WSWXTB微生物学通报Microbiology China0253-2654微生物学通报编辑部中国北京wswxtb-47-11-352710.13344/j.microbiol.china.190991海洋微生物学Marine Microbiology一株中肋骨条藻特异溶藻菌的分离鉴定及溶藻特性Isolation and algicidal characteristics of a specific algicidal bacterium on Skeletonemacostatum石新国SHIXin-Guo112李悦LIYue1郑文煌ZHENGWen-Huang1肖宇淳XIAOYu-Chun1刘乐冕LIULe-Mian12陈剑锋CHENJian-Fengjfchen@fzu.edu.cn12*福州大学生物科学与工程学院 福建 福州 350108College of Biological Science and Engineering, Fuzhou University, Fuzhou, Fujian 350108, China福州大学福建海产品废弃物综合利用工程技术研究中心 福建 福州 350108ujian Engineering Research Center for Comprehensive Utilization of Marine Products Waste, Fuzhou University, Fuzhou, Fujian 350108, China陈剑, E-mail:jfchen@fzu.edu.cn
The frequent occurrence of harmful algae blooms causes serious marine ecological problems, which not only directly affects the stability of marine ecosystems, the sustainable use of marine biological resources, and the healthy development of marine aquaculture, but also lead to a serious threat to human health and even life. Efficient algae-lysing bacteria are one of the effective tools to control harmful algae blooms by biological methods.
Objective
Isolation and molecular identification of algae-lysing bacteria with high algicidal effect on Skeletonemacostatum, and studying the algae-lysing mechanism of this strain and the characteristics of algae-lysing substances secreted by algae-lysing bacteria.
Methods
The 2216E plate dilution method was used to isolate and purify bacteria. The 16S rRNA gene sequence was determined to identify the bacterial species. The cell concentration was monitored using microscope for both control and treatment to calculate algicidal rate. The algicidal progress of challenged S. costatum was observed by scanning electron microscope. The algae-dissolving process of S.costatum was studied by using conventional physiological and biochemical methods to study the characteristics of algae-lysing substances. The molecular weight of algae-lysing substances was detected by dialysis bag retention method.
Results
A high-efficiency alginolytic bacterium FDHY-CJ for S.costatum was isolated, which belonged to the genus Alteromonas (Alteromonas sp. FDHY-CJ). The 72-hour treatment of red tide algae by this strain showed that the algae lysis rate was 95.45% against S. costatum and less than 40% for other common red tide algae. Algae-lysing FDHY-CJ lyses algae through extracellular secretions. The algae-lysing properties of algae-lysing substances are not affected by repeated freezing and thawing, but are sensitive to acidity, alkalinity and temperature. Scanning electron microscopy observations showed that the algae-lysing bacteria's algae the substance directly dissolves the cell wall of S. costatum, causing the siliceous shell to open and the contents to flow out, then the algae was lysed. The algae-lysing active substance has the characteristics of being precipitated by ethanol and ethyl acetate.
Conclusion
The algae-lysing bacteria FDHY-CJ has specific algae-lysing effect on S. costatum. The bacterial algae-lysing was indirect algae-dissolving through secretion, and the molecular weight of this compound was between 3.5 kD and 10.0 kD.
赤潮藻中肋骨条藻特异溶藻菌溶藻特性Harmful algaeSkeletonemacostatumSpecific algicidal bacteriumAlgicidal characteristics国家自然科学基金41976130国家自然科学基金41606121国家自然科学基金(41976130,41606121)National Natural Science Foundation of China41976130National Natural Science Foundation of China41606121National Natural Science Foundation of China (41976130, 41606121)
为了确定溶藻细菌FDHY-CJ的溶藻方式,将对数期的中肋骨条藻分为4组,每组3个平行,每个平行样为50 mL藻液。第1组加入1 mL (2%体积)溶藻细菌菌液;第2组加入1 mL (2%体积)溶藻细菌的胞外产物,即溶藻细菌的培养液经8 000 r/min离心8 min后用0.22 μm微滤膜过滤的上清;第3组加入1 mL (2%体积)溶藻细菌的菌细胞悬液,即溶藻细菌的培养液经8 000 r/min离心8 min后去上清,用等体积无菌海水重悬清洗一次,再次离心去上清,加入等体积无菌海水重悬菌体备用;第4组加入1 mL (2%体积)无菌细菌培养基作为对照组。计算滤液和细菌细胞处理的培养物的藻细胞浓度和溶藻率。
Phylogenetic tree of algicidal bacteria strain FDHY-CJ based on 16S rRNA gene sequence
Gammaproteobacteria; II: Cytophaga-Flavobacterium-Bacteroidetes. The bootstrap values are shown at the node; Bar 0.05 means the nucleotide substitution rate of 0.05.
