Assessing the effects of Bt maize on the non-target pest Rhopalosiphum maidis by demographic and life-history measurement endpoints.

The most commercialized Bt maize plants in Europe were transformed with genes which express a truncated form of the insecticidal delta-endotoxin (Cry1Ab) from the soil bacterium Bacillus thuringiensis (Bt) specifically against Lepidoptera. Studies on the effect of transgenic maize on non-target arthropods have mainly converged on beneficial insects. However, considering the worldwide extensive cultivation of Bt maize, an increased availability of information on their possible impact on non-target pests is also required. In this study, the impact of Bt-maize on the non-target corn leaf aphid, Rhopalosiphum maidis, was examined by comparing biological traits and demographic parameters of two generations of aphids reared on transgenic maize with those on untransformed near-isogenic plants. Furthermore, free and bound phenolics content on transgenic and near-isogenic plants were measured. Here we show an increased performance of the second generation of R. maidis on Bt-maize that could be attributable to indirect effects, such as the reduction of defense against pests due to unintended changes in plant characteristics caused by the insertion of the transgene. Indeed, the comparison of Bt-maize with its corresponding near-isogenic line strongly suggests that the transformation could have induced adverse effects on the biosynthesis and accumulation of free phenolic compounds. In conclusion, even though there is adequate evidence that aphids performed better on Bt-maize than on non-Bt plants, aphid economic damage has not been reported in commercial Bt corn fields in comparison to non-Bt corn fields. Nevertheless, Bt-maize plants can be more easily exploited by R. maidis, possibly due to a lower level of secondary metabolites present in their leaves. The recognition of this mechanism increases our knowledge concerning how insect-resistant genetically modified plants impact on non-target arthropods communities, including tritrophic web interactions, and can help support a sustainable use of genetically modified crops.

Elucidating the Genetic Relationships on the Original Old Sicilian Triticum Spp. Collection by SNP Genotyping.

Several Triticum species spread in cultivation in Sicily and neighboring regions over the centuries, which led to the establishment of a large genetic diversity. Many ancient varieties were widely cultivated until the beginning of the last century before being replaced by modern varieties. Recently, they have been reintroduced in cultivation in Sicily. Here, the genetic diversity of 115 and 11 accessions from Sicily and Calabria, respectively, belonging to Triticum species was evaluated using a high-density SNP array. Einkorn, emmer, and spelta wheat genotypes were used as outgroups for species and subspecies; five modern varieties of durum and bread wheat were used as references. A principal coordinates analysis (PCoA) and an unweighted pair group method with arithmetic mean (UPGMA) showed four distinct groups among Triticum species and T. turgidum subspecies. The population structure analysis distinguished five gene pools, among which three appeared private to the T. aestivum, T. turgidum subsp. Turgidum, and 'Timilia' group. The principal component analysis (PCA) displayed a bio-morphological trait relationship of a subset (110) of ancient wheat varieties and their wide variability within the T. turgidum subsp. durum subgroups. A discriminant analysis of principal components (DAPC) and phylogenetic analyses applied to the four durum wheat subgroups revealed that the improved varieties harbored a different gene pool compared to the most ancient varieties. The 'Russello' and 'Russello Ibleo' groups were distinguished; both displayed higher genetic variability compared to the 'Timilia' group accessions. This research represents a comprehensive approach to fingerprinting the old wheat Sicilian germplasm, which is useful in avoiding commercial fraud and sustaining the cultivation of landraces and ancient varieties.

Phenolic acids of modern and ancient grains: Effect on in vitro cell model.

BACKGROUND: The gastrointestinal tract establishes a barrier between the external and internal compartments. When this barrier is disrupted, an inflammatory cascade promotes intestinal inflammation and the development of several intestinal diseases. Plant-derived polyphenols are health-promoting phytochemicals with a role in the regulation of the intestinal barrier and in the prevention of intestinal inflammatory diseases. Modern wheat-breeding programs have been focused primarily on yield improvement rather than nutritional and functional proprieties. Research that aims to characterize the phytochemical profile of wheat varieties and their healthy proprieties could therefore provide new prospects for the genetic improvement of the genus Triticum. In the present work, the effects of phenolic compounds extracted from nine soft and seven durum wheat varieties were studied for their polyphenol content and antioxidant activity. Experiments were conducted to study their effects on cell proliferation and wound healing in three different cell lines: mouse fibroblasts (L929), intestinal human cells (Caco2), and human monocytes (U937). RESULTS: Discriminant analysis evidenced differences between soft and durum wheat phenolic compounds. Among the soft varieties, it was possible to identify clusters in which ancient wheat varieties showed different properties from modern ones, whereas no evident clusters were detected among durum varieties. CONCLUSION: Taken together, these results suggest that the selection of specific wheat grains based on their nutritional parameters will help in the design of diets with protective effects against chronic and inflammatory diseases. © 2019 Society of Chemical Industry.

