ArticleMolecular plant pathology2008
Extraction of high-quality bacterial RNA from infected leaf tissue for bacterial in planta gene expression analysis by multiplexed fluorescent Northern hybridization.
Article in Molecular plant pathology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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13 citing papers in PubMed, 27 citations in OpenAlex.
- Contribution of mechanosensitive channels to osmoadaptation and ectoine excretion in Halomonas elongata.Extremophiles : life under extreme conditions · 2020Article
- Biological relevance of volatile organic compounds emitted during the pathogenic interactions between apple plants and Erwinia amylovora.Molecular plant pathology · 2018Article
- Expression of extra-cellular levansucrase in Pseudomonas syringae is controlled by the in planta fitness-promoting metabolic repressor HexR.BMC microbiology · 2015Article
- Bacterial N2-fixation in mangrove ecosystems: insights from a diazotroph-mangrove interaction.Frontiers in microbiology · 2015Article
- AraC/XylS family stress response regulators Rob, SoxS, PliA, and OpiA in the fire blight pathogen Erwinia amylovora.Journal of bacteriology · 2014Article
- Characterization and regulation of the resistance-nodulation-cell division-type multidrug efflux pumps MdtABC and MdtUVW from the fire blight pathogen Erwinia amylovora.BMC microbiology · 2014Article
- The conserved upstream region of lscB/C determines expression of different levansucrase genes in plant pathogen Pseudomonas syringae.BMC microbiology · 2014Article
- Characterization of AcrD, a resistance-nodulation-cell division-type multidrug efflux pump from the fire blight pathogen Erwinia amylovora.BMC microbiology · 2014Article
- An optimized method for the extraction of bacterial mRNA from plant roots infected with Escherichia coli O157:H7.Frontiers in microbiology · 2014Article
- Article
- Comparative RNA-seq analysis of early-infected peach leaves by the invasive phytopathogen Xanthomonas arboricola pv. pruni.PloS one · 2013Article
- Coronatine Gene Expression In Vitro and In Planta, and Protein Accumulation During Temperature Downshift in Pseudomonas syringae.Sensors (Basel, Switzerland) · 2009Article
- Characterization of the RND-type multidrug efflux pump MexAB-OprM of the plant pathogen Pseudomonas syringae.Applied and environmental microbiology · 2008Article
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Authors and funding
3 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
Plant pathogenic bacteria possess a large number of genes that allow them to grow and cause disease on plants. In planta gene expression analysis is important to understand the impact of these genes on bacterial virulence. A new mRNA-based approach using multiplexed Northern hybridization was developed. High-quality bacterial and plant total RNA was successfully isolated from leaf tissue infiltrated with Pseudomonas syringae. The procedure employs a new extraction buffer formulation containing glycine, sodium dodecylsulphate, cetyltrimethylammonium bromide, high-molecular-weight polyethylene glycol and beta-mercaptoethanol. Cell lysis and classical acid-phenol extraction steps followed by LiCl precipitation yielded large amounts of total RNA of high purity and integrity. Multiplexing of DIG and chemically fluorescently labelled RNA probes was developed and expression data were normalized using the 23S rRNA gene as reference. The method was validated by studying in planta expression of the P. syringae genes mucD, cmaA, cfl, corR, corS and corP comprising a selection of highly expressed biosynthetic and low-expressed regulatory genes. The method was assessed regarding its sensitivity and might by useful for studying a variety of plant-microbe interactions.
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