荧光示踪技术对土壤中诱变菌的生长研究

发布时间：2018-04-12 00:08

本文选题：荧光标记 + 荧光计数　；参考：《中国环境科学》2017年01期

【摘要】：采用荧光染料DAPI(4',6-Diamidino-2-Phenylindole)对外源微生物Bacillus subtilis 38(B38)的DNA分子进行标记,利用荧光示踪技术对外源微生物的生长繁殖进行有效监测.通过筛选DAPI工作液的使用浓度与菌悬液的稀释比例,得出采用0.1μg/mL浓度的DAPI对稀释10~6倍的菌悬液进行染色,荧光信号强度适宜,菌体分散度良好,可实现准确计数.采用上述染色条件,对60d内土壤B38菌体数量的变化进行阶段性监测.结果发现,接种后菌体浓度显著增加,平衡后菌体浓度达到(1.05±0.01)×10~(10)cells/mL,与初期施加量相比,增长量达到3个数量级.以上结果表明,采用DAPI染料对外源微生物进行荧光示踪计数,为监测外源微生物的生长繁殖状态提供了一种简便、有效的方法.
[Abstract]:The DNA molecule of exogenous microorganism Bacillus subtilis 38 (B38) was labeled with the fluorescent dye DAPI4 (6-Diamidino-2-Phenylindole), and the growth and reproduction of exogenous microorganism was effectively monitored by fluorescence tracer technique.By screening the dilution ratio between the concentration of DAPI working solution and the suspension of bacteria, it is concluded that the DAPI of 0.1 渭 g/mL concentration can be used to dye the bacterial suspension of 10 ~ 6 times dilution, the fluorescence signal intensity is suitable, the bacterial dispersion is good, and the accurate count can be realized.The changes of the number of B38 bacteria in soil within 60 days were monitored by using the above dyeing conditions.The results showed that the bacterial concentration increased significantly after inoculation and reached 1.05 卤0.01 脳 10 ~ (10) cells / mL after inoculation, which was three orders of magnitude higher than that of initial application.The results show that the fluorescent tracer counting of exogenous microorganisms with DAPI dyes provides a simple and effective method for monitoring the growth and reproduction of exogenous microbes.
【作者单位】：南开大学环境科学与工程学院;
【基金】：天津市应用基础与前沿技术研究计划(青年项目)(15JCQNJC15200) 国家科技支撑计划项目(2014BAC23B0205)
【分类号】：X53;X172

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