A Khorasan wheat-based diet improves systemic inflammatory profile in semi-professional basketball players: a randomized crossover pilot study.

BACKGROUND/OBJECTIVES: Khorasan wheat is an ancient grain with widely acclaimed beneficial effects on human health. The objective of the study was to examine the effect of a Khorasan-based diet on the wellbeing and inflammatory profile of young athletes. RESULTS: We conducted a randomized, single-blinded crossover trial involving 20 male young athletes. The participants were randomly assigned to consume products (pasta, bread, biscuits and crackers) made either with Khorasan (KAMUT® brand) or modern semi-whole-grain wheat for 4-weeks with a 4-week washout period before the crossover. Laboratory analyses and fitness tests were performed both at the beginning and end of each diet period. The consumption of Khorasan products was associated with a significant reduction of monocyte chemoattractant protein-1 (MCP-1; mean reduction: -36.15 pg/mL; -25.67%) while the consumption of modern wheat was not associated with significant differences in Interleukin-8 (IL-8) or Interleukin-1 receptor antagonist (IL-1ra). The consumption of the Khorasan-based diet also resulted in a significant improvement in self-rated health status. No statistically significant differences in any athletic performance parameter were observed between the two diets. CONCLUSION: The present results suggest that a Khorasan-based diet could be effective in reducing the inflammatory status in young athletes. © 2019 Society of Chemical Industry.

Determination of phenolic compounds in ancient and modern durum wheat genotypes.

Due to its significant amount of antioxidants, durum wheat (Triticum turgidum ssp. durum) could potentially contribute to the protection against a number of chronic diseases, such as diabetes, cardiovascular disease, and cancer. The increasing interest toward healthy food among both consumers and scientists has moved the focus toward the phytochemical content of whole wheat grains. The aim of this study was to identify the phytochemical composition of 22 cultivars belonging to old and modern durum wheat genotypes, including antioxidant capacity (DPPH and FRAP tests). In addition, five phenolic acids involved in the vanillin biosynthesis pathway and in the defence mechanism of plants were screened using UHPLC-MS/MS. Remarkable quantitative differences in the amount of the five phenolic acids analysed (p < 0.05) were detected among the wheat genotypes investigated. Results showed that among the investigated phenolic compounds, trans-ferulic acid was the most abundant, ranging from 13.28 to 324.69 µg/g; all the other identified compounds were present at lower concentrations. Moreover, significant differences on the antioxidant activity were observed. Collected data suggested possible differences between biosynthetic pathway of secondary metabolites among durum wheat genotypes.

Environment and genotype effects on antioxidant properties of organically grown wheat varieties: a 3-year study.

BACKGROUND: Wheat grain (Triticum aestivum L.) possesses significant amounts of antioxidants that contribute to the dietary antiradical protection against a number of chronic diseases. Despite the increasing interest in organic food among both consumers and scientists, the availability of literature studies concerning the environment effect under organic management is still scarce. The aim of this study was to evaluate the antioxidant properties of wheat varieties by considering the genotype response to different environmental factors under biodynamic management. RESULTS: The soluble fraction of phenolic compounds was mainly determined by the environment, whereas a major genotypic effect was observed for the bound forms, which were present at higher amounts in red grain varieties. Moreover, a predominant effect of genotype was observed for yellow pigment content and antioxidant activity determined by the FRAP method. Despite some changes induced by environment, most genotypes had stable antioxidant properties and different phenolic profiles as determined by high-performance liquid chromatography-mass spectrometry, except for the old variety Inallettabile, which was the most sensitive to environmental fluctuations. CONCLUSION: The red grain varieties Andriolo, Gentil rosso and Verna were identified as the most promising breeding material for the development of varieties with high nutraceutical value under low-input management. © 2016 Society of Chemical Industry.

Bioactive peptides in cereals and legumes: agronomical, biochemical and clinical aspects.

