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Snakin-1 is a member of the Solanum tuberosum Snakin/GASA family. We previously demonstrated that Snakin-1 is involved in plant defense to pathogens as well as in plant growth and development, but its mechanism of action has not been completely elucidated yet. Here, we showed that leaves of Snakin-1 silenced potato transgenic plants exhibited increased levels of reactive oxygen species and significantly reduced content of ascorbic acid. Furthermore, Snakin-1 silencing enhanced salicylic acid content in accordance with an increased expression of SA-inducible PRs genes. Interestingly, gibberellic acid levels were also enhanced and transcriptome analysis revealed that a large number of genes related to sterol biosynthesis were downregulated in these silenced lines. Moreover, we demonstrated that Snakin-1 directly interacts with StDIM/DWF1, an enzyme involved in plant sterols biosynthesis. Additionally, the analysis of the expression pattern of PStSN1::GUS in potato showed that Snakin-1 is present mainly in young tissues associated with active growth and cell division zones. Our comprehensive analysis of Snakin-1 silenced lines demonstrated for the first time in potato that Snakin-1 plays a role in redox balance and participates in a complex crosstalk among different hormones., Competing Interests: The authors have declared that no competing interests exist.

Droëwors are traditional South African salted and dried sausages, made without nitrites/nitrates and non- fermented. Different meat sources (beef, game and ostrich) are traditionally used in droëwors processing, while the use of pork is uncommon, as it is said to lead to rancidity. The first part of the study analysed the physico-chemical composition of commercially available beef, game and ostrich meat droëwors (n = 20). On average, they were composed of 26.3-29.2 g/100 g moisture, 41.3-44.0 g/100 g protein, 26.2-33.1 g/100 g fat and 5.9-6.5 g/100 g ash and 5.0-5.4 pH. Water activity (0.76-0.82) was sufficiently low to ensure shelf stability at ambient temperatures. In the second part beef and pork droëwors were formulated in accordance with these results and with similar fat content, dried for 2 days (30 °C, 40% relative humidity) and stored for 26 days (25 °C, 50% relative humidity); measuring moisture, water activity, pH and lipid oxidative stability (thiobarbituric acid reactive substances (TBARS)) weekly. At day 5, moisture and thus water activity of pork droëwors was slightly higher compared to beef ones and fat and ash content slightly lower ( P  ≤ 0.05) despite similar weight loss. Even with slightly less fat, TBARS in pork droëwors were significantly higher after drying and throughout storage (3.83 vs 0.99 mg MDA equivalents/kg dry matter at a maximum)., Competing Interests: Compliance with ethical standardsThe authors declare no conflict of interest.

Plants suffer from a broad range of abiotic and biotic stresses that do not occur in isolation but often simultaneously. Productivity of natural and agricultural systems is frequently constrained by water limitation, and the frequency and duration of drought periods will likely increase due to global climate change. In addition, phytoviruses represent highly prevalent biotic threat in wild and cultivated plant species. Several hints support a modification of epidemiological parameters of plant viruses in response to environmental changes but a clear quantification of plant-virus interactions under abiotic stresses is still lacking. Here we report the effects of a water deficit on epidemiological parameters of Cauliflower mosaic virus (CaMV), a non-circulative virus transmitted by aphid vectors, in nine natural accessions of Arabidopsis thaliana with known contrasted responses to water deficit. Plant growth-related traits and virus epidemiological parameters were evaluated in PHENOPSIS, an automated high throughput phenotyping platform. Water deficit had contrasted effects on CaMV transmission rate and viral load among A. thaliana accessions. Under well-watered conditions, transmission rate tended to increase with viral load and with CaMV virulence across accessions. Under water deficit, transmission rate and virulence were negatively correlated. Changes in the rate of transmission under water deficit were not related to changes in viral load. Our results support the idea that optimal virulence of a given virus, as hypothesized under the transmission-virulence trade-off, is highly dependent on the environment and growth traits of the host.

