Comparison of drought stress response and gene expression between a GM maize variety and a near-isogenic non-GM variety

Maize MON810, grown and commercialised worldwide, is the only cultivated GM event in the EU. Maize MON810, variety DKC6575, and the corresponding near-isogenic line Tietar were studied in different growth conditions, to compare their behaviour in response to drought. Main photosynthetic parameters were significantly affected by water stress in both GM and non –GM varieties to a similar extents. Though DKC6575 (GM) had a greater sensitivity in the early phase of stress response as compared with Tietar (non GM), after six days of stress they behaved similarly, and both varieties recovered from stress damage. Profiling gene expression in water deficit regimes and in a generalised water stress condition showed an up-regulation of many stress- responsive genes, but a greater number of differentially expressed genes was observed in Tietar, with genes belonging to transcription factor families and genes encoding HSPs, LEAs and detoxification enzymes. Since induction of these genes have been indicated from the literature as typical of stress responses, their activation in Tietar rather than in DKC6575 may be reminiscent of a more efficient response to drought. DKC6575 was also analysed for the expression of the transgene CryIAb (encoding the delta-endotoxin insecticidal protein) in water deficit conditions. In all the experiments, the CryIAb transcript was not influenced by water stress, but was expressed at a constant level.. This suggests that though possessing a different pattern of sensitivity to stress, the GM variety maintains the same expression level for the transgene

New gene functions are involved in the thermotolerance of the wild wheat relative Aegilops umbellulata

Wheat is one of the most important food crops in the world for human consumption, like all plants it is exposed to environmental stresses including high temperatures. The deleterious effect of high temperatures negatively affects plant growth and development, leading to reduced viability and yield. These effects can be reduced by improvement of thermotolerance through innovative breeding strategies, based on the expansion of the genetic pool available, by exploring important genetic functions from wheat wild progenitors. Improving the genetic thermotolerance characteristics of wheat requires greater understanding of genetic bases of thermotolerance, through identification of high temperature stress related genes. A good source of new useful alleles is given by Aegilops species characterized by thermotolerant habits. In this study we have classified as thermotolerant or thermosensitive, on the basis of physiologic tests, some accessions of wheat wild relative species belonging to Aegilops and Triticum genera. A thermotolerant accession of Aegilops umbellulata (AUM5) was selected, subjected to different thermal treatments and analyzed at transcriptional level. By differential display reverse transcriptase polymerase chain reaction (DDRT-PCR), we investigated modulation of gene expression elicited by heat treatments. This approach allowed the identification of various transcript-derived fragments (TDFs) produced by AUM5 in response to different thermal treatments. The functions of the inducible unique genes in the molecular determination of thermotolerance process are discussed

MicroRNA global profiling in cystic fibrosis cell lines reveals dysregulated pathways related with inflammation, cancer, growth, glucose and lipid metabolism, and fertility: an exploratory study

Cystic fibrosis (CF), is due to CF transmembrane conductance regulator (CFTR) loss of function, and is associated with comorbidities. The increasing longevity of CF patients has been associated with increased cancer risk besides the other known comorbidities. The significant heterogeneity among patients, suggests potential epigenetic regulation. Little attention has been given to how CFTR influences microRNA (miRNA) expression and how this may impact on biological processes and pathways

The Diverse Potential of Gluten from Different Durum Wheat Varieties in Triggering Celiac Disease: A Multilevel In Vitro, Ex Vivo and In Vivo Approach

The reasons behind the increasing prevalence of celiac disease (CD) worldwide are still not fully understood. This study adopted a multilevel approach (in vitro, ex vivo, in vivo) to assess the potential of gluten from different wheat varieties in triggering CD. Peptides triggering CD were identified and quantified in mixtures generated from simulated gastrointestinal digestion of wheat varieties (n = 82). Multivariate statistics enabled the discrimination of varieties generating low impact on CD (e.g., Saragolla) and high impact (e.g., Cappelli). Enrolled subjects (n = 46) were: 19 healthy subjects included in the control group; 27 celiac patients enrolled for the in vivo phase. Celiacs were divided into a gluten-free diet group (CD-GFD), and a GFD with Saragolla-based pasta group (CD-Sar). The diet was followed for 3 months. Data were compared between CD-Sar and CD-GFD before and after the experimental diet, demonstrating a limited ability of Saragolla to trigger immunity, although not comparable to a GFD. Ex vivo studies showed that Saragolla and Cappelli activated immune responses, although with great variability among patients. The diverse potential of durum wheat varieties in triggering CD immune response was demonstrated. Saragolla is not indicated for celiacs, yet it has a limited potential to trigger adverse immune response

Detection of allergen coding sequences of kiwi, peach, and apple in processed food by qPCR

