Glutamate dehydrogenase in "Liverworld"-A study in selected species to explore a key enzyme of plant primary metabolism in Marchantiophyta.

In plants, glutamate dehydrogenase (GDH) is an ubiquitous enzyme that catalyzes the reversible amination of 2-oxoglutarate in glutamate. It contributes to both the amino acid homeostasis and the management of intracellular ammonium, and it is regarded as a key player at the junction of carbon and nitrogen assimilation pathways. To date, information about the GDH of terrestrial plants refers to a very few species only. We focused on selected species belonging to the division Marchantiophyta, providing the first panoramic overview of biochemical and functional features of GDH in liverworts. Native electrophoretic analyses showed an isoenzymatic profile less complex than what was reported for Arabidposis thaliana and other angiosperms: the presence of a single isoform corresponding to an α-homohexamer, differently prone to thermal inactivation on a species- and organ-basis, was found. Sequence analysis conducted on amino acid sequences confirmed a high similarity of GDH in modern liverworts with the GDH2 protein of A. thaliana, strengthening the hypothesis that the duplication event that gave origin to GDH1-homolog gene from GDH2 occurred after the evolutionary bifurcation that separated bryophytes and tracheophytes. Experiments conducted on Marchantia polymorpha and Calypogeia fissa grown in vitro and compared to A. thaliana demonstrated through in gel activity detection and monodimensional Western Blot that the aminating activity of GDH resulted in strongly enhanced responses to ammonium excess in liverworts as well, even if at a different extent compared to Arabidopsis and other vascular species. The comparative analysis by bi-dimensional Western Blot suggested that the regulation of the enzyme could be, at least partially, untied from the protein post-translational pattern. Finally, immuno-electron microscopy revealed that the GDH enzyme localizes at the subcellular level in both mitochondria and chloroplasts of parenchyma and is specifically associated to the endomembrane system in liverworts.

Biochar effects on early decomposition of standard litter in a European beech forest (northern Italy).

The release of biochar (BC) on forest soil is a strategy aimed at increasing carbon reserves and forest productivity. The effect of BC amendments on the decomposition of different quality litter is, however, poorly understood. With this study we investigate the effects of wood-derived BC applications on early decomposition in a European beech (Fagus sylvatica L.) forest through the burial of standard material, i.e. green tea and rooibos tea (high- and low-quality litter surrogates, respectively). Two main questions were addressed: 1) Do BC applications influence the decomposition of high- and low-quality standard litter and, if so, in what way? and 2) Does this effect (if measurable) depend on where the sample is placed with respect to the BC application layer? To test BC amendment effects, four application percentages were employed (0, 10, 20 and 100 %), after which standard litter mass loss was recorded. To investigate the effects of sample position, only three BC application percentages were used (0, 10 and 20 %), with teabags buried at three different depths - within the BC amended layer, between this layer and the unamended soil, and below the latter. Results show that early decomposition of high-quality standard litter was not influenced by BC applications, while a significant reduction in mass loss of low-quality standard litter was observed when the percentage of BC application was higher, specifically of litter within the 20 % and 100 % BC amended layers. Decomposition was also affected by sample position relative to the BC layer, exhibiting higher levels of mass loss when samples were placed within the BC amended layer. Overall, BC applications on beech forest soils not only seem to produce negligible effects on the early decomposition rate of high-quality standard litter, but such applications also seem to have the ability to reduce carbon loss following plant material degradation.

Effects of Wood-Derived Biochar on Germination, Physiology, and Growth of European Beech (Fagus sylvatica L.) and Turkey Oak (Quercus cerris L.)

