Inheritance of rol-genes from Agrobacterium rhizogenes through two generations in Kalancho?

Transformation of Kalancho? blossfeldiana ‘Molly’ using the naturally occurring bacterium Agrobacterium rhizogenes is a non-GMO strategy to breed compact plants. In the present study, crosses resembling a commercial breeding strategy were made to determine if the improved ornamental quality observed in the T1 generation was inherited to the offspring. F1 lines were produced by crossing the commercial Kalancho? cultivar ‘Sarah’ with a selected T1 line. Subsequently, F2 populations were produced by self-pollination of individual selected lines. The rol-genes were inherited to the progeny and the presence of rol-genes was confirmed in all F1 and many F2 plant lines exhibiting dwarfism. Screening of F1 and F2 plants showed that the rol-genes were inherited together. Besides decreased plant height, several F1 and F2 lines containing rol-genes exhibited changes in plant diameter, number of branches, flower diameter and time to first open flower and duration of flowering compared to the WT ‘Molly’. Furthermore, increased ethylene tolerance was observed in several lines containing rol-genes compared to the WT ‘Sarah’. Screening of three selected F2 populations derived from self-pollination of F1 lines containing rol-genes indicated a shift in distribution towards a lower mean plant height within the entire plant population compared to a control population of plants without rol-genes. Moreover, one of these F2 populations also exhibited earlier flowering compared to the control population. Compact potted plants and lines without delayed flowering and with improved ethylene tolerance were obtained and are valuable in commercial breeding programmes without using recombinant DNA technology.     

Enterococcus faecalis of human and poultry origin share virulence genes supporting the zoonotic potential of E. faecalis

Enterococcus faecalis is a major cause of nosocomial infections in humans and has been linked to severe extra‐intestinal infections in poultry. A zoonotic potential has been suggested and the aim of the present study was to investigate similarities in virulence gene profiles of E. faecalis originating from infections in humans and poultry respectively. A total of 106 isolates of E. faecalis [26 human clinical isolates, 60 poultry clinical isolates (including two small‐colony variants (SCVs) and 20 poultry cloacal isolates] were investigated for presence of seven virulence‐associated genes: ace, asa1, cylA, efaA, EF0591, esp and gelE. For each gene, the PCR‐amplification product was sequenced from one isolate in each group to explore intragenic variations between genes of human and poultry origin. Haemolytic and protease activities were assessed and isolates were assigned a sequence type (ST). Three of the seven genes investigated (ace, efaA and gelE) were present in all isolates. The asa1 was detected in 63/80 and 13/26 isolates of poultry and human origin respectively. For cylA, the numbers were 46/80 and 14/26 respectively. Among poultry isolates, esp and EF0591 were the least frequently observed genes (1/80 and 20/80 respectively); the prevalences among human isolates were 1/26 and 18/26 respectively. A high degree of similarity between genes in human and poultry isolates were confirmed by sequencing of amplification products. None of the cylA‐positive isolates demonstrated haemolytic activity, while the phenotypic expression of gelatinase varied. The ST16 was the only ST shared by human and poultry isolates. The SCV isolates did not show a unique virulence profile or phylogeny. In conclusion, regardless of the distinct phylogenetic background of most E. faecalis isolates of human and poultry origin, we found major similarities in virulence gene profile and gene sequences in isolates from the two sources, supporting the zoonotic risk associated with this organism.     

Comparative genomics of multiple plasmids from APEC associated with clonal outbreaks demonstrates major similarities and identifies several potential vaccine-targets

