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首页> 外文期刊>Environmental Biosafety Research >Effects of temperature on detection of plasmid or chromosomally encoded gfp- and Iux-labeled Pseudomonas fluorescens in soil
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Effects of temperature on detection of plasmid or chromosomally encoded gfp- and Iux-labeled Pseudomonas fluorescens in soil

机译：温度对土壤中质粒或染色体编码的gfp和Iux标记的荧光假单胞菌检测的影响

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Pseudomonas fluorescens is a normal inhabitant of the soil rhizosphere. The use of genetically altered strains of P. fluorescens in bioremediation has led to the need for effective monitoring of such cells released into the environment. In this study,we present data on the persistence in soil of P. fluorescens harboring gfp (green fluorescent protein) or lux (bioluminescence) genes. Comparisons were made between strains marked chromosomally and strains carrying these markers on a plasmid. Overall effects of plasmid carriage on culturability were also examined. Sterile soil microcosms were inoculated with washed cells to a final concentration of ca. 10~6 CFU.g~(-1) and placed at 5, 23, and 35-37°C. Samples were taken periodically and examined for culturability and viability, using the substrate responsiveness assay. Our results indicated no significant loss of culturability at 5 and 23°C for a period of over one year. In contrast, cells of P. fluorescens incubated at 35-37°C entered the viable but nonculturable state within 7 days. All cells labeled with gfp retained fluorescence regardless of culturability, suggesting that the green fluorescent protein can be of value in monitoring the presence of cells following their release to the environment. Because fluorescence was maintained regardless of the cells' physiological state, this protein may also be an indicator of cell viability.
机译：荧光假单胞菌是土壤根际的正常居民。在生物修复中使用遗传改变的荧光假单胞菌菌株导致需要有效监测释放到环境中的此类细胞。在这项研究中，我们提出了带有gfp（绿色荧光蛋白）或lux（生物发光）基因的荧光假单胞菌在土壤中的持久性数据。在染色体上标记的菌株与在质粒上带有这些标记的菌株之间进行比较。还检查了质粒运输对可培养性的总体影响。用洗涤过的细胞接种无菌土壤微观世界，使其最终浓度约为。 10〜6 CFU.g〜（-1）并置于5、23和35-37°C下。定期采集样品，并使用底物反应性测定法检查其可培养性和生存力。我们的结果表明，在超过5年和23°C的条件下，一年以上的可培养性没有明显降低。相反，在35-37℃下孵育的荧光假单胞菌的细胞在7天内进入存活但不可培养的状态。不论可培养性如何，所有标记有gfp的细胞均保留荧光，这表明绿色荧光蛋白在监测释放到环境中的细胞的存在后可能具有重要的价值。由于无论细胞的生理状态如何都可以维持荧光，因此该蛋白也可能是细胞活力的指标。

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