Cereals and legumes are key components of a healthy and balanced diet. Accordingly, many national nutritional guidelines emphasize their health promoting properties by placing them at the base of nutritional food pyramids. This concept is further validated by the observed correlation between a lower risk and occurrence of chronic diseases and the adherence to dietary patterns, like the Mediterranean diet, in which cereal grains, legumes and derived products represent a staple food. In the search for a dietary approach to control/prevent chronic degenerative diseases, protein derived bioactive peptides may represent one such source of health-enhancing components. These peptides may already be present in foods as natural components or may derive from hydrolysis by chemical or enzymatic treatments (digestion, hydrolysis or fermentation). Many reports are present in the literature regarding the bioactivity of peptides in vitro and a wide range of activities has been described, including antimicrobial properties, blood pressure-lowering (ACE inhibitory) effects, cholesterol-lowering ability, antithrombotic and antioxidant activities, enhancement of mineral absorption/bioavailability, cyto- or immunomodulatory effects, and opioid-like activities. However it is difficult to translate these observed effects to human. In fact, the active peptide may be degraded during digestion, or may not be absorbed or reach the target tissues at a concentration necessary to exert its function. This review will focus on bioactive peptides identified in cereals and legumes, from an agronomical and biochemical point of view, including considerations about requirements for the design of appropriate clinical trials necessary for the assessment of their nutraceutical effect in vivo.

Biostimulants for Sustainable Management of Sport Turfgrass.

Research on the efficacy of innovative, ecofriendly biostimulants in sport turf management is scarce, with less information available from open-field experiments, and even less pertaining to thatch control-related problems. The objective was to investigate the open-field effectiveness of a commercial product, EM-1, and two newly developed products, ExpA and ExpB, in improving both rhizosphere and turfgrass, Agrostis stoloniferous L., characteristics on a golf green. ExpA and ExpB, identical in microbial composition, were equally effective in significantly increasing chlorophyll synthesis and visual turf quality, as well as in resistance to tearing out, compared to the untreated control 56 days after treatment (DAT). EM-1 showed intermediate trends between the control and novel biostimulants. The inclusion of humic acids and mycorrhizal fungi to the microbial composition in ExpB significantly improved some rhizosphere properties 56 DAT relative to the control. Results on ExpB evidenced a significant decrease in the thatch layer thickness and fresh leaf weight, associated with a significant increase in the humus thickness, organic matter decomposition and evapotranspiration efficiency. An increased dry leaf biomass was also shown. ExpA and EM-1 showed either marginal or intermediate improvements relative to the control. ExpB represents a promising alternative to alleviate negative environmental impacts associated with turf maintenance-related activities.

Prebiotic effect of soluble fibres from modern and old durum-type wheat varieties on Lactobacillus and Bifidobacterium strains.

BACKGROUND: Wheat grains are a rich source of dietary fibres, particularly in the western human diet. Many of the health effects attributed to dietary fibres are believed to be related to their microbial fermentation in the gut. This study evaluated the ability of two potentially probiotic strains, Lactobacillus plantarum L12 and Bifidobacterium pseudocatenulatum B7003, to ferment soluble dietary fibres (SDFs) from modern and ancient durum-type wheat grains. RESULTS: Fibre microbial utilisation was highly variable and dependent on the strain. SDFs from the varieties Svevo and Solex supported the growth of L. plantarum L12 the best, whereas those from the varieties Anco Marzio, Solex and Kamut(®) Khorasan were good carbohydrate substrates for B. pseudocatenulatum B7003. The highest prebiotic activity scores (describing the extent to which prebiotics support selective growth of probiotics) for B7003 were obtained with SDFs from the varieties Solex (0.57), Kamut(®) Khorasan (0.56) and Iride (0.55), whereas for L12 the highest scores were achieved with the varieties Orobel (0.63), Kamut(®) Khorasan (0.56) and Solex (0.53). CONCLUSION: The present study has identified some SDFs from durum-type wheat grains as suitable prebiotic substrates for the selective proliferation of B. pseudocatenulatum B7003 and L. plantarum L12 in vitro. The results provide the basis for the potential utilisation of wheat-based prebiotics as a component of synbiotic formulations.

Health-promoting phytochemicals of Italian common wheat varieties grown under low-input agricultural management.

