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The majority of the crops and vegetables of today were domesticated from their wild progenitors within the past 12 000 years. Considerable research effort has been expended on characterizing the genes undergoing positive and negative selection during the processes of crop domestication and improvement. Many studies have also documented how the contents of a handful of metabolites have been altered during human selection, but we are only beginning to unravel the true extent of the metabolic consequences of breeding. We highlight how crop metabolomes have been wittingly or unwittingly shaped by the processes of domestication, and highlight how we can identify new targets for metabolite engineering for the purpose of de novo domestication of crop wild relatives.

The present study was carried out with the aim of (i) evaluating the effect of chitosan (CTS) on the growth of durum wheat under salinity and (ii) examining CTS-regulated mechanisms of salinity tolerance associated with the antioxidant defense system. To achieve these goals, durum wheat seedlings were treated with CTS at different molecular weight, low (L-CTS, 50–190 kDa), medium (M-CTS, 190–310 kDa) and high (H-CTS, 310–375 kDa). The results obtained show that exposure to 200 mM NaCl reduced the shoot and the root dried biomass by 38% and 59%, respectively. The growth impairment induced by salinity was strongly correlated with an increase in the superoxide anion production (5-fold), hydrogen peroxide content (2-fold) and malondialdehyde (MDA) content (4-fold). Seedlings responded to the oxidative stress triggered by salinity with an increase in the total phenolic content (TPC), total flavonoid content (TFC) and total antioxidant activity (TAA) by 67%, 51% and 32%, respectively. A salt-induced increase in the activity of the antioxidant enzymes superoxide dismutase and catalase (CAT) of 89% and 86%, respectively, was also observed. Treatment of salt-stressed seedlings with exogenous CTS significantly promoted seedling growth, with the strongest effects observed for L-CTS and M-CTS, which increased the shoot biomass of stressed seedlings by 32% and 44%, respectively, whereas the root dried biomass increased by 87% and 64%, respectively. L-CTS and M-CTS treatments also decreased the superoxide anion production (57% and 59%, respectively), the hydrogen peroxide content (35% and 38%, respectively) and the MDA content (48% and 56%, respectively) and increased the TPC (23% and 14%, respectively), the TFC (19% and 10%, respectively), the TAA (up to 10% and 7%, respectively) and the CAT activity (29% and 20%, respectively). Overall, our findings indicate that CTS exerts its protective role against the oxidative damages induced by salinity by enhancing the antioxidant defense system. L-CTS and M-CTS were the most effective in alleviating the adverse effect of NaCl, thus demonstrating that the CTS action is strictly related to its molecular weight.

Food legumes are at the crossroads of many societal challenges that involve agriculture, such as climate change and food sustainability and security. In this context, pulses have a crucial role in the development of plant-based diets, as they represent a very good source of nutritional components and improve soil fertility, such as by nitrogen fixation through symbiosis with rhizobia. The main contribution to promotion of food legumes in agroecosystems will come from plant breeding, which is guaranteed by the availability of well-characterized genetic resources. Here, we analyze seeds of 25 American and European common bean purified accessions (i.e., lines of single seed descent) for different morphological and compositional quality traits. Significant differences among the accessions and superior genotypes for important nutritional traits are identified, with some lines showing extreme values for more than one trait. Heritability estimates indicate the importance of considering the effects of environmental growth conditions on seed compositional traits. They suggest the need for more phenotypic characterization in different environments over different years to better characterize combined effects of environment and genotype on nutritional trait variations. Finally, adaptation following the introduction and spread of common bean in Europe seems to have affected its nutritional profile. This finding further suggests the relevance of evolutionary studies to guide breeders in the choice of plant genetic resources.

Key message: The suppression of the HYD-1 gene by a TILLING approach increases the amount of ?-carotene in durum wheat kernel. Abstract: Vitamin A deficiency is a major public health problem that affects numerous countries in the world. As humans are not able to synthesize vitamin A, it must be daily assimilated along with other micro- and macronutrients through the diet. Durum wheat is an important crop for Mediterranean countries and provides a discrete amount of nutrients, such as carbohydrates and proteins, but it is deficient in some essential micronutrients, including provitamin A. In the present work, a targeting induced local lesions in genomes strategy has been undertaken to obtain durum wheat genotypes biofortified in provitamin A. In detail, we focused on the suppression of the ?-carotene hydroxylase 1 (HYD1) genes, encoding enzymes involved in the redirection of ?-carotene toward the synthesis of the downstream xanthophylls (neoxanthin, violaxanthin and zeaxanthin). Expression analysis of genes involved in carotenoid biosynthesis revealed a reduction of the abundance of HYD1 transcripts greater than 50% in mutant grain compared to the control. The biochemical profiling of carotenoid in the wheat mutant genotypes highlighted a significant increase of more than 70% of ?-carotene compared to the wild-type sibling lines, with no change in lutein, ?-carotene and zeaxanthin content. This study sheds new light on the molecular mechanism governing carotenoid biosynthesis in durum wheat and provides new genotypes that represent a good genetic resource for future breeding programs focused on the provitamin A biofortification through non-transgenic approaches.

