Multiplex real-time PCR assays for the detection and identification of Heterobasidion species attacking conifers in Europe

Four species of the destructive forest pathogen Heterobasidion annosum sensu lato (s.l.) are present in Europe: H. annosum sensu stricto (s.s.), H. abietinum and H. parviporum are native species, while H. irregulare is a non-native invasive species currently reported only in Italy, yet recommended for regulation throughout Europe. In this study, real-time PCR detection tests were developed for each of the four species, which can be used simultaneously or individually thanks to probes labelled with species-specific fluorescent dyes. The different performance criteria of each assay were evaluated, and it was determined that they were theoretically capable of detecting amounts of DNA corresponding to 311, 29 and 29 cell nuclei in H. annosum s.s., H. irregulare and H. parviporum, respectively. The specificity of each assay was assessed with a wide set of strains. Real-time PCR tests successfully detected Heterobasidion species from 36 fruiting bodies taken from the forest, as well as from artificially inoculated or naturally infected wood samples. The multiplex real-time PCR assays developed in this study could have practical applications both in forest management and in phytosanitary monitoring.     

Mitigating the impact of selective phenotyping in training populations on the prediction ability by multi-trait pedigree and genomic selection models

Training populations for pedigree and genomic prediction in plant breeding programmes are largely updated with superior genotypes from multi-environment trials, where they are tested with the goal of variety development. Such a selective phenotyping has, however, a negative impact on prediction abilities, especially when only a subset of breeding lines can be tested, as for laborious and costly to phenotype traits. This study focused, thus, on investigating the impact of selective phenotyping in the training population of an applied wheat breeding programme, and assessing the potential to mitigate this impact by pedigree and genomic multi-trait prediction models as well as bi-directionally selected training populations for several baking quality parameters. Combining both pedigree and genomic information in multi-trait prediction models with pre-existing phenotypic information for protein content and sedimentation value compensated for the observed loss in prediction ability, while entering few inferior breeding lines into the training population further mitigated the impact of selective phenotyping and even led to a slight increase of prediction ability in comparison to a randomly chosen training population.     

Successes and limitations of phytotechnologies at field scale: outcomes,assessment and outlook from COST Action 859

Purpose  

Many agricultural and brownfield soils are polluted and more have become marginalised due to the introduction of new, risk-based legislation. The European Environment Agency estimates that there are at least 250,000 polluted sites in the member states that require urgent remedial action. There is also significant volumes of wastewaters and dredged polluted sediments. Phytotechnologies potentially offer a cost-effective in situ alternative to conventional technologies for remediation of low to medium-contaminated matrices, e.g. soils, sediments, tailings, solid wastes and waters. For further development, social and commercial acceptance, there is a clear requirement for up-to-date information on successes and failures of these technologies based on evidence from the field. This review reports the outcomes from several integrated experimental attempts to address this at both field and market level in the 29 countries participating in COST Action 859.     

Compressive properties of Hemp (Cannabis sativa L.) stalks

Compressive behaviour of hemp (Cannabis Sativ L.) stems is important for the design of hemp handling and processing machines. Experiments were carried out to measure the compressive properties of stems from two hemp varieties: Alyssa (grown for fibre only) and Petera (grown for both fibre and seed), produced in Manitoba, Canada. The physical properties of the hemp specimens were measured. For each variety, an air-dried hemp stem was divided into three height sections along the stem: upper, middle and lower. For each section, the hemp stem was further cut into 25.4 mm long specimens. Individual specimens were compressed in the axial and lateral directions using a universal testing machine. The load-displacement curves were recorded, and compressive properties were derived from the load-displacement curves. The outer diameter of the hollow hemp specimens varied from 6 to 17 mm; the linear density varied from 10 to 37 g m⁻¹. Diameter and linear density were greater for Petera than Alyssa, and at the lower height section than the higher section for both varieties. The compression tests showed that the maximum compressive load varied from 58 to 1425 N, and the energy requirement varied from 23 to 1809 mJ, depending on the variety, height section and diameter of stem, and compression direction. In general, the maximum compressive load and the energy requirement were greater in the lower section and for larger diameters; higher loads and energy were observed for Petera than for Alyssa and for the axial compression direction than for the lateral direction.     

Identifying the presence of neonicotinoidresistant peach-potato aphid (Myzus persicae) in the peach-growing regions of southern France and northern Spain

BACKGROUND: The neonicotinoid class of insecticides is a key component of pest management strategies used by stone fruit producers in Europe. Neonicotinoids are currently one of the most important tools for control of the peach‐potato aphid (Myzus persicae). Overreliance on neonicotinoids has led to the development of resistance through a combination of metabolic and target‐site resistance mechanisms in individual aphids. A resistance monitoring project was conducted by Syngenta in 2010 to determine the resistance status of M. persicae populations collected from France and Spain, and to determine the frequency of the target‐site mutation in those populations. RESULTS: Resistance monitoring suggests that resistance to neonicotinoids is relatively widespread in populations of M. persicae collected from peach orchards in the Languedoc‐Roussillon, Provence‐Alpes‐Cote d'Azur and Rhone‐Alpes regions of France, and resistance can be associated with the frequency of the target‐site mutation (R81T). The R81T mutation in its heterozygous form is also present in Spanish populations and is associated with neonicotinoid resistance. CONCLUSION: The widespread nature of neonicotinoid resistance in southern France and the potential for resistance development in northern Spain highlight the need for a coordinated management strategy employing insecticides with different modes of action to reduce the selection pressure with neonicotinoids. Copyright © 2011 Society of Chemical Industry     

Polymerase exchange during Okazaki fragment synthesis observed in living cells

DNA replication machineries have been studied extensively, but the kinetics of action of their components remains largely unknown. We report a study of DNA synthesis during replication in living Escherichia coli cells. Using single-molecule microscopy, we observed repetitive fluorescence bursts of single polymerase IIIs (Pol IIIs), indicating polymerase exchange at the replication fork. Fluctuations in the amount of DNA-bound single-stranded DNA-binding protein (SSB) reflect different speeds for the leading- and lagging-strand DNA polymerases. Coincidence analyses of Pol III and SSB fluctuations show that they correspond to the lagging-strand synthesis and suggest the use of a new Pol III for each Okazaki fragment. Based on exchanges involving two Pol IIIs, we propose that the third polymerase in the replisome is involved in lagging-strand synthesis.     

The centrosome in cells and organisms

The centrosome acts as the main microtubule-nucleating organelle in animal cells and plays a critical role in mitotic spindle orientation and in genome stability. Yet, despite its central role in cell biology, the centrosome is not present in all multicellular organisms or in all cells of a given organism. The main outcome of centrosome reproduction is the transmission of polarity to daughter cells and, in most animal species, the sperm-donated centrosome defines embryo polarity. Here I will discuss the role of the centrosome in cell polarity, resulting from its ability to position the nucleus at the cell center, and discuss how centrosome innovation might have been critical during metazoan evolution.