Cultivation of hitherto-uncultured bacteria belonging to the Verrucomicrobia subdivision 1 from the potato (Solanum tuberosum L.) rhizosphere

The role of dominant bacterial groups in the plant rhizosphere, e.g., those belonging to the phyla Acidobacteria and Verrucomicrobia, has, so far, not been elucidated, and this is mainly due to the lack of culturable representatives. This study aimed to isolate hitherto-uncultured bacteria from the potato rhizosphere by a combination of cultivation approaches. An agar medium low in carbon availability (oligotrophic agar medium) and either amended with potato root exudates or catalase or left unamended was used with the aim to improve the culturability of bacteria from the potato rhizosphere. The colony forming unit numbers based on colonies and microcolonies were compared with microscopically determined fluorescence-stained cell numbers. Taxonomical diversity of the colonies was compared with that of library clones made from rhizosphere DNA, on the basis of 16S rRNA gene comparisons. The oligotrophic media amended or not with catalase or rhizosphere extract recovered up to 33.6% of the total bacterial numbers, at least seven times more than the recovery observed on R2A. Four hitherto-uncultured Verrucomicrobia subdivision 1 representatives were recovered on agar, but representatives of this group were not found in the clone library. The use of oligotrophic medium and its modifications enabled the growth of colony numbers, exceeding those on classical agar media. Also, it led to the isolation of hitherto-uncultured bacteria from the potato rhizosphere. Further improvement in cultivation will certainly result in the recovery of other as-yet-unexplored bacteria from the rhizosphere, making these groups accessible for further investigation, e.g., with respect to their possible interactions with plants.

Endophytic Colonization of Potato (Solanum tuberosum L.) by a Novel Competent Bacterial Endophyte, Pseudomonas putida Strain P9, and Its Effect on Associated Bacterial Communities

Pseudomonas putida strain P9 is a novel competent endophyte from potato. P9 causes cultivar-dependent suppression of Phytophthora infestans. Colonization of the rhizoplane and endosphere of potato plants by P9 and its rifampin-resistant derivative P9R was studied. The purposes of this work were to follow the fate of P9 inside growing potato plants and to establish its effect on associated microbial communities. The effects of P9 and P9R inoculation were studied in two separate experiments. The roots of transplants of three different cultivars of potato were dipped in suspensions of P9 or P9R cells, and the plants were planted in soil. The fate of both strains was followed by examining colony growth and by performing PCR-denaturing gradient gel electrophoresis (PCR-DGGE). Colonies of both strains were recovered from rhizoplane and endosphere samples of all three cultivars at two growth stages. A conspicuous band, representing P9 and P9R, was found in all Pseudomonas PCR-DGGE fingerprints for treated plants. The numbers of P9R CFU and the P9R-specific band intensities for the different replicate samples were positively correlated, as determined by linear regression analysis. The effects of plant growth stage, genotype, and the presence of P9R on associated microbial communities were examined by multivariate and unweighted-pair group method with arithmetic mean cluster analyses of PCR-DGGE fingerprints. The presence of strain P9R had an effect on bacterial groups identified as Pseudomonas azotoformans, Pseudomonas veronii, and Pseudomonas syringae. In conclusion, strain P9 is an avid colonizer of potato plants, competing with microbial populations indigenous to the potato phytosphere. Bacterization with a biocontrol agent has an important and previously unexplored effect on plant-associated communities.

Transgenic tobacco revealing altered bacterial diversity in the rhizosphere during early plant development

The rhizosphere constitutes a complex niche that may be exploited by a wide variety of bacteria. Bacterium-plant interactions in this niche can be influenced by factors such as the expression of heterologous genes in the plant. The objective of this work was to describe the bacterial communities associated with the rhizosphere and rhizoplane regions of tobacco plants, and to compare communities from transgenic tobacco lines (CAB1, CAB2 and TRP) with those found in wild-type (WT) plants. Samples were collected at two stages of plant development, the vegetative and flowering stages (1 and 3 months after germination). The diversity of the culturable microbial community was assessed by isolation and further characterization of isolates by amplified ribosomal RNA gene restriction analysis (ARDRA) and 16S rRNA sequencing. These analyses revealed the presence of fairly common rhizosphere organisms with the main groups Alphaproteobacteria, Betaproteobacteria, Actinobacteria and Bacilli. Analysis of the total bacterial communities using PCR-DGGE (denaturing gradient gel electrophoresis) revealed that shifts in bacterial communities occurred during early plant development, but the reestablishment of original community structure was observed over time. The effects were smaller in rhizosphere than in rhizoplane samples, where selection of specific bacterial groups by the different plant lines was demonstrated. Clustering patterns and principal components analysis (PCA) were used to distinguish the plant lines according to the fingerprint of their associated bacterial communities. Bands differentially detected in plant lines were found to be affiliated with the genera Pantoea, Bacillus and Burkholderia in WT, CAB and TRP plants, respectively. The data revealed that, although rhizosphere/rhizoplane microbial communities can be affected by the cultivation of transgenic plants, soil resilience may be able to restore the original bacterial diversity after one cycle of plant cultivation.