Biochar (BC) soil amendments could partially counteract soil carbon (C) stock decrease in broad-leaved forests in Italy; however, its effects on the growth of representative tree species­Fagus sylvatica L. and Quercus cerris L.­has not yet been addressed. We examine whether seed germination and growth of these species are affected by addition of BC obtained from deciduous broadleaf trees. Seeds were left to germinate in greenhouse conditions under three different BC amendments: 0% (control), 10% and 20% (v/v). Seedlings were then subjected to controlled conditions under the same BC percentage. Biochar effects on seed germination were assessed measuring germination time and percentage, while effects on photosynthesis were assessed using leaf chlorophyll content (mg/m2) and photosynthetic efficiency (FV/FM). Plant growth was estimated by recording leaf number, longest leaf length and plant height. Biochar treatments had no negative effects on germination and early growth stage of the two species. Positive effects were found on the chlorophyll content of both species (ca. +8%) regardless of the treatment and on the leaf number (+30%), leaf length (+14%) and plant height (+48%) of Q. cerris (only with 10% BC). Biochar applications seem, therefore, a suitable method for increasing broad-leaved forest C stock in Italy.

Phenology, Canopy Aging and Seasonal Carbon Balance as Related to Delayed Winter Pruning of Vitis vinifera L. cv. Sangiovese Grapevines.

Manipulating or shifting annual grapevine growing cycle to offset limitations imposed by global warming is a must today, and delayed winter pruning is a tool to achieve it. However, no information is available about its physiological background, especially in relation to modifications in canopy phenology, demography and seasonal carbon budget. Mechanistic hypothesis underlying this work was that very late winter pruning (LWP) can achieve significant postponement of phenological stages so that ripening might occur in a cooler period and, concurrently, ripening potential can be improved due to higher efficiency and prolonged longevity of the canopy. Variability in the dynamics of the annual cycle was created in mature potted cv. Sangiovese grapevines subjected to either standard winter pruning (SWP) or late and very late winter pruning (LWP, VLWP) performed when apical shoots on the unpruned canes were at the stage of 2 and 7 unfolded leaves. Vegetative growth, phenology and canopy net CO2 exchange (NCER) were followed throughout the season. Despite LWP and VLWP induced a bud-burst delay of 17 and 31 days vs. SWP, the delay was fully offset at harvest for LWP and was reduced to 6 days in VLWP. LWP showed notably higher canopy efficiency as shorter time needed to reach maximum NCER/leaf area (22 days vs. 34 in SWP), highest maximum NCER/leaf area (+37% as compared to SWP) and higher NCER/leaf area rates from veraison to end of season. As a result, seasonal cumulated carbon in LWP was 17% higher than SWP. A negative functional relationship was also established between amount of leaf area removed at winter pruning and yield per vine and berry number per cluster. Although retarded winter pruning was not able to postpone late-season phenological stages under the warm conditions of this study, it showed a remarkable potential to limit yield while improving grape quality, thereby fostering the hypothesis that it could be used to replace time-consuming and costly cluster thinning. This preliminary study indicates that proper winter pruning date should be timed so as not to exceed the stage of two unfolded leaves.

Urethral polyp in a 1-month-old child.

Urethral polyps are a rare finding in children, particularly in the very young. They are suspected by the presence of various clinical signs such as obstruction, voiding dysfunction and haematuria. There is an association with other urinary tract congenital anomalies. They are usually benign fibro-epithelial lesions with no tendency to recur and are treated by surgical ablation, fulguration or laser therapy. We report a 1-month-old boy with an antenatally diagnosed left ectopic pelvic kidney, postnatal urinary tract infection and no clinical signs of obstruction. Voiding cystourethrography to exclude vesico-ureteric reflux showed a trabeculated bladder and a mobile filling defect in the posterior urethra. Owing to its large size, cystotomy was necessary to remove the polyp successfully.

Bladder papilloma in a 12-year-old girl.

Epithelial bladder tumours are rare in childhood. A case of bladder papilloma in a 12-year-old girl is reported. Emphasis is given to the important role of ultrasound in diagnosis. Endoscopic treatment was successful and no evidence of recurrence was found at 5-year follow-up. Exposure to a chemical environmental factor (volatile shoe adhesives) is suggested as a possible cause of the tumour.