BACKGROUND: The increasing interest in organic food products and environmental friendly practices has emphasised the importance of selecting crop varieties suitable for the low-input sector. Moreover, in recent years the relationship between diet and human health has gained much attention among consumers. The aim of this study was to comparatively evaluate the agronomic performance and the nutrient and phytochemical composition of old and modern Italian wheat genotypes grown under low-input management. RESULTS: Research highlighted that several old wheat genotypes were comparable to the modern ones in terms of agronomic performance and nutrient content. Genotype and environmental conditions (growing season), as well as their interaction, significantly affected the phytochemical composition of wheat grains for most of the analysed bioactive compounds. High variability was observed among the wheat genotypes for dietary fibre (154.7-183.3 g kg⁻¹), polyphenol (1.94-2.77 mg g⁻¹), tocopherol (9.1-21.2 mg kg⁻¹) and carotenoid (701.4-3243 µg kg⁻¹) content. CONCLUSION: The comparative study of old and modern wheat varieties highlighted that, under low-input conditions, ancient genotypes may equal modern ones in terms of agronomic traits and additionally provide nutraceutical value-added wheat grains. The most promising ancient varieties for the unique phytochemical profiles are Gentil rosso, Marzuolo d'aqui and Verna.

Amylase Trypsin Inhibitors (ATIs) in a Selection of Ancient and Modern Wheat: Effect of Genotype and Growing Environment on Inhibitory Activities.

Wheat amylase-trypsin inhibitors (ATIs) are a family of plant defense proteins with an important role in human health for their involvement in allergies, celiac disease and non-celiac wheat sensitivity. Information about the differences in ATI activities among wheat genotypes and the influence of the growing environment is scarce. Therefore, ten selected wheat accessions with different ploidy level and year of release, previously characterized for their ATI gene sequences, were grown during three consecutive crop years at two growing areas and used for in vitro ATI activities. The contributions of the genotype and the crop year were significant for both activities. The hexaploid wheat genotypes showed the highest inhibitory activities. Einkorn had a peculiar behavior showing the lowest alpha-amylase inhibitory activity, but the highest trypsin inhibitory activity. It was not possible to observe any trend in ATI activities as a function of the release year of the wheat samples. The two inhibitory activities were differently affected by the growing conditions and were negatively correlated with the protein content. This information can be important in understanding the extent of variation of ATI inhibitory properties in relation to the wheat genotype and the growing environment and the impact of ATIs, if any, on human health and nutrition.

Evaluation of the propensity of interspecific hybridization between oilseed rape (Brassica napus L.) to wild-growing black mustard (Brassica nigra L.) displaying mixoploidy.

Potential gene flow from transgenic Brassica napus to widely-distributed, cross-compatible weedy relatives has received significant attention. All previous, albeit scarce, research has shown little to no success in producing viable F1 hybrids between B. napus (n = 38) and B. nigra (n = 16). The present study tested the working premise that the propensity for interspecific hybridization is significantly higher between B. napus and wild-growing, B. nigra displaying mixoploidy (n = 32). Controlled hybridization was performed using local, wild-growing B. nigra (♀) x transgenic (Bt Cry1Ac) B. napus (♂). Spontaneous hybridization was performed using the same B. nigra (♀) population x non-transgenic B. napus (♂) under sympatric open-field and greenhouse conditions. The total hybridization frequency, determined by the functional expression of the Bt Cry1Ac endotoxin, was 1.8 % of the F1 hybrids (n = 35). Gene flow from non-transgenic B. napus to B. nigra ranged from 4 to 29 % in F1 hybrids, with combined wind- and wild-insect-mediated pollen dispersal being the most effective. Successful interspecific hybridization is significantly enhanced using mixoploid B. nigra progenitor material. Gene flow rates in F1 hybrids were equivalent to those previously reported between B. napus with B. rapa and B. juncea, respectively, which are at the forefront of risk assessment concerns.

Field-amplified sample injection and sweeping micellar electrokinetic chromatography in analysis of glyphosate and aminomethylphosphonic acid in wheat.

