Variation of prezygotic barriers in the interspecific hybridization between oilseed rape and wild radish

Escape of transgenes from genetically modified oilseed rape, Brassica napus, into wild radish, Raphanus raphanistrum, depends on sexual compatibility. The variation in prezygotic barriers of two different cultivars for interspecific hybridization with a population of wild radish was investigated by hand crossing and fluorescence microscopy of pistils. Significant differences were observed between oil seed rape cultivars in their ability to accept wild radish pollen germinating onto their stigma and the rate of fertilization of ovules. Some differences among the pollen donor plants were also detected. These results suggest that the rate of interspecific hybridization in the field would depend upon the oilseed rape cultivar and the genotype composition of the local wild populations. The implication of S-related genes, as revealed through identification by pistil tissue prints of class I and II S-types of SLG (S-Locus Glycoprotein) and SLR1 (S-Locus Related),and immuno-IEF, was not significant. This revised version was published online in August 2006 with corrections to the Cover Date.     

Variation of pathotypes and races and their correlations with clonal lineages in Verticillium dahliae

Understanding pathogenic variation in plant pathogen populations is key for the development and use of host resistance for managing verticillium wilt diseases. A highly virulent defoliating (D) pathotype in Verticillium dahliae has previously been shown to occur only in one clonal lineage (lineage 1A). By contrast, no clear association has yet been shown for race 1 with clonal lineages. Race 1 carries the effector gene Ave1 and is avirulent on hosts that carry resistance gene Ve1 or its homologues. The hypothesis tested was that race 1 arose once in a single clonal lineage, which might be expected if V. dahliae acquired Ave1 by horizontal gene transfer from plants, as hypothesized previously. In a diverse sample of 195 V. dahliae isolates from nine clonal lineages, all race 1 isolates were present only in lineage 2A. Conversely, all lineage 2A isolates displayed the race 1 phenotype. Moreover, 900‐bp nucleotide sequences from Ave1 were identical among 27 lineage 2A isolates and identical to sequences from other V. dahliae race 1 isolates in GenBank. The finding of race 1 in a single clonal lineage, with identical Ave1 sequences, is consistent with the hypothesis that race 1 arose once in V. dahliae. Molecular markers and virulence assays also confirmed the well‐established finding that the D pathotype is found only in lineage 1A. Pathogenicity assays indicated that cotton and olive isolates of the D pathotype (lineage 1A) were highly virulent on cotton and olive, but had low virulence on tomato.     

Effects of copper and cadmium on uptake and leakage of K(+) in birch (Betula pendula) roots

Uptake and leakage experiments were performed to study the effects of copper and cadmium on K(+) fluxes in birch (Betula pendula Roth) roots. Labeled rubidium ((86)Rb(+)) was used as a tracer for K(+). Plants were pretreated with Cu or Cd (0, 2, 5 or 25 microM) for 0-300 min and then transferred to radiolabeled nutrient solution (150 microM K(+)) with or without 2,4-dinitrophenol (DNP) to separate the effects of heavy metal on active and passive K(+) influxes. Passive K(+) influx was decreased by pretreatment with Cu but was only slightly affected by pretreatment with Cd. Pretreatment with 2 microM Cu increased active K(+) influx, whereas pretreatment with 25 microM Cu decreased active K(+) influx and intermediate Cu concentrations (5 microM) did not affect active K(+) influx. The pretreatment effects of Cu on active and passive K(+) influxes increased with increasing pretreatment time. During the first hour, pretreatment with Cd decreased active K(+) influx with increasing pretreatment time, whereafter recovery began. To measure K(+) efflux, birch plants were loaded with (86)Rb(+) for 7 days before being exposed to Cu or Cd (0, 1, 3, 5 or 10 microM) in unlabeled nutrient solutions for 24 h. Net efflux of K(+) was measured as (86)Rb(+)-activity in the nutrient solutions 24 h after a heavy metal had been introduced. Efflux of K(+) increased with increasing Cu or Cd concentration in the unlabeled nutrient solution.The data indicate that Cu and Cd affected K(+) influx differently and that recovery mechanism(s), which were induced shortly after heavy metal introduction, counteracted the heavy-metal induced inhibition of active K(+) influx. Efflux of K(+) from plant roots over a 24-h period indicated that Cu and Cd had similar effects on K(+) efflux. There was no evidence of a recovery mechanism counteracting the heavy-metal-induced inhibition of K(+) efflux.     

