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.     

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.     

Effects of feed ingredients on nutrient digestibility,nitrogen/energy balance and morphology changes in the intestine of Nile tilapia (Oreochromis niloticus)

The present study assessed the effect of different feed ingredients on nutrient apparent digestibility coefficients (ADC), nitrogen/energy balance and morphology changes in the intestine of Nile tilapia; using a control diet and six test diets, in which the following six ingredients were included at 30%: hydrolysed feather meal (HFM), soybean meal (SBM), rice bran (RB), rapeseed meal (RM), sunflower meal (SFM) and dried distillers grains with solubles (DDGS). The proximal, middle and distal intestine was processed for quantitative histology counting the number of goblet cells (GC), and measuring the thickness of lamina propria (LP) and submucosa (SM). The study showed that the ADC of protein in raw materials were highest in SBM (92.2%), followed by SFM (90.2%), DDGS (89.2%), RM (87.8%), HFM (86.9%) and RB (84.0%). The nutrient ADCs had no correlation with intestinal morphology changes. Only the SBM diet caused noticeable changes in intestinal morphology such as an increase the thickness of SM and LP and the number of GC. The diet composition, however, altered the protein efficiency and the maintenance energy requirement. Protein retention efficiency was the lowest in fish fed HFM and the highest in RB. The highest maintenance energy requirements were observed in HFM and SBM treatments.     

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.     

HD 181068: a red giant in a triply eclipsing compact hierarchical triple system

Hierarchical triple systems comprise a close binary and a more distant component. They are important for testing theories of star formation and of stellar evolution in the presence of nearby companions. We obtained 218 days of Kepler photometry of HD 181068 (magnitude of 7.1), supplemented by ground-based spectroscopy and interferometry, which show it to be a hierarchical triple with two types of mutual eclipses. The primary is a red giant that is in a 45-day orbit with a pair of red dwarfs in a close 0.9-day orbit. The red giant shows evidence for tidally induced oscillations that are driven by the orbital motion of the close pair. HD 181068 is an ideal target for studies of dynamical evolution and testing tidal friction theories in hierarchical triple systems.     

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.     

Mass mortality of common eiders (Somateria mollissima) in the Dutch Wadden Sea, winter 1999/2000: starvation in a commercially exploited wetland of international importance

Mass mortality of common eiders (Somateria mollissima) was observed in winter 1999/2000 in the Dutch Wadden Sea. Approximately 21,000 common eiders died. Dissected birds were severely emaciated and 94% were infected with the acanthocephalan parasite Profilicollis botulus. Green shore crabs (Carcinus maenas), intermediate hosts of the parasite, were slightly more ‘available’ than in other years, but parasite infections in the eiders were close to normal. Few eiders were oiled (5%), there were no toxicological, bacteriological, or virological explanations for the observed mortality.In the Wadden Sea, a wetland of international importance, mussel (Mytilus edulis) cultures occur in sublittoral areas, while mechanical cockle (Cerastoderma edule) fisheries are licensed annually after evaluation of available resources. The wintering eiders in 1999/2000 required c. 3.1 million kg ash-free dry mass, while information on mussel and cockle stocks (irrespective of accessibility and profitability) suggested a resource 4.7× the requirement of common eiders only. Food shortage is suggested to have caused the observed mortality, involving both principal (mussels and cockles) and secondary (Spisula) prey.Winter census reports showed shifts in wintering distribution of common eiders in the 1990s, indicating the utilisation of Spisula in the North Sea in poor food years in the Wadden Sea. Following particularly intense fisheries in summer 1999, attempts to feed on Spisula in winter 1999/2000 failed. It is hypothesised that overfishing of mussels and cockles in the Wadden Sea in the early 1990s resulted in structurally reduced food resources, contractions of the foraging area of common eiders, and increased use of secondary prey in the North Sea.