The two‐component histidine kinase Fhk1 controls stress adaptation and virulence of Fusarium oxysporum

Fungal histidine kinases (HKs) have been implicated in different processes, such as the osmostress response, hyphal development, sensitivity to fungicides and virulence. Members of HK class III are known to signal through the HOG mitogen‐activated protein kinase (MAPK), but possible interactions with other MAPKs have not been explored. In this study, we have characterized fhk1, encoding a putative class III HK from the soil‐borne vascular wilt pathogen Fusarium oxysporum. Inactivation of fhk1 resulted in resistance to phenylpyrrole and dicarboximide fungicides, as well as increased sensitivity to hyperosmotic stress and menadione‐induced oxidative stress. The osmosensitivity of Δfhk1 mutants was associated with a striking and previously unreported change in colony morphology. The Δfhk1 strains showed a significant decrease in virulence on tomato plants. Epistatic analysis between Fhk1 and the Fmk1 MAPK cascade indicated that Fhk1 does not function upstream of Fmk1, but that the two pathways may interact to control the response to menadione‐induced oxidative stress.     

Ant cues affect the oviposition behaviour of fruit flies (Diptera: Tephritidae) in Africa

Abstract. Although most studies on fruit fly oviposition behaviour focus on horizontal interactions with competitors and cues from host plants, vertical interactions with predators are poorly documented. The present study provides direct evidence indicating that the oviposition behaviour of the two main mango fruit fly species, Ceratitis cosyra (Walker) and Bactrocera invadens Drew‐Tsurata & White, is affected by secretions of the dominant arboreal ant Oecophylla longinoda (Latreille). When offered ant‐exposed and unexposed mangoes in the absence of the ants, both fly species are reluctant to land on ant‐exposed fruits and, when having landed, often take off quickly and fail to oviposit. The number of puparia collected from unexposed mangoes is approximately eight‐fold higher than from ant‐exposed ones. The results obtained from laboratory experiments and field observations confirm that adult fruit flies are more affected through repellence by ant cues than by direct predation. The use of cues by fruit flies in predator avoidance has implications for evolutionary ecology, behavioural ecology and chemical ecology.     

Microtubular Systems of Paramecium in Division: Pattern of Cytospindle Assembly

The pattern of cytospindle assembly and the modifications of the microtubular cytoplasmic network during division of Paramecium are studied by means of indirect immunofluorescence. The assembly of cytospindle starts at two independent areas placed respectively around the proter's and opisthe's buccal overture. The moment of the microtubule bundles’appearance depends on their distance from the buccal opening, with those closest appearing 1st. The existence of microtubule organizing centers that act transiently during division of Paramecium is discussed.     

Characterization of 16 highly variable tetranucleotide microsatellite loci for Oregon chub (Oregonichthys crameri) and cross amplification in Umpqua chub (O. kalawatseti)

We describe the isolation and development of 16 polymorphic tetranucleotide microsatellite loci for the endangered Oregon chub (Oregonichthys crameri). Two loci appear to be duplicated. For the remaining 14 loci, we observed between three and 19 alleles per locus in a sample of 42 fish. Thirteen of these loci were also polymorphic in the closely related Umpqua chub (O. kalawatseti). These loci will aid in our understanding of the molecular ecology and conservation of these two species.     

Testing the role of male–male competition in the evolution of sexual dimorphism: a comparison between two species of porcelain crabs

Theory predicts marked sexual dimorphism in terms of body size and body structures used as weapons (e.g. chelipeds) in gonochoric species with intense male sexual competition for receptive females and reduced or no sexual dimorphism in species where competition among males is trivial. We tested this hypothesis using a pair of closely‐related species of symbiotic porcelain crabs as a model. In one species that inhabits sea anemones solitarily, competition among males for receptive females is unimportant. In a second species that dwells as dense aggregations on sea urchins, male–male competition for sexual partners is recurrent. We expected considerable sexual dimorphism in body size and weaponry in the urchin‐dwelling crab and reduced sexual dimorphism in the anemone‐dwelling crab. In agreement with expectations, in the urchin‐dwelling crab, male body size was, on average, larger than that of females and males invested considerably more to cheliped length than females. Also supporting theoretical considerations, in the anemone‐dwelling crab, sexual dimorphism in terms of body size was not detected and differences between the sexes in investment to cheliped length were minor. Interestingly, chelipeds were more developed both in males and females of the anemone‐dwelling crab than in the urchin‐dwelling crab as a result of the importance of these structures for monopolization of their naturally scarce anemone hosts. Another difference between the studied species was the existence of two clearly distinguishable ontogenetic phases in males of the urchin‐dwelling crab but not in males of the anemone‐dwelling crab. Whether the two different male morphs display different male reproductive strategies in the urchin‐dwelling crab remains to be addressed. Other conditions that might additionally explain the observed differences in sexual dimorphism (e.g. female mate choice) between the studied species remain to be explored. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 548–558.     

OUTLINING ETHICAL ISSUES IN NANOTECHNOLOGIES

Nanotechnologies are an expression of the human ability to control and manipulate matter on a very small scale. Their use will enable an even and constant monitoring of human organisms, in a new and perhaps less invasive way. Debates at all levels – national, European and international – have pointed out the common difficulty of giving a complete, clear definition of nanotechnologies. This is primarily due to the variety of their components, to the fact that there is not just one technology but several.
The most significant medical applications of nanotechnologies are in the diagnostic and the therapeutic fields, eg biosensors and molecular imaging, providing diagnosis and drug delivery with no invasive methods involved.
Like any other emerging field, such technologies imply new possibilities for improving health but, on the other hand, they are still at an experimental stage and therefore should be implemented under rigorous safety testing before going on general release. For this purpose, the ethical, legal and social implications (ELSI) of nanotechnologies have been elaborated by study groups, in order to develop solutions before the results of the tests are diffused into medical practice.
The aim of this paper is to define some of the ethical issues concerning biomedical applications and to evaluate whether there is a need for new or additional guidelines and regulations.     

cpFBPaseII, a novel redox-independent chloroplastic isoform of fructose-1,6-bisphosphatase

A full-length FBPase cDNA has been isolated from Fragaria  ×  ananassa (strawberry) corresponding to a novel putative chloroplastic FBPase but lacking the regulatory redox domain, a characteristic of the plastidial isoenzyme (cpFBPaseI). Another outstanding feature of this novel isoform, called cpFBPaseII, is the absence of the canonical active site. Enzymatic assays with cpFBPaseII evidenced clear Mg²⁺-dependent FBPase activity and a K _m for fructose-1,6-bisphosphate (FBP) of 1.3 m m . Immunolocalization experiments and chloroplast isolation confirmed that the new isoenzyme is located in the stroma. Nevertheless, unlike cpFBPaseI, which is redox activated, cpFBPaseII did not increase its activity in the presence of either DTT or thioredoxin f (TRX f ) and is resistant to H₂O₂ inactivation. Additionally, the novel isoform was able to complement the growth deficiency of the yeast FBP1 deletion fed with a non-fermentable carbon source. Furthermore, orthologues are restricted to land plants, suggesting that cpFBPaseII is a novel and an intriguing chloroplastic FBPase that emerged late in the evolution of photosynthetic organisms, possibly because of a pressing need of land plants.     

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.