Protocol: optimised electrophyiological analysis of intact guard cells from Arabidopsis

ABSTRACT: Genetic resources available for Arabidopsis thaliana make this species particularly attractive as a model for molecular genetic studies of guard cell homeostasis, transport and signalling, but this facility is not matched by accessible tools for quantitative analysis of transport in the intact cell. We have developed a reliable set of procedures for voltage clamp analysis of guard cells from Arabidopsis leaves. These procedures greatly simplify electrophysiological recordings, extending the duration of measurements and scope for analysis of the predominant K+ and anion channels to that achieved previously in work with Vicia and tobacco guard cells.     

The role of oxylipins and antioxidants on off-flavor precursor formation during potato flake processing

The impact of processing on nonenzymatic antioxidant degradation and lipid oxidation leading to off-flavor development in potato flakes during storage was investigated. Lipoxygenase activity measurements in parallel with the analysis of lipid oxidation products (oxylipins) profiles using HPLC showed that the processing conditions used inhibited efficiently enzymatic lipid oxidation. However, nonenzymatic lipid oxidation products were found throughout processing and in fresh potato flakes. Furthermore, these autoxidative processes cannot be inactivated by the main endogenous nonenzymatic antioxidants in potato tubers (ascorbic acid, phenolic compounds and carotenoids), as these antioxidants are degraded during processing. Indeed, leaching and thermal treatments taking place during processing lead to a decrease of about 95%, 82% and 27% in the content of ascorbic acid, phenolic compounds and carotenoids, respectively. Therefore, storage is a critical step to prevent off-flavor development in potato flakes. Specific attention has thus to be paid on the use of efficient exogenous antioxidants as well as on storage conditions.     

Black locust (<Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> L.) as a multi-purpose tree species in Hungary and Romania: a review

Black locust (Robinia pseudoacacia L.) was the first North-American tree species imported to Europe at the beginning of the seventeenth century. It is commonly planted worldwide because of its adaptability to environmental stresses, its valuable wood, easy propagation, frequent and abundant seed production, excellent coppicing, high seedling survival, and relatively high wood yield. In Europe, Romania and Hungary have the most highly-developed black locust growing techniques and experiences. As a result of increasing interest in black locust in many countries, this review aims to provide a comprehensive overview of state-of-the-art site requirements, propagation, improvement and management (including growth and yield as well as use in energy plantations).     

Pesticide diversity in rice growing areas of Northern Vietnam

Pesticide use in developing countries increases rapidly. In many regions, we miss knowledge of how frequently pesticides are applied and which active ingredients are used. We present a new cost-efficient and rapid assessment method of recording pesticides diversity in rice-dominated landscapes and present some evidence of the misuse of active ingredients in our study regions. We investigated 17 rice fields in two regions of Northern Vietnam in 2014 and 2015. At each region, we explore the abundance of pesticides used with three methods including (1) the novel approach of collecting pesticide packages close to our target rice fields, (2) observations of farmers spraying pesticides in the surrounding and (3) interviewing local farmers. By collecting pesticide packages, we found 811 packages containing 74 different active ingredients. On average, 19 active ingredients (ranging from four to 40 active ingredients) were applied with an average content of 275.3 g of active ingredients per site. Insecticide packages (39%) were most abundant followed by those of fungicides (31%), herbicides (16%) and other active ingredients (14%). On all sites, active ingredients banned in the European Union were applied by the farmers. Collecting pesticide packages proved to be an efficient and rapidly implemented method to obtain some baseline information about pesticide application (for Northern Vietnam). We suspect that improved agricultural extension services could contribute to good agricultural practices in pest management. Generally, better information and education for local farmers for appropriate use of pesticides seem a necessity.     

From science to application: field demonstrations to enhance sustainable rice production in the north of Vietnam—lessons from the LEGATO project

Two major human-made problems in rice production systems in the north of Vietnam concern the low plant-available silicon content of soils and the low biodiversity. The results of the LEGATO project suggest a change to an environmentally friendly rice production system that will help to recover biodiversity. We propose here a framework for a demonstration and dissemination model that will be exemplary for the farmers once it has been successfully realized. We advocate local-option models in different districts to demonstrate to farmers. The methods should be adapted to local and ecoregional differences in climate and land-use tradition, and they explicitly take into account soil care, organic fertilizer, manual weeding, native nectar-rich plant bunds, manual pest snail collection, hymenopteran nesting aids, and biodiversity and yield monitoring.     

High HEV presence in four different wild boar populations in East and West Germany

Swine Hepatitis E virus (HEV) can be transmitted from pigs to humans causing hepatitis. A high prevalence of HEV in wild boar populations is reported for several European countries, but actual data for Germany are missing. During the hunting season from October to December 2007 liver, bile and blood samples were collected from wild boars in four different German regions. The samples were tested for HEV RNA by quantitative PCR (qPCR) and anti-HEV IgG antibodies by two different ELISAs and a Line immunoassay. A seroprevalence of 29.9% using ELISA and 26.2% in the Line immunoassay was determined. The seroprevalence rate varied greatly within the analyzed regions. However, qPCR analysis revealed a higher prevalence of 68.2% positive animals with regional differences. Surprisingly, also adult wild sows and wild boars were highly HEV positive by qPCR. Compared to liver and serum samples, bile samples showed a higher rate of positive qPCR results. Sequencing and phylogenetic analysis of a 969 nt fragment within ORF 2 revealed that all isolates clustered within genotype 3 but differed in the subtype depending on the hunting spot. Isolates clustered within genotypes 3i, 3h, 3f and 3e. Within one population HEV isolates were closely related, but social groups of animals in close proximity might be infected with different subtypes. Two full-length genomes of subtypes 3i and 3e from two different geographic regions were generated. The wild boar is discussed as one of the main sources of human autochthonous infections in Germany.     

Contribution of methylated exudate flavonoids to the anti-inflammatory activity of Grindelia robusta

The flavonoid pattern of an acetonic extract of Grindelia robusta Nutt. was investigated in detail. The flavonoids were enriched by CC. In addition to twelve known methylated exudate flavonols four compounds were identified for the first time in G. robusta. Several substances of the flavonoid complex, among them the main compounds quercetin-3-methylether and 6-OH-kaempferol-3,6-dimethylether, were tested for their activity to inhibit neutrophil elastase. Quercetin-3-methylether was shown to be most active with an IC₅₀ of 19 µM, thus obviously contributing to the anti-inflammatory activity of the drug.     

Effect of germination and subsequent oven-drying on folate content in different wheat and rye cultivars

Cereals are recognised as an important food source of folate, and germinated cereals are reported to contain even more folate. This study examined the effects of germination and oven-drying on folate content in different wheat and rye cultivars. The native folate content in four wheat cultivars ranged from 23 to 33 μg/100 g dry matter (DM) and that in six rye cultivars from 31 to 39 μg/100 g DM. Mean folate content in rye was 25% higher than in wheat. Germination of both cereals resulted in a 4- to 6-fold increase in folate content, depending on cultivar and duration of germination. The highest folate content in both cereals was found after 96 h of germination and was 181 μg/100 g DM for cv. Kaskelott (rye) and 155 μg/100 g DM for cv. Kosack (wheat). Germination increased the amount of 5-CH₃–H₄folate in both cereals from 45 to 75%. Oven-drying of germinated wheat grains (for 48 and 72 h) at 50 °C did not affect the folate content. In conclusion, germination increases the folate content in wheat and rye cultivars, while subsequent oven-drying does not affect the folate content. Germination can therefore be recommended for producing bakery ingredients with increased folate content.