Inter- and Intramolecular recombinations in the Cucumber Mosaic Virus genome related to adaptation to Alstroemeria

In four distinct alstroemeria-infecting cucumber mosaic virus (CMV) isolates, additional sequences of various lengths were present in the 3' nontranslated regions of their RNAs 2 and 3, apparently the result of intra- and intermolecular recombination events. Competition experiments revealed that these recombined RNA 2 and 3 segments increased the biological fitness of CMV in alstroemeri

The NS3 protein of rice hoja blanca virus suppresses RNA silencing in mammalian cells

The NS3 protein of the tenuivirus rice hoja blanca virus (RHBV) has previously been shown to represent the viral RNA interference (RNAi) suppressor and is active in both plant and insect cells by binding short interfering RNAs (siRNAs) in vitro. Using a firefly luciferase-based silencing assay it is described here that NS3 is also active in mammalian cells. This activity is independent of the inducer molecule used. Using either synthetic siRNAs or a short hairpin RNA construct, NS3 was able to significantly suppress the RNAi-mediated silencing of luciferase expression in both monkey (Vero) and human (HEK293) cells. These results support the proposed mode of action of NS3 to act by sequestering siRNAs, the key molecules of the RNAi pathway conserved in all eukaryotes. The possible applications of this protein in modulating RNAi and investigating the proposed antiviral RNAi response in mammalian cell systems are discussed

Efficiency of Rz-1 based rhizomania resistance and molecular studies on BNYVV isolates from sugar beet cultivation in Geece

A survey was carried out to investigate the current situation concerning rhizomania disease incidence in sugar beet cultivation of Greece. A systematic field evaluation over locations and years revealed a consistent disease severity pattern according to favourable agroclimatic conditions and pointed to the so far effectiveness of the Rz1 gene-based resistance, as no major disease outbreaks were observed. Molecular analyses aiming at the characterization of the type and genetic diversity of the virus further confirmed the widespread occurrence of BNYVV in the country, as evidenced by RT-PCR amplification of all five known genomic molecules and nested-PCR assays. None of the isolates contained an RNA 5, typically found in pathotype P. On the basis of RFLP patterns, all BNYVV isolates analysed were classified as pathotype A. Sequence determination of the full-length RNA 3-encoded p25 protein, responsible for symptom development, revealed amino acid motifs ACHG/VCHG in the hypervariable region aa67–70. The presence of valine in position 67 did not appear associated with increased pathogenicity and resistance breaking properties, as earlier reported

Binding of small interfering RNA molecules is crucial for RNA interference suppressor activity of rice hoja blanca virus NS3 in plants

The NS3 protein of rice hoja blanca tenuivirus represents a viral suppressor of RNAi that sequesters small interfering (si)RNAs in vitro. To determine whether this siRNA binding property is the critical determinant for the suppressor activity of NS3, an alanine point mutational analysis was performed and the resulting mutant proteins were tested for both siRNA binding ability and RNAi suppressor activity in plants. Alanine substitutions of lysine residues at position 173-175 resulted in mutant proteins that lost both their affinity for siRNAs and their RNAi suppressor activity in planta. This indicates that siRNA binding of NS3 is indeed essential for the suppressor function of NS3 and that residues at position 173-175 are involved in the siRNA binding and suppressor activit

Instrumentarium monitoring mestmarkt en enkele analyses

Het nieuwe mestbeleid met daarin een stelsel van gebruiksnormen en de bijbehorende normen grijpen naar verwachting fors in op de mestmarkt. Dit rapport beschrijft onderdelen van het instrumentarium voor de monitoring van de mestmarkt. De onderdelen zijn analyses van derogatiebedrijven (1), van de acceptatie van dierlijke mest op kleiakkerbouwbedrijven (2) en van het gedrag van melkveehouders op het nieuwe mestbeleid (3). Daarnaast beschrijft het rapport de opzet voor een systeem voor monitoring van mestprijzen (4) en de opzet voor interviews met mestdistributeurs (5)

Selection of authentic modelling practices as contexts for chemistry education

In science education, students should come to understand the nature and significance of models. In case of chemistry education it is argued that the present use of models is often not meaningful from the students’ perspective. A strategy to overcome this problem is to use an authentic chemical modelling practice as a context for a curriculum unit. The theoretical framework for this strategy is activity theory rooted in sociocultural theories on learning. An authentic chemical modelling practice is characterized by a set of motives for model development through a well defined modelling procedure using only relevant issue knowledge. The aim of this study was to explore, analyse and select authentic chemical modelling practices for use in chemistry education. The suitability of the practices was reviewed by applying a stepwise procedure focussed on criteria such as students' interest and ownership, modelling procedure, issue knowledge and feasibility of the laboratory work in the classroom. It was concluded that modelling drinking water treatment and human exposure assessment are both suitable to serve as contexts, because both practices exhibit clear motives for model construction and the applied modelling procedures are in line with students' pre-existing procedural modelling knowledge. The issue knowledge involved is consistent with present Dutch science curriculum and it is possible to carry out experimental work in the classroom for model calibration and validation. The method described here to select and evaluate practices for use as contexts in chemistry education can also be used in other science domains

Afname derogatie: verkenning omvang en beweegredenen ondernemers

Wageningen Economic Research voerde in opdracht van LNV een onderzoek uit met als doelstelling inzicht te geven in ontwikkelingen in de tijd van het aantal bedrijven, inclusief omvang, met derogatie (kwantitatieve analyse), zowel op nationaal als regionaal niveau. Daarnaast is gevraagd inzicht te geven in de beweegredenen van ondernemers die de teruglopende deelname aan derogatie kunnen verklaren (kwalitatieve analyse). Op verzoek zijn ook aanbevelingen voor beleid gedaan die zouden kunnen leiden tot een groter derogatieareaal in de toekomst

Phage display-selected single chain antibodies confer high levels of resistance against Tomato spotted wilt virus

Rational design of antibodies targeting essential viral proteins can complement the palette of antiviral resistance strategies. Here, stable and high expression of single-chain monoclonal antibodies targeting the nucleoprotein of the economically important plant virus Tomato spotted wilt virus, a protein that is involved in multiple steps in the viral infection cycle, is reported. High cytoplasmic expression levels of three selected phage display-derived anti-viral single-chain antibodies were established. Of these antibodies, two led to high levels of resistance against this plant virus. Protoplast experiments provided evidence that the two resistance-conferring antibodies may have a different mode of action and could be combined for higher durability of resistance in the fiel

The influenza A virus NS1 protein binds small interfering RNAs and suppresses RNA silencing in plants

RNA silencing comprises a set of sequence-specific RNA degradation pathways that occur in a wide range of eukaryotes, including animals, fungi and plants. A hallmark of RNA silencing is the presence of small interfering RNA molecules (siRNAs). The siRNAs are generated by cleavage of larger double-stranded RNAs (dsRNAs) and provide the sequence specificity for degradation of cognate RNA molecules. In plants, RNA silencing plays a key role in developmental processes and in control of virus replication. It has been shown that many plant viruses encode proteins, denoted RNA silencing suppressors, that interfere with this antiviral response. Although RNA silencing has been shown to occur in vertebrates, no relationship with inhibition of virus replication has been demonstrated to date. Here we show that the NS1 protein of human influenza A virus has an RNA silencing suppression activity in plants, similar to established RNA silencing suppressor proteins of plant viruses. In addition, NS1 was shown to be capable of binding siRNAs. The data presented here fit with a potential role for NS1 in counteracting innate antiviral responses in vertebrates by sequestering siRNAs