溶藻细菌DC-L5的分离、鉴定及其溶藻特性
ISOLATION, IDENTIFICATION AND CHARACTERIZATION OF A STRA IN OF BLUE2GREEN ALGAE2LYS ING BACTERIUM FROM LAKE D IANCHI
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摘要: 从滇池蓝藻水华集聚区分离获得一株溶藻细菌DC-L5,通过形态及16S rDNA测序分析鉴定为短小芽孢杆菌。用小白鼠进行生物安全实验,小白鼠无中毒症状。研究表明当细菌处于对数生长期时溶藻效果最强,共培养5d使铜锈微囊藻的叶绿素a含量下降83.33%,使惠氏微囊藻、绿色微囊藻、水华束丝藻和水华鱼腥藻4种蓝藻叶绿素a下降率最高为67.6%,最低为58.5%,平均为62.25%。离心沉降后,发现沉淀菌体和无菌上清液对铜锈微囊藻都有溶藻效果,但溶藻效果不及原菌液,推测DC-L5可能是通过直接接触使藻细胞凝聚下沉及进一步的生物降解,同时存在抑制藻细胞生长的胞外分泌物。高温热处理后菌液溶藻作用不明显,推测高温可能使菌体或胞外分泌物质失活。Abstract: In this study,one high-effect algae-lysing bacterium was isolated and purified from Lake Dianchi,which wasabled as the strain DC-L5. The effect,host range,biosafety,lysing mechanism and lysing phenomenon of the algae-ly-ing bacteria were analyzed. Resultswere as the followings: The strain was identified asB acillus pum ilus bymeans ofmor-hological observation and 16S rDNA sequence analysis in the CCTCC (China Center for Type Culture Collection). Toxic-ty test with mouse primarily showed that the strain was non-toxic. 4 kinds of DC-L5 cultures of different growth phasesere added to M icrocystis aeruginosa,and the strongest algae-lytic ability was found in the log phase of DC-L5 cultures,hlorophyll a contents in culture ofM icrocystis aeruginosa reduced by 83. 33% in 5 days. For the host range experiment,kinds of dominant cyanobacteria in Lake Dianchi was researched,and average reduction rates of Chlorophyll a of-. -5% in cultures ofM icrocystis viridis,M icrocystis wesenbergii,Aphanizomenon flos-aquae and Anabaena flos-aquaewasbserved with the highest 67. 6% and the lowest 58. 5% in 4 days respectively. After centrifugation and sedimentation,oth the cells of bacteria and the cell-free suspension had strong algae-lytic activity,but the bacteria culture had the stron-est algae-lytic activity,so it maybe showed the strain DC-L5 could directly cause the cyanobacterial cells agglomerated,inked down and further lysed while the strain could excrete extra-cellular substances able to lyse cyanobacterial cells. Inrder to improve the algae-lytic activity of the strain DC-L5 to nature bloom,the bacteria cultures were added to natureyanobacterial bloom dominant with M icrocystis aeruginosa,which were samp led from Lake Dianchi and cultured in theab. The result of nature bloom experiment showed the strain DC-L5 firstly disaggregated the cyanobacterial colony,educed the floatability,degraded the p igments and then released inclusions of algal cells.
