Transcriptional and functional responses of Escherichia coli O157:H7 growing in the lettuce rhizoplane

Hou, Zhe; Fink, Ryan C.; Sugawara, Masayuki; Diez-Gonzalez, Francisco; Sadowsky, Michael J.

doi:10.1016/j.fm.2013.03.002

Cited by 20 publications

(27 citation statements)

References 32 publications

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“…Similar to other stress-related genes, upregulation of this gene confers resistance to these stresses [48]. Upregulation was also observed by Kyle et al (2010) [4] and Fink et al (2012) [4] on lettuce leaves and Hou et al (2013) [6] on lettuce roots. The last two authors also showed that a deletion mutant showed reduced attachment to both lettuce roots and lettuce leaves.…”

Section: Flagella Motility Fimbrial Expression Biofilmmentioning

confidence: 65%

“…Fink et al (2012) [4] however, found no significant regulation of the generic E. coli and E. coli O157:H7 csgAB genes on lettuce roots, although the curli regulator crl was induced in generic E. coli. However, they could demonstrate that the deletion of the csgA genes (but not of the csgB gene), resulting in a reduced capacity of generic E. coli to attach to roots [6]. Curli-related genes were also expressed during the exponential growth of E. coli O157 EDL933 in radish seedlings [7].…”

Section: Flagella Motility Fimbrial Expression Biofilmmentioning

confidence: 99%

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Microarray-Based Screening of Differentially Expressed Genes of E. coli O157:H7 Sakai during Preharvest Survival on Butterhead Lettuce

Linden

Cottyn²,

Uyttendaele

et al. 2016

Agriculture

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Numerous outbreaks of Escherichia coli O157:H7 have been linked to the consumption of leafy vegetables. However, up to the present, little has been known about E. coli O157:H7's adaptive responses to survival on actively growing (and thus responsive) plants. In this study, whole genome transcriptional profiles were generated from E. coli O157:H7 cells (isolate Sakai, stx-) one hour and two days after inoculation on the leaves of growing butterhead lettuce, and compared with an inoculum control. A total of 273 genes of E. coli O157:H7 Sakai (5.04% of the whole genome) were significantly induced or repressed by at least two-fold (p < 0.01) in at least one of the analyzed time points in comparison with the control. Several E. coli O157:H7 genes associated with oxidative stress and antimicrobial resistance were upregulated, including the iron-sulfur cluster and the multiple antibiotic resistance (mar) operon, whereas the Shiga toxin virulence genes were downregulated. Nearly 40% of the genes with significantly different expression were poorly characterized genes or genes with unknown functions. These genes are of special interest for future research as they may play an important role in the pathogens' adaptation to a lifestyle on plants. In conclusion, these findings suggest that the pathogen actively interacts with the plant environment by adapting its metabolism and responding to oxidative stress.

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Section: Flagella Motility Fimbrial Expression Biofilmmentioning

confidence: 65%

Section: Flagella Motility Fimbrial Expression Biofilmmentioning

confidence: 99%

Microarray-Based Screening of Differentially Expressed Genes of E. coli O157:H7 Sakai during Preharvest Survival on Butterhead Lettuce

Linden

Cottyn²,

Uyttendaele

et al. 2016

Agriculture

View full text Add to dashboard Cite

show abstract

“…In contrast, investigation of bacteria gene expression on living plant roots presents challenges in obtaining similar weights of material. A hydroponic system for high-throughput propagation of lettuce plants was described recently (Hou et al, 2012, 2013), where sterilized seeds were germinated in 96-well pipette tip boxes. Whole transcriptome analysis of E. coli was examined three days after inoculation with the plant roots, although the report does not state the weight of material required for RNA extraction.…”

Section: Discussionmentioning

confidence: 99%

“…However, roots are known to support relatively high densities of bacteria, providing a more favorable habitat than the phyllosphere. In general, published reports describe that a high number of plant roots is required to retrieve sufficient bacterial RNA (Matilla et al, 2007; Hou et al, 2012, 2013; Zyśko et al, 2012). Further to this, the application of techniques described for RNA purification from infected leaves cannot always be successfully applied to roots.…”

Section: Introductionmentioning

confidence: 99%

An optimized method for the extraction of bacterial mRNA from plant roots infected with Escherichia coli O157:H7

Holmes¹,

Birse²,

Jackson³

et al. 2014

Front. Microbiol.

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Analysis of microbial gene expression during host colonization provides valuable information on the nature of interaction, beneficial or pathogenic, and the adaptive processes involved. Isolation of bacterial mRNA for in planta analysis can be challenging where host nucleic acid may dominate the preparation, or inhibitory compounds affect downstream analysis, e.g., quantitative reverse transcriptase PCR (qPCR), microarray, or RNA-seq. The goal of this work was to optimize the isolation of bacterial mRNA of food-borne pathogens from living plants. Reported methods for recovery of phytopathogen-infected plant material, using hot phenol extraction and high concentration of bacterial inoculation or large amounts of infected tissues, were found to be inappropriate for plant roots inoculated with Escherichia coli O157:H7. The bacterial RNA yields were too low and increased plant material resulted in a dominance of plant RNA in the sample. To improve the yield of bacterial RNA and reduce the number of plants required, an optimized method was developed which combines bead beating with directed bacterial lysis using SDS and lysozyme. Inhibitory plant compounds, such as phenolics and polysaccharides, were counteracted with the addition of high-molecular-weight polyethylene glycol and hexadecyltrimethyl ammonium bromide. The new method increased the total yield of bacterial mRNA substantially and allowed assessment of gene expression by qPCR. This method can be applied to other bacterial species associated with plant roots, and also in the wider context of food safety.

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“…It is well established that bacterial cells that are previously exposed to chlorine stress (cells adapted to chlorine treatment) acquire increased resistance to other stresses such as high temperature and oxidative stresses (Vidovic & Korber, 2014). Additionally, YcfR is known to be responsive to chlorine stress and plays a role in the attachment and colonization of E. coli O157:H7 in lettuce rhizoplane (Hou, Fink, Sugawara, Diez-Gonzalez, & Sadowsky, 2013). Another study also showed D ycfR mutant was unable to attach or colonize lettuce leaves (Fink et al, 2012).…”

Section: Growth Of Chlorine-stressed Stecmentioning

confidence: 99%

Growth characteristics of Shiga toxin-producing Escherichia coli (STEC) stressed by chlorine, sodium chloride, acid, and starvation on lettuce and cantaloupe

et al. 2015

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Transcriptional and functional responses of Escherichia coli O157:H7 growing in the lettuce rhizoplane

Cited by 20 publications

References 32 publications

Microarray-Based Screening of Differentially Expressed Genes of E. coli O157:H7 Sakai during Preharvest Survival on Butterhead Lettuce

Microarray-Based Screening of Differentially Expressed Genes of E. coli O157:H7 Sakai during Preharvest Survival on Butterhead Lettuce

An optimized method for the extraction of bacterial mRNA from plant roots infected with Escherichia coli O157:H7

Growth characteristics of Shiga toxin-producing Escherichia coli (STEC) stressed by chlorine, sodium chloride, acid, and starvation on lettuce and cantaloupe

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