BACKGROUND: Food traceability becomes lifesaving for persons suffering severe allergy or intolerance, and therefore need a complete avoidance of the immune-trigger food. This paper describes how to fingerprint the presence of some allergenic species (kiwi, peach, and apple) in foods by quantitative Real-Time PCR (qPCR). RESULTS: Five DNA extraction procedures were tested on fruits and foods. The results were statistically evaluated, and discussed. Analysis by qPCR with SYBR Green was developed to detect traces of these allergenic species in foods. Plasmids containing the target sequences of kiwi, peach and apple were employed as Internal Reference Standard. Analysis of spiked food samples showed a Limit of Detection of 25 mg kg-1 for kiwi, 20 mg kg-1 for peach and 50 mg kg-1 for apple. CONCLUSION: The qPCR method here developed, combined with the use of Internal Plasmid Reference Standard, represent a specific system for the quick detection of allergenic species in complex food matrices, with a Limit of Detection comparable with those reported using more timeconsuming methods

Gluten aggregation properties as a tool for durum wheat quality assessment: A chemometric approach

The study aims at assessing the gluten aggregation properties of wholemeal semolina for the differentiation of durum wheat cultivars with different amounts of gluten-quality related fractions. To this end, a calibration set of 56 durum wheat cultivars harvested in 2015–2016 was categorized into four quality classes based on the total gluten protein content and the glutenin to gliadin ratio. Quality class distribution was carried out by Principal Component Analysis (PCA) performed on either gluten aggregation parameters alone or with gluten fractions. PCA differentiated high and medium-high quality varieties from those of low and medium-low quality. Furthermore, PCA confirmed the relationship between gluten fractions and their aggregation kinetics, by means of a validation step consisting of 22 cultivars harvested in 2016–2017. In conclusion, gluten aggregation properties proved to be good indicators of durum wheat semolina quality, and thus a reliable tool for blend preparation in the pasta-making process

Utilisation and limitations of pseudocereals (quinoa, amaranth, and buckwheat) in food production: a review

Background Pseudocereals, especially quinoa, amaranth, and buckwheat, have attracted an increasing amount of attention because of their nutritive and health-benefiting properties, and their suitability for people suffering from coeliac disease and gluten intolerance. However, the utilisation of pseudocereals is hampered by the presence of anti-nutritional compounds (phytates and saponins) and/or substances that yield a bitter taste in the seeds, the latter of which must be minimised before or during food processing, consequently increasing the cost and risk of environmental contamination. Scope and approach The objective of this review is to analyse issues relating to the use of pseudocereals in food production, including: i ) technological limits in the food industry; ii ) agronomic limitations to pseudocereal cultivation and distribution; iii ) technological and biotechnological tools for addressing these issues; and iv ) socio-economic and ethical implications of extensive cultivation. Key findings and conclusions Although pseudocereals have great potential for use in the food industry, they cannot completely replace true cereals due to the presence of compounds that confer undesirable organoleptic and technological characteristics to their products. As the growth in pseudocereal cultivation, especially that of quinoa, remains largely restricted to the nations in which the pseudocereals originated, it is imperative that the excessive exploitation of resources be avoided in these areas. Moreover the improvement of the socio-economic conditions of small farmers is necessary, since they manage the germplasm of these species. Biotechnologies are valuable tools for exploiting the considerable diversity of these species for breeding programs aimed to improve palatable, technological and agronomic characteristics

A stress-related transcription factor belonging to the YL-1 family is differently regulated in durum wheat cultivars differing in drought sensitivity

The Mediterranean area is characterised by unfavorable environmental conditions such as heat stress and drought responsible for yield loss of crops like durum wheat, widely cultivated in this area. The response of plants to stressing environments is mediated by activation of a complex gene network, strictly related to the genetic background. Among the genes induced by drought, those coding for proteins acting as key regulators of signal transduction are of great interest. Characterization of these genes is a crucial point to understand their potential roles in plant stress response, also in view of their possible use in molecular breeding. In this work we have characterised a Triticum durum gene, named TdDRG1, in two commercial cultivars, Primadur and Svevo, differing for drought stress resistance. TdDRG1 codes for a putative transcription factor belonging to the VPS72/YL-1 family, highly conserved in plants and animals. The expression analysis indicates that this gene is expressed at higher level in roots of the resistant cultivar Svevo, than in the susceptible Primadur. The gene structure was determined in both cultivars and the regulatory activity of 5′ upstream regions was analysed by transient expression analysis using tobacco protoplasts. Dissimilar expression level of TdDRG1 in the two cultivars can be explained by the differences observed in gene structure. In particular, differences in 5’ upstream regions could account for contrasting ability to cope with drought of the two cultivars. The data obtained in this study provide indications for further insight into the molecular basis of differences in drought stress response