Debranning was applied to durum wheat to the study the relationship between kernel shape and size, and ash and mineral distribution having implications for semolina yield. To this aim four durum wheat genotypes carried out over three environments were selected to determine the morphological and yield traits as well as the distribution along the kernel of the ash, macro- (Na, K, P, Ca, and Mg), and micro-elements (Mn, Fe, Cu, Zn, and Mo). A descendent ash gradient within the kernel reflects the decreases in the minerals that occurred during debranning. Perciasacchi with high seed weight (TKW) and greater thickness followed by Cappelli showed a more uniform distribution of ash content along the kernels. High r Pearson coefficient (p &lt, 0.01) showed an inverse relationship between thickness and ash decay. Since thickness was strongly affected by the genotype, it could represent a useful trait for breeding programs to predict the milling quality.

The granary weevil, Sitophilus granarius (L.) (Coleoptera, Dryophthoridae), is a primary pest of stored cereals worldwide. To identify possible sources of wheat resistance toward this pest, two commercial durum and bread wheat varieties with yellow pericarp were compared with anthocyanin-pigmented durum and bread wheat genotypes that have never entered commercial production. The composition of the kernel head-space volatile organic compounds (VOCs) and the olfactory responses of granary weevil adults to these kernel VOCs were investigated. Head-space solid-phase micro-extraction and gas chromatography–mass spectrometry analysis highlighted 17 and 13 kernel VOCs from durum and bread wheats, respectively. These compounds mainly included aldehydes and alcohols, and to a lesser extent, terpenes and benzene derivatives. Quantitative and qualitative differences were seen between the odor profiles of yellow and pigmented wheat kernels. In two-choice behavioral bioassays, granary weevil adults were significantly attracted by the kernel odors from the yellow commercial wheat varieties and their hexane extracts, but not by those of the pigmented wheat genotypes and their hexane extracts. Electroantennography confirmed the presence of VOCs in all of the hexane extracts that stimulated the olfactory system of both sexes of the granary weevil in a dose-dependent manner. Thus, differences among the odor blends were responsible for the different olfactory responses of granary weevils to the yellow and pigmented wheat kernels. These differences in VOC emissions and olfactory responses induced in granary weevils by the yellow and pigmented wheat kernels can be exploited to characterize resistance mechanisms associated with different genotypes and to incorporate resistance into improved varieties.

Metabolomics is increasingly being applied in various fields offering a highly informative tool for high-throughput diagnostics. However, in plant sciences, metabolomics is underused, even though plant studies are relatively easy and cheap when compared to those on humans and animals. Despite their importance for human nutrition, cereals, and especially wheat, remain understudied from a metabolomics point of view. The metabolomics of durum wheat has been essentially neglected, although its genetic structure allows the inference of common mechanisms that can be extended to other wheat and cereal species. This review covers the present achievements in durum wheat metabolomics highlighting the connections with the metabolomics of other cereal species (especially bread wheat). We discuss the metabolomics data from various studies and their relationships to other "-omics" sciences, in terms of wheat genetics, abiotic and biotic stresses, beneficial microbes, and the characterization and use of durum wheat as feed, food, and food ingredient.

SummaryMounting evidence indicates the key role of Nitrogen (N) on diverse processes in plant, including not only yield but also development and defense. Using a combined transcriptomics and metabolomics approach, we studied the response of seedlings to N starvation of two different tetraploid wheat genotypes from the two main domesticated subspecies, emmer (Triticum turgidum ssp. dicoccum) and durum wheat (Triticum turgidum ssp. durum). We found that durum wheat exhibits broader and stronger response in comparison to emmer as evidenced by the analysis of the differential expression pattern of both genes and metabolites and gene enrichment analysis. Emmer and durum wheat showed major differences in the responses to N starvation for transcription factor families. While emmer showed differential reduction in the levels of primary metabolites to N starvation, durum wheat exhibited increased levels of most metabolites, including GABA as an indicator of metabolic imbalance. The correlation-based networks including the differentially expressed genes and metabolites revealed tighter regulation of metabolism in durum wheat in comparison to emmer, as evidenced by the larger number of significant correlations. We also found that glutamate and GABA had highest values of centrality in the metabolic correlation network, suggesting their critical role in the genotype-specific response to N starvation of emmer and durum wheat, respectively. Moreover, this finding indicates that there might be contrasting strategies associated to GABA and Glutamate signaling modulating shoot vs root growth in the two different wheat subspecies.

