and the respective defensive roles of its constitutive and inducible volatiles. This Eurasian plant has invaded other continents, where it evolved for many generations in the absence of specialized herbivores and their natural enemies. We found that, compared to native ragworts, invasive plants release higher levels of constitutive volatiles but considerably lower levels of herbivore-induced volatiles. As a consequence, invasive ragwort is more attractive to a specialist moth but avoided by an unadapted generalist moth. Importantly, conforming to the indirect defense hypothesis, a specialist parasitoid was much more attracted to caterpillar-damaged native ragwort, which was reflected in higher parasitism rates in a field trial. The evolution of foliar volatile emissions appears to be indeed driven by their direct and indirect roles in defenses against insects.

Background Tumor associated macrophages (TAM) stimulate tumor proliferation and facilitate immune escape via production of immunosuppressive cytokines. We hypothesize that non-random spatial clustering of TAMs within the tumor are associated with poor survival in ccRCC patients. Methods Tumor specimens were obtained from 41 patients with metastatic ccRCC who received immunotherapy (IT). Sections from the tumor core underwent multiplex immunofluorescence staining for CD68, CD163, and CD206. Digital pathologic analysis was used to convert the digital images to spatial point pattern plots (PPP). Ripley’s K function, the current standard metric for spatial heterogeneity, was utilized. Novel metrics were developed using a probability density function (PDF) for distances between cells, assuming that cells can be located anywhere with equal probability. Empirical histograms were generated from the PPPs. Deviation from the PDF demonstrates a non-random distribution. Deviations were quantified with the Kolmogorov-Smirnov (KS) test and Cramer-von Mises (CVM) criterion. Overall survival (OS) was assessed between groups stratified by the median value for each metric using Kaplan-Meier and log-rank analysis. Figure 1A. Results 75 slides were analyzed from the 41 patients. The three metrics for measuring spatial heterogeneity had moderate and statistically significant correlation with each other (Spearman’s R: Ripley/KS=0.68, p Conclusions We describe CVM and KS as novel metrics for measuring spatial heterogeneity of immune cells. Increased spatial heterogeneity of CD68+ TAMs and tumor cells was associated with worse OS in patients with metastatic ccRCC who received IT. These findings corroborate prior reports of TAMs eliciting an immunosuppressive effect on the tumor-immune microenvironment, and demonstrate the novel finding of a clinically significant effect of TAM spatial clustering on OS.

The most evident trade-off in specialized herbivores is the reduction of host range. However, trade-offs that limit the adaptation of specialized herbivores to an evolving host could exist and have been scarcely explored. Here, we tested whether the specialized leaf beetle species Lema daturaphila expresses trade-offs related to its co-adaptation with the host plant species Datura stramonium. Firstly, we found that increases in the concentration of scopolamine, a tropane alkaloid that is produced by the plant species, reduce beetle survival, showing that scopolamine is still a defensive trait against the specialized herbivores. In addition, we performed a selection experiment to increase survival of the beetles on the normal host and explored the consequences for life history and host-related traits and the corresponding G-matrix. After three selection events, we observed an improvement of survival in the selection line, but no correlational selection on other traits. There was also evidence that the G-matrix structure changed after selection. The genetic correlations between fecundity and relative growth rate and consumption efficiency were negative in the selection line, but neutral in the control line. The change in genetic correlations after selection suggests that there are trade-offs between survival and host utilization traits, which have the potential to limit the beetles’ co-adaptation to evolving plant defenses.

Although the cotton bollworm Helicoverpa armigera has traditionally been controlled by application of chemical pesticides, chemical control selects for resistance, pollutes the environment, and endangers human health. New methods for controlling H. armigera are therefore needed. Heliothis virescens ascovirus 3i (HvAV-3i) is a recently identified virus of the lepidopteran larvae. We tested the effects of HvAV-3i on H. armigera larvae following oral ingestion of HvAV-3i-containing hemolymph (about 1.0 × 1010 virus genome copies per larvae) and following injection of HvAV-3i-containing hemolymph by insertion of a needle. Following oral ingestion, first-instar to fifth-instar larvae grew and developed normally. Following needle injection, in contrast, the corrected mortality of third and fourth instars was 88.9 ± 2.1 and 93.7 ± 3.4%, respectively. Food intake was significantly lower for larvae injected with virus-containing hemolymph than with virus-free hemolymph. Larvae injected with virus-containing hemolymph had extended survival times and could not complete the pre-pupal stage. These results indicate that inoculation of HvAV-3i via needle injection, but not via oral ingestion, significantly reduced the growth and development of H. armigera larvae.

