Synthesis and characterization of a potential prebiotic trisaccharide from cheese whey permeate and sucrose by Leuconostoc mesenteroides dextransucrase

The production of new bioactive oligosaccharides is currently garnering much attention for their potential use as functional ingredients. This work addresses the enzymatic synthesis and NMR structural characterization of 2-α-D-glucopyranosyl-lactose derived from sucrose:lactose and sucrose:cheese whey permeate mixtures by using a Leuconostoc mesenteroides B-512F dextransucrase. The effect of synthesis conditions, including concentration of substrates, molar ratio of donor/acceptor, enzyme concentration, reaction time, and temperature, on the formation of transfer products is evaluated. Results indicated that cheese whey permeate is a suitable material for the synthesis of 2-α-D-glucopyranosyl-lactose, giving rise to yields around 50% (in weight respect to the initial amount of lactose) under the optimum reaction conditions. According to its structure, this trisaccharide is an excellent candidate for a new prebiotic ingredient, due to the reported high resistance of α-(1→2) linkages to the digestive enzymes in humans and animals, as well as to its potential selective stimulation of beneficial bacteria in the large intestine mainly attributed to the two linked glucose units located at the reducing end that reflects the disaccharide kojibiose (2-α-D-glucopyranosyl-D-glucose). These findings could contribute to broadening the use of important agricultural raw materials, such as sucrose or cheese whey permeates, as renewable substrates for enzymatic synthesis of oligosaccharides of nutritional interest.     

Arabidopsis ethylene receptors have different roles in Fumonisin B1-induced cell death

Ethylene is a central signalling agent in mediating plant defence against pathogens. Mutations to the ethylene receptor ETR1 have been shown to alter susceptibility of plants to mycotoxin-induced cell death. Using Fumonisin B₁ (FB₁) to induce cell death, we demonstrate that the receptor mutant ein4-1 has a reduced rate of necrosis, potentially due to an upregulation of ETHYLENE RESPONSE FACTOR1. Mutations in other ethylene receptors differentially affected the expression of genes in the jasmonic and salicylic acid defence pathways. Together these data indicate that ethylene receptors do not have redundant roles in mediating FB₁-induced cell death.     

Correlation between hydrolytic enzyme activities measured in bean seedlings afterTrichoderma koningii treatment combined with pregermination and the protective effect againstPythium splendens

An experimental protocol consisting in the colonisation of pregerminated bean seeds dressed withTrichoderma sp. was used in order to study the mechanisms correlated with the protective effect againstPythium splendens. Seed dressed with TH-11 (T. koningii) for 24 h presented a higher protective effect and a higher level of seed colonisation as compared to those dressed with TH-13 (T. longibranchiatum). The levels of seed coat colonisation by TH-11 and TH-13 was shown to be correlated with the carboxymethylcellulase activity, as measured in the seed coats retreived from germinating dressed bean seeds. The seed coat colonisation was also associated with an increased activities of endo-1,3--glucanase and endochitinase measured in seed extracts, and an inhibitory effect of seed extracts onPythium sporangia germination. Pretreatment of TH-13-dressed seeds with a commercial cellulase improved all parameters mentioned above, thus suggesting a role of cellulase activity in the colonisation process and the linked protective effect. The possible role of hydrolytic enzymes in the protective effects is discussed.     

Heat treatments and expansin gene expression in strawberry fruit

Heat treatments have been applied in fruit postharvest technology for insect disinfestations, decay control, ripening delay and modification of fruit responses to other stresses. Heat treatment affects several aspects of fruit ripening, such as ethylene production and cell wall degradation probably through changes in gene expression and protein synthesis. In this paper, strawberries (Fragaria × ananassa Duch., cv Camarosa) at 50–75% red stage of ripening were heat-treated at 45 °C during 3 h in an air oven and then stored at 20 °C for 0, 4, 18, 24 and 48 h. Fruit firmness was determined and the expression of a set of expansin genes (FaEXP1, FaEXP2, FaEXP4, FaEXP5 and FaEXP6) was analyzed. The firmness of treated fruit was higher than that of control fruit 24 h after treatment, though the differences disappeared after 48 h at 20 °C. The analysis by northern-blot indicated that heat treatments affected differently the expression of expansin genes. The expression of FaEXP1, FaEXP2 and FaEXP6 was lower in treated fruit during the following 24 h post-treatment. The lower expression of these expansin genes could contribute to delay softening after heat treatment.     