Recently, the interest in improving the content of bioactive compounds for enhancing the nutritional value of cereal-based products has largely increased, and several strategies, both genetic and agronomic, were proposed. Here, we report the preliminary results of the effect of two sowing periods (winter and spring) on the accumulation of bioactive compounds (i.e., anthocyanins, carotenoids, minerals) and grain morphology in three pigmented cereal species (barley, bread and durum wheat). The results showed that with delayed sowing time, a significant reduction in yield was observed in all species, less in barley (50.4%) than in wheat (70.3% and 66.5% for bread and durum, respectively). On the contrary, the carotenoids accumulation was favored (increased by 12.2%, 15.7% and 27.8% for barley, bread and durum wheat, respectively), as well as those of anthocyanins although strongly dependent on the genotype (i.e., considering only the purple genotypes, the anthocyanins increase by 4.6%, 35.4% and 72.8% in barley, bread and durum wheat, respectively). For minerals compounds, our study highlighted a strong environmental influence even when the pigmented genotypes accumulated more minerals than commercial ones, while the morphometric seed traits were less influenced by this agronomic practice. Therefore, by exploiting the genetic variability for the accumulation of anthocyanins and carotenoids in the grain and by optimizing the sowing time, it could be possible to increase the nutritional value of the final cereal-based products.

A multidisciplinary approach was adopted to investigate multiple relationships between root morphology, rhizosphere soil exudates and soil microbial community in eight durum wheat [Triticum turgidum ssp. durum (Desf.)] cultivars previously characterized in terms of their ability to interact with mycorrhizal fungi. Six plants from each cultivar were grown to the five-leaf stage in a growth chamber. Root system architecture was characterized by root scans, root exudates were identified and quantified by gas chromatography–mass spectrometry (GC–MS), and the rhizosphere bacterial, archaeal and fungal community structure was assessed using terminal-restriction fragment length polymorphism (T-RFLP). Wheat cultivars were very different in terms of the morphological root traits. Cappelli, Grecale, Ofanto and PR22D89 cultivars showed the most developed root system. Root exudates included sugars, amino acids, polyalcohols and organic acids, with carbohydrates accounting for 87.3% of the whole data set. All the compounds showed significant differences among cultivars, with Grecale and Claudio showing the highest exudation ability of metabolites. T-RFLP profiles showed a greater relative abundance of bacterial taxa (55.8% on average) than those of archaea and fungi (35.6% and 8.6%, on average, respectively) in all cultivars, and responded positively to root exudation. Principal component analysis identified root dry weight, surface area and volume, carbohydrate exudation, bacterial and archaeal Shannon–Weaver diversity indices and the fungal evenness index as the traits that mainly contributed to the cultivar variability. A larger root system did not always promote a higher diversity of the rhizosphere microbial biomass. This then suggests complex interactions between traits, probably due to the differences in the quality and/or composition of the root exudates. Based on our results, we propose that durum wheat cultivars could be classified according to the different plant resource-use strategies employed, such as conservative (i.e. Cappelli, with a large root system but low exudation ability) and explorative/competitive (i.e. Grecale and Claudio, with a high exudation ability) cultivars. The results are discussed with reference to the complex relationships between the plant and the rhizospheric microorganisms in order to determine the best combinations that would enhance root efficiency.

Whole grains were characterised using a metabolite profiling approach for 11 genotypes that belonged to five modern and ancient Triticum sub-species, briefly distinguished as: monococcum, dicoccum, spelta, durum and aestivum. Hydrophilic and lipophilic metabolites were extracted separately, and about 51 compounds of different classes were analysed: sugars, alcohols, amino acids, organic acids, plant sterols, phenolic lipids, tocopherols and fatty acids. Differences in the metabolite content among sub-species were statistically significant for 22 compounds out of 51. On the basis of multivariate analysis, the metabolites that provide the greatest contributions to discrimination among these five sub-species were identified. Galacturonic acid (precursor of ascorbic acid) was detected only in dicoccum. Alkylresorcinol homologues AR17 and AR19 were greatly accumulated by aestivum and spelta. Aspartic acid and asparagine were highest in monococcum. Asparagine was not detected in durum and aestivum, but these instead had the highest levels of the correlated essential amino acid tryptophan. The alkylresorcinol homologue composition, and in particular the AR17/AR23 ratio, can be used to distinguish between the monococcum, hexaploid and tetraploid species. Vice versa, the distinctions (in terms of these metabolites), between the two tetraploid species and between the hexaploid sub-species were difficult to clearly sort out.