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Keywords:
- Algaelysing bacterium /
- Biological control /
- B acillus /
- Water bloom /
- M icrocystis
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[1] Yin L Y, Huang J Q, Liu Y D. Microcystin2RR up take and itsffects on the growth of submerged macrophyte Vallisneria natans lour. ) hara [J]. Environm ental Toxicology, 2005, 3 ( 20 ) :08-313
[2] Hu ZQ, Liu YD, LiD H, et al. Growth and antioxidant systemf the cyanobacterium Synechococcus elongtus in response to mi2rocystin2RR [J]. Hydrobiologia, 2005, 534: 23-29
[3] Li X Y, Wang J, Liang J B, et al. Toxicity ofmicrocystins in thesolated hepatocytes of common carp (Cyprinus carpio L. ) [J].cotoxicology and Environm ental Safety, 2007, 67: 447-451
[4] Zhao C P, Pu Y P, Yin L H. Development of research on algi2idal bacteria and its evaluation [J]. Journal of Southeast Uni2ersity (Medical Science Edition), 2005, 24 ( 3 ) : 202-206[赵传鹏, 浦跃朴, 尹立红. 溶藻细菌及其测定评价方法的究进展. 东南大学学报(医学版), 2005, 24 ( 3 ) : 202-06] Zhao C P, Pu Y P, Yin L H. Development of research on algi2idal bacteria and its evaluation [J]. Journal of Southeast Uni2ersity (Medical Science Edition), 2005, 24 ( 3 ) : 202-206[赵传鹏, 浦跃朴, 尹立红. 溶藻细菌及其测定评价方法的究进展. 东南大学学报(医学版), 2005, 24 ( 3 ) : 202-06]
[5] Studies on the Ecophysiological Effects andMechanisms ofAlgae2ysing Bacteria against the Algae Thesis for doctor of Science[D]. Institute of Hydrobiology, Chinese Academy of Sciences,uhan. 2006 [史顺玉. 溶藻细菌对藻类的生理生态效应及用机理研究. 博士学位论文. 中国科学院水生生物研究, 武汉. 2006]. Studies on the Ecophysiological Effects andMechanisms ofAlgae2ysing Bacteria against the Algae Thesis for doctor of Science[D]. Institute of Hydrobiology, Chinese Academy of Sciences,uhan. 2006 [史顺玉. 溶藻细菌对藻类的生理生态效应及用机理研究. 博士学位论文. 中国科学院水生生物研究, 武汉. 2006].
[6] Li X C, Pei H Y, HuW R. Development of the research on algi2idal bacteria and its algicidal substances [J]. IndustrialWaterreatm ent, 2007, 27 ( 6) : 10-13 [李小彩,裴海燕,胡文容.藻细菌及溶藻物质研究进展. 工业水处理, 2007, 27 ( 6) :0-13] Li X C, Pei H Y, HuW R. Development of the research on algi2idal bacteria and its algicidal substances [J]. IndustrialWaterreatm ent, 2007, 27 ( 6) : 10-13 [李小彩,裴海燕,胡文容.藻细菌及溶藻物质研究进展. 工业水处理, 2007, 27 ( 6) :0-13]
[7] Li X Y, SongL R, Liu YD. The p roduction, detection and toxi2ology of microcystins [J]. Acta Hydrobiologica S inica, 1999,3 (5) : 517-522 [李效宇, 宋立荣, 刘永定. 微囊藻毒素的生、检测和毒理学研究. 水生生物学报, 1999, 23 ( 5 ) :17-522] Li X Y, SongL R, Liu YD. The p roduction, detection and toxi2ology of microcystins [J]. Acta Hydrobiologica S inica, 1999,3 (5) : 517-522 [李效宇, 宋立荣, 刘永定. 微囊藻毒素的生、检测和毒理学研究. 水生生物学报, 1999, 23 ( 5 ) :17-522]
[8] Zhao Y J, Liu YD. Possiblemicrobial control on the adverse im2acts of algae 2 current information about the relationship betweenlgae and microbes [J]. Acta Hydrobiologica S inica, 1996, 202) : 173-181 [赵以军, 刘永定. 有害藻类及其微生物防的基础-藻菌关系的研究动态. 水生生物学报, 1996, 202) : 173-181] Zhao Y J, Liu YD. Possiblemicrobial control on the adverse im2acts of algae 2 current information about the relationship betweenlgae and microbes [J]. Acta Hydrobiologica S inica, 1996, 202) : 173-181 [赵以军, 刘永定. 有害藻类及其微生物防的基础-藻菌关系的研究动态. 水生生物学报, 1996, 202) : 173-181]
[9] Shi S Y, Shen YW, LiD H, et al. Isolation, identification andlgae2lytic characteristic of a bacterium Staphylococcus sp. [J].hina Environm ental Science, 2006, 26 ( 5) : 587-590 [史顺,沈银武,李敦海,等. 溶藻细菌DC221的分离、鉴定及其溶特性. 中国环境科学, 2006, 26 (5) : 587-590] Shi S Y, Shen YW, LiD H, et al. Isolation, identification andlgae2lytic characteristic of a bacterium Staphylococcus sp. [J].hina Environm ental Science, 2006, 26 ( 5) : 587-590 [史顺,沈银武,李敦海,等. 溶藻细菌DC221的分离、鉴定及其溶特性. 中国环境科学, 2006, 26 (5) : 587-590]