Avian pathogenic Escherichia coli (APEC) is associated with several types of extraintestinal infections, collectively known as colibacillosis. A heterogeneous population structure has hindered development of vaccines protective against all APEC. Recently, however, the existence of different APEC subpathotypes have been suggested, which are defined by specific disease syndromes and associated virulence genes. A collection of 14 APEC isolates representing clonal outbreaks of salpingitis accompanied by peritonitis and sepsis were characterized in the present study. All the strains carried large plasmids and the aim of the study was to investigate the similarity of these by sequencing, annotating and comparative analysis to identify potential vaccine targets. In addition, a comparison with gene content of human extraintestinal E. coli (ExPEC) subtypes was conducted. Results obtained demonstrated highly similar plasmid contents of the 14 APEC strains, despite the diversity of their chromosomal background. All 14 APEC carried the colicin V operon and numerous virulence genes. These included iss, traT, hlyF, eitABC, ompT, iroBCDEN, sitABCD, iutA and lucABCD. Several of these are shared with human ExPEC, implicating a possible zoonotic potential. Despite a diverse chromosomal background, it was concluded that the plasmid content of virulence genes are highly similar for the investigated APEC subpathotype. Based on their frequency, protein uniformity and subcellular localization iroN, iutA, iss, traT, ompT and etsC are suggested as vaccine-candidates. Experimental studies are, however, necessary to determine the protective potential of the candidates against the APEC subpathotype characterized by salpingitis, peritonitis and possibly septicaemia.     

Does canopy mean nitrogen concentration explain variation in canopy light use efficiency across 14 contrasting forest sites?

The maximum light use efficiency (LUE?=?gross primary production (GPP)/absorbed photosynthetic photon flux density (aPPFD)) of plant canopies has been reported to vary spatially and some of this variation has previously been attributed to plant species differences. The canopy nitrogen concentration [N] can potentially explain some of this spatial variation. However, the current paradigm of the N-effect on photosynthesis is largely based on the relationship between photosynthetic capacity (A(max)) and [N], i.e., the effects of [N] on photosynthesis rates appear under high PPFD. A maximum LUE-[N] relationship, if it existed, would influence photosynthesis in the whole range of PPFD. We estimated maximum LUE for 14 eddy-covariance forest sites, examined its [N] dependency and investigated how the [N]-maximum LUE dependency could be incorporated into a GPP model. In the model, maximum LUE corresponds to LUE under optimal environmental conditions before light saturation takes place (the slope of GPP vs. PPFD under low PPFD). Maximum LUE was higher in deciduous/mixed than in coniferous sites, and correlated significantly with canopy mean [N]. Correlations between maximum LUE and canopy [N] existed regardless of daily PPFD, although we expected the correlation to disappear under low PPFD when LUE was also highest. Despite these correlations, including [N] in the model of GPP only marginally decreased the root mean squared error. Our results suggest that maximum LUE correlates linearly with canopy [N], but that a larger body of data is required before we can include this relationship into a GPP model. Gross primary production will therefore positively correlate with [N] already at low PPFD, and not only at high PPFD as is suggested by the prevailing paradigm of leaf-level A(max)-[N] relationships. This finding has consequences for modelling GPP driven by temporal changes or spatial variation in canopy [N].     

Plant availability and leaching of 15N-labelled mineral fertilizer residues retained in agricultural soil for 25 years: A lysimeter study

Background

A high use-efficiency of fertilizer N remains essential to sustain high crop productivity with low environmental impact. However, little is known on the long-term lability of mineral fertilizer N.

Aims

To quantify crop uptake and leaching of ¹⁵N-labelled mineral fertilizer that has been retained in an agricultural soil for 25–30 years in crops with variable growing season.

Methods

A field plot received ¹⁵N-labelled mineral fertilizers over a period of 5 years and was then kept under arable cropping for 12 years. After relocation to 16 lysimeters, the topsoil grew set-aside grassland for the next 13 years. Then crop uptakes and leaching losses of ¹⁵N remaining in soil was tested over a 2-year period by either converting set-aside grass to production grassland, or by replacing it with spring barley (+/− autumn cover crop) or vegetation-free fallow. All treatments received unlabelled mineral N fertilizers.

Results

Crop uptake and leaching of ¹⁵N were generally highest in the first test year after termination of the set-aside. The leaching of residual ¹⁵N in soil declined in the order: vegetation-free soil (4.7%), spring barley (1.9%), spring barley + cover crop (0.7%) and production grassland (0.2%). Corresponding losses for the second leaching period were 2.7%, 0.9%, 0.4% and 0.06%. There was a fixed relationship between leaching losses of ¹⁵N and total N.

