粘着剑菌溶藻效能与溶藻过程藻胞内物质释放
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摘要
粘着剑菌菌株r23(Ensifer adhaerens)是从微囊藻水体中筛选出的溶藻菌。文章研究了不同温度和投加方式下r23溶解铜绿微囊藻(Microcystis aeruginosa)的效果,以及不同投加方式下胞内外藻毒素-LR(MC-LR)的变化和胞内有机物质的释放。结果显示,r23菌液的EC_(50,72 h)是2.04×10~6CFU/m L,能够在温度低至15℃时仍有95%的溶藻效果。在没有培养基条件下,菌体细胞不具有溶藻能力,菌液上清液能够实现70%的溶藻效果。菌液5%(V/V)的投加剂量会刺激藻细胞产生更多MC-LR,总MC-LR达到256.7 mg/L,高于对照样的232.1 mg/L,而10%和20%的投加剂量会抑制MC-LR产生,总MC-LR浓度在溶藻30 d后会降至36.8 mg/L和12.24 mg/L。三维荧光测定结果显示,菌液的投入以及藻细胞的死亡会导致水溶液中类蛋白质物质(λ_(Ex)/λ_(Em)=280 nm/350 nm;λ_(Ex)/λ_(Em)=230 nm/330 nm)含量增加,而类腐殖质物质(λ_(Ex)/λ_(Em)=340 nm/430 nm)变化不大。
Ensifer adhaerens(r23)is an algicidal bacterium screened from the Microcystis bloom water.This paper studied the algicidal effects of temperature and different dosing modes of strain r23 on Microcystis aeruginosa,as well as the variation of intracellular/extracellular microcystin-LR and intracellular organic matters.The results showed that the EC_(50,72 h)of strain r23 was 2.04×10~6CFU/m L.The strain r23 could preformed the algicidal activity even if the temperature was as low as 15℃,achieving 95%of algae-lysing efficiency.The strain r23 could not inhibit the growth of M.aeruginosa without the presence of media.The supernatant of bacterial culture reached 70%of algae-lysing efficiency.The dosage of 5%(V/V)bacterial culture stimulated algal cells to produce more MC-LR with the total(intra-and extra-cellular)MC-LR of 256.7μg/L after 30 days,slightly higher than 232.1μg/L of control samples.However,the higher dosage of 10%and 20%compressed the MC-LR production and the total MC-LR were apparently decreased to 36.8μg/L and 12.2μg/L after 30 days,respectively.The three-dimensional fluorescence matrix showed that the addition of strain r23 culture and the lysing of algae cells resulted in the increase of protein-like substances(λ_(Ex)/λ_(Em)=280 nm/350 nm;λ_(Ex)/λ_(Em)=230 nm/330 nm),but humus substances(λ_(Ex)/λ_(Em)=340 nm/430 nm)with unchanged.
Ensifer adhaerens(r23)is an algicidal bacterium screened from the Microcystis bloom water.This paper studied the algicidal effects of temperature and different dosing modes of strain r23 on Microcystis aeruginosa,as well as the variation of intracellular/extracellular microcystin-LR and intracellular organic matters.The results showed that the EC_(50,72 h)of strain r23 was 2.04×10~6CFU/m L.The strain r23 could preformed the algicidal activity even if the temperature was as low as 15℃,achieving 95%of algae-lysing efficiency.The strain r23 could not inhibit the growth of M.aeruginosa without the presence of media.The supernatant of bacterial culture reached 70%of algae-lysing efficiency.The dosage of 5%(V/V)bacterial culture stimulated algal cells to produce more MC-LR with the total(intra-and extra-cellular)MC-LR of 256.7μg/L after 30 days,slightly higher than 232.1μg/L of control samples.However,the higher dosage of 10%and 20%compressed the MC-LR production and the total MC-LR were apparently decreased to 36.8μg/L and 12.2μg/L after 30 days,respectively.The three-dimensional fluorescence matrix showed that the addition of strain r23 culture and the lysing of algae cells resulted in the increase of protein-like substances(λ_(Ex)/λ_(Em)=280 nm/350 nm;λ_(Ex)/λ_(Em)=230 nm/330 nm),but humus substances(λ_(Ex)/λ_(Em)=340 nm/430 nm)with unchanged.
引文
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[20]朱叶飞,唐婉莹.1株溶藻真菌的分离鉴定及其作用机理研究[].广东化工,2016,43(7):3-4.Zhu Yefei,Tang Wanying.Isolation,identification and algicidal mechanism of an algae-lysing fungus[J].Guangdong Chemical Industry,2016,43(7):3-4.
