一株新型鱼腥藻溶藻细菌的分离鉴定及其溶藻特性
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摘要
蓝藻水华在全球范围频繁暴发,鱼腥藻(Anabaena)水华出现频度仅次于微囊藻(Microcystis)水华,其中的一些鱼腥藻种类能产生多样的藻毒素或异味物质,鱼腥藻水华去除技术的研发备受关注.本研究以从武汉市新洲区土壤中分离筛选到的一株细菌WP为材料,通过分析其生理生化特征及16S r DNA序列对其进行分类地位鉴定,考察了该菌株对真紧密鱼腥藻(Anabaena eucompacta)CHAB 1799的溶藻特性.结果表明,菌株WP与Lysinibacillus fusiformis NRS-350的同源性达到了99.6%,故鉴定WP属于Lysinibacillus属.菌株WP对真紧密鱼腥藻CHAB 1799发生溶藻作用的细菌阈值为6×10~3cfu/ml,其生理状态不会明显影响溶藻效果,且对不同生长时期的藻液均有较强的溶藻作用.该菌株的溶藻方式为间接溶藻.本研究结果丰富了溶藻细菌的资源库,也为治理鱼腥藻水华提供了更多的技术支持.
Occurrence of cyanobacterial blooms is becoming a global environmental issue. Next to Microcystis bloom,Anabaena bloom is also the main bloom type with more diversity in production of cyanotoxins and odour substances,and removal of Anabaena blooms is also much concerned. One strain encoded as WP with strong algicidal activity against Anabaena eucompacta CHAB 1799 was isolated from a soil environment in Wuhan city. The 16 S rRNA gene sequence of the WP strain showed 99.6% similarity with that of Lysinibacillus fusiformis NRS-350. Based on morphological,physiological and molecular phylogenetic features,algicidal bacterium WP was identified as Lysinibacillus sp. Its algicidal characteristics on A. eucompacta was also tested. Lysinibacillus sp. WP lysed A. eucompacta CHAB 1799 at lowcell density( 6×10~3 cfu/ml),and lysis effects of the Lysinibacillus sp. WP on A. eucompacta were related to growth stages of A. eucompacta CHAB 1799 rather than the growth stages of the strain WP. The strain WP lysed algae by secreting metabolites. Such results in the present study increase the resource bank of algicidal bacteria,providing more potential to treat the newtype blooms such as A. eucompacta dominated bloom in China.
Occurrence of cyanobacterial blooms is becoming a global environmental issue. Next to Microcystis bloom,Anabaena bloom is also the main bloom type with more diversity in production of cyanotoxins and odour substances,and removal of Anabaena blooms is also much concerned. One strain encoded as WP with strong algicidal activity against Anabaena eucompacta CHAB 1799 was isolated from a soil environment in Wuhan city. The 16 S rRNA gene sequence of the WP strain showed 99.6% similarity with that of Lysinibacillus fusiformis NRS-350. Based on morphological,physiological and molecular phylogenetic features,algicidal bacterium WP was identified as Lysinibacillus sp. Its algicidal characteristics on A. eucompacta was also tested. Lysinibacillus sp. WP lysed A. eucompacta CHAB 1799 at lowcell density( 6×10~3 cfu/ml),and lysis effects of the Lysinibacillus sp. WP on A. eucompacta were related to growth stages of A. eucompacta CHAB 1799 rather than the growth stages of the strain WP. The strain WP lysed algae by secreting metabolites. Such results in the present study increase the resource bank of algicidal bacteria,providing more potential to treat the newtype blooms such as A. eucompacta dominated bloom in China.
引文
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[11]Mu R,Fan Z,Pei H et al.Isolation and algae-lysing characteristics of the algicidal bacterium B5.Journal of Environmental Sciences,2007,19(11):1336-1340.DOI:10.1016/S1001-0742(07)60218-6.
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[13]Ahmed I,Yokota A,Yamazoe A et al.Proposal of Lysinibacillus boronitolerans gen.nov.sp.nov.,and transfer of Bacillus fusiformis to Lysinibacillus fusiformis comb.nov.and Bacillus sphaericus to Lysinibacillus sphaericus comb.nov..International Journal of Systematic and Evolutionary Microbiology,2007,57(5):1117-1125.DOI:10.1099/ijs.0.63867-0.