Plants that synthesize and store terpenes in specialized cells accumulate large concentrations of these compounds while avoiding autotoxicity. Stored terpenes may influence the quantity and profile of volatile compounds that are emitted into the environment and the subsequent role of those volatiles in mediating the activity of herbivores. The Australian medicinal tea tree, Melaleuca alternifolia, occurs as several distinct terpene chemotypes. We studied the profile of its terpene emissions to understand how variations in stored foliar terpenes influenced emissions, both constitutive and when damaged either by herbivores or mechanically. We found that foliar chemistry influenced differences in the composition of terpene emissions, but those emissions were minimal in intact plants. When plants were damaged by herbivores or mechanically, the emissions were greatly increased and the composition corresponded to the constitutive terpenes and the volatility of each compound, suggesting the main origin of emissions is the stored terpenes and not de novo biosynthesized volatiles. However, herbivores modified the composition of the volatile emissions in only one chemotype, probably due to the oxidative metabolism of 1,8-cineole by the beetles. We also tested whether the foliar terpene blend acted as an attractant for the specialized leaf beetles Paropsisterna tigrina and Faex sp. and a parasitoid fly, Anagonia zentae. None of these species responded to extracts of young leaves in an olfactometer, so we found no evidence that these species use plant odor cues for host location in laboratory conditions.

Somatic mutations can have important effects on the life history, ecology, and evolution of plants, but the rate at which they accumulate is poorly understood and difficult to measure directly. Here, we develop a method to measure somatic mutations in individual plants and use it to estimate the somatic mutation rate in a large, long-lived, phenotypically mosaic Eucalyptus melliodora tree. Despite being 100 times larger than Arabidopsis, this tree has a per-generation mutation rate only ten times greater, which suggests that this species may have evolved mechanisms to reduce the mutation rate per unit of growth. This adds to a growing body of evidence that illuminates the correlated evolutionary shifts in mutation rate and life history in plants.

Simple Summary Immune modulation is considered a hallmark of cancer initiation and progression, and has offered promising opportunities for therapeutic manipulation. Multiplex immunofluorescence (mIF) technology has enabled the tumor immune microenvironment (TIME) to be studied at an increased scale, in terms of both the number of markers and the number of samples. Another benefit of mIF technology is the ability to measure not only the abundance but also the spatial location of multiple cells types within a tissue sample simultaneously, allowing for assessment of the co-localization of different types of immune markers. Thus, the use of mIF technologies have enable researchers to characterize patient, clinical, and tumor characteristics in the hope of identifying patients whom might benefit from immunotherapy treatments. In this review we outline some of the challenges and opportunities in the statistical analyses of mIF data to study the TIME. Abstract Immune modulation is considered a hallmark of cancer initiation and progression. The recent development of immunotherapies has ushered in a new era of cancer treatment. These therapeutics have led to revolutionary breakthroughs; however, the efficacy of immunotherapy has been modest and is often restricted to a subset of patients. Hence, identification of which cancer patients will benefit from immunotherapy is essential. Multiplex immunofluorescence (mIF) microscopy allows for the assessment and visualization of the tumor immune microenvironment (TIME). The data output following image and machine learning analyses for cell segmenting and phenotyping consists of the following information for each tumor sample: the number of positive cells for each marker and phenotype(s) of interest, number of total cells, percent of positive cells for each marker, and spatial locations for all measured cells. There are many challenges in the analysis of mIF data, including many tissue samples with zero positive cells or “zero-inflated” data, repeated measurements from multiple TMA cores or tissue slides per subject, and spatial analyses to determine the level of clustering and co-localization between the cell types in the TIME. In this review paper, we will discuss the challenges in the statistical analysis of mIF data and opportunities for further research.

The microbial loop has been suggested as an alternative route for better utilization of phytate, a poorly available P source to plants. We hypothesized that bacterial grazer activity might dramatically enhance bacterial migration and proliferation, increasing the probability of phytate hydrolysis by bacterial phytases and, thus, phytate mineralization and release of free phosphate. We tested this hypothesis in a two-compartment system with a solid medium containing phytate or free phosphate as the source of P. Two bacterial species, B. subtilis 168 or Bradyrhizobium sp., with or without bacterial grazing nematodes belonging to Acrobeloides sp. previously fed on each of the bacterial species, were inoculated at a single point in the medium. Whatever the P source, nematode migration within both zones allowed the proliferation of bacteria. However, B. subtilis 168 was more efficient in using phytate than Bradyrhizobium sp. since the highest bacterial cell density and free phosphate concentrations were reached by Acrobeloides sp. fed on B. subtilis 168. The grazer activity seemed to be crucial to enhance phytate mineralization, despite Acrobeloides sp. showing a higher preference to feed on Bradyrhizobium sp. This study provides new insights into the effects of bacterial grazer activity on phytate mineralization.

