Cultivable fungal diversity in two karstic caves in Italy: under-investigated habitats as source of putative novel taxa.

Microbial diversity of caves is largely understudied and its possible applications are still unknown. Autochthonous fungi, in particular, may have the potential to biomineralize metals and may be used as promising agents for bioremediation of polluted sites; thus, unearthing the fungal diversity in hypogean ecosystems is nowadays of utmost importance. To start addressing this knowledge gap, the cultivable mycobiota of two neighbouring caves-one natural and one exploited for touristic purposes-were characterised and compared by studying fungi isolated from sediments collected at increasing distances from the entrance. Overall, 250 fungal isolates ascribable to 69 taxa (mainly Ascomycota) were found, a high percentage of which was reported in caves for the first time. The sediments of the touristic cave displayed a richer and more diversified community in comparison with the natural one, possibly due to visitors carrying propagules or organic material. Considering that these environments are still poorly explored, chances to detect new fungal lineages are not negligible.

Local versus landscape spatial influence on biodiversity: a case study across five European industrialized areas.

Land use change-mostly habitat loss and fragmentation-has been recognized as one of the major drivers of biodiversity loss worldwide. According to the habitat amount hypothesis, these phenomena are mostly driven by the habitat area effect. As a result, species richness is a function of both the extent of suitable habitats and their availability in the surrounding landscape, irrespective of the dimension and isolation of patches of suitable habitat. In this context, we tested how the extent of natural areas, selected as proxies of suitable habitats for biodiversity, influences species richness in highly anthropogenic landscapes. We defined five circular sampling areas of 5 km radius, including both natural reserves and anthropogenic land uses, centred in five major industrial sites in France, Italy and Germany. We monitored different biodiversity indicators for both terrestrial and aquatic ecosystems, including breeding birds, diurnal butterflies, grassland vegetation, odonata, amphibians, aquatic plants and benthic diatoms. We studied the response of the different indicators to the extent of natural land uses in the sampling area (local effect) and in the surrounding landscape (landscape effect), identified as a peripheral ring encircling the sampling area. Results showed a positive response of five out of seven biodiversity indicators, with aquatic plants and odonata responding positively to the local effect, while birds, vegetation and diatoms showed a positive response to the landscape effect. Diatoms also showed a significant combined response to both effects. We conclude that surrounding landscapes act as important biodiversity sources, increasing the local biodiversity in highly anthropogenic contexts.

Environmental drivers of phototrophic biofilms in an Alpine show cave (SW-Italian Alps).

The proliferation of lampenflora is a major threat for the conservation of show caves, since phototrophic organisms cause physical, chemical and aesthetic damage to speleothems. In this paper we examine the environmental factors influencing the presence and the growth of the three main photosynthetic groups composing phototrophic biofilms in the Bossea show cave (SW-Italian Alps). The presence and the primary production of cyanobacteria, diatoms and green algae were detected with BenthoTorch®, an instrument for in situ measurement of chlorophyll a concentration that has never been used before in caves. By means of different techniques of regression analysis, we highlighted the response of the three photosynthetic groups to different environmental factors. Illuminance proved to be the main factor influencing positively both the probability of the presence and the productivity of the three groups. The presence of seeping water on the substrate and the distance from the cave entrance proved to play an important role in determining patterns of colonization. By means of GIS techniques, we provide thematic maps of the cave, providing a representation of pattern of the density of the three examined photosynthetic groups within different areas of the cave. The same approach may apply to other show caves, aiming at providing suggestions for the cave management (i.e. cleaning of the cave walls and positioning of artificial lights) and reduce impact caused by tourism.

Production of transgenic barrel medic (Medicago truncatula Gaernt.) using the ipt-type MAT vector system and impairment of Recombinase-mediated excision events.

