Pathogenicity of <Emphasis Type="Italic">Stemphylium vesicarium</Emphasis> from different hosts causing brown spot in pear

Stemphylium vesicarium (teleomorph: Pleospora herbarum) is the causal agent of brown spot disease in pear. The species is also able to cause disease in asparagus, onion and other crops. Saprophytic growth of the fungus on plant debris is common. The objective of this study was to investigate whether isolates of S. vesicarium from different hosts can be pathogenic to pear. More than hundred isolates of Stemphylium spp. were obtained from infected pear fruits, dead pear leaves, dead grass leaves present in pear orchard lawns as well as from necrotic leaf parts of asparagus and onion. Only isolates originating from pear orchards, including isolates from dead grass leaves, were pathogenic on pear leaves or fruits in bioassays. Non-pathogenic isolates were also present in pear orchards. Stemphylium vesicarium from asparagus or onion, with one exception, were not pathogenic to pear. Analysis of the genetic variation between isolates using Amplified Fragment Length Polymorphism (AFLP) showed significant concordance with host plants. Isolates from asparagus or onion belonged to clusters separate from the cluster with isolates from pear or grass leaves collected in pear orchards. Multilocus sequencing of a subset of isolates showed that such isolates were similar to S. vesicarium.     

Partial stem and leaf resistance against the fungal pathogen <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in wild relatives of tomato

Tomato (Solanum lycopersicum) is one of many greenhouse crops that can be infected by the necrotrophic ascomycete Botrytis cinerea. Commercial cultivation of tomato is hampered by the lack of resistance. Quantitative resistance has been reported in wild tomato relatives, mostly based on leaf assays. We aimed to identify wild tomato relatives with resistance to B. cinerea based on quantitative assays both on leaves and stem segments, monitoring infection frequency and disease expansion rate as parameters. A quantitative tomato stem segment assay was developed. This stem assay and a previously described leaf assay were used to screen a collection of 22 Solanum accessions. Significant differences in disease parameters were observed among accessions. Resistance to B. cinerea was observed in a number of wild Solanum species, including accessions of S. chilense, S. habrochaites and S. neorickii, both in the leaf assay and the stem segment assay. A number of resistant and susceptible accessions were evaluated as adult plants under greenhouse conditions. The data obtained in greenhouse assays confirmed the leaf and stem disease data. The expression of several defence-related genes was studied in a subset of accessions. There was no apparent correlation between the expression levels of the genes tested and the quantitative resistance level to B. cinerea. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Cluster analysis of 36Globodera pallida field populations using two sets of molecular markers

Thirty-six populations of the potato cyst nematodeGlobodera pallida, all collected in the Netherlands, were analysed twice: by two-dimensional gel electrophoresis of proteins (2-DGE) and by random amplified polymorphic DNA fingerprinting (RAPD). Two-DGE revealed frequencies of 21 alleles at eight putative loci in each population. The same populations were subjected to RAPD analysis. This qualitative technique revealed 38 polymorphic DNA fragments. Both datasets were independently processed to determine the intraspecific variation. UPGMA analysis resulted in a 2-DGE- and a RAPD-based dendrogram with cophenetic correlation coefficients of 0.755 and 0.838 respectively. The correlation between the genetic similarity values for the populations was 0.572. Comparison between the 2-DGE- and the RAPD-based dendrogram revealed that only thirteen of the 36 populations analysed were clustered identically. It is concluded that the gene pool similarity concept is only in some instances applicable to Dutch populations ofG. pallida. For populations that could not be differentiated unequivocally on the basis of molecular markers, markers closely linked to avirulence genes should be identified. Approaches that will lead to the identification of such markers are discussed.     

Immunity to Babesia in mice. I. Adoptive transfer of immunity to Babesia rodhaini with immune spleen cells and the effect of irradiation on the protection of immune mice

Immunisation of Balb/c mice against Babesia rodhaini by an amicarbalide-controlled infection resulted in a solid immunity which lasted for 216 days. With spleen cells of immune mice protection could be transferred both to naive mice pretreated with cyclophosphamide. Treatment of naive mice with cyclophosphamide (300 mg/kg) five days before a lethal B. rodhaini inoculation resulted in over 50% survival. This protective effect of cyclophosphamide is explained by its inhibiting effect on suppressor T-cells. The protection against B. rodhaini challenge infection afforded to immune Balb/c mice was completely resistant to a sublethal irradiation of 400 rad. Since B-lymphocyte function in antibody production is suppressed by this dose, the role of antibodies in the effector phase of the immunity appears to be of minor if any importance. A considerable degree of protection was still preserved after irradiation of immune animals with 875 rad. Sensitivity to this irradiation dose of all immunocompetent cells except macrophages and a small fraction of T-lymphocytes indicates the involvement of these cell types in the effector phase of the specific immunity. Highly radioresistant macrophages are therefore considered to play the major role but T-lymphocytes are also required for complete protection.     

Social interactions in urban parks: Stimulating social cohesion?