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溶藻细菌FDHY-CJ扫描电镜图片
Scanning electron microscopy image of algicidal bacterium FDHY-CJ
van Dolah FM. Diversity of marine and freshwater algal toxins[A]//Botana L. Seafood and Freshwater Toxins: Pharmacology, Physiology, and Detection[M]. New York: CRC Press, 2000: 173-186
LandsbergJHThe effects of harmful algal blooms on aquatic organisms200210211339010.1080/20026491051695
Landsberg JH. The effects of harmful algal blooms on aquatic organisms[J]. Reviews in Fisheries Science, 2002, 10(2): 113-390
Anderson DM, Cembella AD, Hallegraeff GM. Physiological Ecology of Harmful Algal Blooms[M]. Berlin: Springer, 1998: 662
HuangXQJiangXSWangGLPrincipal environmental factors during red tide outbreak of Skeletonema costatum in Yangtse estuary Ⅲ: water temperature, salinity, DO and pH19941343540
Huang XQ, Jiang XS, Wang GL, et al. Principal environmental factors during red tide outbreak of Skeletonema costatum in Yangtse estuary Ⅲ: water temperature, salinity, DO and pH[J]. Marine Science Bulletin, 1994, 13(4): 35-40 (in Chinese)
BuchanALeCleirGRGulvikCAMaster recyclers: features and functions of bacteria associated with phytoplankton blooms2014121068669810.1038/nrmicro3326
Buchan A, LeCleir GR, Gulvik CA, et al. Master recyclers: features and functions of bacteria associated with phytoplankton blooms[J]. Nature Reviews Microbiology, 2014, 12(10): 686-698
PokrzywinskiKLTilneyCLModlaSEffects of the bacterial algicide IRI-160AA on cellular morphology of harmful dinoflagellates20176212713510.1016/j.hal.2016.12.004
Pokrzywinski KL, Tilney CL, Modla S, et al. Effects of the bacterial algicide IRI-160AA on cellular morphology of harmful dinoflagellates[J]. Harmful Algae, 2017, 62: 127-135
LiDXZhangHChenXHMetaproteomics reveals major microbial players and their metabolic activities during the blooming period of a marine dinoflagellate Prorocentrum donghaiense201820263264410.1111/1462-2920.13986
Li DX, Zhang H, Chen XH, et al. Metaproteomics reveals major microbial players and their metabolic activities during the blooming period of a marine dinoflagellate Prorocentrum donghaiense[J]. Environmental Microbiology, 2018, 20(2): 632-644
WichelsAHummertCElbrächterMErratum to bacterial diversity in toxic Alexandrium tamarense blooms off the Orkney isles and the firth of forth20045829310310.1007/s10152-004-0174-6
Wichels A, Hummert C, Elbrächter M, et al. Erratum to bacterial diversity in toxic Alexandrium tamarense blooms off the Orkney isles and the firth of forth[J]. Helgoland Marine Research, 2004, 58(2): 93-103
TilneyCLPokrzywinskiKLCoyneKJEffects of a bacterial algicide, IRI-160AA, on dinoflagellates and the microbial community in microcosm experiments20143921022210.1016/j.hal.2014.08.001
Tilney CL, Pokrzywinski KL, Coyne KJ, et al. Effects of a bacterial algicide, IRI-160AA, on dinoflagellates and the microbial community in microcosm experiments[J]. Harmful Algae, 2014, 39: 210-222
ShiXGLiuLMLiYIsolation of an algicidal bacterium and its effects against the harmful-algal-bloom dinoflagellate Prorocentrum donghaiense (Dinophyceae)201880727910.1016/j.hal.2018.09.003
Shi XG, Liu LM, Li Y, et al. Isolation of an algicidal bacterium and its effects against the harmful-algal-bloom dinoflagellate Prorocentrum donghaiense (Dinophyceae)[J]. Harmful Algae, 2018, 80: 72-79
ZhangFXYeQChenQLAlgicidal activity of novel marine bacterium Paracoccus sp. strain Y42 against a harmful algal-bloom-causing dinoflagellate, Prorocentrum donghaiense20188419e0101518
Zhang FX, Ye Q, Chen QL, et al. Algicidal activity of novel marine bacterium Paracoccus sp. strain Y42 against a harmful algal-bloom-causing dinoflagellate, Prorocentrum donghaiense[J]. Applied and Environmental Microbiology, 2018, 84(19): e01015-18
