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

Linden, Inge Van der; Cottyn, Bart; Uyttendaele, Mieke; Vlaemynck, Geertrui; Heyndrickx, Marc; Maes, Martine; Holden, Nicola

doi:10.3390/agriculture6010006

Cited by 15 publications

(16 citation statements)

References 67 publications

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“…Similarly, transient cold shock in romaine lettuce leaf lysate resulted in upregulation of stx1 but downregulation of eae [50] . However, eae was not differentially regulated during preharvest survival on butterhead lettuce [14] concurring with the results presented by Xicohtencatl-Cortez et al [51] who reported that this gene was not involved in attachment to fresh produce. Again, no direct comparison can take place with the results obtained in the present study due to different quantification strategies.…”

Section: Figure 1 Effect Of Substrate Type and Temperature On The Resupporting

confidence: 90%

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Transcription of Escherichia Coli O157:H7 Key Virulence Genes During Growth in Liquid Medium and Rocket (Eruca Sativa) at 4 and 10oC

Paramithiotis

Hadjilouka

Panagiotopoulou

et al. 2020

Applied Microbiology: Theory ＆ Technology

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The aim of the present study was to comparatively assess the transcription of key virulence genes, during growth of an Escherichia coli O157:H7 isolate in liquid medium and rocket (Eruca sativa) at different temperatures and time. For that purpose, rocket leaves and BHI broth were inoculated with 7.5 log CFU/g or CFU/mL, respectively, and incubated at 4 and 10oC. Sampling took place upon inoculation and after 0.5, 6 and 24 h of incubation; the RNA was stabilized and the transcription of stx1, stx2, eae, ehxA and etpD was assessed by RT-qPCR. Downregulation of stx1 and ehxA during growth on rocket surface at 4oC, upregulation of stx2 and eae during growth on rocket surface at both 4 and 10oC and the upregulation of etpD and ehxA during growth on rocket surface at 4 and 10oC, respectively, were observed. In addition, increase of incubation temperature, i.e. from 4 to 10oC resulted in upregulation of stx1 and downregulation of stx2 and eae. These results suggest that through differential regulation of genes encoding for virulence factors, the virulence potential of E. coli O157:H7 may be altered when it resides on the surface of rocket, at least for short time and at temperature relevant to its storage. However, further research is still necessary in order to improve our understanding of the physiology of this foodborne pathogen and the responses during exposure to food-related stresses.

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Section: Figure 1 Effect Of Substrate Type and Temperature On The Resupporting

confidence: 90%

“…Mahmoudzadeh et al [13] studied the transcription of stx1A and stx2A during growth on ground beef stored at 4 o C for up to 7 days. Finally, Van der Linden et al [14] used a microarray-based screening to assess the transcriptomic response during preharvest survival on butterhead lettuce.…”

Section: Introductionmentioning

confidence: 99%

Transcription of Escherichia Coli O157:H7 Key Virulence Genes During Growth in Liquid Medium and Rocket (Eruca Sativa) at 4 and 10oC

Paramithiotis

Hadjilouka

Panagiotopoulou

et al. 2020

Applied Microbiology: Theory ＆ Technology

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“…Plant cell recognition of Microbe-Associated Molecular Patterns (MAMPs) of human bacterial pathogens can trigger the production of Pattern-Triggered Immunity (PTI)-associated defense responses (Garcia et al, 2014), including a decrease in stomatal aperture width (Melotto et al, 2006;Roy et al, 2013). On the other hand, bacteria might counter-attack the plant responses by subverting the stomatal closure defense (Roy et al, 2013) or activating genes associated with oxidative stress tolerance and antimicrobial resistance (Van der Linden et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Human Pathogen Colonization of Lettuce Dependent Upon Plant Genotype and Defense Response Activation

Jacob

Melotto

2020

Front. Plant Sci.