Glyphosate Sublethal Effects on the Population Dynamics of the Earthworm Eisenia fetida (Savigny, 1826)

Pesticides’ sublethal effects are not regularly taken into account when assessing agrochemical’s toxicity. With the objective of detecting chronic, sublethal effects of the widely used herbicide glyphosate, an experiment was performed using the earthworm Eisenia fetida as model organism. Earthworm adults were randomly assigned to three glyphosate treatments: control (no glyphosate), regular dose for perennial weeds, and double dose. Six E. fetida individuals were placed in each pot. Two random pots were taken weekly from each treatment and the number of adults, individual weight, number of cocoons, and presence and number of young earthworms were recorded. A matrix analysis was performed with the data. The matrix population model built showed that while the control population had a positive growth rate, both glyphosate treatments showed negative growth rates. The results suggest that under these sublethal effects, non-target populations are at risk of local extinction, underscoring the importance of this type of studies in agrochemical environmental risk assessment.     

Protein-lipid interactions during liposome oxidation with added anthocyanin and other phenolic compounds

Oxidation of bovine serum albumin, casein, and lactalbumin and the effect of different procyanidins, anthocyanins, and their aglycons (10 and 20 microM) on lactalbumin oxidation were investigated in a liposome system. Samples were incubated in the dark at 37 degrees C with copper, and the extent of oxidation was measured by determining the loss of tryptophan fluorescence and the formation of protein carbonyls, conjugated diene hydroperoxides, and hexanal. The correlation between different protein and lipid oxidation measurements was good and statistically significant. Casein was the most stable protein in the liposome model, and it was also the best inhibitor of liposome oxidation. All tested anthocyanins and other phenolic compounds inhibited both lipid and protein oxidation. There were no systematic differences with anthocyanins and their aglycons in relation to the concentrations used or glycosylation with either glucose or rutinose. Procyanidins B1 and B2 and ellagic acid were potentially better antioxidants than anthocyanins due to their several hydroxyl groups as measured by both protein and lipid oxidation. In conclusion, oxidative deterioration of liposomes due to protein-lipid interaction is inhibited by anthocyanins, procyanidins, and ellagitannin present, for example, in berries.     

Control of potato powdery scab caused by Spongospora subterranea by foliage cover and soil application of chemicals under field conditions with naturally infested soil

Powdery scab (PS), caused by Spongospora subterranea, reduces the quality and marketability of potatoes worldwide. Disease symptoms include lesions on the tuber surface and root galling, which may lead to yield losses. In the current study we report a sustainable approach to reduce PS by manipulating soil temperature during tuber initiation. Plant cover with nonwoven fabric significantly reduced PS on tubers by 54%–69% in 2017 and 84%–93% in 2019, compared to the control, and root galling by 96% in 2019, due to an increased average minimum and maximum soil temperature of 1.8 and 4.2 °C in respective years. Additional preplanting soil treatments were also evaluated in naturally infested soil. In 2017, disease incidence and severity were significantly reduced using 2.5 or 5 L a.i./ha fluazinam in broadcast application or in-furrow, and by 0.375 or 0.75 L a.i./ha flusulphamide applied in-furrow. In 2019, disease incidence and severity were significantly reduced by the broadcast application of fluazinam, and 75 kg/ha calcium cyanamide, but the latter had a negative impact on yield. Soil fumigation with metam sodium resulted in a 98% reduction in PS. Root galling was significantly reduced by calcium cyanamide, metam sodium, and fluazinam in the 2019 trial only. Foliar application of resistance-inducing phosphonates combined with fluazinam application had no additive effect on PS incidence and severity. Integrated approaches such as tolerant cultivars, soil testing, preplanting fungicide application, and sustainable means of control such as foliage cover for a short period may be implemented in order to control the disease and minimize damage.