DRASTIC--INSIGHTS: querying information in a plant gene expression database.

DRASTIC--Database Resource for the Analysis of Signal Transduction In Cells (http://www.drastic.org.uk/) has been created as a first step towards a data-based approach for constructing signal transduction pathways. DRASTIC is a relational database of plant expressed sequence tags and genes up- or down-regulated in response to various pathogens, chemical exposure or other treatments such as drought, salt and low temperature. More than 17700 records have been obtained from 306 treatments affecting 73 plant species from 512 peer-reviewed publications with most emphasis being placed on data from Arabidopsis thaliana. DRASTIC has been developed by the Scottish Crop Research Institute and the University of Abertay Dundee and allows rapid identification of plant genes that are up- or down-regulated by multiple treatments and those that are regulated by a very limited (or perhaps a single) treatment. The INSIGHTS (INference of cell SIGnaling HypoTheseS) suite of web-based tools allows intelligent data mining and extraction of information from the DRASTIC database. Potential response pathways can be visualized and comparisons made between gene expression patterns in response to various treatments. The knowledge gained informs plant signalling pathways and systems biology investigations.

Characterisation of early transcriptional changes involving multiple signalling pathways in the Mla13 barley interaction with powdery mildew ( Blumeria graminis f. sp. hordei).

Suppression subtractive hybridisation was used to isolate 21 cDNAs ( bmi1- bmi21) up-regulated 1-5 h post-inoculation (hpi) in a barley ( Hordeum vulgare L. cv. Pallas) near-isogenic line (NIL) P11 ( Mla13) challenged with either avirulent or virulent isolates of Blumeria graminis f. sp. hordei. Transcriptional changes at these time-points are crucial for the Mla-mediated hypersensitive response [W.R. Bushnell and Z. Liu (1994) Physiol Mol Plant Pathol 44:389-402]. Seven sequences were up-regulated by 1 hpi, when the pathogen has formed only the primary germ tube. Some transcripts were similar to genes with a role in regulating programmed cell death in animals, including NF kappaB and oxysterol-binding protein. Moreover, bmi7, similar to rice resistance gene Xa21, was rapidly up-regulated in both compatible and incompatible interactions, but was then down-regulated by 5 hpi in the virulent interaction. Only nine of the transcripts were up-regulated in mlo5 resistance in cv. Pallas NIL P22, confirming differential pathway induction between Mla13 and mlo5. However, eight sequences up-regulated in the Mla13 response in P11 were already highly elevated in uninoculated mlo5 mutant P22, suggesting that they may be negatively regulated by wild-type Mlo. Regulation of bmi sequences was investigated using salicylic acid, methyl jasmonate, ethylene, H(2)O(2), abscisic acid, wounding and a glucan elicitor. No single stimulus up-regulated all genes, suggesting either combinations of these stimuli, or additional stimuli, are involved in early Mla13 and mlo5 resistances. Whereas H(2)O(2) up- or down-regulated 17 of the transcripts detected in Northern analyses, salicylic acid stimulated only down-regulation of 5 transcripts.

The need for a standard nomenclature for gene classification (a Nucleotide Function Code) and an automated data-based tool to assist in understanding the molecular associations in cell signalling in plant-pathogen interactions.

summary Despite the adoption of Arabidopsis thaliana as a model plant system and the plethora of molecular information being obtained from its use, it is disappointing that the scientific community has not devised a cell signalling model integrating and visualizing these data. Lack of common systems of nomenclature and the sheer size and complexity of the task inhibit any individual from bringing together the knowledge into a unified structure. There are clearly many aspects of cell biology that are similar, even between plants and animals, that could facilitate development of a generic model. A gene-coding or nucleotide classification system which is 'user-friendly' would be beneficial to building such a model and enable rapid identification of orthologues of genes from different organisms. Whilst some international projects seek to address the problem of assigning unique numbers to genes, none suggest a nucleotide classification system that provides biological information that is transparent within the code. This paper discusses these issues and identifies the need for a more formal, semi-automated approach to modelling signal transduction utilizing the strengths of the proposed classification approach. By way of illustration, an example of a possible nucleotide function code is suggested, to demonstrate more clearly the benefits of such a system. Further discussion of this topic will be encouraged on websites ( and ).

cDNA-AFLP analysis of differential gene expression in the prokaryotic plant pathogen Erwinia carotovora.

For studies of differential gene expression in prokaryotes, methods for synthesizing representative cDNA populations are required. Here, a technique is described for the synthesis of cDNA from the potato pathogens Erwinia carotovora subsp. atroseptica (Eca) and Erwinia carotovora subsp. carotovora (Ecc) using a combination of short oligonucleotide (11-mer) primers that were known to anneal to conserved sequences in the 3' regions of enterobacterial genes. Specific PCR amplifications with primers designed to anneal to 14 known genes from either Eca or Ecc revealed the presence of the corresponding transcripts in cDNA, suggesting that the cDNA represented a broad genomic coverage. cDNA-amplified fragment length polymorphism (cDNA-AFLP) was used to identify differentially expressed genes in Eca, including one that shows significant similarity, at the protein level, to an avirulence gene from Xanthomonas campestris pv. raphani. Northern analysis was used to confirm that differentially amplified cDNA fragments were derived from differentially expressed genes. This is the first report of the use of cDNA-AFLP to study differential gene expression in prokaryotes.