The key mechanisms of salinity tolerance (ST) in durum wheat were investigated, with five genotypes used to determine changes in morpho-physiological traits and mineral and metabolite contents after exposure to 50, 100 and 200 mM NaCl. Plant growth impairment was evident at the highest salt level. Under this condition, a wide range of shoot Na+ contents and ST were observed within genotypes. However, no significant correlation was seen between ST and Na+ exclusion from the shoots, which indicates that tissue tolerance also has a role. Consistent with this, there was significant correlation between ST and the Na+:K+ ratio in the shoots. Indeed, the maintenance of the shoot Na+ and K+ homeostasis was found to be essential to achieve osmotic adjustment, which relied substantially on inorganic osmolytes, and to avoid toxicity symptoms, such as chlorophyll loss, which appeared only at the highest salinity level. Consistently, the metabolite changes occurred mainly in the shoots, with a dual response to salinity: (i) a conserved response that was common to all the genotypes and resulted in the accumulation of proline and in the depletion of organic acids, including some intermediates of the Krebs cycle; and (ii) a genotype-specific response that involved the accumulation of GABA, threonine, leucine, glutamic acid, glycine, mannose and fructose and appeared related to the different tolerance of genotypes to salinity. The lower magnitude of response to salinity detected in the roots confirmed the major role of the shoots in the determination of ST of durum wheat.

Macro- and micronutrients, essential for the maintenance of human metabolism, are assimilated daily through the diet. Wheat and other major cereals are a good source of nutrients, such as carbohydrates and proteins, but cannot supply a sufficient amount of essential micronutrients, including provitamin A. As vitamin A deficiency (VAD) leads to several serious diseases throughout the world, the biofortification of a major staple crop, such as wheat, represents an effective way to preserve human health in developing countries. In the present work, a key enzyme involved in the branch of carotenoids pathway producing &beta, carotene, lycopene epsilon cyclase, has been targeted by a Targeting Induced Local Lesions in Genomes (TILLING) approach in a &ldquo, block strategy&rdquo, perspective. The null mutant genotype showed a strong reduction in the expression of the lcyE gene and also interesting pleiotropic effects on an enzyme (&beta, ring hydroxylase) acting downstream in the pathway. Biochemical profiling of carotenoids in the wheat mutant lines showed an increase of roughly 75% in &beta, carotene in the grains of the complete mutant line compared with the control. In conclusion, we describe here the production and characterization of a new wheat line biofortified with provitamin A obtained through a nontransgenic approach, which also sheds new light on the molecular mechanism governing carotenoid biosynthesis in durum wheat.

The concentrations of 10 minerals were investigated in the grain of 12 durum wheat genotypes grown under free air CO 2 enrichment conditions, and in four of their derived pasta samples, using inductively coupled plasma mass spectrometry. Compared to ambient CO 2 (400 ppm; AMB), under elevated CO 2 (570 ppm; ELE), the micro-element and macro-element contents showed strong and significant decreases in the grain: Mn, −28.3%; Fe, −26.7%; Zn, −21.9%; Mg, −22.7%; Mo, −40.4%; K, −22.4%; and Ca, −19.5%. These variations defined the 12 genotypes as sensitive or non-sensitive to ELE. The pasta samples under AMB and ELE showed decreased mineral contents compared to the grain. Nevertheless, the contributions of the pasta to the recommended daily allowances remained relevant, also for the micro-elements under ELE conditions (range, from 18% of the recommended daily allowance for Zn, to 70% for Mn and Mo).

Domestication has induced major genetic changes in crop plants to satisfy human needs and as a consequence of adaptation to agroecosystems. This adaptation might have affected root exudate composition, which can influence the interactions in the rhizosphere. Here, using two different soil types (sand, soil), we provide an original example of the impact of domestication and crop evolution on root exudate composition through metabolite profiling of root exudates for a panel of 10 wheat genotypes that correspond to the key steps in domestication of tetraploid wheat (wild emmer, emmer, durum wheat). Our data show that soil type can dramatically affect the composition of root exudates in the rhizosphere. Moreover, the composition of the rhizosphere metabolites is associated with differences among the genotypes of the wheat domestication groups, as seen by the high heritability of some of the metabolites. Overall, we show that domestication and breeding have had major effects on root exudates in the rhizosphere, which suggests the adaptive nature of these changes.

