Early detection of sugar beet pathogen Ramularia beticola in leaf and air samples using qPCR

A quantitative PCR method (qPCR) was developed for the detection and quantification of Ramularia beticola causing Ramularia leaf spot in sugar beet. R. beticola specific primers were designed based on the internal transcribed spacer region 2 (ITS2). The assay was applied on DNA extracted from spores trapped on tape from Burkard spore traps placed in an artificially inoculated sugar beet field trial and in two sugar beet fields with natural infections. R. beticola DNA was detected at variable amounts in the air samples 14 to 16 days prior to first visible symptoms. R. beticola DNA was detected in air samples from fields with natural infection at significant and increasing levels from development of the first symptoms, indicating that spore production within the crop plays a major role in the epidemic development of the disease. Sugar beet leaves sampled from the inoculated field trial were also tested with the qPCR assay. It was possible to detect the presence of R. beticola in the leaves pre-symptomatic at least 10 days before the occurrence of the visible symptoms of Ramularia leaf spot. This is the first report of a molecular assay, which allows screening for the presence of R. beticola in plant material and in air samples prior to the appearance of visible symptoms. An early detection has potential as a tool, which can be part of a warning system predicting the onset of the disease in the sugar beet crop and helping to optimise fungicide application.     

Assessing the consequences of habitat fragmentation for two migratory salmonid fishes

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Reduction of early sexual maturation in male S0 Atlantic salmon (Salmo salar L.) by dietary supplementation of tetradecylthioacetic acid (TTA)

Effects of dietary tetradecylthioacetic fatty acid (TTA) on muscle fat, development of gonads and early sexual maturation in S0 Atlantic salmon during the first year in sea were investigated. TTA (0.5% w/w) was added to the feeds for 8 weeks in the spring. In May, at the end of the TTA‐feeding period, the fish in the TTA group had significantly (P < 0.05) less fat (10.1%) stored in muscle compared with the control group (10.8%). In September, mean male gonadosomatic index (GSI) in maturing fish in the TTA group was found to be lower compared with the maturing fish in the control group (P = 0.05). On the basis of GSI values, male sexual maturation in September was 10.0% vs. 14.4% for the TTA and the control group respectively. Thus, relative to the control group, the incidence of male sexual maturation in the TTA group was reduced by about 1/3 (P = 0.002). Production data was not affected by dietary supplementation of TTA. This study reveals that TTA significantly reduces the incidence of male sexual maturation in S0 Atlantic salmon. A significant elimination model of TTA in fish muscle that takes into account, the growth rate of the fish was further developed in this study.     

An investigation to enhance understanding of the stimulation of weed seedling emergence by soil disturbance

Enhanced understanding of soil disturbance effects on weed seedling recruitment will help guide improved management approaches. Field experiments were conducted at 16 site‐years at 10 research farms across Europe and North America to (i) quantify superficial soil disturbance (SSD) effects on Chenopodium album emergence and (ii) clarify adaptive emergence behaviour in frequently disturbed environments. Each site‐year contained factorial combinations of two seed populations (local and common, with the common population studied at all site‐years) and six SSD timings [0, 50, 100, 150, 200 day‐degrees (d°C, base temperature 3°C) after first emergence from undisturbed soil]. Analytical units in this study were emergence flushes. Flush magnitudes (maximum weekly emergence per count flush) and flush frequencies (flushes year^?1) were compared between disturbed and undisturbed seedbanks. One year after burial, SSD promoted seedling emergence relative to undisturbed seedbanks by increasing flush magnitude rather than increasing flush frequency. Two years after burial, SSD promoted emergence through increased flush magnitude and flush frequency. The promotional effects of SSD on emergence were strongest within 500 d°C following SSD; however, low levels of SSD‐induced emergence were detected as late as 3000 d°C following SSD. Accordingly, stale seedbed practices that eliminate weed seedlings should occur within 500 d°C of disturbance, because few seedlings emerge after this time. However, implementation of stale seedbed practices will probably cause slight increases in weed population densities throughout the year. Compared with the common population, local populations exhibited reduced variance in total emergence measured within sites and across SSD treatments, suggesting that C. album adaptation to local pedo‐climatic conditions involves increased consistency in SSD‐induced emergence.     

<Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> occurs asymptomatically on apple leaves

Fungi within the Colletotrichum acutatum species complex occur asymptomatically on plant parts of many different plant species. Leaves from apple orchards in southern Norway were sampled, frozen for five hours and incubated for six days to reveal presence of asymptomatic infections of C. acutatum. Number of leaves (incidence) and leaf area covered (severity) with conidial masses of C. acutatum were assessed biweekly on cv. Aroma from late May to late September during three growing seasons. The first finding of conidial masses occurred in the second half of July, and there was a higher incidence occurring in August and September. Sampling of leaves from fruit spurs and vegetative shoots of cvs. Aroma and Elstar showed that conidial masses of C. acutatum developed on leaves on both shoot types, and there was no difference in incidence between these two types. The fungus was detected on leaves from six of eight commercial orchards of cv. Aroma over three years, with a mean incidence of 5.5 %. After storage, bitter rot was found on apple fruit from all eight orchards. There was no correlation between incidence of conidial masses of C. acutatum on leaves and on fruit. In all orchards and seasons investigated, incidence and severity on leaves varied from 0 to 67 % and 0 to 85 %, respectively. The discovery of apple leaves containing conidial masses of C. acutatum clearly indicate for leaves as a potential source of inoculum for fruit infections.     

Between- and within-site variation of density and bending properties of Picea abies structural timber from Norway

This study provides an analysis on the variability of structural timber of Norway spruce (Picea abies) grown in Norway. Density, modulus of elasticity (MOE) and bending strength were measured on 1188 boards from 205 trees, sampled from 14 sites throughout Southern Norway, Eastern Norway and Trøndelag. The area represents the procurement area for the majority of Norwegian sawmills. The variability of the timber properties was analysed in a linear mixed model where the random variance was divided into variance due to site, variance due to trees and within-tree variance. Models describing variance due to site based on site index, altitude and latitude were developed, and combined with data from the Norwegian National Forest Inventory to estimate mean values and variability of the timber properties. The results showed that major parts of the variance due to site are explained by altitude and site index, and for density also by latitude. Major parts of the variance due to site and the variance due to trees in bending strength and MOE were explained by density.     

Selection in a provenance trial of Norway spruce (Picea abies L. Karst) produced a land race with desirable properties

Selections were made within East European Norway spruce provenances in a provenance trial in northern Sweden and the selected clones were grafted in a Norwegian seed orchard mixed with plus trees of local origin. Full-sib families were generated by controlled crosses, producing sets of both inter- and intraprovenance families, which were planted in two short-term and in long-term trials. Measurements were made of frost damage, tree height, annual shoot growth rhythm and lammas shoots, and genetic parameters were estimated. The intraprovenance families from selected trees of East European origin showed the least variation in all traits and had the lowest heritability values. The variation among the intraprovenance families of local origin was large and with the highest estimates of heritability. The means of the provenance hybrid families were intermediate between those of the intraprovenance families. Strong genetic correlations were present between the growth rhythm traits measured, and in the family group of local origin also between these traits and frost damage, as well as for lammas shoots and height. The progenies of the selected parents of East European origin seem to be a desirable land race with less damage, lower percentage of lammas shoots and less variability in growth capacity.     

Multi-locus Sequence Analysis (MLSA) of Edwardsiella tarda isolates from fish

Edwardsiella tarda is an enteric fish pathogen that has caused significant economic losses in a range of fish species residing in diverse ecological conditions. Several molecular methods relying on DNA fingerprinting (RAPD, RFLP and ERIC-PCR) and the gyrB gene marker have been used to characterize E. tarda isolates. However, all had drawbacks in resolving power and reproducibility. The present study was aimed at developing a novel Multi-locus Sequence Analysis (MLSA) scheme for genetic characterization of E. tarda isolates originating from multiple sources. MLSA has been described as an effective molecular tool with superior discriminatory power and reproducibility for exploring intra-species genetic diversity of several bacterial species. Nucleotide sequence fragments of eight protein coding housekeeping genes (gyrB, mdh, adk, dnaK, phoR, metG, pyrG and aroE2) were obtained from 23 fish pathogenic E. tarda isolates of different geographical origins, one human isolate and 3 reference strains. The phylogenetic relationships between isolates in individual gene analyses were not consistent, although some common patterns were apparent. Phylogenetic analysis based on concatenated sequences of seven gene loci, however, buffered the conflicting phylogenetic signals and resolved isolates according to their geographical origin and/or fish host. The MLSA revealed two major genetically diverging clusters in E. tarda isolates examined, one cluster representing isolates from fish and the other representing (in the main) human isolates, with E. ictaluri cluster situated in between. The results suggest, therefore, that the fish pathogenic E. tarda isolates may have been previously misclassified and probably represent one or more as yet unrecognized taxa within the genus Edwardsiella. The MLSA described here was robust enough in discriminating E. tarda isolates not only with respect to their geographical origins but also within different hosts from the same geographical location, high-lighting its potential application in tracing the source of infection and understand the epidemiological relationships among isolates of environmental, fish, other domestic animals or human origins.