[10] Shi S Y, Liu Y D, Shen YW, et al. The algae2lytic ability ofacterium DC10 and the influence of environmental factors on thebility [J]. Science in China (Life Sciences), 2004, 6 ( 34 ) :64-568
[11] Shi S Y, LiuYD, Shen YW. Preliminary research on the algae2ysing characteristics of bacterium ( strain DC23 )[J]. Actaydrobiologica S inica, 2004, 28 ( 2) : 219-221 [史顺玉, 刘定, 沈银武. 细菌溶藻的初步研究. 水生生物学报, 2004,8 (2) : 219-221] Shi S Y, LiuYD, Shen YW. Preliminary research on the algae2ysing characteristics of bacterium ( strain DC23 )[J]. Actaydrobiologica S inica, 2004, 28 ( 2) : 219-221 [史顺玉, 刘定, 沈银武. 细菌溶藻的初步研究. 水生生物学报, 2004,8 (2) : 219-221]
[12] Shi S Y, Liu Y D, Shen YW, et al. Lysis of Aphanizomenonlos2aquae ( Cyanobacterium ) by a bacterium Bacillus cereus[J]. B iological Control, 2006, 3 (39) : 345-351
[13] Liu YM, ChenW, Liu YD, et al. First report of aphantoxins inhina-waterblooms of toxigenic Aphanizom enon flos2aquae inake Dianchi [J]. Ecotoxicology and Environm ental Safety,006, 65: 84-92
[14] Pan X J, Chang F Y, Liu YD, et al. Mouse toxicity ofAnabae2a flos2aquae from Lake Dianchi, China. Environm ental Toxicol2gy. Published online in W iley InterScience. DO I 10. 1002/tox.0385. 2008
[15] Shen P, Fan X R, Li GW. Microbiology test [M]. Beijing:igh Education Publishers. 2002 [沈萍, 范秀容, 李广武. 微物学试验(第三版). 北京: 高等教育出版社. 2002]. Shen P, Fan X R, Li GW. Microbiology test [M]. Beijing:igh Education Publishers. 2002 [沈萍, 范秀容, 李广武. 微物学试验(第三版). 北京: 高等教育出版社. 2002].
[16] Department of classification of bacteria, Institute ofMicrobiology,AS. Common methods of identification of bacteria [M]. Bei2ing: Science Publishers. 1987 [中国科学院微生物研究所细分类组. 一般细菌常用鉴定方法. Department of classification of bacteria, Institute ofMicrobiology,AS. Common methods of identification of bacteria [M]. Bei2ing: Science Publishers. 1987 [中国科学院微生物研究所细分类组. 一般细菌常用鉴定方法.
[17] Wintermans J F GM, DeMotsA. Spectrophotometric character2stics of chlorophylls a and b and their pheophytins in ethanol[J]. B iochim B iophys Acta, 1965, 109: 448-453
[18] Liu J, PanW B, Qin Y J. Isolation and identification of two al2ae2lysing Bacterie and their lytic character [J]. Environm entalcience and Technology, 2007, 2 ( 30) : 17-21 [刘晶, 潘伟, 秦玉洁. 两株溶藻细菌的分离鉴定及其溶藻特性. 环境学与技术, 2007, 2 (30) : 17-21] Liu J, PanW B, Qin Y J. Isolation and identification of two al2ae2lysing Bacterie and their lytic character [J]. Environm entalcience and Technology, 2007, 2 ( 30) : 17-21 [刘晶, 潘伟, 秦玉洁. 两株溶藻细菌的分离鉴定及其溶藻特性. 环境学与技术, 2007, 2 (30) : 17-21]
[19] Zheng T, Tian Y, Su J Q, et al. Study on the ecological rela2ionship between a red tide causative alga and three strains ofacteria isolated from Xiamen Harbor [J]. Acta Ecologica S ini2a, 2002, 12 (22) : 2063-2070 [郑天凌, 田蕴, 苏建强,等.洋赤潮生物与厦门海域几种细菌的生态关系研究. 生态学, 2002, 12 (22) : 2063-2070] Zheng T, Tian Y, Su J Q, et al. Study on the ecological rela2ionship between a red tide causative alga and three strains ofacteria isolated from Xiamen Harbor [J]. Acta Ecologica S ini2a, 2002, 12 (22) : 2063-2070 [郑天凌, 田蕴, 苏建强,等.洋赤潮生物与厦门海域几种细菌的生态关系研究. 生态学, 2002, 12 (22) : 2063-2070]
[20] Liu L P, Zhang X M, Zhao X H. App roach on comp
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