BACKGROUND Several studies have demonstrated that metabolomics has a definite place in food quality, nutritional value, and safety issues. The aim of the present study was to determine and compare the metabolites in different pasta samples with fibre, and to investigate the modifications induced in these different kinds of pasta during cooking, using a gas chromatography–mass spectrometry-based metabolomics approach. RESULTS Differences were seen for some of the amino acids, which were absent in control pasta, while were present both in the commercially available high-fibre pasta (samples A–C) and the enriched pasta (samples D–F). The highest content in reducing sugars was observed in enriched samples in comparison with high-fibre pasta. The presence of stigmasterol in samples enriched with wheat bran was relevant. Cooking decreased all of the metabolites: the high-fibre pasta (A–C) and Control showed losses of amino acids and tocopherols, while for sugars and organic acids, the decrease depended on the pasta sample. The enriched pasta samples (D–F) showed the same decreases with the exception of phytosterols, and in pasta with barley the decrease of saturated fatty acids was not significant as for tocopherols in pasta with oat. Principal component analysis of the metabolites and the pasta discrimination was effective in differentiating the enriched pasta from the commercial pasta, both uncooked and cooked. CONCLUSIONS The study has established that such metabolomic analyses provide useful tools in the evaluation of the changes in nutritional compounds in high-fibre and enriched pasta, both before and after cooking. © 2015 Society of Chemical Industry

Smell and aroma are important determinants of consumer acceptance, so gaining deeper insight into bread smell and aroma perception is a research goal. Sixteen combinations of four variables were investigated, to evaluate the contributions of bread chemical and rheological properties and volatile organic compounds (VOCs) towards sensory acceptability of breads: genotypes (landrace vs. modern); types of flour (wholemeal vs. semolina); leavening agents (brewing yeast vs. sourdough starter); and baking modes (gas-fired vs. wood-fired). Milling had the greatest impact over the other treatments for the rheological and chemical properties, including for VOCs, with great impact on the sensory traits of the flours and breads. The processing phases had great impact on smell and aroma, as defined through formation of alcohols, aldehydes, terpenes, and other compounds (e.g., ethylbenzene, 2-pentylfuran, methoxyphenyl oxime). Leavening agent had great impact on sensory perception, although breads from the sourdough starter were perceived as with lower taste and colour than the brewing yeast. Baking mode had no relevant role on sensory perception. These data strongly undermine the belief of a ‘better product’ that is frequently attributed to old genotypes versus modern cultivars, and indicate that the milling and the bread-making processes determine the quality of the end product.

The concentrations of 10 minerals were investigated in the grain of 12 durum wheat genotypes grown under free air CO

The aim of this study was to assess the relative roles of genotype, environment and genotype-by-environment interactions in determining the metabolite profile of durum wheat grain. Four durum wheat cultivars were grown under conventional and organic farming systems over three consecutive years. The use of a high-throughput gas chromatography–mass spectrometry platform allowed the analysis of sets of different polar and non-polar compounds, including amino acids, sugars, organic acids, fatty acids (saturated and unsaturated), and sterols. Statistical analysis of the data showed a small impact of genotype and large effects of both year and genotype-by-environment interaction on the metabolite composition and quality of the wheat grain. Overall, the data from this study highlight the potential role of metabolic profiling in the analysis of durum wheat quality and production.

Domestication and breeding have influenced the genetic structure of plant populations due to selection for adaptation from natural habitats to agro-ecosystems. Here, we investigate the effects of selection on the contents of 51 primary kernel metabolites and their relationships in three Triticum turgidum L. subspecies (i.e., wild emmer, emmer, durum wheat) that represent the major steps of tetraploid wheat domestication. We present a methodological pipeline to identify the signature of selection for molecular phenotypic traits (e.g., metabolites and transcripts). Following the approach, we show that a reduction in unsaturated fatty acids was associated with selection during domestication of emmer (primary domestication). We also show that changes in the amino acid content due to selection mark the domestication of durum wheat (secondary domestication). These effects were found to be partially independent of the associations that unsaturated fatty acids and amino acids have with other domestication-related kernel traits. Changes in contents of metabolites were also highlighted by alterations in the metabolic correlation networks, indicating wide metabolic restructuring due to domestication. Finally, evidence is provided that wild and exotic germplasm can have a relevant role for improvement of wheat quality and nutritional traits.

Durum wheat (Triticum turgidum (L.) subsp. turgidum (L.) convar. durum (Desf.)) is momentous for human nutrition, and environmental stresses can strongly limit the expression of yield potential and affect the qualitative characteristics of the grain. The aim of this study was to determine how heat stress (five days at 37 °C) applied five days after flowering affects the nutritional composition, antioxidant capacity and metabolic profile of the grain of two durum wheat genotypes: “Primadur”, an elite cultivar with high yellow index, and “T1303”, an anthocyanin-rich purple cultivar. Qualitative traits and metabolite evaluation (by gas chromatography linked to mass spectrometry) were carried out on immature (14 days after flowering) and mature seeds. The effects of heat stress were genotype-dependent. Although some metabolites (e.g., sucrose, glycerol) increased in response to heat stress in both genotypes, clear differences were observed. Following the heat stress, there was a general increase in most of the analyzed metabolites in “Primadur”, with a general decrease in “T1303”. Heat shock applied early during seed development produced changes that were observed in immature seeds and also long-term effects that changed the qualitative and quantitative parameters of the mature grain. Therefore, short heat-stress treatments can affect the nutritional value of grain of different genotypes of durum wheat in different ways.