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[23]Duan Y L,Su R G,Shi X Y,et al.Research on using 3-D fluorescence spectroscopy-wavelet transform-PSO for rapid discrimination of compositions of phytoplankton population[J].Spectroscopy and Spectral Analysis,2012,32(6):1562-1569.
[24]Kong Y,Zhu L,Meile L V,et al.Research advances in water environment remediation and wastewater treatment based on three dimensional fluorescence spectroscopy technology[J].Ecology&Environmental Sciences,2012,7(7):1647-1654.
[25]Wang J X,Luo G Y,Xu Xiaoyi,et al.Lytic effect of environmental factors on algae-lysing bacteria strain s7 by fluorescence spectroscopy[J].Journal of Shanghai Jiaotong University,2012,46(5):780-784.
[26]Wang Z,Chen Y,Xie P,et al.Removal of Microcystis aeruginosa by UV-activated persulfate:performance and characteristics[J].Chemical Engineering Journal,2016(300):245-253.
[27]丰桂珍,董秉直.水中藻类溶解性有机物特性研究[J].环境科学与技术,2016,39(11):144-149.Feng Guizhen,Dong Bingzhi.Study on the characteristics of algogenic organic matters[J].Environmental Science&Technology,2016,39(11):144-149.
[28]陈诗雨,李燕,李爱民.溶解性有机物研究中三维荧光光谱分析的应用[J].环境科学与技术,2015,38(5):64-68.Chen Shiyu,Li Yan,Li Aimin.Application of three-dimensional fluorescence spectroscopy in the study of dissolved organic matter[J].Environmental Science&Technology,2015,38(5):64-68.
[2]李媛,张家卫,魏杰,等.我国蓝藻水华的发生机理、危害及防控利用研究进展[J].微生物学杂志,2015(4):93-97.Li Yuan,Zhang Jiawei,Wei Jie,et al.Advances in mechanism of the occurrence,hazard,and prevention/control utilization of cyanophytic blooms in China[J].Journal of Microbiology,2015(4):93-97.
[3]Liao C,Liu X,Liu R,et al.Characterization and effects of two algicidal isolates on antioxidase activities of Chlorella pyrenoidosa[J].Environmental Progress&Sustainable Energy,2016,34(6):1647-1651.
[4]Lin S,Pan J,Li Z,et al.Characterization of an algicidal bacterium Brevundimonas J4 and chemical defense of Synechococcus sp.BN60 against bacterium J4[J].Harmful Algae,2014,37(4):1-7.
[5]Li Y,Liu L,Xu Y,et al.Stress of algicidal substances from a bacterium Exiguobacterium sp.h10 on Microcystis aeruginosa[J].Letters in Applied Microbiology,2016,64(1):57-65.
[6]Sun R,Sun P,Zhang J,et al.Microorganisms-based methods for harmful algal blooms control:a review[J].Bioresource Technology,2018,248(part B):12-20.
[7]刘征宇,宁喜斌,李文利,等.3株太湖溶藻细菌的分离及溶藻特性的初步研究[J].食品与发酵工业,2016,42(2):59-64.Liu Zhengyu,Ning Xibin,Li Wenli,et al.Isolation and primary study of three algicidal bacteria strains of Taihu Lake[J].Food and Fermentation Industries,2016,42(2):59-64.
[8]马健荣,邓建明,秦伯强,等.湖泊蓝藻水华发生机理研究进展[J].生态学报,2013,33(10):3020-3030.Ma Jianrong,Deng Jianming,Qin Boqiang,et al.Progress and prospects on cyanobacteria bloom-forming mechanism in lakes[J].Acta Ecologica Sinica,2013,33(10):3020-3030.
[9]Loisa O,K??ri?J,Laaksonlaita J,et al.From phycocyanin fluorescence to absolute cyanobacteria biomass:an application using in-situ fluorometer probes in the monitoring of potentially harmful cyanobacteria blooms[J].Water Practice&Technology,2015,10(4):695-698.
[10]Harke M J,Steffen M M,Gobler C J,et al.A review of the global ecology,genomics,and biogeography of the toxic cyanobacterium,Microcystis spp[J].Harmful Algae,2016(54):4-20.
[11]陈向东,沈萍.微生物学实验[M].第4版.北京:高等教育出版社,2010.Chen Xiangdong,Shen Ping.Experiment of Microbiology[M].4th Edition.Beijing:Higher Education Press,2010.
