绿色荧光蛋白标记检测枯草芽孢杆菌在水体中的存活动态  

Survival Dynamics of Bacillus Subtilis in Water by Green Fluorescent Protein Labeling

作　　者：尹敬芳[1] 周向阳[2] 李健强[1] 李永红[2] 侯红利[2] 张文华[1] 

机构地区：[1]中国农业大学农学与生物技术学院植物病理学系,北京100094 [2]深圳市农作物良种引进中心,深圳518040

出　　处：《分析化学》2007年第10期1405-1409,共5页Chinese Journal of Analytical Chemistry

基　　金：国家攻关项目(No.2004BA521B04);深圳市科技与信息局计划项目(No.S040161);教育部"新世纪优秀人才支持计划"(No.NCET-04-0128)资助

摘　　要：基于生物-化学协同控制植物病害的原理,构建了一种由枯草芽孢杆菌和烯酰吗啉组成的对辣椒等作物疫病具有较好的防治效果的菌药合剂(DMBS)。按照农药降解研究基本规则,采用绿色荧光蛋白(greenfluorescent protein,GFP)标记技术和细菌学研究方法研究了DMBS在去离子水、地下水、自来水、河水和雨水中的降解动态。结果表明,GFP标记可以用于枯草芽孢杆菌在5种水环境中的存活检测。在(25±1)℃条件下,枯草芽孢杆菌菌剂和DMBS中的枯草芽孢杆菌数量主要表现为前12 d迅速下降,此后则随时间的延长在一定的范围内呈变动的上升或下降趋势。在(50±1)℃灭菌和不灭菌的条件下,均表现为前12 d迅速下降,12 d后趋于稳定或缓慢下降。枯草芽孢杆菌在5种水中的降解速度较慢,在(25±1)℃和(50±1)℃条件下存放86 d后,其含量均在104cfu/mL以上。培养温度和灭菌条件对枯草芽孢杆菌在不同水体中存活动态有一定的影响,菌药合剂中的烯酰吗啉对该菌的存活则没有显著影响。A cocktail formulation （Bacillus subtilis-dimethomorph, DMBS） was developed with good control of Phytophthora blight of pepper and other crops based on the integrating of biological and chemical control, which was composed of Bacillus subtilis （BS） and dimethomorph. Based on the general principles of the hydrolysis research of agro-chemicals, the survival kinetics of BS in DMBS in de-ionized water, ground water, tap water, river water, and rain water were studied by using green fluorescent protein （GFP） labeling and bacteriological methods. The results showed that GFP was an efficient method for detecting the survival of Bacillus subtilis in the five kinds of water environment. At 25 ℃±1）℃, the quantity of the bacteria in its single formulation （SF） and in DMBS was decreased rapidly within 12 d of their storage. After 12 d, the amount of the bacteria exhibited fluctuant ascend or descend with the extending of their storage. At 50℃ ±1℃ , the quantity of bacteria in both formulations was declined quickly during 12 d within incubation in autoclaved and unautoclaved treatments. After that, the amount of the bacteria tend to stabilize or descend mildly. The bacteria was decreased slowly in the five kinds of waters and kept above 104 cfu/mL after incubated 86 d under both conditions of （25 ± 1 ） ℃and （50 ± 1 ）℃. The incubation temperature and autoclaved state had certain effects to the survival dynamic of B. subtilis in SF and in DMBS, while dimethomorph in DMBS had no remarkable effect.

关 键 词：绿色荧光蛋白标记 枯草芽孢杆菌 存活数量 水环境 

分 类 号：S476[农业科学—农业昆虫与害虫防治]