Biochemical Traits, 1H NMR Profile and Residual DNA Content of 'Asprinio', White Wine from Campania Region (Southern Italy).

'Asprinio' is a white dry wine characteristic for its acidity and aromatic flavour, known as emerging DOP wine in Southern Italy. Nevertheless, little information is available on the metabolomic profile of this wine. Thus, in this paper we evaluated the colourimetric parameters, 1H NMR profiles and free amino acids content of 'Asprinio' wines, bottled by two different wineries (hereafter 'Asprinio_A' and 'Asprinio_B') collected in 2019 and 2020, using 'Greco di Tufo' for comparison. The colourimetric parameters are similar for both 'Asprinio' wines and differ from 'Greco di Tufo' wines. On the other hand, both 1H NMR and free amino acid content profiles show different chemometric profiles among the three wines analysed, although the profiles are similar for both vintages. Moreover, the multivariate analyses carried out highlight differences between 'Asprinio_A' and 'Asprinio_B', which exbibit also different residual yeast and plant DNA. Overall, considering that the two-manufacturing wineries use 100% 'Asprinio' grape, the difference retrieved between the two 'Asprinio' wines could be explained by the different grapevine training systems: 'vite maritata' (training system inherited from Etruscans) for 'Asprinio_A' and 'guyot' for 'Asprinio_B'.

Salinity Duration Differently Modulates Physiological Parameters and Metabolites Profile in Roots of Two Contrasting Barley Genotypes.

Hordeum maritimum With. is a wild salt tolerant cereal present in the saline depressions of the Eastern Tunisia, where it significantly contributes to the annual biomass production. In a previous study on shoot tissues it was shown that this species withstands with high salinity at the seedling stage restricting the sodium entry into shoot and modulating over time the leaf synthesis of organic osmolytes for osmotic adjustment. However, the tolerance strategy mechanisms of this plant at root level have not yet been investigated. The current research aimed at elucidating the morphological, physiological and biochemical changes occurring at root level in H. maritimum and in the salt sensitive cultivar Hordeum vulgare L. cv. Lamsi during five-weeks extended salinity (200 mM NaCl), salt removal after two weeks of salinity and non-salt control. H. maritimum since the first phases of salinity was able to compartmentalize higher amounts of sodium in the roots compared to the other cultivar, avoiding transferring it to shoot and impairing photosynthetic metabolism. This allowed the roots of wild plants to receive recent photosynthates from leaves, gaining from them energy and carbon skeletons to compartmentalize toxic ions in the vacuoles, synthesize and accumulate organic osmolytes, control ion and water homeostasis and re-establish the ability of root to grow. H. vulgare was also able to accumulate compatible osmolytes but only in the first weeks of salinity, while soon after the roots stopped up taking potassium and growing. In the last week of salinity stress, the wild species further increased the root to shoot ratio to enhance the root retention of toxic ions and consequently delaying the damages both to shoot and root. This delay of few weeks in showing the symptoms of stress may be pivotal for enabling the survival of the wild species when soil salinity is transient and not permanent.

Enhancing Sustainability by Improving Plant Salt Tolerance through Macro- and Micro-Algal Biostimulants.

Algal biomass, extracts, or derivatives have long been considered a valuable material to bring benefits to humans and cultivated plants. In the last decades, it became evident that algal formulations can induce multiple effects on crops (including an increase in biomass, yield, and quality), and that algal extracts contain a series of bioactive compounds and signaling molecules, in addition to mineral and organic nutrients. The need to reduce the non-renewable chemical input in agriculture has recently prompted an increase in the use of algal extracts as a plant biostimulant, also because of their ability to promote plant growth in suboptimal conditions such as saline environments is beneficial. In this article, we discuss some research areas that are critical for the implementation in agriculture of macro- and microalgae extracts as plant biostimulants. Specifically, we provide an overview of current knowledge and achievements about extraction methods, compositions, and action mechanisms of algal extracts, focusing on salt-stress tolerance. We also outline current limitations and possible research avenues. We conclude that the comparison and the integration of knowledge on the molecular and physiological response of plants to salt and to algal extracts should also guide the extraction procedures and application methods. The effects of algal biostimulants have been mainly investigated from an applied perspective, and the exploitation of different scientific disciplines is still much needed for the development of new sustainable strategies to increase crop tolerance to salt stress.

Hordeum vulgare and Hordeum maritimum respond to extended salinity stress displaying different temporal accumulation pattern of metabolites.