Genomics of pyrrolnitrin biosynthetic loci: evidence for conservation and whole-operon mobility within Gram-negative bacteria

Pyrrolnitrin (PRN) is a tryptophan-derived secondary metabolite produced by a narrow range of Gram-negative bacteria. The PRN biosynthesis by rhizobacteria presumably has a key role in their life strategies and in the biocontrol of plant diseases. The biosynthetic operon that encodes the pathway that converts tryptophan to PRN is composed of four genes, prnA through D, whose diversity, genomic context and spread over bacterial genomes are poorly understood. Therefore, we launched an endeavour aimed at retrieving, by in vitro and in silico means, diverse bacteria carrying the prnABCD biosynthetic loci in their genomes. Analysis of polymorphisms of the prnD gene sequences revealed a high level of conservation between Burkholderia, Pseudomonas and Serratia spp. derived sequences. Whole-operon- and prnD-based phylogeny resulted in tree topologies that are incongruent with the taxonomic status of the evaluated strains as predicted by 16S rRNA gene phylogeny. The genomic composition of c. 20 kb DNA fragments containg the PRN operon varied in different strains. Highly conserved and distinct transposase-encoding genes surrounding the PRN biosynthetic operons of Burkholderia pseudomallei strains were found. A prnABCD-deprived genomic region in B. pseudomallei strain K96243 contained the same gene composition as, and shared high homology with, the flanking regions of the PRN operon in B. pseudomallei strains 668, 1106a and 1710b. Our results strongly suggest that the PRN biosynthetic operon is mobile. The extent, frequency and promiscuity of this mobility remain to be understood.

The 2008 update of the Aspergillus nidulans genome annotation: A community effort

The identification and annotation of protein-coding genes is one of the primary goals of whole-genome sequencing projects, and the accuracy of predicting the primary protein products of gene expression is vital to the interpretation of the available data and the design of downstream functional applications. Nevertheless, the comprehensive annotation of eukaryotic genomes remains a considerable challenge. Many genomes submitted to public databases, including those of major model organisms, contain significant numbers of wrong and incomplete gene predictions. We present a community-based reannotation of the Aspergillus nidulans genome with the primary goal of increasing the number and quality of protein functional assignments through the careful review of experts in the field of fungal biology. (C) 2009 Elsevier Inc. All rights reserved.

Learning for fun: The unique contribution of educational leisure experiences

What do visitors want or expect from an educational leisure activity such as a visit to a museum, zoo, aquarium or other such experience? Is it to learn something or to experience learning? This paper uses the term 'learning for fun' to refer to the phenomenon in which visitors engage in a learning experience because they value and enjoy the process of learning itself. Five propositions regarding the nature of learning for fun are discussed, drawing on quantitative and qualitative data from visitors to a range of educational leisure activities. The commonalities between learning for fun and other theoretical constructs such as 'experience,' 'flow', 'intrinsic motivation', and 'curiosity' are explored. It is concluded that learning for fun is a unique and distinctive offering of educational leisure experiences, with implications for future research and experience design.

It's all very well, in theory: Theoretical perspectives and their applications in contemporary pedagogical research

Current debates about educational theory are concerned with the relationship between knowledge and power and thereby issues such as who possesses a truth and how have they arrived at it, what questions are important to ask, and how should they best be answered. As such, these debates revolve around questions of preferred, appropriate, and useful theoretical perspectives. This paper overviews the key theoretical perspectives that are currently used in physical education pedagogy research and considers how these inform the questions we ask and shapes the conduct of research. It also addresses what is contested with respect to these perspectives. The paper concludes with some cautions about allegiances to and use of theories in line with concerns for the applicability of educational research to pressing social issues.

On a semilinear Schrodinger equation with critical Sobolev exponent

We consider the semilinear Schrodinger equation -Deltau+V(x)u= K(x) \u \ (2*-2 u) + g(x; u), u is an element of W-1,W-2 (R-N), where N greater than or equal to4, V, K, g are periodic in x(j) for 1 less than or equal toj less than or equal toN, K>0, g is of subcritical growth and 0 is in a gap of the spectrum of -Delta +V. We show that under suitable hypotheses this equation has a solution u not equal 0. In particular, such a solution exists if K equivalent to 1 and g equivalent to 0.

Compact tiled display with uniform illumination

The demand for more pixels is beginning to be met as manufacturers increase the native resolution of projector chips. Tiling several projectors still offers a solution to augment the pixel capacity of a display. However, problems of color and illumination uniformity across projectors need to be addressed as well as the computer software required to drive such devices. We present the results obtained on a desktop-size tiled projector array of three D-ILA projectors sharing a common illumination source. A short throw lens (0.8:1) on each projector yields a 21-in. diagonal for each image tile; the composite image on a 3×1 array is 3840×1024 pixels with a resolution of about 80 dpi. The system preserves desktop resolution, is compact, and can fit in a normal room or laboratory. The projectors are mounted on precision six-axis positioners, which allow pixel level alignment. A fiber optic beamsplitting system and a single set of red, green, and blue dichroic filters are the key to color and illumination uniformity. The D-ILA chips inside each projector can be adjusted separately to set or change characteristics such as contrast, brightness, or gamma curves. The projectors were then matched carefully: photometric variations were corrected, leading to a seamless image. Photometric measurements were performed to characterize the display and are reported here. This system is driven by a small PC cluster fitted with graphics cards and running Linux. It can be scaled to accommodate an array of 2×3 or 3×3 projectors, thus increasing the number of pixels of the final image. Finally, we present current uses of the display in fields such as astrophysics and archaeology (remote sensing).

