Differences in internalization and growth of <i><scp>E</scp>scherichia coli</i> O157:H7 within the apoplast of edible plants, spinach and lettuce, compared with the model species <i><scp>N</scp>icotiana benthamiana</i>

Wright, Kathryn M.; Crozier, Louise; Marshall, J.; Merget, Bernhard; Holmes, Ashleigh; Holden, Nicola

doi:10.1111/1751-7915.12596

Cited by 56 publications

(60 citation statements)

References 74 publications

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“…Seeds were stratified for 48h at 4°C and germinated under a 16/8 h light/dark cycle at 20°C in MLR-350 growth chamber (Panasonic). Nicotiana benthamiana plants expressing mOrange-LTI6b fluorescent protein as a plasma membrane marker (mOrg-LTI; [32]) were used for confocal imaging. Transgenic and wild-type Nicotiana benthamiana plants were grown in commercial compost and maintained under glasshouse conditions of 16/8 h light/ dark cycle, daytime temperature 26°C maximum, nighttime 22°C for 4-5 weeks before use.…”

Section: Plant Lines and Growth Conditionsmentioning

confidence: 99%

“…Leaves of 4-5 week old wild type and mOrg-LTI [32] expressing Nicotiana benthamiana plants were syringe infiltrated with Agrobacterium tumefaciens GV3101 pMP90 containing expression plasmids for each receptor-like kinase alongside p19, each at an OD600 of 0.1. Plants were maintained under standard growth conditions for 48-60h before use in experiments.…”

Section: Transient Expression In N Benthamianamentioning

confidence: 99%

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Juxta-membrane S-acylation of plant receptor-like kinases – fortuitous or functional?

Wright

Turnbull

et al. 2019

Preprint

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S-acylation is a common post-translational modification of membrane protein cysteine residues with many regulatory roles. S-acylation adjacent to transmembrane domains has been described in the literature as affecting diverse protein properties including turnover, trafficking and microdomain partitioning. However, all of these data are derived from mammalian and yeast systems. Here we examine the role of S-acylation adjacent to the transmembrane domain of the plant pathogen perceiving receptor-like kinase FLS2. Surprisingly, S-acylation of FLS2 adjacent to the transmembrane domain is not required for either FLS2 trafficking or signalling function. Expanding this analysis to the wider plant receptor-like kinase superfamily we find that S-acylation adjacent to receptor-like kinase domains is common but poorly conserved between orthologues through evolution. This suggests that S-acylation of receptor-like kinases at this site is likely the result of chance mutation leading to cysteine occurrence. As transmembrane domains followed by cysteine residues are common motifs for Sacylation to occur, and many S-acyl transferases appear to have lax substrate specificity, we propose that many receptor-like kinases are fortuitously S-acylated once chance mutation has introduced a cysteine at this site. Interestingly some receptor-like kinases show conservation of S-acylation sites between orthologues suggesting that S-acylation has come to play a role and has been positively selected for during evolution. The most notable example of this is in the ERECTA-like family where Sacylation of ERECTA adjacent to the transmembrane domain occurs in all ERECTA orthologues but not in the parental ERECTA-like clade. This suggests that ERECTA S-acylation occurred when ERECTA emerged during the evolution of angiosperms and may have contributed to the neo-functionalisation of ERECTA from ERECTA-like proteins.

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Section: Plant Lines and Growth Conditionsmentioning

confidence: 99%

Section: Transient Expression In N Benthamianamentioning

confidence: 99%

Juxta-membrane S-acylation of plant receptor-like kinases – fortuitous or functional?

Wright

Turnbull

et al. 2019

Preprint

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“…More recently various plant foods such as spinach, tomato, lettuce and fresh fruits have been identified as sources (Grant et al 2008;Herman et al 2015;Denis et al 2016). Initially these foods were thought to be fecally contaminated, but recent reports suggest growth of E. coli O157:H7 (Brandl 2008;Wright et al 2013;Wright et al 2017) in tissues of salad leaves and tomatoes. Upon inoculation from an unknown source, the enteric bacteria multiply inside the growing plant, and cannot be removed through surface treatment such as washing.…”

Section: Introductionmentioning

confidence: 99%

Growth and extended survival ofEscherichia coliO157:H7 in soil organic matter

NandaKafle

Christie

Vilain

et al. 2017

Preprint

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Enterohaemorrhagic Escherichia coli such as serotype O157:H7 are a leading cause of food-associated outbreaks. While the primary reservoir is associated with cattle, plant foods have been associated as sources of human infection. E. coli is able to grow in the tissue of food plants such as spinach. While fecal contamination is the primary suspect, soil has been underestimated as a potential reservoir. Persistence of bacterial populations in open systems is the product of growth, death, predation, and competition. Here we report that E. coli O157:H7 can grow using the soluble compounds in soil, and characterize the effect of soil growth in the stationary phase proteome.