Hordeum maritimum With. (= H. marinum Huds. subsp. marinum, 2n=14) is a wild cereal present in the saline depressions of the Soliman and Kelbia Sebkhas, which contributes significantly to annual biomass production in Tunisia. This species is able to tolerate high NaCl concentrations at the seedling stage without showing symptoms of toxicity; however, the tolerance strategy mechanisms of this plant have not yet been unravelled. Our metabolite analysis, performed on leaves of H. maritimum during extended stress in comparison with Hordeum vulgare L. cv. Lamsi, has revealed an adaptive response of the wild species based on a different temporal accumulation pattern of ions and compatible metabolites. Further, wild and cultivated genotypes with contrasting salt-tolerant behaviour display different pattern of metabolites when salt stress is prolonged over 2 weeks. In particular, when exposed to up to 3 weeks of 200mM NaCl salt stress, H. maritimum is able to maintain lower leaf concentrations of sodium and chloride, and higher concentrations of potassium compared with H. vulgare. This likely restricts sodium entry into plants at the root level, and uses the toxic ions, glycine betaine and low levels of proline for osmotic adjustment. Under prolonged stress, the accumulation of proline increases, reaching the highest levels in concomitance with the decrease of potassium to sodium ratio, the increase of hydrogen peroxide and decrease of chlorophylls. The modulation of proline accumulation over time can be interpreted as an adaptive response to long-term salinity. Moreover, once synthetised glycine betaine is transported but not metabolised, it can contribute together with proline to osmotically balance H. maritimum leaves and protect them from oxidative stress. The 2-3 week delay of H. maritimum in showing the symptoms of stress and damages compared with H. vulgare could be important in the survival of plants when soil salinity is not a permanent condition, but just a transient state of stress.

Metabolic characterization and antioxidant activity in sweet cherry (Prunus avium L.) Campania accessions: Metabolic characterization of sweet cherry accessions.

The failure of the antioxidant scavenging system in advanced ripening stages, causing oxidative stress, is one of the most important factor of fruit decay. Production of rich antioxidant fruit could represent a way to delay fruit senescence and preserve its characteristics. We investigated the antioxidant metabolites (ascorbate, glutathione, tocopherols, and polyphenols) and enzymes (ascorbic peroxidases, peroxidases and polyphenol oxidases) involved in the antioxidant response in forty-three accessions of sweet cherry fruits from Campania region. Our results highlight accessions with high antioxidant metabolites contents but low enzymatic activities. These represent important factors in both pre- and post-harvest on the qualitative and nutritional characteristics of sweet cherry. Observed differences are probably due to endogenous characteristics making these accessions particularly interesting for breeding programs aimed to improve fruit quality and shelf-life and for addressing the cultivation of a specific characterized cultivar based on the intended use, fresh consumption or processed products.

Dataset on antioxidant metabolites and enzymes activities of freshly harvested sweet cherries (Prunus avium L.) of Campania accessions.

In this article, we reported the original data obtained by the study of metabolites and enzymes involved in sweet cherry antioxidant system. We measured hydrogen peroxide (H2O2) and malondialdehyde (MDA), which are indicator of oxidative stress. Moreover, we measured the concentration of reduced and oxidized ascorbate and glutathione that are involved in ROS detoxification together with phenolics, anthocyanins and tocopherols. Among antioxidant enzymes, we analyzed the activities of ascorbate peroxidase (APX; EC 1.11.1.11), and the soluble and bound forms of polyphenol oxidase (PPO; EC 1.10.3.1) and guaiacol peroxidase (POD; EC 1.11.1.7). The data reported in this paper are related to the research article "Metabolic characterization and antioxidant activity in sweet cherry (Prunus avium L.) Campania accessions", authored by Mirto et al. (2018) [1].

Durum wheat seedling responses to simultaneous high light and salinity involve a fine reconfiguration of amino acids and carbohydrate metabolism.

Durum wheat plants are extremely sensitive to drought and salinity during seedling and early development stages. Their responses to stresses have been extensively studied to provide new metabolic targets and improving the tolerance to adverse environments. Most of these studies have been performed in growth chambers under low light [300-350 µmol m-2 s-1 photosynthetically active radiation (PAR), LL]. However, in nature plants have to face frequent fluctuations of light intensities that often exceed their photosynthetic capacity (900-2000 µmol m-2 s-1 ). In this study we investigated the physiological and metabolic changes potentially involved in osmotic adjustment and antioxidant defense in durum wheat seedlings under high light (HL) and salinity. The combined application of the two stresses decreased the water potential and stomatal conductance without reducing the photosynthetic efficiency of the plants. Glycine betaine (GB) synthesis was inhibited, proline and glutamate content decreased, while γ-aminobutyric acid (GABA), amides and minor amino acids increased. The expression level and enzymatic activities of Δ1-pyrroline-5-carboxylate synthetase, asparagine synthetase and glutamate decarboxylase, as well as other enzymatic activities of nitrogen and carbon metabolism, were analyzed. Antioxidant enzymes and metabolites were also considered. The results showed that the complex interplay seen in durum wheat plants under salinity at LL was simplified: GB and antioxidants did not play a main role. On the contrary, the fine tuning of few specific primary metabolites (GABA, amides, minor amino acids and hexoses) remodeled metabolism and defense processes, playing a key role in the response to simultaneous stresses.