The carotenoid concentration of durum wheat is a criterion for the assessment of semolina quality, and it is of particular importance in determining the color of pasta. Thus, the development of a rapid screening method for kernel and semolina color has facilitated increasing the yellow color in durum wheat cultivars. However, the distribution of the pigment might vary within the ear of the wheat. The micromethod for the determination of yellow pigment concentration (YPC) of single wheat kernels now allows the discovery of whether this variation is the same for cultivars with different carotenoid concentrations. All of the cultivars investigated showed similar trends in the arrangements of the kernels within the ears, with the central positions of the ears being the most stable for YPC. Indeed, the best combination of higher YPC and larger kernel size is seen for the basal-central region of the ear, which can be used for the selection during wheat-breeding programs of cultivars with a more intense...

Colour is an important component of durum wheat quality affecting consumer’s choice. Pasta colour is the result of the carotenoid pigments accumulated in the seed endosperm and of the enzymatic activities responsible for their degradation. Thus, increasing yellow colour is a valuable goal for improving the commercial and nutritional values of durum wheat products. This work reports the set up of a micro-method for the determination of Yellow Pigment Content (YPC) in durum wheat. The novel method, although similar in principle to the Official Method AACC 14-50, allows the use of micro-amount of samples (10–100 mg), a condition that for durum wheat corresponds to a single kernel, and micro-amount of extraction solvent (water-saturated 1-butanol, 250–500 μL). These improvements together with a short extraction time lead to significant benefits from the practical and the economic point of view. The improved method has been validated using nine durum wheat cultivars extracted in 15 min with the ultrasonic bath. The YPC was calculated by reading the absorbance at 435 nm after the manual injection of an aliquot of extract directly into a UV–Vis HPLC detector. The results obtained were comparable and not significantly different from the Official AACC Method 14-50.

Using a polyphasic approach, we have isolated and identified, lactic acid bacteria (LAB) in samples directly collected from an artisanal pasta-making manufactory located in Puglia (South Italy). Samples were collected during several steps of pasta processing and LAB were isolated on MRS and M17 plates. Furthermore, strains were grouped in a total of eight species by means of randomly amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) typing and 16S rDNA sequencing. The majority of strains were identified as belonging to Pediococcus pentosaceus and Enterococcus faecium species. The remaining strains were characterized and assigned to Weissella confusa, Pediococcus acidilactici, Leuconostoc mesenteroides, Leuconostoc citreum, Lactobacillus fermentum and Lactobacillus plantarum species. Strains were identified from all the analysed samples, and growth of LAB was monitored from extrusion to pre-heating steps.

Mineral deficiencies are prevalent in human populations and the improvement of the mineral content in cereal products represents a possible strategy to increase the human mineral intake. Nevertheless, most of the inorganic phosphorus (Pi) present in mature cereal seeds (40–80%) is stored as phytate, an anti-nutritional factor that forms complexes with minerals such as Ca, Mg, Zn and Fe reducing their bioavailability. The present study was undertaken: (i) to determine the variation in phytate and mineral concentrations in the whole grains of 84 Italian durum wheat (Triticum durum Desf.) cultivars representative of old and modern germplasm; (ii) to estimate the magnitude of genotype × environment interaction effects; and (iii) to examine the interrelationships among mineral concentrations in durum wheat with the final aim to identify superior durum wheat cultivars that possess low phytate content and high concentration of mineral elements in their whole-wheat flour. The cultivars were grown in field trials during 2004–2005 at Foggia, Italy and during 2005–2006 at Foggia and Fiorenzuola d’Arda—Southern and Northern Italy. The phytate content was estimated indirectly by using a microtitre plate assay evaluating the Pi absorbance at 820 nm, while the Cu, Fe, Mn, Ca, K, Mg, Na and Zn mineral contents were determined by ICP/OES. The contents of Zn and Fe across years and locations ranged from 28.5 to 46.3 mg/kg for Zn with an average of 37.4 mg/kg and from 33.6 to 65.6 mg/kg for Fe with an average of 49.6 mg/kg. Pi grain content was between 0.46 and 0.76 mg/g showing a positive correlation with all minerals except Cu and Zn. Although breeding activity for Fe and Zn would be difficult because G × E interaction is prevalent, multi-location evaluation of germplasm collection help to identify superior genotypes to achieve this objective. The results here reported open the possibility of designing a specific breeding program for improving the nutritional value of durum wheat through the identification of parental lines with low-Pi and high minerals concentration in whole grains.