[14]Lu LL,Li GB,Shen YW et al.Isolation,identification and characterization of blue-green algae-lysing strain DC-L14 from Lake Dianchi,China.Chinese Journal of Applied and Environmental Biology,2009,15(1):106-109.DOI:10.3724/SP.J.1145.2009.00106.[卢兰兰,李根保,沈银武等.溶藻细菌DC-L14的分离、鉴定与溶藻特性.应用与环境生物学报,2009,15(1):106-109.]
[15]Liu L,Chen X,Li Q et al.Isolation,identification and algicidal effect of an algicidal bacterium Lysinibacillus fusiformis TL.Fresenius Environmental Bulletin,2016,25(12):5842-5848.
[16]Zhang J,Wei KF,Li CC et al.Identification and algae-lytic characteristics of endophyte algicidal bacteria ZB 1 strain isolated from wheat.Southwest China Journal of Agricultural Sciences,2017,30(5):1068-1073.[张菊,韦坤逢,李灿灿等.小麦内生溶藻细菌ZB1的分离鉴定及其溶藻特性.西南农业学报,2017,30(5):1068-1073.]
[17]Liu GY,Hu YP,Shi XD et al.Isolation,identification and characterization of algae-lysing strain H5 from Xiangxi Bay of Three Gorges Reservoir.Journal of Anhui Agricultural Sciences,2012,40(28):13955-13956,13959.DOI:0517-6611(2012)28-13955-02.[刘国勇,胡亚平,石小丹等.溶藻细菌H5的分离、鉴定及溶藻特性研究.安徽农业科学,2012,40(28):13955-13956,13959.]
[18]Xuan H,Dai X,Li J et al.A Bacillus sp.strain with antagonistic activity against Fusarium graminearum kills Microcystis aeruginosa selectively.Science of the Total Environment,2017,583:214-221.DOI:10.1016/j.scitotenv.2017.01.055.
[19]Kang YH,Kim JD,Kim BH et al.Isolation and characterization of a bio-agent antagonistic to diatom,Stephanodiscus hantzschii.Journal of Applied Microbiology,2005,98(5):1030-1038.DOI:10.1111/j.1365-2672.2005.02533.x.
[20]Jung SW,Kim BH,Katano T et al.Pseudomonas fluorescens HYK0210-SK09 offers species-specific biological control of winter algal blooms caused by freshwater diatom Stephanodiscus hantzschii.Journal of Applied Microbiology,2008,105(1):186-195.DOI:10.1111/j.1365-2672.2008.03733.
[2]Zhang H,Huang Y.Character of algae bloom in Chaohu Lake base on NOAA/AVHRR.China Environmental Science,2009,29(7):727-732.DOI:1000-6923(2009)07-0727-06.[张红,黄勇.基于NOAA/AVHRR卫星资料的巢湖水华规律分析.中国环境科学,2009,29(7):727-732.]
[3]Zhang YH,Li WF,Chen QW.Spatial-temporal variance of the intensity of algal bloom and related environmental and ecological factors in Lake Taihu.Acta Ecologica Sinica,2016,36(14):4337-4345.DOI:10.5846/stxb201412112468.[张艳会,李伟峰,陈求稳.太湖水华程度及其生态环境因子的时空分布特征.生态学报,2016,36(14):4337-4345.]
[4]Liu X,Lu XH,Chen YW.The effects of temperature and nutrient ratios on Microcystis blooms in Lake Taihu,China:an11-year investigation.Harmful Algae,2011,10(3):337-343.DOI:10.1016/j.hal.2010.12.002.
[5]Huang CM,Pan WB,Li Y et al.Effect of three algicidal bacteria strains on antioxidase activities of Anabaena flos-aquae.Environmental Science and Technology,2009,22(6):10-13.DOI:1674-4829(2009)06-0010-04.[黄昌妙,潘伟斌,李燕等.三株溶藻菌对水华鱼腥藻抗氧化酶活性的影响.环境科技,2009,22(6):10-13.]
[6]Ye JY,Zhong YR,Yu L et al.Identification of an algae-lysing bacterium of Anabaena flos-aquae and primary research on their relationship.Journal of Anhui Agricultural Sciences,2011,39(29):18121-18124.DOI:0517-6611(2011)29-18121-04.[叶姜瑜,钟以蓉,俞岚等.一株水华鱼腥藻溶藻菌的分离鉴定及菌藻关系初探.安徽农业科学,2011,39(29):18121-18124.]
[7]Li RH,Watanabe MW.Anabaena eucompacta sp.nov.(Nostocales,Cyanobacteria),a new planktonic species with tightly spiraled filaments from Japan.Bulletin of the National Science Museum,Tokyo Serie B,1999,25(3):89-94.