Durum Wheat Roots Adapt to Salinity Remodeling the Cellular Content of Nitrogen Metabolites and Sucrose.

Plants are currently experiencing increasing salinity problems due to irrigation with brackish water. Moreover, in fields, roots can grow in soils which show spatial variation in water content and salt concentration, also because of the type of irrigation. Salinity impairs crop growth and productivity by inhibiting many physiological and metabolic processes, in particular nitrate uptake, translocation, and assimilation. Salinity determines an increase of sap osmolality from about 305 mOsmol kg-1 in control roots to about 530 mOsmol kg-1 in roots under salinity. Root cells adapt to salinity by sequestering sodium in the vacuole, as a cheap osmoticum, and showing a rearrangement of few nitrogen-containing metabolites and sucrose in the cytosol, both for osmotic adjustment and oxidative stress protection, thus providing plant viability even at low nitrate levels. Mainly glycine betaine and sucrose at low nitrate concentration, and glycine betaine, asparagine and proline at high nitrate levels can be assumed responsible for the osmotic adjustment of the cytosol, the assimilation of the excess of ammonium and the scavenging of ROS under salinity. High nitrate plants with half of the root system under salinity accumulate proline and glutamine in both control and salt stressed split roots, revealing that osmotic adjustment is not a regional effect in plants. The expression level and enzymatic activities of asparagine synthetase and Δ1-pyrroline-5-carboxylate synthetase, as well as other enzymatic activities of nitrogen and carbon metabolism, are analyzed.

Analysis of different European hazelnut (Corylus avellana L.) cultivars: authentication, phenotypic features, and phenolic profiles.

Hazelnuts exhibit functional properties due to their content in fatty acids and phenolic compounds that could positively affect human health. The food industry requires precise traits for morphological, chemical, and physical kernel features so that some cultivars could be more suitable for specific industrial processing. In this study, agronomical and morphological features of 29 hazelnut cultivars were evaluated and a detailed structural characterization of kernel polyphenols was performed, confirming the presence of protocatechuic acid, flavan-3-ols such as catechin, procyanidin B2, six procyanidin oligomers, flavonols, and one dihydrochalcone in all the analyzed cultivars. In addition, an innovative methodology based on the MALDI-TOF mass spectrometric analysis of peptide/protein components extracted from kernels was developed for the authentication of the most valuable cultivars. The proposed method is rapid, simple, and reliable and holds the potential to be applied in quality control processes. These results could be useful in hazelnut cultivar evaluation and choice for growers, breeders, and food industry.

Ty1-copia group retrotransposons and the evolution of retroelements in several angiosperm plants: evidence of horizontal transmission.

The phylogenetic relationships among thirty-seven new Ty1-copia group retrotransposons in seven angiosperm plants were examined by reverse transcriptase and ribonuclease H sequence analysis. Distribution pattern of the retrotransposons of closely related plant species generally reflects a close phylogenetic relationship. In contrast, we found that several retrotransposon sequences from the same genome exhibited a high degree of divergence and had a relatively high degree of identity versus retrotransposon sequences from widely divergent species, including an ancestral phytopathogen fungus. This finding supports the hypothesis that the horizontal transmission from phytopatogen organism to the host flowering plants could have played a role in the evolutionary dynamics of Ty1-copia group retrotransposons.

Isolation of Ty1-copia retrotransposon in myrtle genome and development of S-SAP molecular marker.

Long terminal repeat (LTR)-retrotransposons are mobile genetic elements that are ubiquitous in plants and constitute a major portion of their nuclear genomes. LTR- retrotransposons possess unique properties that make them appropriate for investigating relationships between populations, varieties and closely related species. Myrtus communis L. is an evergreen shrub growing spontaneously throughout the Mediterranean area. Accessions show significant variations for agriculturally important traits, so the development of specific molecular markers for conservation and characterization of myrtle germplasm is desirable to conserve biodiversity. In this study, we isolated the first retrotransposon Ty1-copia-like element (Tmc1) in Myrtus communis L. genome and used this as a molecular marker. We successfully employed the S-SAP marker system to specifically characterize four myrtle accessions belonging to different areas in the province of Caserta (Italy). The high level of polymorphism detected in isolated LTRs, make Tmc1 a good molecular marker for this species. Our findings confirm that retrotransposon-based molecular markers are particularly valuable tools for plant molecular characterization studies.