PaulCPohnertGInteractions of the algicidal bacterium Kordia algicida with diatoms: regulated protease excretion for specific algal lysis201166e2103210.1371/journal.pone.0021032
Paul C, Pohnert G. Interactions of the algicidal bacterium Kordia algicida with diatoms: regulated protease excretion for specific algal lysis[J]. PLoS One, 2011, 6(6): e21032
Guillard RRL, Ryther JH. 1962. Studies of marine planktonic diatoms: I. Cyclotella nana Hustedt, and Detonula confervacea (Cleve) gran[J]. Canadian Journal of Microbiology, 1962, 8(2): 229-239
Lin SJ, Cheng SF, Song B, et al. The Symbiodinium kawagutii genome illuminates dinoflagellate gene expression and coral symbiosis[J]. Science, 2015, 350(6261): 691-694
DongXZCaiMYBeijingScience Press2001353390
Dong XZ, Cai MY. Manual of Bacterial Culture Identification[M]. Beijing: Science Press, 2001: 353-390 (in Chinese)
东秀珠蔡妙英北京科学出版社2001353390
东秀珠, 蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社, 2001: 353-390
Bacterial Taxonomy Group, Institute of Microbiology, Chinese Academy of SciencesBeijingScience Press1978150
Bacterial Taxonomy Group, Institute of Microbiology, Chinese Academy of Sciences. Common Identification Methods for Common Bacteria[M]. Beijing: Science Press, 1978: 1-50 (in Chinese)
DeLongEFArchaea in coastal marine environments199289125685568910.1073/pnas.89.12.5685
DeLong EF. Archaea in coastal marine environments[J]. Proceedings of the National Academy of Sciences of the United States of America, 1992, 89(12): 5685-5689
TeelingHFuchsBMBecherDSubstrate-controlled succession of marine bacterioplankton populations induced by a phytoplankton bloom2012336608160861110.1126/science.1218344
Teeling H, Fuchs BM, Becher D, et al. Substrate-controlled succession of marine bacterioplankton populations induced by a phytoplankton bloom[J]. Science, 2012, 336(6081): 608-611
KimJDKimJYParkJKSelective control of the Prorocentrum minimum harmful algal blooms by a novel algal-lytic bacterium Pseudoalteromonas haloplanktis AFMB-0080412009114463472
Kim JD, Kim JY, Park JK, et al. Selective control of the Prorocentrum minimum harmful algal blooms by a novel algal-lytic bacterium Pseudoalteromonas haloplanktis AFMB-008041[J]. Marine Biotechnology, 2009, 11(4): 463-472
RuffSEProbandtDZinkannACIndications for algae-degrading benthic microbial communities in deep-sea sediments along the Antarctic Polar Front201410861610.1016/j.dsr2.2014.05.011
Ruff SE, Probandt D, Zinkann AC, et al. Indications for algae-degrading benthic microbial communities in deep-sea sediments along the Antarctic Polar Front[J]. Deep Sea Research Part Ⅱ: Topical Studies in Oceanography, 2014, 108: 6-16
MeyerNBigalkeAKaulfußAStrategies and ecological roles of algicidal bacteria201741688089910.1093/femsre/fux029
Meyer N, Bigalke A, Kaulfuß A, et al. Strategies and ecological roles of algicidal bacteria[J]. FEMS Microbiology Reviews, 2017, 41(6): 880-899
AndersonCRBerdaletEKudelaRMScaling up from regional case studies to a global harmful algal bloom observing system2019625010.3389/fmars.2019.00250
Anderson CR, Berdalet E, Kudela RM, et al. Scaling up from regional case studies to a global harmful algal bloom observing system[J]. Frontiers in Marine Science, 2019, 6: 250
YangXRLiXYZhouYYNovel insights into the algicidal bacterium DH77-1 killing the toxic dinoflagellate Alexandrium tamarense2014482-48311612410.1016/j.scitotenv.2014.02.125
Yang XR, Li XY, Zhou YY, et al. Novel insights into the algicidal bacterium DH77-1 killing the toxic dinoflagellate Alexandrium tamarense[J]. Science of the Total Environment, 2014, 482-483: 116-124
KristyantoSKimJIsolation of marine algicidal bacteria from surface seawater and sediment samples associated with harmful algal blooms in Korea20165214048