Intraspecific plant diversity can modify the properties of associated arthropod communities and plant fitness. However, it is not well understood which plant traits determine these ecological effects. We explored the effect of intraspecific chemical diversity among neighbouring plants on the associated invertebrate community and plant traits. In a common garden experiment, intraspecific diversity among neighbouring plants was manipulated using three plant populations of wild cabbage that differ in foliar glucosinolates. Plants were larger, harboured more herbivores, but were less damaged when plant diversity was increased. Glucosinolate concentration differentially correlated with generalist and specialist herbivore abundance. Glucosinolate composition correlated with plant damage, while in polycultures, variation in glucosinolate concentrations among neighbouring plants correlated positively with herbivore diversity and negatively with plant damage levels. The results suggest that intraspecific variation in secondary chemistry among neighbouring plants is important in determining the structure of the associated insect community and positively affects plant performance.

Establishing the putative links between sylvatic and domestic transmission cycles of Trypanosoma cruzi, the etiological agent of Chagas disease, is of public health relevance. We conducted three surveys to assess T. cruzi infection in wild mammals from a rural and a preserved area in Misiones Province, Northeastern Argentina, which had recently been declared free of vector- and blood-borne transmission of human T. cruzi infection. A total of 200 wild mammals were examined by xenodiagnosis (XD) and/or polymerase chain reaction (PCR) amplification of the hyper-variable region of kinetoplast DNA minicircles of T. cruzi (kDNA-PCR). The overall prevalence of T. cruzi infection was 8%. Nine (16%) of 57 Didelphis albiventris opossums and two (7%) of 29 Desmodus rotundus vampire bats were positive by both XD and kDNA-PCR. Additionally, one D. rotundus positive for T. cruzi by kDNA-PCR tested positive by satellite-DNA-PCR (SAT-DNA-PCR). The T. cruzi-infected bats were captured indoors and in the yard of a vacant dwelling. All D. albiventris were infected with TcI and both XD-positive D. rotundus by TcII. Fifty-five opossum cubs within the marsupium were negative by XD. The mean infectiousness to the vector was 62% in D. albiventris and 50% in D. rotundus. Mice experimentally infected with a parasite isolate from a vampire bat displayed lesions typically caused by T. cruzi. Our study documents the presence of the genotype TcII in a sylvatic host for the first time in Argentina, and the occurrence of two transmission cycles of T. cruzi in a district free of domestic vector-borne transmission. Fil: Argibay, Hernán Darío. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Orozco, Maria Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Cardinal, Marta Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina Fil: Rinas, Miguel A.. Gobierno de Misiones. Ministerio de Ecología y Recursos Naturales Renovables. Parque Ecológico "El Puma"; Argentina Fil: Arnaiz, Maria Rosa. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina Fil: Mena Segura, Carlos. Gobierno de la Ciudad de Buenos Aires. Instituto de Zoonosis Luis Pasteur; Argentina Fil: Gurtler, Ricardo Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentina

Insect desaturases are known to play an important role in chemical communication between individuals. However, their roles in insect growth, development and fecundity, and in regulating interactions of insects with plants, remain largely unknown. In this study, we explored the functions of Nlug-desatA2, a desaturase gene of the brown planthopper (BPH), Nilaparvata lugens (St&aring, l). The RNA interference-based knockdown of Nlug-desatA2 decreased the ratio of monounsaturated fatty acids to saturated fatty acids, and the level of fatty acids and triglycerides in BPH. Nlug-desatA2-knockdown also reduced the food intake, body mass and fecundity of female BPH adults, and led to abdomen atrophy and ovarian agenesis. Nlug-desatA2-knockdown suppressed the transcription of TOR (target of rapamycin), Lpp (Lipophorin) and AKHR (adipokinetic hormone receptor) in female adults. Moreover, the corrected survival rate of BPH with Nlug-desatA2-knockdown fed an artificial diet was higher than the survival rate of those fed on rice plants. Higher levels of salicylic acid in rice infested by Nlug-desatA2-knockdown female BPH adults than in rice infested by control BPH may be the reason. These findings demonstrate that Nlug-desatA2 has an essential role in lipid metabolism and is involved in the food intake, survival, development and fecundity of BPH. In addition, this gene is likely involved in regulating the responses of rice to BPH infestation.