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“…Colonisation of the E. coli O157:H7 isolate (ZAP1589) was quantified on spinach and lettuce, and for both isolates (ZAP1589 and Sakai) on sprouted seeds. Our previous in planta data for lettuce and spinach plants showed that the highest levels of E. coli isolate Sakai occurred on spinach roots (73). Inoculation of spinach and lettuce with the high dose (10 7 cfu ml −1 ) of E. coli isolate ZAP1589 also resulted in higher levels of bacteria on the roots compared to leaves, with similar levels on spinach and lettuce roots, e.g.…”

Section: Resultsmentioning

confidence: 94%

“…The apoplast contains metabolites, such as solutes, sugars, proteins and cell wall components (54) and as such provides a rich environment for many bacterial species, both commensal bacteria and human pathogens (20, 28). The rate of STEC internalisation is dependent on multiple factors including the plant species and tissue (73) and how plants are propagated (17-19). Specificity in the response of STEC to different plant species and tissue types has been demonstrated at the transcriptional level (9, 38).…”

Section: Introductionmentioning

confidence: 99%

Relating growth potential and biofilm formation of ShigatoxigenicEscherichia colitoin plantacolonisation and the metabolome of ready-to-eat crops

Merget

Forbes

Brennan

et al. 2019

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/ 250 words)1 Contamination of fresh produce with pathogenic Escherichia coli, including Shigatoxigenic E. 2 coli (STEC), represents a serious risk to human health. Colonisation is governed by multiple 3 bacterial and plant factors that can impact on the probability and suitability of bacterial growth. 4 Thus, we aimed to determine whether the growth potential of STEC for plants associated with 5 foodborne outbreaks (two leafy vegetables and two sprouted seed species), is predictive for 6 colonisation of living plants as assessed from growth kinetics and biofilm formation in plant 7 extracts. Fitness of STEC was compared to environmental E. coli, at temperatures relevant to 8 plant growth. Growth kinetics in plant extracts varied in a plant-dependent and isolate-9 dependent manner for all isolates, with spinach leaf lysates supporting the fastest rates of 10 growth. Spinach extracts also supported the highest levels of biofilm formation. Saccharides 11 were identified as the major driver of bacterial growth, although no single metabolite could be 12 correlated with growth kinetics. The highest level of in planta colonisation occurred on alfalfa 13 sprouts, though internalisation was 10-times more prevalent in the leafy vegetables than in 14 sprouted seeds. Marked differences in in planta growth meant that growth potential could only 15 be inferred for STEC for sprouted seeds. In contrast, biofilm formation in extracts related to 16 spinach colonisation. Overall, the capacity of E. coli to colonise, grow and internalise within 17 plants or plant-derived matrices were influenced by the isolate type, plant species, plant tissue 18 type and temperature, complicating any straight-forward relationship between in vitro and in 19 planta behaviours. 20 3 Importance (149 / 150 word) 21 Fresh produce is an important vehicle for STEC transmission and experimental evidence 22shows that STEC can colonise plants as secondary hosts, but differences in the capacity to 23 colonise occur between different plant species and tissues. Therefore, an understanding of the 24 impact of these plant factors have on the ability of STEC to grow and establish is required for 25 food safety considerations and risk assessment. Here, we determined whether growth and the 26 ability of STEC to form biofilms in plants extracts could be related to specific plant metabolites 27 or could predict the ability of the bacteria to colonise living plants. Growth rates for sprouted 28 seeds (alfalfa and fenugreek) exhibited a positive relationship between plant extracts and living 29 plants, but not for leafy vegetables (lettuce and spinach). Therefore, the detailed variations at 30 the level of the bacterial isolate, plant species and tissue type all need to be considered in risk 31 assessment. 32 33 34Contamination of fresh produce from Shigatoxigenic Escherichia coli (STEC) presents a 35 serious hazard as a cause of food-borne illnesses, diarrhoea and enterohemorrhagic disease. 36Fresh produce is a major vehicle of transmission of STEC, with foods of...

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Differences in internalization and growth of Escherichia coli O157:H7 within the apoplast of edible plants, spinach and lettuce, compared with the model species Nicotiana benthamiana

Cited by 56 publications

References 74 publications

Juxta-membrane S-acylation of plant receptor-like kinases – fortuitous or functional?

Juxta-membrane S-acylation of plant receptor-like kinases – fortuitous or functional?

Growth and extended survival ofEscherichia coliO157:H7 in soil organic matter

Relating growth potential and biofilm formation of ShigatoxigenicEscherichia colitoin plantacolonisation and the metabolome of ready-to-eat crops

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