NMR sugar-profile in genuine grape must.

The wine market has always faced the problem of fraud, including the addition of exogenous sugar solutions to grape musts to increase the final alcohol content. Since in some countries the practice of chaptalization is prohibited (except by adding concentrated must) it is necessary to broaden the analytical techniques that allow the identification of this type of fraud. The aim of this study was to define an NMR-based sugar profile of genuine grape must to set concentration limits for each sugar as parameters of authenticity. Glucose, fructose, together with eleven minor sugars were quantified in 82 genuine Italian grape musts, developing an analytical procedure based on highly selective chemical shift filters followed by TOCSY. Alongside the characteristic myo- and scyllo-inositol, significant contents of mannose, galactose, and trehalose were also found. Otherwise, maltose, rhamnose, arabinose, sucrose and lactose are present in lower concentrations and show great concentration variability.

Myo-Inositol, Scyllo-Inositol, and Other Minor Carbohydrates as Authenticity Markers for the Control of Italian Bulk, Concentrate, and Rectified Grape Must.

Myo-inositol polyalcohol is a characteristic component of natural and concentrated grape musts (CMs), and Regulation (EU) no. 1308/2013 prescribes its presence as a marker of the authenticity of rectified concentrated must (RCM). Other polyalcohols besides myo-inositol, such as scyllo-inositol or minor sugars, could be considered authenticity markers, but an extensive search in the literature yielded no exhaustively investigated study of their concentration variability in genuine products. The aim of this study was to create an extensive national data bank of minor carbohydrates profiles and investigate the impact of the geographical origin and the different vintages on the concentration of these compounds; to this end, 450 authentic Italian grape musts of different varieties were sampled and analyzed during the harvest season in 2019, 2020, and 2021. The grape musts from the Italian wine-growing areas CII and CIIIb had myo- and scyllo-inositol contents always higher than 756 and 39 mg/kg of sugar, respectively. Conversely, also considering other mono- and disaccharides, sucrose, sorbitol, lactose, maltose, and isomaltose showed contents always lower than 534, 1207, 390, 2222, and 1639 mg/kg of sugar, respectively. The general applicability to the CM and RCM of the proposed authenticity thresholds, established in the must, was demonstrated by studying the influence of must concentration on the myo- and scyllo-inositol content. Inter-laboratory comparison experiments were also conducted to harmonize and characterize laboratory methods and validate the analytical dataset. Based on the obtained results, the text of the EU legislation (Reg. (EU) 1308/2013), which defines the characteristics of the must and the CRM product, should be revised.

Authentication and Geographical Characterisation of Italian Grape Musts through Glucose and Fructose Carbon Isotopic Ratios Determined by LC-IRMS.

The authenticity of grape musts is normally checked through a time-consuming stable isotopic analysis of carbon (δ13C) after fermentation and distillation by following the official OIV MA AS-312-06 method. In this study, the alternative use of a technique based on δ13C isotopic analysis of the major sugars of the grape must by liquid chromatography coupled with isotope ratio mass spectrometry (LC-IRMS) is provided. It allows not only the detection of the fraudulent addition to grape must of exogenous glucose and fructose deriving from C4 plants but also the characterisation of it based on its geographical origin. In order to discriminate between musts from different areas of Italy, a preliminary dataset was considered; the δ13C isotopic ratios of glucose and fructose of around 100 authentic samples were analysed. The two analysed parameters, ranging from -29.8‱ to -21.9‱, are well correlated (R2 = 0.7802) and the northern regions showed significantly more negative δ13C values for both sugars than the rest of the dataset.

Time-Resolved Investigation of Molecular Components Involved in the Induction of [Formula: see text] High Affinity Transport System in Maize Roots.

The induction, i.e., the rapid increase of nitrate ([Formula: see text]) uptake following the exposure of roots to the anion, was studied integrating physiological and molecular levels in maize roots. Responses to [Formula: see text] treatment were characterized in terms of changes in [Formula: see text] uptake rate and plasma membrane (PM) H+-ATPase activity and related to transcriptional and protein profiles of NRT2, NRT3, and PM H+-ATPase gene families. The behavior of transcripts and proteins of ZmNRT2s and ZmNRT3s suggested that the regulation of the activity of inducible high-affinity transport system (iHATS) is mainly based on the transcriptional/translational modulation of the accessory protein ZmNRT3.1A. Furthermore, ZmNRT2.1 and ZmNRT3.1A appear to be associated in a ∼150 kDa oligomer. The expression trend during the induction of the 11 identified PM H+-ATPase transcripts indicates that those mainly involved in the response to [Formula: see text] treatment are ZmHA2 and ZmHA4. Yet, partial correlation between the gene expression, protein levels and enzyme activity suggests an involvement of post-transcriptional and post-translational mechanisms of regulation. A non-denaturing Deriphat-PAGE approach allowed demonstrating for the first time that PM H+-ATPase can occur in vivo as hexameric complex together with the already described monomeric and dimeric forms.

Nitrate induction triggers different transcriptional changes in a high and a low nitrogen use efficiency maize inbred line.

In higher plants, NO3(-) can induce its own uptake and the magnitude of this induction is positively related to the external anion concentration. This phenomenon has been characterized in both herbaceous and woody plants. Here, different adaptation strategies of roots from two maize (Zea mays L., ZmAGOs) inbred lines differing in nitrogen use efficiency (NUE) and exhibiting different timing of induction were discussed by investigating NO3(-) -induced changes in their transcriptome. Lo5 line (high NUE) showing the maximum rate of NO3(-) uptake 4 h after the provision of 200 µmol/L NO3(-) treatment modulated a higher number of transcripts relative to T250 (low NUE) that peaked after 12 h. The two inbred lines share only 368 transcripts that are modulated by the treatment with NO3(-) and behaved differently when transcripts involved in anion uptake and assimilation were analyzed. T250 line responded to the NO3(-) induction modulating this group of genes as reported for several plant species. On the contrary, the Lo5 line did not exhibit during the induction changes in this set of genes. Obtained data suggest the importance of exploring the physiological and molecular variations among different maize genotypes in response to environmental clues like NO3(-) provision, in order to understand mechanisms underlying NUE.

Induction of high-affinity NO3- uptake in grapevine roots is an active process correlated to the expression of specific members of the NRT2 and plasma membrane H+-ATPase gene families.

The phenomenon of NO3- induction in plant roots has been characterised both in herbaceous and woody plants. Grapevine (Vitis vinifera L.) plants, hydroponically grown, showed an increase in NO3- uptake rate in response to anion treatment for different periods in the nutrient solution after 1 week of NO3- deprivation. The expression profile of the two high-affinity NO3- transporters VvNRT2.4A and VvNRT2.4B, and the gene encoding the accessory protein VvNAR2.2 exhibits a similar trend to that of the anion uptake. The induction, also involving the increase in activity and protein levels of plasma membrane H+-ATPase, is correlated with the expression profile of two (VvHA2 and VvHA4) out of eight putative plasma membrane H+-ATPase genes identified in grapevine genome.