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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2013, Vol. 39 Issue (6): 613-620    DOI: 10.3785/j.issn.1008-9209.2012.11.301
生物科学与技术     
柠檬酸杆菌N10的分离及其溶藻特性
孙朋飞， 卢丽玲， 王冠， 赵宇华*
（浙江大学生命科学学院微生物研究所，杭州 310058）
Isolation and algicidal characteristics of Citrobacter sp. strain N10
SUN Pengfei, LU Liling, WANG Guan, ZHAO Yuhua*
(Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou 310058, China)
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摘要： 以铜绿微囊藻（Microcystis aeruginosa）为溶解对象，利用液体感染分离技术从浙江大学华家池校区湖水样品中分离到1株溶藻菌N10，对其生长及溶藻相关特性进行了研究.结果表明：经过16S rDNA核苷酸序列分析，该菌株属于柠檬酸杆菌属（Citrobacter sp.）；当N10无细胞培养物（cellfree culture filtrate，CCF）与铜绿微囊藻液按照体积比为1∶5混合后，在24 h内N10 CCF对铜绿微囊藻的溶藻率可达到86.55%，并且溶藻率随着溶藻时间的延长而增强，在72 h内溶藻率可达到97.08%；对N10生长量影响最大的是pH值，其次是NaCl质量分数，最后是温度，但N10菌株对铜绿微囊藻的溶藻率受培养温度和培养基中NaCl质量分数的影响很小；N10菌株通过分泌一种对高温（115 ℃）敏感但却能抗蛋白酶K的物质来溶解铜绿微囊藻；透射电子显微镜观察表明，经过N10 CCF处理后的藻细胞出现了细胞膜、细胞壁破裂，伪空胞结构、细胞质中磷酸颗粒以及蓝色体等颗粒性物质消失，细胞中的光合片层排列趋于松散且混乱的现象.
Abstract: In recent decades, harmful algal blooms (HABs), which are natural phenomena that occur across all the world, have posed threats to decrease dissolved oxygen and have had a negative effect on fisheries, aquaculture, drinking water, tourism and human health. It is increasingly urgent to develop useful strategies for predicting and reducing the negative impacts of HABs. Actually, many methods have been devoted to controlling HABs, including copper sulfate, hydrogen peroxide, ozonization, ultrasonication, collection of algae from water surface using nets, iron powder and magnets, centrifugal separation, or ultraviolet radiation. As an effective and environment friendly strategy to control harmful algal bloom outbreaks, biologic control methods such as using algicidal bacteria against M. aeruginosa have been more and more attractive. It has been demonstrated that algicidal bacteria could serve as potential ways in reducing the impacts of HABs. In this study, a Citrobacter sp. N10 with algicidal activity against the toxic algae M. aeruginosa was isolated and identified from a lake of Zhejiang University using liquid infection technology and 16S rDNA sequence analysis. The growth and algicidal characters of Citrobacter sp. N10 were then studied in order to offer some useful information for HABs controlling. The results showed that the bacterium was gramnegative and its colonies were smooth, convex, low moisture, translucent or opaque and gray, and its colony surfaces were glossy. Sequence analysis of 16S rDNA demonstrated that strain N10 belonged to Citrobacter sp. When inoculation time was 2 to 12 h, N10 grew in logarithmicgrowth period, and when inoculation time was beyond 12 h, N10 was in stationary phase. Strain N10 showed a high lysis of M. aeruginosa with an algicidal rate of 86.55% within 24 h. The algicidal rate of N10 increased with increasing incubation time and reached 97.08% within 72 h. pH value had the most significant effect on the growth of strain N10, followed by NaCl concentration and culture temperature, while these three conditions had different effects on algicidal activities of strain N10 against M. aeruginosa. A kind of substance against M. aeruginosa was secreted by strain N10, and the substance was sensitive to high temperature (121 ℃) but resistant to protease K. Relatively speaking, the thalli of strain N10 had hardly algicidal effect against M. aeruginosa. Transmission electron microscope was subsequently used to investigate algal cells’ structures after 3 d with treatment of strain N10 cellfree culture filtrate (CCF). The results indicated that the cell membranes and cell walls of M. aeruginosa were broken, while the gas vesicles and some particulate matters such as the cytoplasm phosphate particles and blue bodies disappeared. In addition, the cellular ultrastructure of the photosynthetic lamellae was also found to be disordered and disappeared. In conclusion, the research on the characteristics of N10 against M. aeruginosa can provide a tool in controlling the bloom of M. aeruginosa. The findings are of theoretical significance and potential in reducing the impacts of HABs.
出版日期: 2013-11-20
CLC:  Q 939.123  
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孙朋飞
卢丽玲
王冠
赵宇华*

引用本文:

孙朋飞， 卢丽玲， 王冠， 赵宇华*. 柠檬酸杆菌N10的分离及其溶藻特性[J]. 浙江大学学报(农业与生命科学版), 2013, 39(6): 613-620.

SUN Pengfei, LU Liling, WANG Guan, ZHAO Yuhua*. Isolation and algicidal characteristics of Citrobacter sp. strain N10. Journal of Zhejiang University (Agriculture and Life Sciences), 2013, 39(6): 613-620.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2012.11.301    或    http://www.zjujournals.com/agr/CN/Y2013/V39/I6/613

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