Specific responses of Salmonella enterica to tomato varieties and fruit ripeness identified by in vivo expression technology.
Specific responses of Salmonella enterica to tomato varieties and fruit ripeness identified by in vivo expression technology.
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DOI：
10.1371/journal.pone.0012406
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发表时间：
2010-08-31
期刊：
影响因子：
3.7
通讯作者：
Teplitski M
中科院分区：
文献类型：
--
作者：
Noel JT;Arrach N;Alagely A;McClelland M;Teplitski M
最近爆发的与蔬菜有关的胃肠炎表明，肠道病原体在植物中定植、繁殖和持续很长一段时间，最终感染人类。肠道菌在植物中的遗传和生理途径仍然知之甚少。为了更好地了解肠道沙门氏菌sv.鼠伤寒沙门氏菌和番茄，在红色成熟果实内筛选GFP标记的沙门氏菌启动子文库。确定了51个独特的构建体，这些构建体在番茄中相对于体外生长可能受到差异调节。使用基于重组酶的体内表达技术（RIVET）在植物中测试这些启动子的子集的表达，并测试相应突变体的适合性。沙门氏菌的基因表达受果实成熟度和番茄品种的影响。一个假定的fadH启动子上调最强烈的未成熟的番茄。fadH构建体的表达依赖于亚油酸的存在，这与该化合物在成熟番茄果实中的积累减少一致。cysB构建体在cv的果实中被激活。夏威夷7997（抗青枯菌的一个种族）更强烈的比普遍敏感的番茄品种。邦妮·贝斯特已知的沙门氏菌运动性和动物毒力基因（hilA，flhDC，fliF和那些编码的peptide毒力质粒）没有显着贡献的健身番茄内的细菌，即使sirA和motA的删除适度增加健身番茄内的沙门氏菌。本研究揭示了沙门氏菌与植物宿主相互作用的遗传基础。沙门氏菌依赖于一组独特的代谢和调控基因，这些基因在植物中响应于宿主基因型和果实成熟度而受到差异调节。这种肠道病原体与植物病原体不同地定植在番茄的组织中，并且很少依赖于其动物毒力基因在果实中的持久性。
Recent outbreaks of vegetable-associated gastroenteritis suggest that enteric pathogens colonize, multiply and persist in plants for extended periods of time, eventually infecting people. Genetic and physiological pathways, by which enterics colonize plants, are still poorly understood. To better understand interactions between Salmonella enterica sv. Typhimurium and tomatoes, a gfp-tagged Salmonella promoter library was screened inside red ripe fruits. Fifty-one unique constructs that were potentially differentially regulated in tomato relative to in vitro growth were identified. The expression of a subset of these promoters was tested in planta using recombinase-based in vivo expression technology (RIVET) and fitness of the corresponding mutants was tested. Gene expression in Salmonella was affected by fruit maturity and tomato cultivar. A putative fadH promoter was upregulated most strongly in immature tomatoes. Expression of the fadH construct depended on the presence of linoleic acid, which is consistent with the reduced accumulation of this compound in mature tomato fruits. The cysB construct was activated in the fruit of cv. Hawaii 7997 (resistant to a race of Ralstonia solanacearum) more strongly than in the universally susceptible tomato cv. Bonny Best. Known Salmonella motility and animal virulence genes (hilA, flhDC, fliF and those encoded on the pSLT virulence plasmid) did not contribute significantly to fitness of the bacteria inside tomatoes, even though deletions of sirA and motA modestly increased fitness of Salmonella inside tomatoes. This study reveals the genetic basis of the interactions of Salmonella with plant hosts. Salmonella relies on a distinct set of metabolic and regulatory genes, which are differentially regulated in planta in response to host genotype and fruit maturity. This enteric pathogen colonizes tissues of tomatoes differently than plant pathogens, and relies little on its animal virulence genes for persistence within the fruit.
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