Through a mutualistic relationship with woody plant roots, ectomycorrhizal fungi provide growth-limiting nutrients, including inorganic phosphate (Pi), to their host. Reciprocal trades occur at the Hartig net, which is the symbiotic interface of ectomycorrhizas where the two partners are symplasmically isolated. Fungal Pi must be exported to the symbiotic interface, but the proteins facilitating this transfer are unknown. In the present study, we combined transcriptomic, microscopy, whole plant physiology, X-ray fluorescence mapping, 32 P labeling and fungal genetic approaches to unravel the role of HcPT2, a fungal Pi transporter, during the Hebeloma cylindrosporum-Pinus pinaster ectomycorrhizal association. We localized HcPT2 in the extra-radical hyphae and the Hartig net and demonstrated its determinant role for both the establishment of ectomycorrhizas and Pi allocation towards P. pinaster. We showed that the host plant induces HcPT2 expression and that the artificial overexpression of HcPT2 is sufficient to significantly enhance Pi export towards the central cylinder. Together, our results reveal that HcPT2 plays an important role in ectomycorrhizal symbiosis, affecting both Pi influx in the mycelium and efflux towards roots under the control of P. pinaster.

Plant chemical polymorphisms, or plant chemotypes, are characterized by intraspecific discrete differences of plant secondary metabolites in the same plant tissue. Chemotypes that differ in foliar terpene composition are found commonly in Myrtaceae. In this study, we focused on terpene chemotypes of medicinal tea tree, Melalecua alternifolia, to explore whether this variation affects two specialist herbivores Paropsisterna tigrina and Faex sp. (Coleoptera: Chrysomelidae), and if this could explain the maintenance of this variation. We tested whether insect performance, oviposition preference, and plant damage were associated with different chemotypes. We found that larval growth rate of Faex sp. was higher in chemotypes with high concentrations of 1,8-cineole, and that oviposition preference depended on the chemotype of the larval diet. Although performance traits and preference for oviposition of P. tigrina did not vary among chemotypes, adults inflicted less damage on plants with a high concentration of terpinolene. Additionally, we tested whether different chemotypes showed different levels of susceptibility by myrtle rust (Puccinia psidii). We found that plants with a high concentration of 1,8-cineole were more likely to be infected under controlled conditions. Although there is evidence that terpene chemotypes are a mediator of the interaction with natural enemies, the most detrimental pest of this plant, P. tigrina, does not seem to be affected by variation in plant terpenes.

Plant chemotypes or chemical polymorphisms are defined by discrete variation in secondary metabolites within a species. This variation can have consequences for ecological interactions or the human use of plants. Understanding the molecular basis of chemotypic variation can help to explain how variation of plant secondary metabolites is controlled. We explored the transcriptomes of the 3 cardinal terpene chemotypes of Melaleuca alternifolia in young leaves, mature leaves, and stem and compared transcript abundance to variation in the constitutive profile of terpenes. Leaves from chemotype 1 plants (dominated by terpinen-4-ol) show a similar pattern of gene expression when compared to chemotype 5 plants (dominated by 1,8-cineole). Only terpene synthases in young leaves were differentially expressed between these chemotypes, supporting the idea that terpenes are mainly synthetized in young tissue. Chemotype 2 plants (dominated by terpinolene) show a greater degree of differential gene expression compared to the other chemotypes, which might be related to the isolation of plant populations that exhibit this chemotype and the possibility that the terpinolene synthase gene in M. alternifolia was derived by introgression from a closely related species, Melaleuca trichostachya. By using multivariate analyses, we were able to associate terpenes with candidate terpene synthases.

Both theoretical and empirical works have highlighted the difference in the evolutionary implications of host resistance and tolerance against their enemies. However, it has been difficult to show evolutionary changes in host defences in natural populations; thus, evaluating theoretical predictions of simultaneous evolution of defences remains a challenge. We studied the evolutionary changes in traits related to resistance and tolerance against herbivory in a natural plant population using seeds from two collections made in a period of 20 years. In a common garden experiment, we compared defensive traits of ancestral (1987) and descendant (2007) subpopulations of the annual plant Datura stramonium that shows genetic variation for tolerance and to which the specialist herbivore Lema daturaphila is locally adapted. We also examined the effects of different plant genotypes on the herbivore for testing the plant genetic variation in resistance. Based on the response to the contemporary herbivore populations, results revealed a nonsignificant response in plant resistance traits (herbivore consumption, foliar trichomes and tropane alkaloids), but a significant one in tolerance. The survival of herbivores in laboratory experiments depended on the plant genotype, which suggests genetic variation in plant resistance. Although we cannot identify the selective agent for the change nor exclude genetic drift, the results are consistent with the expectation that when resistance fails to control herbivory, tolerance should play a more important role in the evolution of the interaction.