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Fresh produce contaminated with human pathogens may result in foodborne disease outbreaks that cause a significant number of illnesses, hospitalizations, and death episodes affecting both public health and the agribusiness every year. The ability of these pathogens to survive throughout the food production chain is remarkable. Using a genetic approach, we observed that leaf colonization by Salmonella enterica serovar Typhimurium 14028s (S. Typhimurium 14028s) and Escherichia coli O157:H7 was significantly affected by genetic diversity of lettuce (Lactuca sativa L. and L. serriola L.). In particular, there was a significant variation among 11 lettuce genotypes in bacterial attachment, internalization, and apoplastic persistence after surface-and syringeinoculation methods. We observed a significant correlation of the bacterial leaf internalization rate with stomatal pore traits (width and area). Moreover, bacterial apoplastic populations significantly decreased in 9 out of 11 lettuce genotypes after 10 days of surface inoculation. However, after syringe infiltration, populations of E. coli O157: H7 and S. Typhimurium 14028s showed positive, neutral, or negative net growth in a 10day experimental period among seedlings of different lettuce types. The relative ability of the bacteria to persist in the apoplast of lettuce genotypes after syringe inoculation was minimally altered when assessed during a longer period (20 days) using 3.5-to 4-weekold plants. Interestingly, contrasting bacterial persistence in the lettuce genotypes Red Tide and Lollo Rossa was positively correlated with significant differences in the level of reactive oxygen species burst and callose deposition against S. Typhimurium 14028s and E. coli O157:H7 which are related to plant defense responses. Overall, we characterized the genetic diversity in the interaction between lettuce genotypes and enterobacteria S. Typhimurium 14028s and E. coli O157:H7 and discovered that this genetic diversity is linked to variations in plant immune responses towards these bacteria. These results provide opportunities to capitalize on plant genetics to reduce pathogen contamination of leaves.

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“…Spinach was selected as it is relevant to large-scale STEC outbreaks [18], and we have previously shown specific adherence to spinach roots [14, 15, 6]. High-throughput screening enables wholesale analysis and previous global transcriptomic analysis has shown induction of STEC fimbrial and afimbrial adhesins in lettuce leaf lysates [19–21]. In a similar manner, high-throughput negative- and positive-selection approaches have identified colonisation factors, e.g.…”

Section: Introductionmentioning

confidence: 99%

A high-throughput genomic screen identifies a role for the plasmid-borne Type II secretion system ofEscherichiacoli O157:H7 (Sakai) in plant-microbe interactions

Holmes

Pritchard

Hedley

et al. 2020

Preprint

Self Cite

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Food-borne illness arising from Shiga-toxigenic Escherichia coli (STEC) is often linked to consumption of fruit and vegetables as the bacteria have the ability to interact with plants and use them as alternative or secondary hosts. The initial stages of the interaction involve chemotaxis, attachment and potentially, responding to the early stages of microbe perception by the plant host. We used a high-throughput positive-selection approach to identify early interaction factors of E. coli O157:H7 isolate Sakai to spinach. A bacterial artificial chromosome (BAC) clone library was quantified by microarray hybridisation, and gene loci enrichment measured using a Bayesian hierarchical model. The screen of four successive rounds of short-term (2 hour) interaction with spinach roots produced in 115 CDS credible candidates, comprising seven contiguous genomic regions. Two candidate regions were selected for functional assessment: a chaperone-usher fimbrial gene cluster (loc6) and the pO157 plasmid-encoded type two secretion system (T2SS). Interaction of bacteria with spinach tissue was reduced in the absence of the pO157 plasmid, which was appeared to involve the T2SS EtpD secretin protein, whereas loss of loc6 did not impact interactions. The T2SS genes, etpD and etpC, were expressed at a plant-relevant temperature of 18 °C, and etpD expressed in planta by E. coli Sakai on spinach plants. Thus, a whole genome screening approach using a combination of computational modelling and functional assays has identified a novel function for STEC T2SS in interactions with plant tissue.

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

Cited by 15 publications

References 67 publications

Transcription of Escherichia Coli O157:H7 Key Virulence Genes During Growth in Liquid Medium and Rocket (Eruca Sativa) at 4 and 10oC

Transcription of Escherichia Coli O157:H7 Key Virulence Genes During Growth in Liquid Medium and Rocket (Eruca Sativa) at 4 and 10oC

Human Pathogen Colonization of Lettuce Dependent Upon Plant Genotype and Defense Response Activation

A high-throughput genomic screen identifies a role for the plasmid-borne Type II secretion system ofEscherichiacoli O157:H7 (Sakai) in plant-microbe interactions

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