Conclusions

After residing in soil for 25–30 years, the lability of labelled mineral N fertilizer residues appeared slightly higher than the lability of bulk soil N. Autumn vegetation was crucial for reducing leaching losses.     

Random amplification of polymorphic DNA and phenotypic typing of Zimbabwean isolates of Pasteurella multocida

Eighty-one isolates presumptively identified as Pasteurella multocida from a variety of diseases in animals in Zimbabwe were subjected to biochemical characterization, capsular typing and RAPD analysis. The majority of isolates (over 80%) were assigned into named taxa and were predominantly P. multocida subsp. multocida and P. multocida subsp. septica, whilst the remainder were unassigned. Serogroup A was predominant among the three capsular types (A, B and D) of P. multocida detected. Three main RAPD clusters and three subclusters were observed among the majority of isolates (93.8%), whilst the remainder was found to be weakly related. Nine different groups of strains with similar RAPD profiles (100% similarity) were also observed. The reference strain of capsular serogroup F clustered with the reference strain of P. multocida subsp. septica, whilst all other serogroups clustered with reference strains of subsp. multocida and gallicida. Notably, serogroups A and D were observed to be closely related to the reference strain of subsp. multocida. The relationship between biotype, capsular type, host origin and disease manifestation was not clear-cut. However, most pig isolates of subsp. multocida clustered together as did most cattle isolates of subsp. multocida. RAPD tended to separate subsp. multocida from septica.     

Herd prevalence of Salmonella spp. in Danish pig herds after implementation of the Danish Salmonella Control Program with reference to a pre-implementation study

The problems addressed are: (1) comparison of prevalences of Salmonella spp. in different herd types in the Danish pig population after implementation of the Danish Salmonella Control Program (DSCP), and (2) to make a reference to a study from 1993/1994 (pre-implementation) with a discussion of possible biases when diagnostic methods differ slightly. The objectives were to present the prevalences of Salmonella spp., Salmonella Typhimurium, and multiresistant S. Typhimurium DT104 in Danish pig herds in 1998. Further, to discuss how herd prevalences may be compared to a previous study.A bacteriological study in 1998 comprised: (a) a random sample of slaughter pig producing herds (N=1962); (b) a random sample of farrow-to-grower (sow) herds (N=305); and (c) all breeding and multiplying (genetic) herds (N=366). A previous bacteriological study on Salmonella presence in 1993/1994 served as a model for the present study. The results of the study were that multiresistant S. Typhimurium DT104 was detected in one herd producing slaughter pigs. The herd apparent prevalences (HAPs) of Salmonella spp. were 11.7, 16.7 and 11.4% in genetic, sow, and slaughter pig herds, respectively. The conclusion of the study was that prevalence of multiresistant S. Typhimurium DT104 was low in the examined slaughter pig herds. The herd true prevalence (HTP) of Salmonella spp. in pigs had declined from before the start of the DSCP in 1993/1994 to 4 years later (1998).     

A correction factor for,and its application to,visual censuses of littoral fish

Visual estimation techniques were used between October 1976 and May 1977 on the fish fauna of an enclosed mid-intertidal pool in the eastern Cape, South Africa, to test the usefulness and accuracy of this non-consumptive census method. In January, 1977, two pools were censused for fish, then poisoned using the ichthyocide ‘pronoxfish’ and all fish collected. Forty species in 21 families were obtained from the two stations. The accuracy of visual estimates varied with the species of fish. Secretive and cryptically coloured types, as well as species which Inhabit crevices, were underestimated, between 0 and 86% being counted. Other fish with protective colours but that occur more openly and do not react adversely to divers, as well as schooling species, were also underestimated, but up to 100% of the total were observed. Those which occur singly or in small groups in the water column were counted more accurately, with 57 to 100% of the actual number being seen. Correction factors were calculated from these data and applied to a census taken of one pool in May. Comparisons of corrected and actual numbers Indicate that these factors were relatively accurate for non-secretive species. The factors will, however, vary for each species, from observer to observer, depending on their personal experience, and from area to area.