[8]Salvatore M,Marisa C,Francesca T et al.Comparison of plate count agar and R2A medium for enumeration of heterotrophic bacteria in natural mineral water.World Journal of Microbiology and Biotechnology,1998,14(5):727-730.
[9]Ichimura T.Media for freshwater cyanobacteria.In:Nishizawa K,Chihara M eds.Methods in phycology.Tokyo:Kyouritsu Shuppan,1979:295-296.
[10]Shao J,Jiang Y,Wang Z et al.Interactions between algicidal bacteria and the cyanobacterium Microcystis aeruginosa:lytic characteristics and physiological responses in the cyanobacteria.International Journal of Environmental Science and Technology,2014,11(2):469-476.DOI:10.1007/s13762-013-0205-4.
[11]Mu R,Fan Z,Pei H et al.Isolation and algae-lysing characteristics of the algicidal bacterium B5.Journal of Environmental Sciences,2007,19(11):1336-1340.DOI:10.1016/S1001-0742(07)60218-6.
[12]Li SH,Zhang QY.Partial biological characteristics and algicidal activity of an algicidal bacterium.Environmental Science,2013,34(2):583-588.DOI:10.13227/j.hjkx.2013.02.001.[李三华,张奇亚.1株溶藻菌的部分生物学特性及溶鱼腥藻作用.环境科学,2013,34(2):583-588.]
[13]Ahmed I,Yokota A,Yamazoe A et al.Proposal of Lysinibacillus boronitolerans gen.nov.sp.nov.,and transfer of Bacillus fusiformis to Lysinibacillus fusiformis comb.nov.and Bacillus sphaericus to Lysinibacillus sphaericus comb.nov..International Journal of Systematic and Evolutionary Microbiology,2007,57(5):1117-1125.DOI:10.1099/ijs.0.63867-0.
[14]Lu LL,Li GB,Shen YW et al.Isolation,identification and characterization of blue-green algae-lysing strain DC-L14 from Lake Dianchi,China.Chinese Journal of Applied and Environmental Biology,2009,15(1):106-109.DOI:10.3724/SP.J.1145.2009.00106.[卢兰兰,李根保,沈银武等.溶藻细菌DC-L14的分离、鉴定与溶藻特性.应用与环境生物学报,2009,15(1):106-109.]
[15]Liu L,Chen X,Li Q et al.Isolation,identification and algicidal effect of an algicidal bacterium Lysinibacillus fusiformis TL.Fresenius Environmental Bulletin,2016,25(12):5842-5848.
[16]Zhang J,Wei KF,Li CC et al.Identification and algae-lytic characteristics of endophyte algicidal bacteria ZB 1 strain isolated from wheat.Southwest China Journal of Agricultural Sciences,2017,30(5):1068-1073.[张菊,韦坤逢,李灿灿等.小麦内生溶藻细菌ZB1的分离鉴定及其溶藻特性.西南农业学报,2017,30(5):1068-1073.]
[17]Liu GY,Hu YP,Shi XD et al.Isolation,identification and characterization of algae-lysing strain H5 from Xiangxi Bay of Three Gorges Reservoir.Journal of Anhui Agricultural Sciences,2012,40(28):13955-13956,13959.DOI:0517-6611(2012)28-13955-02.[刘国勇,胡亚平,石小丹等.溶藻细菌H5的分离、鉴定及溶藻特性研究.安徽农业科学,2012,40(28):13955-13956,13959.]
[18]Xuan H,Dai X,Li J et al.A Bacillus sp.strain with antagonistic activity against Fusarium graminearum kills Microcystis aeruginosa selectively.Science of the Total Environment,2017,583:214-221.DOI:10.1016/j.scitotenv.2017.01.055.
[19]Kang YH,Kim JD,Kim BH et al.Isolation and characterization of a bio-agent antagonistic to diatom,Stephanodiscus hantzschii.Journal of Applied Microbiology,2005,98(5):1030-1038.DOI:10.1111/j.1365-2672.2005.02533.x.
[20]Jung SW,Kim BH,Katano T et al.Pseudomonas fluorescens HYK0210-SK09 offers species-specific biological control of winter algal blooms caused by freshwater diatom Stephanodiscus hantzschii.Journal of Applied Microbiology,2008,105(1):186-195.DOI:10.1111/j.1365-2672.2008.03733.