Glyphosate, a widely used herbicide, has been classified as probably carcinogenic to humans by the International Agency for Research on Cancer (IARC). In the present study a method based on Field-Amplified Sample Injection and Sweeping Micellar Electrokinetic Chromatography (FASI sweep-MEKC) has been developed and validated for determination of glyphosate and its microbial metabolite aminomethylphosphonic acid (AMPA) in wheat flour. The method involved a preliminary solid phase extraction for cleanup of the aqueous extracts from wheat flour, based sequentially on C18 and strong anion exchange cartridges, followed by derivatization using 9-fluorenylmethylchloroformate. Optimization of sample cleanup and derivatization procedure was carried out by a HPLC-UV method, whereas FASI sweep-MEKC was applied for achieving the sensitivity necessary for analysis of real samples. To this regard, optimum conditions involved the use of an extended path fused-silica capillary (80 cm total length, 50 µm, i.d.) filled with a high concentration buffer (sodium phosphate 100 mM, pH 2.2). Electrokinetic sampling was carried out at -10 kV with injection time of 700 s and the separation of the loaded analytes was performed under MEKC conditions using sodium phosphate buffer 50 mM at pH 2.2, supplemented with sodium dodecyl sulfate, 100 mM. The method was validated for linearity, precision, accuracy and sensitivity, showing that using conventional UV detection (210 nm) the achieved limit of quantitation (LOQ) values for both the analytes were widely lower than those set by Authorities. In particular, LOQ for glyphosate and AMPA were found to be 5 and 2.5 ng/mL, respectively, corresponding to 0.1 and 0.05 mg/kg, in wheat flour. The method, applied to commercially available real samples (wheat flour from different manufacturers) and to an experimental sample obtained by cv. Svevo wheat, can be considered as a convenient alternative to the existing approaches in analysis of complex matrices.

Evaluation of the potential exposure of butterflies to genetically modified maize pollen in protected areas in Italy.

Environmental impacts of genetically modified crops are mandatorily assessed during their premarket phase. One of the areas of concern is the possible impact on nontarget organisms. Crops expressing Cry toxins might affect Lepidoptera larvae living outside cultivated fields, through pollen deposition on wild plants, which constitute their food source. While pollen toxicity varies among different events, possible exposure of nontarget species depends on the agro-environmental conditions. This study was conducted in two protected areas in Italy, characterized by different climatic conditions, where many Lepidoptera species thrive in proximity to maize cultivations. To estimate the possible exposure in absence of the actual stressor (e.g., Cry1-expressing maize plants), we conducted a two-year field survey of butterflies and weeds. Indicator species were selected-Aglais (Inachis) io in the Northern site and Vanessa cardui in the Southern site-and their phenology was investigated. Pollen dispersal from maize fields was measured by collection in Petri dishes. Duration and frequency of exposure was defined by the overlap between pollen emission and presence of larvae on host plants. Different risk scenarios are expected in the two regions: highest exposure is foreseen for A. io in the Northern site, while minimal exposure is estimated for V. cardui in the Southern site. In the latter case, locally grown maize cultivars flower in mid-summer in coincidence with an aestivation period for several butterfly species due to hot and dry conditions. Moreover, host plants of V. cardui are at the end of their life cycle thus limiting food availability.

Differentiation of modern and ancient varieties of common wheat by quantitative capillary electrophoretic profile of phenolic acids.

Phenolic compounds have received great attention among the health promoting phytochemicals in common wheat (Triticum aestivum L.), mainly because of their strong antioxidant properties. In the present study a simple Capillary Zone Electrophoresis (CZE) method with UV detection was optimized and validated for the quantitation of six of the most important phenolic acids in whole grain i.e., sinapic, ferulic, syringic, p-coumaric, vanillic and p-hydroxybenzoic acid. The separation was achieved in a running buffer composed of sodium phosphate solution (50 mM) in water/methanol 80:20 (v/v) at pH 6.0 and using a fused-silica capillary at the temperature of 30 °C under application of 27 kV. By means of diode array detector, and made possible by the favorable characteristic UV spectra, the quantitation of the solutes was carried out at 200, 220 and 300 nm, in the complex matrices represented by the soluble and bound fractions of wheat flours. The validation parameters of the method i.e., linearity, sensitivity, precision, accuracy and robustness were in line with those obtained by consolidated separation techniques applied for the same purposes (e.g., HPLC-UV), with a significant advantage in term of analysis time (less than 12 min). Ten varieties of soft wheat (five modern Italian and five old Italian genotypes) were analysed and the data were subjected to Principal Components Analysis (PCA). Interestingly, significant differences of the quantitative phenolic acids profile were observed between the modern and the ancient genotypes, with the latter showing higher amount of the main represented phenolic acids.

Kombucha Beverage from Green, Black and Rooibos Teas: A Comparative Study Looking at Microbiology, Chemistry and Antioxidant Activity.