Genetic diversity of livestock-associated MRSA isolates obtained from piglets from farrowing until slaughter age on four farrow-to-finish farms

During a previous longitudinal study, performed on four farrow-to-finish farms (A to D), samples were taken from twelve sows, their offspring, and the environment on various occasions over six months to study the MRSA presence. During the present study, a selection of the obtained MRSA isolates were typed by multiple-locus variable-number tandem-repeat analysis (MLVA), Pulsed Field Gel Electrophoresis (PFGE), spa typing, and SCCmec typing to study the genetic diversity of LA-MRSA isolates and to determine possible MRSA sources for pig(let)s. PFGE, spa typing, and SCCmec typing revealed the presence of one or few dominant genotype(s) per farm. In contrast, 212 MLVA types were detected on the four farms, forming one cluster on farm A, three on farm B, four on farm C and two on farm D. The genotype, found on farm A was unique for this farm. Farms B, C and D shared one cluster. In general, MLVA types from these clusters were isolated from piglets, sows, and the environment on various sampling events. Piglets carried MLVA types both related and unrelated to their mother sows’ MLVA types at farrowing and onwards. In conclusion, molecular typing revealed that within a farm one or a few dominant strain(s) are widespread. Potential MRSA sources for piglets were mother sows, the environment and other piglets.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0089-4) contains supplementary material, which is available to authorized users.     

Failure to spread bovine virus diarrhoea virus infection from primarily infected calves despite concurrent infection with bovine coronavirus

Previous reports on the spread of bovine virus diarrhoea virus (BVDV) from animals primarily infected with the agent are contradictory. In this study, the possibility of transmission of BVDV from calves simultaneously subjected to acute BVDV and bovine coronavirus (BCV) infection was investigated. Ten calves were inoculated intranasally with BVDV Type 1. Each of the 10 calves was then randomly allocated to one of two groups. In each group there were four additional calves, resulting in five infected and four susceptible calves per group. Virulent BCV was actively introduced in one of the groups by means of a transmitter calf. Two calves, susceptible to both BVDV and BCV, were kept in a separate group, as controls. All ten calves actively inoculated with BVDV became infected as shown by seroconversions, and six of them also shed the virus in nasal secretions. However, none of the other eight calves in the two groups (four in each) seroconverted to this agent. In contrast, it proved impossible to prevent the spread of BCV infection between the experimental groups and consequently all 20 study calves became infected with the virus. Following infection, BCV was detected in nasal secretions and in faeces of the calves and, after three weeks in the study, all had seroconverted to this virus. All calves, including the controls, showed at least one of the following clinical signs during days 3-15 after the trial started: fever (> or =40 degrees C), depressed general condition, diarrhoea, and cough. The study showed that BVDV primarily infected cattle, even when co-infected with an enteric and respiratory pathogen, are inefficient transmitters of BVDV. This finding supports the principle of the Scandinavian BVDV control programmes that elimination of BVDV infection from cattle populations can be achieved by identifying and removing persistently infected (PI) animals, i.e. that long-term circulation of the virus without the presence of PI animals is highly unlikely.     

Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences

Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions.     

Contrasting phenology and female cone characteristics of the two Macaronesian island endemic cedars (<Emphasis Type="Italic">Juniperus cedrus</Emphasis> and <Emphasis Type="Italic">J. brevifolia</Emphasis>)

Phenology and female cone characteristics of the two endemic cedars (Juniperus cedrus and J. brevifolia) from the Macaronesian islands were studied. Despite their closely taxonomic affinity and their evolution under insular conditions, different trends were recorded. Mature J. cedrus female cones were present throughout the year, while those from J. brevifolia were only present in summer and autumn. J. cedrus female cone size was significantly larger than that of J. brevifolia, a trend consistent with the presence of larger vertebrates (lizards and birds) in the Canary Islands. However, water content was four times higher in J. brevifolia female cones, which can be related with the higher rainfall existing in the Azores. J. cedrus has two or three seeds per cone, whereas J. brevifolia frequently had three. Seeds from J. cedrus were clearly larger and heavier, coinciding with the female cone size trend. However, tetrazolium tests revealed higher viability values in J. brevifolia. The relatively low percentage of filled seeds in J. cedrus could be a consequence of the climatic stress and limits to pollination due to fragmented populations as described for other Juniperus species. In summary, our results reveal that some environmental factors such as the harsh conditions, high population fragmentation and the dependence on large dispersers have compromised the fitness of J. cedrus in the Canary Islands.     

Traces of human migrations in Helicobacter pylori populations

Helicobacter pylori, a chronic gastric pathogen of human beings, can be divided into seven populations and subpopulations with distinct geographical distributions. These modern populations derive their gene pools from ancestral populations that arose in Africa, Central Asia, and East Asia. Subsequent spread can be attributed to human migratory fluxes such as the prehistoric colonization of Polynesia and the Americas, the neolithic introduction of farming to Europe, the Bantu expansion within Africa, and the slave trade.