Expression of the uidA reporter gene was tested in transformation experiments of barrel medic (Medicago truncatula Gaertn.) with the ipt-type control vectors pIPT5, pIPT10 and pIPT20 and distinct in vitro culture conditions. The highest GUS expression levels were obtained with the pIPT10 construct carrying the ipt gene under the control of the native ipt promoter and using kanamycin as selective agent. The ipt-shooty transformants, characterized by the absence of both rooting ability and apical dominance associated with vitrification, were easily identified by visual selection. Using only the ipt gene as selectable marker, we obtained a stable transformation frequency of 9.8% with pIPT10 construct. The ipt-type MAT vector pEXM2 was then used to monitor the excision events mediated by the yeast Recombinase and the consequent production of ipt marker-free transgenic plants. Transgenic ipt-shooty lines were recovered at a frequency of 7.9% in the absence of kanamycin-based selection. The ipt-shooty phenotype was maintained in all the transgenic lines and no reversion to the normal phenotype occurred. PCR analysis revealed the presence of the 'hit and run' cassette in the genome of all the regenerated ipt-shooty lines while RT-PCR experiments confirmed the expression of the R gene, encoding the yeast Recombinase. A detailed molecular investigation, carried out to verify the integrity of the RS sites, revealed that these regions were intact in most cases. Our results with barrel medic suggest that the MAT system must be carefully evaluated and discussed on a case by case basis.

Use of artificial environments to reproduce and exploit genotype x location interaction for lucerne in northern Italy.

Genotype x environment interaction effects can be exploited by breeding for specific adaptation to well-defined subregions within a target region. Previous work showed that genotype x location interaction for dry matter (DM) yield of lucerne (Medicago sativa L. subsp. sativa) cultivars in northern Italy is large and associated with soil type and level of summer drought stress of locations, suggesting the presence of two contrasting subregions. Thirteen farm landraces collected across the region and four control varieties were evaluated for DM yield in four artificial environments created at one site by the factorial combination of soil type (sandy loam or silty clay) and drought stress level (almost nil or high) for: (1) exploring the possibility to reproduce in artificial environments the adaptation patterns occurring across the region; (2) investigating the adaptation pattern of landraces and its relationship with environmental factors at collecting sites; and (3) providing a preliminary comparison of wide- versus specific-adaptation strategies based on yield gains predicted from selection of populations. Different soils filled large (24.0x1.6x0.8-m deep), bottomless containers in concrete. Water amounts were controlled by irrigation under a moving rain shelter. Cultivars varied largely for adaptation pattern across the artificial environments, mainly due to cultivar x stress interaction. Better response to stress conditions of landraces was closely associated with the level of summer drought at collecting sites (r=0.82), highlighting the importance of evolutionary adaptation. The additive main effects and multiplicative interaction-modelled responses of control cultivars successfully reproduced those observed across locations, candidating the artificial environments as a cheaper alternative to more selection locations when breeding for wide or specific adaptation. The latter implied about 40-50% greater estimated gains relative to breeding for wide adaptation.

Lensless digital holographic microscope with light-emitting diode illumination.

We demonstrate the operation of a digital in-line microscope with LED illumination. We show with a practical example that, for typical setups, the limited temporal coherence and the spatial incoherence of the source do not affect the resolving power of the system. On the contrary, important advantages are obtained in terms of signal-to-noise ratio and alignment simplification.

Interference effects in white clover genotypes grown as pure stands and binary mixtures with different grass species and varieties.

Six white clover genotypes and eight grass varieties belonging to four different species were grown both in monoculture and as grass-legume binary mixtures in dense swards for two years under a mowing regime and a management including N fertilization. Dry matter yield and yield-related traits were recorded to investigate some aspects of inter-specific interference in white clover-based mixtures and to define a methodology for selecting genotypes of this clover suited to conditions of association. Clover was at a competitive disadvantage in most mixtures. Differences among grasses for aggressiveness were related more to variety vigour than to species. Clover compatibility proved specific only in relation to grass vigour. Variation among clovers for tolerance to competitive stress involved significant cross-over interactions passing from monoculture to severe stress conditions for clover yield and other traits, and was related positively to stolon density and negatively to yield and leaf gigantism traits recorded in monoculture. Clover selection for high levels of competitive stress seems possible either by genotype assessment in stress conditions or by a combination of high yield and stolon density assessed in monoculture.

The value and limitations of the CO2 laser in neurosurgery.

A report is made of 42 patients operated on by the 100 W CO2 laser from May 1979 to May 1980 and the effects of the continuous and pulsed emission of the beam are discussed. The usefulness of the Laser-microscope coupling is also reported. One estimates the average amount of blood transfused during and soon after the operation and this is compared with blood transfused in 42 patients operated by a 50 W CO2 laser and 42 patients operated by traditional techniques. The authors have been using the CO2 laser routinely since May 1977. Its use has been sometimes useful, sometimes very useful or indispensable, sometimes useless and/or dangerous.