Identification of virulence genes in Fusarium oxysporum f. sp. lycopersici by large-scale transposon tagging

Forward genetic screens are efficient tools for the dissection of complex biological processes, such as fungal pathogenicity. A transposon tagging system was developed in the vascular wilt fungus Fusarium oxysporum f. sp. lycopersici by inserting the novel modified impala element imp160::gfp upstream of the Aspergillus nidulans niaD gene, followed by transactivation with a constitutively expressed transposase. A collection of 2072 Nia⁺ revertants was obtained from reporter strain T12 and screened for alterations in virulence, using a rapid assay for invasive growth on apple slices. Seven strains exhibited reduced virulence on both apple slices and intact tomato plants. Five of these were true revertants showing the re-insertion of imp160::gfp within or upstream of predicted coding regions, whereas the other two showed either excision without re-insertion or no excision. Linkage between imp160::gfp insertion and virulence phenotype was determined in four transposon-tagged loci using targeted deletion in the wild-type strain. Knockout mutants in one of the genes, FOXG_00016 , displayed significantly reduced virulence, and complementation of the original revertant with the wild-type FOXG_00016 allele fully restored virulence. FOXG_00016 has homology to the velvet gene family of A. nidulans . The high rate of untagged virulence mutations in the T12 reporter strain appears to be associated with increased genetic instability, possibly as a result of the transactivation of endogenous transposable elements by the constitutively expressed transposase.     

The powdery mildew fungus Podosphaera fusca (synonym Podosphaera xanthii), a constant threat to cucurbits

Numerous vegetable crops are susceptible to powdery mildew, but cucurbits are arguably the group most severely affected. Podosphaera fusca (synonym Podosphaera xanthii ) is the main causal agent of cucurbit powdery mildew and one of the most important limiting factors for cucurbit production worldwide. Although great efforts have been invested in disease control, by contrast, many basic aspects of the biology of P. fusca remain unknown.
Taxonomy: Podosphaera fusca (Fr.) Braun & Shishkoff. Kingdom Fungi; Phylum Ascomycota; Subdivision Pezizomycotina; Class Leotiomycetes; Order Erysiphales; Family Erysiphaceae; genus Podosphaera ; species fusca.
Identification: Superficial persistent mycelium. Conidia in chains, hyaline, ellipsoid to ovoid or doliform, about 24–40 × 15–22 µm, with cylindrical or cone-shaped fibrosin bodies, which often germinate from a lateral face and produce a broad, clavate germ tube and cylindrical foot-cells. Unbranched erect conidiophores. Cleistothecia globose, mostly 70–100 µm in diameter, dark brown/black. One ascus per cleistothecium with eight ascospores.
Host range: Angiosperm species that include several families, such as Asteracea, Cucurbitaceae, Lamiaceae, Scrophulariaceae, Solanaceae and Verbenaceae.
Disease symptoms: White colonies develop on leaf surfaces, petioles and stems. Under favourable environmental conditions, the colonies coalesce and the host tissue becomes chlorotic and usually senesces early.
Control: Chemical control and the use of resistant cultivars. Resistance has been documented in populations of P. fusca to some of the chemicals registered for control.     

Effects of solar radiation on the endemic Mediterranean red alga Rissoella verruculosa: photosynthetic performance, pigment content and the activities of enzymes related to nutrient uptake

The effects of full-spectrum solar radiation and of solar radiation deprived of total u.v. radiation (λ<395 nm) on the endemic Mediterranean red alga Rissoella verruculosa (Bertoloni) J. Agardh were studied in situ in early summer, from sunrise to sunset. Photosynthetic performance, pigment content and the activities of enzymes related to nutrient uptake, were monitored under both radiation conditions throughout a daylight period.
The doses of solar radiation, measured on the day during which the experiments were carried out (24 June 1997), were 9228·25, 1109·70 and 13·03 kJ m⁻² for PAR (λ=400–700 nm), u.v.-A (λ=315–400 nm) and u.v.-B (λ=280–315 nm), respectively. Under these conditions, a clear daily variation in photosynthetic performance was found. However, no significant differences were detected between the two radiation conditions. At noon, strong sunlight impaired O₂ evolution by 75%, but complete recovery occurred during the afternoon, reaching similar values to those measured in the early morning. By contrast, photoinhibition of optimal quantum yield ( F _v/ F _m), and the relative electron transport rate was followed by only a slight recovery during the afternoon. The rate of photosynthesis in air (simulating emersion conditions), estimated by CO₂ exchange, showed a negative balance at noon, which was accentuated in plants exposed to the full-solar spectrum.
Significant changes in the activities of nitrate reductase and carbonic anhydrase were found throughout the day under both radiation conditions. In thalli receiving solar radiation deprived of u.v., total CA activity decreased throughout the day. However, in thalli exposed to full-spectrum solar radiation, the activity of the enzyme tended to increase in the afternoon, correlating with an increase in NR activity.     

