S1.20 Divalent metal binding to bovine heart F1 ATPase: An FT-ESEEM study

The divalent metal binding sites of beef heart mitochondria F1ATPase were studied by FT-ESEEM spectroscopy, using Mn(II) as a paramagnetic probe, which replaces the naturally occurring Mg(II) and maintains the enzyme catalytic activity. Purified F1ATPase still containing three endogenous tightly bound nucleotides, named MF1(1,2), was obtained under mild conditions, whereas a harsher treatment gave a fully nucleotide depleted enzyme, named MF1(0,0). When MF1(1,2) was loaded with Mn(II) in 1:0.8 ratio, the spectrum showed evidence of a nitrogen interacting with the metal, while this interaction was not present in the spectrum of MF1(0,0) loaded with Mn(II) in the same ratio. However, when MF1(0,0) was loaded with 2.4 Mn(II), the spectrum showed metal-nitrogen interaction resembling that of MF1(1,2) loaded with Mn(II) in 1:0.8 ratio. When MF1(1,2) was loaded with 2.4 Mn(II) the metal-nitrogen interaction signal remained and a phosphorous coordination to the metal was also evident, indicating a binding of Mn2+ to a site containing a tightly bound nucleotide but metal free. These results strongly support the role of the metal alone in structuring the catalytic sites of the enzyme while ESEEM technique appears to be a sensitive and suitable spectroscopic method for conformational studies of MF1 with the advantage of using proteins in frozen solution

ATP synthase complex from beef heart mitochondria. Role of the thiol group of the 25-kDa subunit of Fo in the coupling mechanism between Fo and F1.

In order to assess the role of thiol groups in the Fo part of the ATP synthase in the coupling mechanism of ATP synthase, we have treated isolated Fo, extracted from beef heart Complex V with urea, with thiol reagents, primarily with diazenedicarboxylic acid bis-(dimethylamide) (diamide) but also with Cd2+ and N-ethylmaleimide. FoF1 ATP synthase was reconstituted by adding isolated F1 and the oligomycin-sensitivity-conferring-protein (OSCP) to Fo. The efficiency of reconstitution was assessed by determining the sensitivity to oligomycin of the ATP hydrolytic activity of the reconstituted enzyme. Contrary to Cd2+, incubation of diamide with Fo, before the addition of F1 and OSCP, induced a severe loss of oligomycin sensitivity, due to an inhibited binding of F1 to Fo. This effect was reversed by dithiothreitol. Conversely, if F1 and OSCP were added to Fo before diamide, no effect could be detected. These results show that F1 (and/or OSCP) protects Fo thiols from diamide and are substantiated by the finding that the oligomycin sensitivity of ATP hydrolysis activity of isolated Complex V was also unaltered by diamide. Gel electrophoresis of FoF1 ATP synthase, reconstituted with diamide-treated Fo, revealed that the loss of oligomycin sensitivity was directly correlated with diminution of band Fo 1 (or subunit b). Concomitantly a band appeared of approximately twice the molecular weight of subunit Fo 1. As this protein contains only 1 cysteine residue (Walker, J. E., Runswick, M. J., and Poulter, L. (1987) J. Mol. Biol. 197, 89-100), the effect of diamide is attributed to the formation of a disulfide bridge between two of these subunits. These results offer further evidence for the proposal, based on aminoacid sequence and structural analysis, that subunit Fo 1 of mammalian Fo is involved in the binding with F1 (Walker et al. (1987]. N-Ethylmaleimide affects oligomycin sensitivity to a lesser extent than diamide, suggesting that the mode of action of these reagents (and the structural changes induced in Fo) is different

Nicodemia madagascariensis (Lam.) R. Parker (Family Scrophulariaceae), a casual alien plant new to Italy

In this note the authors report the first case of naturalization of the Malagasy smoke bush within Italian territory. Along with a description of the abiotic and biotic characteristics of the invaded site, the current demographic and dynamic trends of the detected population by means of growth-ring analysis was also assessed. Moreover, an overview on the history of its introduction in Europe is provided, underlining the key role played by the Mediterranean cities with respect to the acclimatization and spread of alien tropical and subtropical plants. Due to its low frequency under cultivation, the Malagasy smoke bush should not behave as an invader in the future; on the other hand, its ability to cover and smother neighbouring trees in very short times, make its spread within the whole Mediterranean area worth regular monitoring

Monitoring the invasion of an exotic tree (Ailanthus altissima) (Mill.) Swingle with Landsat satellite time series imagery in urban forest.