In order to quantify P accumulation and P efflux in the ectomycorrhizal basidiomycete fungus Hebeloma cylindrosporum, we supplied (32)P to mycelia previously grown in vitro in liquid medium. The culture had four main steps that are 1) growing the mycelium on complete medium with P, 2) transfer the mycelia into new culture solution with or without P, 3) adding a solution containing (32)P and 4) rinsing the mycelia before incubation with or without plant. The main point is to rinse very carefully the mycelia after (32)P supply in order to avoid overestimation of (32)P efflux into the medium.

In ectomycorrhizal plants, the fungal cells colonize the roots of their host plant to create new organs called ectomycorrhizae. In these new organs, the fungal cells colonize the walls of the cortical cells, bathing in the same apoplasm as the plant cells in a space named the ‘Hartig net’, where exchanges between the two partners take place. Finally, the efficiency of ectomycorrhizal fungi to improve the phosphorus nutrition of their host plants will depend on the regulation of phosphate transfer from the fungal cells to plant cells in the Hartig net through as yet unknown mechanisms. In order to investigate these mechanisms, we developed an in vitro experimental device mimicking the common apoplasm of the ectomycorrhizae (the Hartig net) to study the phosphorus metabolism in the ectomycorrhizal fungus Hebeloma cylindrosporum when the fungal cells are associated or not with the plant cells of the host plant Pinus pinaster. This device can be used to monitor (32)Phosphate efflux from the fungus previously incubated with (32)P-orthophosphate.

International audience; In response to cancer, AIDS, sepsis and other systemic diseases inducing muscle atrophy, the E3 ubiquitin ligase Atrogin1/MAFbx (MAFbx) is dramatically upregulated and this response is necessary for rapid atrophy. However, the precise function of MAFbx in muscle wasting has been questioned. Here, we present evidence that during muscle atrophy MAFbx targets the eukaryotic initiation factor 3 subunit 5 (eIF3-f) for ubiquitination and degradation by the proteasome. Ectopic expression of MAFbx in myotubes induces atrophy and degradation of eIF3-f. Conversely, blockade of MAFbx expression by small hairpin RNA interference prevents eIF3-f degradation in myotubes undergoing atrophy. Furthermore, genetic activation of eIF3-f is sufficient to cause hypertrophy and to block atrophy in myotubes, whereas genetic blockade of eIF3-f expression induces atrophy in myotubes. Finally, eIF3-f induces increasing expression of muscle structural proteins and hypertrophy in both myotubes and mouse skeletal muscle. We conclude that eIF3-f is a key target that accounts for MAFbx function during muscle atrophy and has a major role in skeletal muscle hypertrophy. Thus, eIF3-f seems to be an attractive therapeutic target

Plants show a high degree of intraspecific variation in several traits including plant secondary metabolites. This variation can be influenced by genetic and environmental factors that result in geographical structure in their distribution. By growing plants from several populations in a controlled environment, we studied variation in foliar terpenes in Eucalyptus camaldulensis, which is the widest distributed eucalypt, with a large range both latitudinally and longitudinally. We found that the concentration of terpenes is highly variable among subspecies. We identified four chemotypes dominated by 1,8-cineole, γ-terpinene, α- and β-phellandrene. While the 1,8-cineole chemotype is abundant in all populations, the other three chemotypes are rare in the central area and the north-east of Australia. The γ-terpinene chemotype is mainly restricted to the north and west of Australia, whereas the α- and β-phellandrene chemotypes show an opposite distribution in the north and south of the continent. The annual mean temperature and humidity of the source populations correlate with the abundance of the dominant terpenes. We also tested the effects of elevated CO2 concentrations on the terpene concentration and found that elevated CO2 atmosphere reduces the overall accumulation of foliar terpenes. The results suggest that variation in terpene composition in E. camaldulensis can be influenced by environmental variables, mainly favouring the 1,8-cineole chemotype in arid locations.