[12]孙秀敏.水华鱼腥藻降解菌及其溶藻效应研究[D].上海:上海交通大学,2013.Sun Xiumin.Algicidal Bacteria against Anabaena flosaquae and Their Algae-lysing Characteristics[D].Shanghai:Shanghai Jiaotong University,2013.
[13]阮铃铃.植物多酚抑藻效能与其作用下藻细胞生理特征的研究[D].北京:北京林业大学,2011.Ruan Lingling.Study on the Algal Inhibition Effect of Plant Polyphenols and the Algal Cell Physiological Characteristics Treated by Them[D].Beijing:Beijing Forestry University,2011.
[14]刘菲菲,冯慕华,尚丽霞,等.温度对铜绿微囊藻(Microcystis aeruginosa)和鱼腥藻(Anabaena sp.)生长及胞外有机物产生的影响[J].湖泊科学,2014,26(5):780-788.Liu Feifei,Feng Muhua,Shang,Lixia,et al.Effects of temperature on the growth and generation of extracellular organic matter of Microcystis aeruginosa and Anabaena sp[J].Journal of Lake Science,2014,26(5):780-788.
[15]Dziallas C,Grossart H P.Microbial interactions with the cyanobacterium Microcystis aeruginosa and their dependence on temperature[J].Marine Biology,2012,159(11):2389-2398.
[16]Zeng G,Wang P,Wang Y.Algicidal efficiency and mechanism of Phanerochaete chrysosporium against harmful algal bloom species[J].Algal Research,2015(12):182-190.
[17]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[J].International Journal of Environmental Science&Technology,2014,11(2):469-476.
[18]Yang L L,Pan W B,Yan L I.Preliminary study on components and its algicidal mechanism of algicidal substance secreted by bacteria[J].Environmental Science and Technology,2010,33(3):72-74.
[19]Fei Y,Yan W H,Qin L X,et al.Isolation and characterization of an algicidal bacterium indigenous to Lake Taihu with a red pigment able to lyse Microcystis aeruginosa[J].Biomedical and Environmental Sciences,2013,26(2):148-154.
[20]朱叶飞,唐婉莹.1株溶藻真菌的分离鉴定及其作用机理研究[].广东化工,2016,43(7):3-4.Zhu Yefei,Tang Wanying.Isolation,identification and algicidal mechanism of an algae-lysing fungus[J].Guangdong Chemical Industry,2016,43(7):3-4.
[21]Lu Y,Wang J,Yu Y,et al.Changes in the physiology and gene expression of Microcystis aeruginosa under EGCG stress[J].Chemosphere,2014,117(1):164-169.
[22]郝瑞霞,曹可心,邓亦文.三维荧光光谱法表征污水中溶解性有机污染物[J].分析试验室,2007,26(10):41-44.Hao Ruixia,Cao Kexin,Deng Yiwen.Characterization of dissolved organic pollutants in wastewater by three-dimensional fluorescent spectroscopy[J].Chinese Journal of Analysis Laboratory,2007,26(10):41-44.
[23]Duan Y L,Su R G,Shi X Y,et al.Research on using 3-D fluorescence spectroscopy-wavelet transform-PSO for rapid discrimination of compositions of phytoplankton population[J].Spectroscopy and Spectral Analysis,2012,32(6):1562-1569.
[24]Kong Y,Zhu L,Meile L V,et al.Research advances in water environment remediation and wastewater treatment based on three dimensional fluorescence spectroscopy technology[J].Ecology&Environmental Sciences,2012,7(7):1647-1654.
[25]Wang J X,Luo G Y,Xu Xiaoyi,et al.Lytic effect of environmental factors on algae-lysing bacteria strain s7 by fluorescence spectroscopy[J].Journal of Shanghai Jiaotong University,2012,46(5):780-784.
[26]Wang Z,Chen Y,Xie P,et al.Removal of Microcystis aeruginosa by UV-activated persulfate:performance and characteristics[J].Chemical Engineering Journal,2016(300):245-253.
[27]丰桂珍,董秉直.水中藻类溶解性有机物特性研究[J].环境科学与技术,2016,39(11):144-149.Feng Guizhen,Dong Bingzhi.Study on the characteristics of algogenic organic matters[J].Environmental Science&Technology,2016,39(11):144-149.
[28]陈诗雨,李燕,李爱民.溶解性有机物研究中三维荧光光谱分析的应用[J].环境科学与技术,2015,38(5):64-68.Chen Shiyu,Li Yan,Li Aimin.Application of three-dimensional fluorescence spectroscopy in the study of dissolved organic matter[J].Environmental Science&Technology,2015,38(5):64-68.