Kombucha is usually obtained from the fermentation of black or green tea by a consortium of acetic acid bacteria and yeasts. In this study, kombucha was prepared from the same starter consortium using green and black teas as well as, for the first time, an infusion of rooibos leaves (Aspalathus linearis). Microbial diversity was analysed during fermentation both in the biofilm and in the corresponding kombuchas, using culture-dependent and -independent methods. Polyphenols, flavonoids, ethanol, and acids were quantified and anti-oxidant activities were monitored. All of the Kombuchas showed similarity in bacterial composition, with the dominance of Komagataeibacter spp. Beta diversity showed that the yeast community was significantly different among all tea substrates, between 7 and 14 days of fermentation and between biofilm and kombucha, indicating the influence of the substrate on the fermenting microbiota. Kombucha from rooibos has a low ethanol concentration (1.1 mg/mL), and a glucuronic acid amount that was comparable to black tea. Although antioxidant activity was higher in black and green kombucha compared to rooibos, the latter showed an important effect on the recovery of oxidative damage on fibroblast cell lines against oxidative stress. These results make rooibos leaves interesting for the preparation of a fermented beverage with health benefits.

Isolation and Characterization of Wheat Derived Nonspecific Lipid Transfer Protein 2 (nsLTP2).

Numerous studies support the protective role of bioactive peptides against cardiovascular diseases. Cereals represent the primary source of carbohydrates, but they also contain substantial amounts of proteins, therefore representing a potential dietary source of bioactive peptides with nutraceutical activities. The analysis of wheat extracts purified by chromatographic techniques by means of HPLC-UV/nanoLC-nanoESI-QTOF allowed the identification of a signal of about 7 kDa which, following data base searches, was ascribed to a nonspecific lipid-transfer protein (nsLTP) type 2 from Triticum aestivum (sequence coverage of 92%). For the first time nsLTP2 biological activities have been investigated. In particular, in experiments with human umbilical vein endothelial cells (HUVEC), nsLTP2 displayed antioxidant and cytoprotective activities, being able to significantly decrease reactive oxygen species (ROS) levels and to reduce lactate dehydrogenase (LDH) release, generated following oxidative (hydrogen peroxide) and inflammatory (tumor necrosis factor α, interleukin-1ß, and lipopolysaccharide) stimulation. The obtained promising results suggest potential protective role of nsLTP2 in vascular diseases prevention. PRACTICAL APPLICATION: nsLTP 2 peptide is resistant to proteases throughout the gastrointestinal tract and exerts antioxidant and cytoprotective activities. These characteristics could be exploited in vascular diseases prevention.

Transcriptome Profiling of Wheat Seedlings following Treatment with Ultrahigh Diluted Arsenic Trioxide.

Plant systems are useful research tools to address basic questions in homeopathy as they make it possible to overcome some of the drawbacks encountered in clinical trials (placebo effect, ethical issues, duration of the experiment, and high costs). The objective of the present study was to test the hypothesis whether 7-day-old wheat seedlings, grown from seeds either poisoned with a sublethal dose of As2O3 or unpoisoned, showed different significant gene expression profiles after the application of ultrahigh diluted As2O3 (beyond Avogadro's limit) compared to water (control). The results provided evidence for a strong gene modulating effect of ultrahigh diluted As2O3 in seedlings grown from poisoned seeds: a massive reduction of gene expression levels to values comparable to those of the control group was observed for several functional classes of genes. A plausible hypothesis is that ultrahigh diluted As2O3 treatment induced a reequilibration of those genes that were upregulated during the oxidative stress by bringing the expression levels closer to the basal levels normally occurring in the control plants.

Lunasin in wheat: a chemical and molecular study on its presence or absence.

Lunasin is a peptide whose anticancer properties are widely reported. Originally isolated from soybean seeds, lunasin was also found in cereal (wheat, rye, barley and Triticale), Solanum and amaranthus seeds. However, it was recently reported that searches of transcript and DNA sequence databases for wheat and other cereals failed to identify sequences with similarity to those encoding the lunasin peptide in soy. In order to clarify the presence or absence of lunasin in wheat varieties, a broad investigation based on chemical (LC-ESI-MS) and molecular (PCR) analyses was conducted. Both approaches pointed out the absence of lunasin in the investigated wheat genotypes; in particular no compounds with a molecular weight similar to that of lunasin standard and no lunasin-related sequences were found in the analysed wheat samples. These findings confirm the hypothesis, reported in recent researches, that lunasin is not a wheat-derived peptide.