摘要
本研究选用前期从受污染土壤中筛选出来的多环芳烃高效降解菌BDG-3为研究对象,以^(13)C^-葡萄糖为底物对其进行培养,并利用Nano SIMS亚细胞成像技术对稳定同位素13 C标记的化合物在菌体内的代谢活性进行分析.将降解菌BDG-3养在含有13 C标记的葡萄糖培养基中,对选中的研究区域内质量数为12、13、16、26的信号进行收集,分别用于表征^(12)C^-、^(13)C^-、^(16)O^-和^(26)(^(12)C1^(14)N)^-离子的信号及分布特征.利用Image J软件处理并分析数据.单菌体切面内各元素的分布有一定规律,存在明显的峰值点.以选定的线域为ROI研究对象,^(13)C^-、^(16)O^-和^(26)(^(12)C^(14)N)^-离子变化趋势接近,在横坐标为10附近达到峰值,含量最高的是^(26)(^(12)C^(14)N)^-.以单一菌体为ROI研究对象,以13 C^-葡萄糖为底物生长出的BDG^-3内单菌体内部^(13)C含量平均值(54.38)远高于以^(12)C葡萄糖为底物的单菌内部^(13)C的信号(5.59).研究结果表明菌体可以吸收13 C^-葡萄糖,用于自身的生长,代谢活性良好.通过对该种方法学的探索,为进一步研究更为复杂的环境污染物质在微生物体内的降解代谢过程提供了一种新的可靠的分析手段.
This work aims to quantitate ion distribution in a single cell and to identify physiological and biochemical functions of carbon compounds in single cells.Nano SIMS was used for subcellar imaging of isotopically labeled carbon compounds.Bacteria BDG-3with PAH degrading ability was isolated from crude oil contaminated soil.Experiments were designed to detect and image signal of different mass and to represent distribution of ^12C、^13C、^16O和^26（^12C^14N）.Software Image J was used to reveal element distribution on a subcellular scale.Quantitative analysis of ion distribution was done in single cells.Line ROI scan revealed that quantity of ^13C、^16O^-and ^26（^12C^14N）changed similarly,the peak value was near 10 on the abscissa.The number of ^26（^12C^14N） was the highest.With single cell as ROI,average ^13C in a single cell BDG-3grown with 13 Cglucose was 54.38 much higher than BDG-3on 12 C glucose.This provides a reliable approach to examine metabolic activity of identified single cells.BDG-3seems to have the ability to absorb ^13C-glucose This affords a new and reliable way to analyze metabolism process when studying degradation of more complex pollutants.
出处
《北京师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2016年第2期223-227,共5页
Journal of Beijing Normal University(Natural Science)
基金
国家自然科学基金资助项目(41372232)