A PCR based SNPs marker for specific characterization of English walnut (Juglans regia L.) cultivars.

English walnut (Juglans regia L.) is the most economically important species from all the 21 species belonging to the genus Juglans and is an important and healthy food as well as base material for timber industry. The aim of this study was to develop a simple technique for specific characterization of English walnut using DNA method. The first and second internal transcribed spacers (ITS1 and ITS2) as well as the intervening 5.8S coding region of the rRNA gene for 18 cultivars of J. regia L. isolated from different geographic origins were characterized. The size of the spacers sequences ranged from 257 to 263 bases for ITS1 and from 217 to 219 bases for ITS2. Variation of GC contents has also been observed and scored as 55-56.7 and 57.1-58.9% for ITS1 and ITS2, respectively. This data exhibited the presence of polymorphism among cultivars. Alignment of the ITS1-5.8S-ITS2 sequences from 18 walnut cultivars showed that there were 244 single nucleotide polymorphisms (SNPs) and 1 short insertion-deletion (indel) at 5' end ITS1. Amplification refractory mutation system strategy was successfully applied to the SNP markers of the ITS1 and ITS2 sequences for the fingerprinting analysis of 17 on 18 walnut cultivars. The prediction of ITS1 and ITS2 RNA secondary structure from each cultivar was improved by detecting key functional elements shared by all sequences in the alignments. Phylogenetic analysis of the ITS1-5.8S-ITS2 region clearly separated the isolated sequences into two clusters. The results showed that ITS1 and ITS2 region could be used to discriminate these walnut cultivars.

Ttd1a promoter is involved in DNA-protein binding by salt and light stresses.

Stress modulation of retrotransposons may play a role in generating host genetic plasticity in response to environmental stress. Transposable elements have been suggested to contribute to the evolution of genes, by providing cis-regulatory elements leading to changes in expression patterns. Indeed, their promoter elements are similar to those of plant defence genes and may bind similar defence-induced transcription factors. We previously isolated a new Ty1-copia retrontrasposon named Ttd1a and showed its activation and mobilization in salt and light stresses. Here, using a retard mobility assay in Triticum durum L. crude extracts, we showed that the CAAT motif present in the Ttd1a retrotransposon promoter, is involved in DNA-protein binding under salt and light stresses and therefore in the regulation of Ttd1a activity. Data presented in this paper suggest that nuclear proteins can interact with the CAAT motif either directly or indirectly and enhance Ttd1a by a specific ligand-dependent activation under stress.

Molecular analysis of a novel tandemly organized repetitive DNA sequence in Citrus limon (L.) Burm.

Repetitive sequences constitute a significant component of most eukaryotic genomes, and the isolation and characterization of repetitive DNA sequences provide an insight into the organization of the genome of interest. Here, we report the isolation and molecular analysis of a novel tandemly organized repetitive DNA sequence from the genome of Citrus limon. Digestion of C. limon DNA with Hinf I produced a prominent fragment of approximately 300 bp. Southern blotting revealed a ladder composed of DNA fragments that were multimers of the 300-bp Hinf I band. Thus, Hinf I digestion revealed a novel satellite, which we have called the C. limon satellite DNA 300 (CL300). Sequence analysis shows significant homology between a portion of the CL300 monomer and the transposase box of an En/Spm-like element. The CL300 satellite was also detected in grapefruit, sour orange, trifoliate orange and kumquat. These results suggest that the CL300 repeat is an ancient satellite, and we propose that a significant portion originated by amplification of a genomic region containing the En/Spm-like transposase element.

Identification of a cDNA clone encoding DIP1-binding protein in Drosophila melanogaster.

The Drosophila melanogaster L27a gene encodes a ribosomal protein which is a member of the L15 family of ribosomal proteins. D.m. L27a is closely related to the mammalian protein that has been found differentially expressed in lung cancer tissues and therefore could be involved in the control of cell proliferation such as the ribosomal protein S6. Our work elucidates the role of DIP1 which is a novel protein that we found in Drosophila. We performed a two-hybrid system assay and identified the L27a protein as an interactor of DIP1. The interaction was then validated by in vitro binding assays. DIP1, similar to other nuclear proteins in eukaryotes, is localized to the nuclear periphery and chromatin domain in all nuclei, but disappears at the metaphase. It is possible that in D.m. L27a protein, via interaction with DIP1, could be involved in protein synthesis as well as in cell cycle regulation.