Transcriptome Analysis of Escherichia coli O157:H7 Exposed to Lysates of Lettuce Leaves

被引:90
作者
Kyle, Jennifer L. [1 ]
Parker, Craig T. [1 ]
Goudeau, Danielle [1 ]
Brandl, Maria T. [1 ]
机构
[1] USDA, ARS, Western Reg Res Ctr, Produce Safety & Microbiol Res Unit, Albany, CA 94710 USA
关键词
VIRULENCE GENE-EXPRESSION; NITRIC-OXIDE; POLYNUCLEOTIDE PHOSPHORYLASE; ANTIBIOTIC-RESISTANCE; HYDROGEN-PEROXIDE; CHROMOSOMAL GENES; OXIDATIVE STRESS; O157-H7; SECRETION; SUPEROXIDE;
D O I
10.1128/AEM.02461-09
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Harvesting and processing of leafy greens inherently cause plant tissue damage, creating niches on leaves that human pathogens can exploit. We previously demonstrated that Escherichia coli O157:H7 (EcO157) multiplies more rapidly on shredded leaves than on intact leaves (M. T. Brandl, Appl. Environ. Microbiol. 74:5285-5289, 2008). To investigate how EcO157 cells adapt to physicochemical conditions in injured lettuce tissue, we used microarray-based whole-genome transcriptional profiling to characterize gene expression patterns in EcO157 after 15- and 30-min exposures to romaine lettuce lysates. Multiple carbohydrate transport systems that have a role in the utilization of substrates known to be prevalent in plant cells were activated in EcO157. This indicates the availability to the human pathogen of a variety of carbohydrates released from injured plant cells that may promote its extensive growth in leaf lysates and, thus, in wounded leaf tissue. In addition, microarray analysis revealed the upregulation of numerous genes associated with EcO157 attachment and virulence, with oxidative stress and antimicrobial resistance (including the OxyR and Mar regulons), with detoxification of noxious compounds, and with DNA repair. Upregulation of oxidative stress and antimicrobial resistance genes in EcO157 was confirmed on shredded lettuce by quantitative reverse transcription-PCR. We further demonstrate that this adaptation to stress conditions imparts the pathogen with increased resistance to hydrogen peroxide and calcium hypochlorite. This enhanced resistance to chlorinated sanitizers combined with increased expression of virulence determinants and multiplication at sites of injury on the leaves may help explain the association of processed leafy greens with outbreaks of EcO157.
引用
收藏
页码:1375 / 1387
页数:13
相关论文
共 76 条
[41]   Transcriptional adaptation of Shigella flexneri during infection of macrophages and epithelial cells:: Insights into the strategies of a cytosolic bacterial pathogen [J].
Lucchini, S ;
Liu, H ;
Jin, Q ;
Hinton, JCD ;
Yu, J .
INFECTION AND IMMUNITY, 2005, 73 (01) :88-102
[42]   Effects of feeding Spodoptera littoralis on lima bean leaves.: III.: Membrane depolarization and involvement of hydrogen peroxide [J].
Maffei, ME ;
Mithöfer, A ;
Arimura, GI ;
Uchtenhagen, H ;
Bossi, S ;
Bertea, CM ;
Cucuzza, LS ;
Novero, M ;
Volpe, V ;
Quadro, S ;
Boland, W .
PLANT PHYSIOLOGY, 2006, 140 (03) :1022-1035
[43]   Distribution of the secondary type III secretion system locus found in enterohemorrhagic Escherichia coli O157:H7 isolates among Shiga toxin-producing E-coli strains [J].
Makino, SI ;
Tobe, T ;
Asakura, H ;
Watarai, M ;
Ikeda, T ;
Takeshi, K ;
Sasakawa, C .
JOURNAL OF CLINICAL MICROBIOLOGY, 2003, 41 (06) :2341-2347
[44]  
Mandrell R. E., 2009, Microbial safety of fresh produce, P5
[45]   A highly sensitive fluorescent micro-assay of H2O2 release from activated human leukocytes using a dihydroxyphenoxazine derivative [J].
Mohanty, JG ;
Jaffe, JS ;
Schulman, ES ;
Raible, DG .
JOURNAL OF IMMUNOLOGICAL METHODS, 1997, 202 (02) :133-141
[46]   Expression analysis of the nrdHIEF operon from Escherichia coli -: Conditions that trigger the transcript level in vivo [J].
Monje-Casas, F ;
Jurado, J ;
Prieto-Alamo, MJ ;
Holmgren, A ;
Pueyo, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (21) :18031-18037
[47]   Exploring the antimicrobial action of a carbon monoxide-releasing compound through whole-genome transcription profiling of Escherichia coli [J].
Nobre, Ligia S. ;
Al-Shahrour, Fatima ;
Dopazo, Joaquin ;
Saraiva, Ligia M. .
MICROBIOLOGY-SGM, 2009, 155 :813-824
[48]   Ribonucleotide reductases [J].
Nordlund, Par ;
Reichard, Peter .
ANNUAL REVIEW OF BIOCHEMISTRY, 2006, 75 :681-706
[49]   Hydrogen peroxide is generated systemically in plant leaves by wounding and systemin via the octadecanoid pathway [J].
Orozco-Cardenas, M ;
Ryan, CA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (11) :6553-6557
[50]   Identification and characterization of a novel ABC iron transport system, fit, in Escherichia coli [J].
Ouyang, Zhiming ;
Isaacson, Richard .
INFECTION AND IMMUNITY, 2006, 74 (12) :6949-6956