Kristyanto S, Kim J. Isolation of marine algicidal bacteria from surface seawater and sediment samples associated with harmful algal blooms in Korea[J]. Korean Journal of Microbiology, 2016, 52(1): 40-48
SuJQYangXRZhengTLIsolation and characterization of a marine algicidal bacterium against the toxic dinoflagellate Alexandrium tamarense20076679981010.1016/j.hal.2007.04.004
Su JQ, Yang XR, Zheng TL, et al. Isolation and characterization of a marine algicidal bacterium against the toxic dinoflagellate Alexandrium tamarense[J]. Harmful Algae, 2007, 6(6): 799-810
Guan CW. Study on algicidal characteristics and mechanism of a novel algicidal bacterium Bacillus sp. LP-10 isolated from East China Sea[D]. Xiamen: Master's Thesis of Xiamen University, 2014 (in Chinese)
PokrzywinskiKLPlaceARWarnerMEInvestigation of the algicidal exudate produced by Shewanella sp. IRI-160 and its effect on dinoflagellates201219232910.1016/j.hal.2012.05.002
Pokrzywinski KL, Place AR, Warner ME, et al. Investigation of the algicidal exudate produced by Shewanella sp. IRI-160 and its effect on dinoflagellates[J]. Harmful Algae, 2012, 19: 23-29
ParkJHYoshinagaINishikawaTAlgicidal bacteria in particle-associated form and in free-living form during a diatom bloom in the Seto Inland Sea, Japan2010602151161
Park JH, Yoshinaga I, Nishikawa T, et al. Algicidal bacteria in particle-associated form and in free-living form during a diatom bloom in the Seto Inland Sea, Japan[J]. Aquatic Microbial Ecology, 2010, 60(2): 151-161
MayaliXAzamFAlgicidal bacteria in the sea and their impact on algal blooms200451213914410.1111/j.1550-7408.2004.tb00538.x
Mayali X, Azam F. Algicidal bacteria in the sea and their impact on algal blooms[J]. Journal of Eukaryotic Microbiology, 2004, 51(2): 139-144
ChoJYAlgicidal activity of marine Alteromonas sp. KNS-16 and isolation of active compounds20127681452145810.1271/bbb.120102
Cho JY. Algicidal activity of marine Alteromonas sp. KNS-16 and isolation of active compounds[J]. Bioscience, Biotechnology, and Biochemistry, 2012, 76(8): 1452-1458
TakamotoSYamadaKEzuraYProduction of bacteriolytic enzymes during the growth of a marine bacterium Alteromonas sp. No. 8-R1994406499508
Takamoto S, Yamada K, Ezura Y. Production of bacteriolytic enzymes during the growth of a marine bacterium Alteromonas sp. No. 8-R[J]. The Journal of General and Applied Microbiology, 1994, 40(6): 499-508
LongRAQureshiAFaulknerDJ2-n-pentyl-4-quinolinol produced by a marine Alteromonas sp. and its potential ecological and biogeochemical roles2003691568576
Long RA, Qureshi A, Faulkner DJ, et al. 2-n-pentyl-4-quinolinol produced by a marine Alteromonas sp. and its potential ecological and biogeochemical roles[J]. Applied and Environmental Microbiology, 2003, 69(1): 568-576
WangHButtLRooksPCharacterisation of algicidal bacterial exometabolites against the lipid-accumulating diatom Skeletonema sp2016131610.1016/j.algal.2015.11.012
Wang H, Butt L, Rooks P, et al. Characterisation of algicidal bacterial exometabolites against the lipid-accumulating diatom Skeletonema sp.[J]. Algal Research, 2016, 13: 1-6
ImaiIKimuraSResistance of the fish-killing dinoflagellate Cochlodinium polykrikoides against algicidal bacteria isolated from the coastal sea of Japan20087336036710.1016/j.hal.2007.12.010
Imai I, Kimura S. Resistance of the fish-killing dinoflagellate Cochlodinium polykrikoides against algicidal bacteria isolated from the coastal sea of Japan[J]. Harmful Algae, 2008, 7(3): 360-367
ImaiIIshidaYSakaguchiKAlgicidal marine bacteria isolated from northern Hiroshima Bay, Japan199561462863610.2331/fishsci.61.628
Imai I, Ishida Y, Sakaguchi K, et al. Algicidal marine bacteria isolated from northern Hiroshima Bay, Japan[J]. Fisheries Science, 1995, 61(4): 628-636