This work describes the formulation of a functional yogurt-like product based on fermented maize with added phytosterols and its oxidative stability during cold storage. The technological challenge was to stabilize 3.5% esterified phytosterols (between 2 and 3 g of free sterols) in a low-fat emulsion and to preserve the obtained product throughout processing and storage. The natural bioactive compounds: lutein, zeaxanthin, β-cryptoxanthin, β-carotene and γ-tocopherol were detected in the yogurt, and remained stable during 12 days of refrigeration. Higher content of C18:1 n-9 and C18:3 n-3 (six and ninefold, respectively) were obtained in samples with phytosterols. This was desirable from a nutritional point of view, but at the same time it induced lipid oxidation that was 1.4-fold higher in the product with phytosterols than in the controls. The use of a multivariate approach served to find descriptors which were related to treatments, and to explain their behavior over time.

Background: Snakin-1 (StSN1) is a broad-spectrum antimicrobial cysteine-rich peptide isolated from Solanum tuberosum. Its biotechnological potential has been already recognized since it exhibits in vivo antifungal and antibacterial activity. Most attempts to produce StSN1, or homologous peptides, in a soluble native state using bacterial, yeast or synthetic expression systems have presented production bottlenecks such as insolubility, misfolding or low yields., Results: In this work, we successfully expressed a recombinant StSN1 (rSN1) in Spodoptera frugiperda (Sf9) insect cells by optimizing several of the parameters for its expression in the baculovirus expression system. The recombinant peptide lacking its putative signal peptide was soluble and was present in the nuclear fraction of infected Sf9 cells. An optimized purification procedure allowed the production of rSN1 that was used for immunization of mice, which gave rise to polyclonal antibodies that detect the native protein in tissue extracts of both agroinfiltrated plants and stable transgenic lines. Our results demonstrated that this system circumvents all the difficulties associated with recombinant antimicrobial peptides expression in other heterologous systems., Conclusions: The present study is the first report of a successful protocol to produce a soluble Snakin/GASA peptide in baculovirus-infected insect cells. Our work demonstrates that the nuclear localization of rSN1 in insect cells can be exploited for its large-scale production and subsequent generation of specific anti-rSN1 antibodies. We suggest the use of the baculovirus system for high-level expression of Snakin/GASA peptides, for biological assays, structural and functional analysis and antibody production, as an important step to both elucidate their accurate physiological role and to deepen the study of their biotechnological uses.