Although pasta is generally not considered for its aromatic properties, some evidence proves that cereal flours release volatile compounds and they might have an effect on the aroma of the transformed products. This work reports on the characterization of the volatile components of semolina and pasta obtained from four durum wheat cultivars (Triticum durum Desf., cvs. PR22D89, Creso, Cappelli, Trinakria). Semolina samples were characterized through polar metabolite profiling and fatty acid analysis to identify potential precursors of the volatile components. The results show significant differences among the samples tested with cv. Trinakria characterized by the highest content of sugars and fatty acids. Volatile composition was investigated both in semolina and in cooked pasta using headspace solid-phase micro-extraction (HS-SPME) and identified by GC–MS. Thirty-five volatile compounds including aldehydes, ketones, alcohols, terpenes, esters, hydrocarbons and a furan were identified. Significant differences were observed between semolina and pasta samples in terms of composition and amount of the volatile compounds. During cooking an increase in aldehyde content, the appearance of ketones and a decrease in alcohol content were observed. Correlations between metabolites and volatiles demonstrate that the flavour of cooked pasta may differ significantly depending on the durum wheat cultivar employed.

The anatomy of strawberry (Fragaria × ananassa) fruit, in which the achene is found on the outer part of the fruit, makes it an excellent species for studying the regulation of fruit development. It can provide a model for the cross talk between primary and secondary metabolism, whose role is of pivotal importance in the process. By combining gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry with the aim of addressing the metabolic regulation underlying fruit seed development, we simultaneously analyzed the composition of primary and secondary metabolites, separately, in achene and receptacle during fruit ripening of strawberry cultivar Herut. The results from these analyses suggest that changes in primary and secondary metabolism reflect organ and developmental specificities. For instance, the receptacle was characterized by increases in sugars and their direct derivatives, while the achene was characterized by a major decrease in the levels of carbon- and nitrogen-rich compounds, with the exception of storage-related metabolites (e.g. raffinose). Furthermore, the receptacle, and to a lesser extent the achene, exhibited dynamic fluctuations in the levels and nature of secondary metabolites across the ripening process. In the receptacle, proanthocyanidins and flavonol derivatives characterized mainly early developmental stages, while anthocyanins were abundant in the mature red stage; in the achene, ellagitannin and flavonoids were abundant during early and late development, respectively. Correlation-based network analysis suggested that metabolism is substantially coordinated during early development in either organ. Nonetheless, a higher degree of connectivity within and between metabolic pathways was measured in the achenes. The data are discussed within the context of current models both of the interaction of primary and secondary metabolism and of the metabolic interaction between the different plant organs.

Several studies have demonstrated that metabolomics has a definite place in food quality, nutritional value, and safety issues. The aim of the present study was to determine and compare the metabolites in different pasta samples with fibre, and to investigate the modifications induced in these different kinds of pasta during cooking, using a gas chromatography-mass spectrometry-based metabolomics approach.Differences were seen for some of the amino acids, which were absent in control pasta, while were present both in the commercially available high-fibre pasta (samples A-C) and the enriched pasta (samples D-F). The highest content in reducing sugars was observed in enriched samples in comparison with high-fibre pasta. The presence of stigmasterol in samples enriched with wheat bran was relevant. Cooking decreased all of the metabolites: the high-fibre pasta (A-C) and Control showed losses of amino acids and tocopherols, while for sugars and organic acids, the decrease depended on the pasta sample. The enriched pasta samples (D-F) showed the same decreases with the exception of phytosterols, and in pasta with barley the decrease of saturated fatty acids was not significant as for tocopherols in pasta with oat. Principal component analysis of the metabolites and the pasta discrimination was effective in differentiating the enriched pasta from the commercial pasta, both uncooked and cooked.The study has established that such metabolomic analyses provide useful tools in the evaluation of the changes in nutritional compounds in high-fibre and enriched pasta, both before and after cooking. © 2015 Society of Chemical Industry.

The process of domestication has led to dramatic morphological and physiological changes in crop species due to adaptation to cultivation and to the needs of farmers. To investigate the phenotypic architecture of shoot- and root-related traits and quantify the impact of primary and secondary domestication, we examined a collection of 36 wheat genotypes under optimal and nitrogen-starvation conditions. These represented three taxa that correspond to key steps in the recent evolution of tetraploid wheat (i.e. wild emmer, emmer, and durum wheat). Overall, nitrogen starvation reduced the shoot growth of all genotypes, while it induced the opposite effect on root traits, quantified using the automated phenotyping platform GROWSCREEN-Rhizo. We observed an overall increase in all of the shoot and root growth traits from wild emmer to durum wheat, while emmer was generally very similar to wild emmer but intermediate between these two subspecies. While the differences in phenotypic diversity due to the effects of primary domestication were not significant, the secondary domestication transition from emmer to durum wheat was marked by a large and significant decrease in the coefficient of additive genetic variation. In particular, this reduction was very strong under the optimal condition and less intense under nitrogen starvation. Moreover, although under the optimal condition both root and shoot traits showed significantly reduced diversity due to secondary domestication, under nitrogen starvation the reduced diversity was significant only for shoot traits. Overall, a considerable amount of phenotypic variation was observed in wild emmer and emmer, which could be exploited for the development of pre-breeding strategies.