Teaching in rural and remote schools: a literature review

In this paper. the authors examine a wide range of recent research into the preparation and support for teachers working in rural and remote schools. The paper reviews many preservice and inservice initiatives which highlight issues affecting:teaching and learning in schools outside the major metropolitan centres. The work is reviewed from an Australian perspective but evaluates research from throughout the world. The paper concludes that despite a large body of research (Gibson, 1994), that has identified the need for specialised pre-service preparation which accommodates the social and professional differences associated with work in rural and remote areas, the implementation of such programs by teacher training institutions has been sparse, lacking in cohesion and in many cases non-existent. (C) 1998 Elsevier Science Ltd. All rights reserved.

Characteristics of long-term cannabis users in Sydney, Australia

Two hundred long-term cannabis users (58% male) were interviewed on their characteristics and experience of use. Respondents had been regularly using cannabis for an average of 11 years and more than half used daily (56%). The most common route of administration was in a waterpipe, and nearly all (93%) smoked the flowering heads ot the plant. One in 5 (21%) had a cannabis-related conviction. The benefits of use were perceived to be its relaxing, mood-enhancing effects, and its ability to alter consciousness. The most commonly cited negative aspects of use were cost, negative psychological effects and legal status. Polydrug use was common, with alcohol and tobacco almost universally used on a regular basis. More than half the drinkers in the sample were consuming alcohol at hazardous or harmful levels.

Researching University Teaching in Australia: themes and issues in academics' reflections

University teaching is a diverse enterprise which encompasses a range of disciplines, cirricula, teaching methods, learning tasks and learning approaches. Within this diversity, common themes and issues which reflect academics' understanding of effective teaching may be discerned. Drawing on written data collected from 708 practising teachers who were nominated by their Heads or Deans as exhibiting exemplary teaching practice, and from interviews conducted with 44 of these, a number of these themes and issues are identified and illustrated The findings offer insights which may stimulate further reflection on, and discussion of, the quality of teaching in higher education.

Heroin-assisted treatment as a response to the public health problem of opiate dependence

Injection drug use (involving the injection of illicit opiates) poses serious public health problems in many countries. Research has indicated that injection drug users are at higher risk for morbidity in the form of HIV/AIDS and Hepatitis B and C, and drug-related mortality, as well as increased criminal activity. Methadone maintenance treatment is the most prominent form of pharmacotherapy treatment for illicit opiate dependence in several countries, and its application varies internationally with respect to treatment regulations and delivery modes. In order to effectively treat those patients who have previously been resistant to methadone maintenance treatment, several countries have been studying and/or considering heroin-assisted treatment as a complementary form of opiate pharmacotherapy treatment. This paper provides an overview of the prevalence of injection drug use and the opiate dependence problem internationally, the current opiate dependence treatment landscape in several countries, and the status of ongoing or planned heroin-assisted treatment trials in Australia, Canada and certain European countries.

Twelve-month Outcomes After Coronary Stenting With the Genous (TM) Bio-Engineered R Stent (TM) in Diabetic Patients from the e-HEALING Registry

Objectives: We compared 12-month outcomes, regarding ischemic events, repeat intervention, and ST, between diabetic and nondiabetic patients treated with the Genous (TM) EPC capturing R stent (TM) during routine nonurgent percutaneous coronary intervention (PCI) using data from the multicenter, prospective worldwide e-HEALING registry. Background: Diabetic patients have an increased risk for restenosis and stent thrombosis (ST). Methods: In the 4,996 patient e-HEALING registry, 273 were insulin requiring diabetics (IRD), 963 were non-IRD (NIRD), and 3,703 were nondiabetics. The 12-month primary outcome was target vessel failure (TVF), defined as target vessel-related cardiac death or myocardial infarction (MI) and target vessel revascularization. Secondary outcomes were the composite of cardiac death, MI or target lesion revascularization (TLR), and individual outcomes including ST. Cumulative event rates were estimated with the Kaplan-Meier method and compared with a log-rank test. Results: TVF rates were respectively 13.4% in IRD, 9.0% in NIRD, and 7.9% in nondiabetics (P < 0.01). This was mainly driven by a higher mortality hazard in IRD (P < 0.001) and NIRD (P = 0.07), compared with nondiabetics. TLR rates were comparable in NIRD and nondiabetics, but significantly higher in IRD (P = 0.04). No difference was observed in ST. Conclusion: The 1-year results of the Genous stent in a real-world population of diabetics show higher TVF rates in diabetics compared with nondiabetics, mainly driven by a higher mortality hazard. IRD is associated with a significant higher TLR hazard. Definite or probable ST in all diabetic patients was comparable with nondiabetics. (J Interven Cardiol 2011;24:285-294)