Wide Screening of Phage-Displayed Libraries Identifies Immune Targets in Planta

Microbe-Associated Molecular Patterns and virulence effectors are recognized by plants as a first step to mount a defence response against potential pathogens. This recognition involves a large family of extracellular membrane receptors and other immune proteins located in different sub-cellular compartments. We have used phage-display technology to express and select for Arabidopsis proteins able to bind bacterial pathogens. To rapidly identify microbe-bound phage, we developed a monitoring method based on microarrays. This combined strategy allowed for a genome-wide screening of plant proteins involved in pathogen perception. Two phage libraries for high-throughput selection were constructed from cDNA of plants infected with Pseudomonas aeruginosa PA14, or from combined samples of the virulent isolate DC3000 of Pseudomonas syringae pv. tomato and its avirulent variant avrRpt2. These three pathosystems represent different degrees in the specificity of plant-microbe interactions. Libraries cover up to 26107 different plant transcripts that can be displayed as functional proteins on the surface of T7 bacteriophage. A number of these were selected in a bio-panning assay for binding to Pseudomonas cells. Among the selected clones we isolated the ethylene response factor ATERF-1, which was able to bind the three bacterial strains in competition assays. ATERF-1 was rapidly exported from the nucleus upon infiltration of either alive or heat-killed Pseudomonas. Moreover, aterf-1 mutants exhibited enhanced susceptibility to infection. These findings suggest that ATERF-1 contains a microbe-recognition domain with a role in plant defence. To identify other putative pathogen-binding proteins on a genome-wide scale, the copy number of selected-vs.-total clones was compared by hybridizing phage cDNAs with Arabidopsis microarrays. Microarray analysis revealed a set of 472 candidates with significant fold change. Within this set defence-related genes, including well-known targets of bacterial effectors, are over-represented. Other genes non-previously related to defence can be associated through this study with general or strain-specific recognition of Pseudomonas.

Libraries and alternative formats research

Good practice pointers

Libraries and alternative formats research

This series of five documents is a result of research carried out with a grant from CLAUD (a group of HE librarians in South West England who are working to help create libraries accessible to users with a disability). The research involved over 60 responses representing 49 different Higher Education Institutes (HEIs).

"Future fit your catalog" Panel [Presentation]:World Cat Local Implementation as part of a Consortia of Libraries within the University System of Maryland and Affiliated Institutitons

Power Point from Panel presentation giving implementation and search result displays and linking (17 slides)

The use of aerial photographs for estimating school sizes of cetaceans

ENGLISH: The accuracy and precision of dolphin school size estimates based on aerial photograph counts were examined using data collected on recent aerial and ship surveys. These estimates were found to be accurate during a 1979research cruise aboard a tuna purse-seiner; dolphin schools were photographed from the ship’s helicopter, encircled with the purse-seine, and then counted as the dolphins were released from the net. A comparison of photographic estimates with these counts indicated that the relationship was fairly close and gave no indication of significantly differing from 1:1. During a 1980 aerial study, photographic estimates from different schools, passes, and camera formats were compared and were found to be quite precise with a standard deviation of approximately 60/0 of school size. Photographic estimates were also compared with estimates made by aerial observers. Most observers tended to underestimate school size, particularly for large schools. The variability among observers was high, indicating that observers should be individually calibrated. SPANISH: Se examinó la exactitud y la precisión de las estimaciones de la magnitud de los cardúmenes de delfines basadas en el cálculo de las fotografías aéreas, usando los datos obtenidos en los últimos reconocimientos aéreos y de los barcos. En 1979, durante un crucero de investigación en un cerquero atunero, se encontró que estas estimaciones eran acertadas; se fotografiaron los cardúmenes de delfines desde un helicóptero del barco, cercados con la red y luego se contaron a medida que se libraban los delfines de la red. Una comparación de las estimaciones fotográficas con estos cálculos indicó que la relación era bastante aproximada y no hubo indicación que se diferenció significativamente de la razón 1:1. Durante un estudio aéreo en 1980, se compararon las estimaciones fotográficas de diferentes del cardúmenes, en los pases y los formatos de las cámaras y se encontró que eran bastante precisos, con una desviación normal de cerca del 60/0 de la magnitud cardumen. Se compararon también las estimaciones fotográficas con las estimaciones realizadas por los observadores aéreos. La mayoría de los observadores tienden a subestimar la magnitud de los cardúmenes, especialmente los cardúmenes grandes. La variabilidad entre los observadores fue elevada, lo que indica que se deben calibrar individualmente los datos de observadores. (PDF contains 39 pages.)