Effects of resource availability and distribution on autumn movements of the Nutcracker Nucifraga caryocatactes in the Alps

Radiotracking was used to study the ranging behaviour and movements of the Nutcracker Nucifraga caryocatactes in the Alevé forest (northwest Italian Alps), from September to December 1994-96, when Nutcrackers collected seeds of Arolla Pines Pinus cembra. Home range size and overlap decreased from September to December. Autumn ranging behaviour was not consistent with territoriality. Nutcrackers stored seeds throughout the Alevé; many apparently stored seeds in scree at high altitudes (above 2200 m) where the forest comes to an end, and also in Larch forests opposite the Alevé (probably to avoid competition with rodents and conspecifics). Estimates of Nutcracker abundance, derived from transect counts, varied significantly among habitats, years and months, which possibly reflected differences in seed availability and abundance. The ranging behaviour observed at the Alevé was compared with that monitored at Mont Avic, another area in the northwest Italian Alps, where resources (in this case Hazel Corylus avellana nuts) were concentrated in a few zones. Home range size was significantly larger at Mont Avic, especially during late autumn. There, the home range size increased and overlap decreased from September to November, when birds abandoned Hazels and dispersed to the coniferous woods. Differences between Alevé and Mont Avic implied that the availability and distribution of resources are the major determinants of the ranging behaviour of the Nutcracker during autumn. However, differences between adults and juveniles were also found.     

Protein and oil concentration of soybean seed cultured in vitro using nutrient solutions of differing glutamine concentration

Oil and protein are the most valuable components of soybean seed. Evidence indicates that growth and composition of soybean seed are controlled by supplies of carbon and nitrogen provided by the maternal plant to the seed, but it is difficult experimentally to control and quantify the precise amount of carbon and nitrogen provided to the seed by the whole plant. To examine whether oil and protein concentrations are affected by the supply of nitrogen to the seed, immature soybean seeds (Glycine max cv. Williams 82) were grown in vitro in nutrient solutions containing 20, 40, 60 or 80 mM of glutamine. The seeds were incubated in Erlenmeyer flasks for 8 days at 25°C. The rate of dry matter accumulation changed from 7.2 to 8.3 mg seed⁻¹ day⁻¹ as the glutamine concentration increased from 20 to 80 mM but the differences were not significant (P 0.05). Seed protein concentration increased as glutamine concentration increased from 294 mg g⁻¹ at 20 mM glutamine to as high as 445 mg g⁻¹ at 80 mM glutamine. Typical in vivo protein concentration of mature soybean seeds is about 400 mg g⁻¹. Oil and protein concentrations were negatively correlated (r²= 0.44), which indicates that oil and protein synthesis are interrelated. Protein synthesis was favoured over oil synthesis when nitrogen became more abundant. The seeds used in this study clearly demonstrated a capacity to respond to nitrogen availability with changes in seed protein concentration.     

A new Spanish species of Linaria Miller (Scrophulariaceae)

Linaria arabiniana sp. nov. is described from coastal sand dunes of a very reduced area in Alicante Province (south-eastern part of the Iberian Peninsula). It is a perennial herb characterized by its 3–4-verticillate leaves, glabrous stems, large violet or rarely yellow flowers, and bicoloured usually flat seeds. Morphological characteristics and taxonomic affinities of this taxon are discussed, as are data on its ecology, phytosociology, and eventual conservation features.     

Water Uptake and Protein Synthesis in Germinating Wheat Embryos under the Osmotic Stress of Polyethylene Glycol

The effect of osmotic stress on wheat-seed germination was testedby imbibition in aqueous polyethylene glycol solutions at differentconcentrations. The experiments were designed to allow blockingand the subsequent recovery of germination by 12 h or 24 h pre-imbibitionof seeds in osmoticum, followed by transfer to water. Seedswere alternatively presoaked in water for 12 or 24 h, then transferredto polyethylene-glycol solutions to study the induced blockingof germination. Water content and [³H]leucine incorporationinto embryo tissues (as a measure of in vivo protein synthesis)were determined over a 48-h imbibition period. A close relationshipwas established overall between hydration status and proteinsynthesis rate. Osmotic stress seems to have a strong influenceupon the quantitative synthesis of proteins, suggesting thatthis biochemical activity is associated with the regulationof the germination process. Triticum durum, embryo, osmotic stress, water uptake, protein synthesis     

The Surface Charge of Tritrichomonas foetus

The surface charge of Tritrichomonas foetus was evaluated by means of the binding of colloidal iron hydroxide particles at pH 1.8 and cationized ferritin particles at pH 7.2 to the cell surface, as visualized by electron microscopy and by direct measurements of the electrophoretic mobility (EPM), of cells suspended in solutions of different ionic strength and pH. At pH 7.2, T. foetus has a negative surface charge with a mean EPM of ?1.03 μmμs^?1μV^?1μcm. At lower pH, there is a decrease in the negative surface charge with an isoelectric point at pH 1.2. At higher pH (> 9.0), there is an increase in the surface charge reaching an EPM of ?2.5 μmμs^?1μV^?1μcm. These results indicate that the surface of T. foetus contains both negatively and positively charged dissociating groups. Binding of colloidal iron hydroxide and cationized ferritin particles throughout the cell surface of the protozoon was observed. Treatment of T. foetus with neuraminidase or trypsin reduced significantly the EPM of the cells. Enzyme-treated cells recovered their normal EPM when incubated for 6 h in fresh culture medium by a process that is inhibited by puromycin.