In the Mediterranean area, one the most threat tree to various ecosystems is Ailanthus altissima (Mill.) Swingle. This is an aggressive invasive species common in natural and semi-natural habitat. Monitoring and mapping of invasive species is an important information for the conservation and management of ecosystems. The study of distribution and diffusion of invasive species are useful to assess their environmental impacts, formulate effective control strategies, and forecast potential spread. The main target of this work is to examine the feasibility of mapping the expansion of A. altissima using remote sensing techniques in a highly complex urban forest setting. Remote sensing has been a useful tool to map the invasive plant. We mapped the pattern of ailanthus expansion from 1990 to 2015 in a suburban area of Palermo, the Favorita park, using time series of Landsat image. This images are nowadays available at no cost. We used that images to analyze larger areas but the 30 m resolution does not permit mapping of individual trees; the combinations of dates and medium spatial resolution with the phenology information allowed the detection of the species. Indeed, the ailanthus is a deciduous tree that we compared with the other evergreen vegetation. Time series of Normalized difference vegetation index (NDVI) and supervised classification were used as a dataset in the classification process. These results provide a basis for more detailed investigations on invasive species and the possibility to increase the spatial resolution with the new platforms might lead to further improvements plant species identification and their distribution patterns recognition

Possibile origine e consistenza di un popolamento di Quercus trojana Webb subsp. trojana (Fagaceae) nel Bosco della Ficuzza (Palermo, Sicilia

Possible origin and demographic asset of a population of Quercus trojana Webb subsp. trojana (Fagaceae) in the wood of Ficuzza (Palermo, Sicily). The first record of Macedonian oak in Sicily is reported. The finding consists of 73 adult and 4 young individuals. A morphometric analysis of all of them, along with an assessment of their phytosanitary status was carried out. They all grow between 700 and 850 m a.s.l. on a 2,30 ha area, characterized by a land mosaic of shrubland and forest patches. Notwithstanding two years of researches, no ultimate proof has been found on their alien status. On this purpose, several alternative hypothesis are formulated and their strengths and weaknesses are discussed. Considering that the environmental conditions of Ficuzza fit very well the ecological requirements of Quercus trojana, even the native status of local population cannot be totally rejected. On the other hand, many data support its recent introduction. In fact, according to interviews to old forest workers, archive documents and aerial photos, most part of the trees of this area have been planted just few years after the end of the Second World War, and dendrochronological analyses have pointed out that the biggest Macedonian oaks of Sicily are no more than 50-60 years old. Whatever the origin of local Macedonian oak population is, the presence of natural renovation testifies, at least, its full naturalisation within the Sicilian territory

Carbon stock increases up to old growth forest along a secondary succession in Mediterranean island ecosystems

The occurrence of old-growth forests is quite limited in Mediterranean islands, which have been subject to particularly pronounced human impacts. Little is known about the carbon stocks of such peculiar ecosystems compared with different stages of secondary succession. We investigated the carbon variation in aboveground woody biomass, in litter and soil, and the nitrogen variation in litter and soil, in a 100 years long secondary succession in Mediterranean ecosystems. A vineyard, three stages of plant succession (high maquis, maquis-forest, and forest-maquis), and an old growth forest were compared. Soil samples at two soil depths (0–15 and 15–30 cm), and two litter types, relatively undecomposed and partly decomposed, were collected. Carbon stock in aboveground woody biomass increased from 6 Mg ha-1 in the vineyard to 105 Mg ha-1 in old growth forest. Along the secondary succession, soil carbon considerably increased from about 33 Mg ha-1 in the vineyard to about 69 Mg ha-1 in old growth forest. Soil nitrogen has more than doubled, ranging from 4.1 Mg ha-1 in the vineyard to 8.8 Mg ha-1 in old growth forest. Both soil parameters were found to be affected by successional stage and soil depth but not by their interaction. While the C/N ratio in the soil remained relatively constant during the succession, the C/N ratio of the litter strongly decreased, probably following the progressive increase in the holm oak contribution. While carbon content in litter decreased along the succession, nitrogen content slightly increased. Overall, carbon stock in aboveground woody biomass, litter and soil increased from about 48 Mg ha-1 in the vineyard to about 198 Mg ha-1 in old growth forest. The results of this study indicate that, even in Mediterranean environments, considerable amounts of carbon may be stored through secondary succession processes up to old growth forest