People from all ethnic backgrounds spend some of their leisure time in green areas. This study found that urban parks are more inclusive green places than non-urban green areas, and that urban parks can promote social cohesion. The objective of the research was to establish the extent to which urban parks facilitate social cohesion and how social interaction and place attachment can contribute to such cohesion. Quantitative research (a survey) and qualitative research (observations and interviews) carried out in five urban parks in the Netherlands revealed that there are many similarities in the ways that ethnic groups use urban parks and in the meanings of such parks to these groups. Urban parks are sites where different ethnic groups mingle and where informal and cursory interactions can stimulate social cohesion. Furthermore, being involved and concerned with parks can facilitate attachment to these places. Urban parks can provide a vital locality where everyday experiences are shared and negotiated with a variety of people. The design of a park, its location and people's image of the park in combination with the cultural characteristics of various ethnic groups inform the opportunities for intercultural interactions.     

From Hiking Through Farmland to Farming in a Leisure Landscape: Changing Social Perceptions of the European Landscape

The idea that landscape has been created by human activities on a biophysical basis allows for clear cause–effect reasoning. However, landscape planning and management practice learns that it is impossible to neglect the social perception of landscape, i.e. the ways people think about nature and landscape. It is the result of social research and human sciences of the last decade that a differentiation in views of nature and landscape can be identified in the different groups of social actors in the landscape. Case studies from France and the Netherlands show a marked change in values attributed to nature and landscape in the end of the last century. Social demand for landscape is growing and a shift from a functional image of nature and landscape to a more hedonistic image like the Arcadian and wilderness images has taken place. Comparing the Netherlands with France and rural with urban inhabitants, the influence of urbanisation is evident in this process. It is further shown that images of nature vary considerably between for example farmers, urban residents, hunters and conservationists. The way people perceive landscape seems determined by their functional ties with the landscape and the social praxis in which they encounter the landscape. It is concluded that the concept of landscape is nearer to the lifeworld of people than the abstract notions of nature and biodiversity. This implies a big challenge both for national and international landscape policies and for local landscape management initiatives to be developed, taking into due consideration both the material and immaterial nature of landscape.     

Screening of bacterial isolates from various European soils for in vitro antagonistic activity towards Rhizoctonia solani and Fusarium oxysporum: Site-dependent composition and diversity revealed

A cultivation-based approach was used to determine the in vitro antagonistic potential of soil bacteria towards Rhizoctonia solani AG3 and Fusarium oxysporum f. sp. lini (Foln3). Four composite soil samples were collected from four agricultural sites with previous documentation of disease suppression, located in France (FR), the Netherlands (NL), Sweden (SE) and the United Kingdom (UK). Similarly, two sites from Germany (Berlin, G-BR; and Braunschweig, G-BS) without documentation of disease suppression were sampled. Total bacterial counts were determined by plating serial dilutions from the composite soil samples onto R2A, AGS and King's B media. A total of 1,788 isolates (approximately 100 isolates per medium and site) was screened for antifungal activity, and in vitro antagonists (327 isolates) were found amongst the dominant culturable bacteria isolated from all six soils. The overall proportion of antagonists and the number of isolates with inhibitory activity against F. oxysporum were highest in three of the suppressive soils (FR, NL and SE). Characterization of antagonistic bacteria revealed a high phenotypic and genotypic diversity. Siderophore and protease activity were the most prominent phenotypic traits amongst the antagonists. The composition and diversity of antagonists in each soil was site-specific. Nevertheless, none of the antimicrobial traits of bacteria potentially contributing to soil suppressiveness analyzed in this study could be regarded as specific to a given site.     

A major genome region underlying artemisinin resistance in malaria

Evolving resistance to artemisinin-based compounds threatens to derail attempts to control malaria. Resistance has been confirmed in western Cambodia and has recently emerged in western Thailand, but is absent from neighboring Laos. Artemisinin resistance results in reduced parasite clearance rates (CRs) after treatment. We used a two-phase strategy to identify genome region(s) underlying this ongoing selective event. Geographical differentiation and haplotype structure at 6969 polymorphic single-nucleotide polymorphisms (SNPs) in 91 parasites from Cambodia, Thailand, and Laos identified 33 genome regions under strong selection. We screened SNPs and microsatellites within these regions in 715 parasites from Thailand, identifying a selective sweep on chromosome 13 that shows strong association (P = 10(-6) to 10(-12)) with slow CRs, illustrating the efficacy of targeted association for identifying the genetic basis of adaptive traits.     

Fungal and plant gene expression during synchronized infection of tomato leaves by Botrytis cinerea

An inoculation procedure was developed to obtain efficient and synchronous infection on detached tomato leaves by Botrytis cinerea. In spray-inoculated leaves incubated at 20 °C, the infection process consisted of three phases: the formation of primary necrotic lesions (until 20 hpi), a quiescent phase (20-72 hpi), and the expansion of a proportion of the primary lesions (from 72 hpi onwards), resulting in full tissue maceration. At 4 °C, the infection progressed slowly but steadily without inducing necrotic responses in the host. The actin and -tubulin genes of B. cinerea were cloned, characterized and used as probes on blots containing RNAs from leaves at various stages of the infection. The genes displayed a similar expression pattern throughout the infection and the hybridization signal reflected the amount of fungal biomass. The actin mRNA accumulated to higher levels than the -tubulin mRNA. Tomato PR protein mRNAs (chitinase, -1,3-glucanase and PR-1) were induced during the infection, albeit with different kinetics and to different levels. At 20 °C, -1,3-glucanase and PR-1 mRNAs were induced more rapidly than chitinase mRNAs. At 4 °C, mRNAs encoding extracellular -1,3-glucanase and intracellular, as well as extracellular chitinase were hardly induced.