The process of domestication has led to dramatic morphological and physiological changes in crop species due to adaptation to cultivation and to the needs of farmers. To investigate the phenotypic architecture of shoot- and root-related traits and quantify the impact of domestication and post-domestication, we examined a collection of 36 wheat genotypes under optimal and nitrogen-starved conditions. These represented three taxa that correspond to key steps in the recent evolution of tetraploid wheat (i.e., wild emmer, emmer, and durum wheat). Overall, nitrogen starvation reduced shoot growth of all genotypes, while it induced the opposite effect on root traits quantified using the automated phenotyping platform GROWSCREEN-Rhizo. We observed an overall increase in all shoot and root growth traits from wild emmer to durum wheat, while emmer was generally very similar to wild emmer, but intermediate between these two subspecies. While the differences in phenotypic diversity due to the effects of domestication were not significant, the post-domestication transition from emmer to durum wheat was marked by a large and significant decrease in the coefficient of variation. In particular, this reduction was very strong at optimal condition, and less intense under nitrogen starvation. Moreover, while at optimal condition both root and shoot traits showed significantly reduced diversity due to post-domestication, under nitrogen starvation the reduced diversity was significant only for shoot traits. Overall, a considerable amount of phenotypic variation was observed in wild emmer and emmer which could be exploited for the development of pre-breeding strategies.

Tetrabutylammonium fluoride (TBAF) is an efficient catalyst in the [3 + 2] cycloaddition reaction of organic nitriles 1 with trimethylsilyl azide (TMSN(3)) in solventless conditions. The corresponding 5-substituted 1H-tetrazoles 2 were obtained under mild conditions and in 80-97% yields.

To investigate the regulation of seed metabolism and to estimate the degree of metabolic natural variability, metabolite profiling and network analysis were applied to a collection of 76 different homozygous tomato introgression lines (ILs) grown in the field in two consecutive harvest seasons. Factorial ANOVA confirmed the presence of 30 metabolite quantitative trait loci (mQTL). Amino acid contents displayed a high degree of variability across the population, with similar patterns across the two seasons, while sugars exhibited significant seasonal fluctuations. Upon integration of data for tomato pericarp metabolite profiling, factorial ANOVA identified the main factor for metabolic polymorphism to be the genotypic background rather than the environment or the tissue. Analysis of the coefficient of variance indicated greater phenotypic plasticity in the ILs than in the M82 tomato cultivar. Broad-sense estimate of heritability suggested that the mode of inheritance of metabolite traits in the seed differed from that in the fruit. Correlation-based metabolic network analysis comparing metabolite data for the seed with that for the pericarp showed that the seed network displayed tighter interdependence of metabolic processes than the fruit. Amino acids in the seed metabolic network were shown to play a central hub-like role in the topology of the network, maintaining high interactions with other metabolite categories, i.e., sugars and organic acids. Network analysis identified six exceptionally highly co-regulated amino acids, Gly, Ser, Thr, Ile, Val, and Pro. The strong interdependence of this group was confirmed by the mQTL mapping. Taken together these results (i) reflect the extensive redundancy of the regulation underlying seed metabolism, (ii) demonstrate the tight co-ordination of seed metabolism with respect to fruit metabolism, and (iii) emphasize the centrality of the amino acid module in the seed metabolic network. Finally, the study highlights the added value of integrating metabolic network analysis with mQTL mapping., Author Summary Seeds represent 70% of the food source for man and livestock. However, as a result of millennia of domestication, crop plants have undergone major genetic deterioration, leading to a loss of important quality traits. Thus, the reintroduction of these quality traits is the key to the improvement of crops in modern agriculture. Seed quality traits include nutritional components, such as proteins and amino acids, and seed germination and storability, which are, in turn, inherently related to metabolism. To understand the genetic basis of seed metabolism—a strategic need in the improvement of seed crops—we studied a collection of offspring plants stemming from the cross between a domesticated tomato cultivar Solanum lycopersicum cv M82 and its distant wild relative S. pennellii. We monitored the changes in metabolism and studied the mode of regulation of the concentration of metabolites in the seeds as a result of genetic introgression, by taking advantage of state-of-the-art technologies and methods of data elaboration such as network-based analysis. We identified a number of candidate genes that may be useful in manipulations to enhance nutritional values in seeds. Finally, in an effort to study the relation among the seed, the fruit, and the mother plant, we determined potential yield-associated metabolic markers.