This study was carried out to assess the impact of heat processing of a complex emulsion on the behavior of fat soluble micronutrients (FSM) in a traditional Tunisian dish. A simplified recipe involved, dried mucilage-rich jute leaves, tomato paste and olive oil, followed by a cooking treatment (150min). Hydrothermal pattern and viscosity were monitored along with the changes of FSM content and the bioaccessibility (called micellarization, using an in vitro digestion model). Partitioning of carotenoids differed according to their lipophilicity: lycopene, β-carotene and lutein diffused to the oil phase (100%, 70% and 10% respectively). In contrast with the poor carotenes/tocopherol bioaccessibility (0.9-1%), the highest micellarization was observed for lutein (57%) and it increased with heating time and viscosity change. Domestic culinary cooking practices probably increase the bioavailability of carotenes mainly by their diffusion to the oil phase, facilitating their in vivo transfer into micelles., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The quality of functional food products designed for the prevention of degenerative diseases can be affected by the incorporation of bioactive compounds. In many types of cheese, the performance of starter microorganisms is critical for optimal elaboration and for providing potential probiotic benefits. Phytosterols are plant lipophilic triterpenes that have been used for the design of functional dairy products because of their ability to lower serum cholesterol levels in humans. However, their effect on the starter culture behavior during cheesemaking has not yet been studied. Here, we followed DNA and RNA kinetics of the bacterium Streptococcus thermophilus, an extensively used dairy starter with probiotic potential, during industrial production of a functional, semi-soft, reduced-fat cheese containing phytosterol esters and alpha-tocopherol as bioactive compounds. For this purpose, real-time quantitative PCR (qPCR) and reverse transcription-qPCR (RT-qPCR) assays were optimized and applied to samples obtained during the manufacture and ripening of functional and control cheeses. An experimental set-up was used to evaluate the detection threshold of free nucleic acids for extraction protocols based on pelleted microorganisms. To our knowledge, this straight-forward approach provides the first experimental evidence indicating that DNA is not a reliable marker of cell integrity, whereas RNA may constitute a more accurate molecular signature to estimate both bacterial viability and metabolic activity. Compositional analysis revealed that the bioactive molecules were effectively incorporated into the cheese matrix, at levels considered optimal to exert their biological action. The starter S. thermophilus was detected by qPCR and RT-qPCR during cheese production at the industrial level, from at least 30min after its inoculation until 81days of ripening, supporting the possible role of this species in shaping organoleptic profiles. We also showed for the first time that the addition of phytosterols at functional concentrations, not only did not affect starter performance but also correlated with a significant increase in target DNA and cDNA levels in most of the time points evaluated throughout cheesemaking. Therefore, these findings suggest that the growth and metabolism of S. thermophilus may be enhanced by the incorporation of these biologically active molecules during cheese production, providing important information for the industrial design of novel fermented foods., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Beef samples from grazing steers finished with different seed-supplemented diets were vacuum packaged for 3, 14 and 56days (VC) and subsequently exposed to aerobic conditions (AE) for 0 and 5days. Different fluorescent compounds of interest in the oxidation process were detected in meat, namely tryptophan residues, Schiff bases and porphyrins. Tryptophan intensity fluorescence increased with 14days of VC; while Schiff bases intensity increased (P<0.05) in beef samples stored under VC-56 and in all samples after AE-5days. Porphyrins increased (P<0.05) gradually with the extension of vacuum storage time, but were degraded in beef with long vacuum storage and 5days of AE. Higher levels of porphyrins in beef under VC were correlated (P<0.05) with lower redness and higher TBARS after AE-5. This study revealed the potential of fluorescence signals to detect oxidative changes in beef under different storage conditions using a fast and nondestructive method such as fluorescence spectroscopy., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Angus steers were grazed on unsupplemented pasture (CNTRL), pasture supplemented with 0.7% BW cracked corn (FLAX-0), FLAX-0 with 0.125% and 0.250% BW of whole flaxseed (FLAX-1 and FLAX-2). Six steers were grazed per treatment for 70 days, with start and finish weights of 458 and 508 kg. At 24 h post slaughter, longissimus thoracis were harvested, and steaks assigned to treatments of postmortem aging time under vacuum (PM; 3, 14 and 56 days) with or without five days of aerobic exposure (AE). Meat antioxidant status was higher (P<0.05) when feeding CNTRL and FLAX-1 than FLAX-0 and FLAX-2. Under AE, lipid oxidation was highest for FLAX-2 (P<0.05), and lowest for FLAX-1. Greatest TBARs and lowest antioxidant capacity and redness values were obtained with AE and the longer PM (P<0.05). Beef oxidative stability through AE improved by adding a low flaxseed level to supplemented corn grain, but deteriorated by adding a high flaxseed level or by extending PM., (Published by Elsevier Ltd.)

The livestock sector faces the challenge to respond to the growing demand for animal protein from an expanding population while reducing environmental impact through GHG emissions. Globally about 2.836 million tons of CO2-eq were emitted by the beef production sector equivalent to 46,2 kg CO2-eq per kg carcass weight (CW). From the 1.485 million cattle head spread out over the world, 82% are on extensive grazing systems while only 18% are on high productive intensive systems. Among the top ten beef exporter countries, five are located in Latin America accounting a quarter of the global stock and two of them, Argentina and Uruguay, produce on temperate pastures under grazing systems. In Argentina, the livestock area was reduced in favor of increasing the grain cropping area, which took place in the last two decades. Production systems were intensified to maintain cattle stock. Cattle programs changed from 100% pasture to pasture supplemented with cereal grains and conserved forages, and confinement on grain feeding for fattening was incorporated. Due to land sharing competition with cash crops, no increment of cattle stock is expected therefore improving production efficiency appears as the only way to increase beef production while reducing methane emissions intensity. Beef produced on intensive grazing systems on supplemented pastures maintained organoleptic, nutritional and lipid profile than that of beef produced on pure grazing systems., (Copyright © 2014 Elsevier Ltd. All rights reserved.)