我国北部湾球形棕囊藻(Phaeocystis globosa)的表面形态和细胞超微结构的电镜观察
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摘要
本文首次对分离于我国广西北部湾的一株棕囊藻(Phaeocystis)纯培养进行了形态学研究。通过对其核糖体大亚基序列(LSU rDNA)进行了系统进化分析,并运用光学显微镜、扫描电镜和透射电镜等,对该藻的游动单细胞和囊体细胞表面结构和超微结构进行了详细的观察。结果表明,通过分子序列和形态学特征确证了该纯培养为球形棕囊藻(Phaeocystisglobosa)。通过分子系统进化分析,从北部湾获得的球形棕囊藻与同属的P. antarctica和P. rex的亲缘关系较近,但与P. jahnii的亲缘关系相对较远。我们发现该藻至少有两种类型的细胞:具鞭毛游动细胞和无鞭毛囊体细胞,其大小分别为2.30—3.98μm和3.69—6.49μm。具鞭毛游动细胞表面具有1—2个位置不固定但形状较为规则的圆盘状凸起,细胞表面的鳞片大小无明显差别;囊体细胞均匀地分布在囊体上,囊体细胞表面光滑,有三个较短的附属物。具鞭毛游动细胞和囊体细胞有2个或4个叶绿体,具鞭毛游动细胞的细胞核位于细胞的中下部,而囊体细胞的细胞核则位于细胞的中上部。细胞分泌物在扫描电镜样品处理过程后形成多数与细胞相连的丝状物,且丝状物形成较为规则的五角星形结构,每个角与一根丝相连,有些丝状体上有"瘤状"结构。研究结果填补了我国在球形棕囊藻形态学和超微结构研究上的空白,为深入认识其生物学特征及其成囊机理提供基础。
A strain of Phaeocystis sp was isolated from the Beibu Gulf, Guangxi, China, and its fine morphology was reported first time in the country. The flagellate and colonial cells were observed by light microscope, scanning and transmission electron microscopy, and molecular phylogeny using partial large subunit ribosomal RNA gene(LSU rDNA)sequence was also analyzed. Results show that this species is Phaeocystis globosa identified from its molecular sequence and morphological characterization. The LSU rDNA sequence and phylogenetic tree reveal that this species is more closely related to P. antarctica and P. rex, but is diverged from P. jahnii. Two cell types are observed in P. globosa: small flagellates and colonial cells, and their sizes are 2.30—3.98μm and 3.69—6.49μm, respectively. The small flagellate cell has 1—2 circular bulges, and the cell surface is covered with an external layer of scales without difference in size. Colonial cells are randomly distributed at the periphery of spherical colonies, which have smooth surface without scales and have three short appendages. Two or four chloroplasts are observed and the nucleus locates in the middle and upper parts. The filaments extruded by the cells form pentagonal star-like structures, the structure connected five filaments at their proximal ends, and a tuberculiform structure appears in some filaments. This study fills the blank in fine morphology and ultrastructure of P. globosa in China and provides a base on understanding its biological characterization and mechanisms of colony formation.
A strain of Phaeocystis sp was isolated from the Beibu Gulf, Guangxi, China, and its fine morphology was reported first time in the country. The flagellate and colonial cells were observed by light microscope, scanning and transmission electron microscopy, and molecular phylogeny using partial large subunit ribosomal RNA gene(LSU rDNA)sequence was also analyzed. Results show that this species is Phaeocystis globosa identified from its molecular sequence and morphological characterization. The LSU rDNA sequence and phylogenetic tree reveal that this species is more closely related to P. antarctica and P. rex, but is diverged from P. jahnii. Two cell types are observed in P. globosa: small flagellates and colonial cells, and their sizes are 2.30—3.98μm and 3.69—6.49μm, respectively. The small flagellate cell has 1—2 circular bulges, and the cell surface is covered with an external layer of scales without difference in size. Colonial cells are randomly distributed at the periphery of spherical colonies, which have smooth surface without scales and have three short appendages. Two or four chloroplasts are observed and the nucleus locates in the middle and upper parts. The filaments extruded by the cells form pentagonal star-like structures, the structure connected five filaments at their proximal ends, and a tuberculiform structure appears in some filaments. This study fills the blank in fine morphology and ultrastructure of P. globosa in China and provides a base on understanding its biological characterization and mechanisms of colony formation.
引文
丁立,王小冬,王艳,2018.不同摄食压力下球形棕囊藻的防御策略.海洋科学进展,36(1):139-144,145
王艳,王小冬,李韶山,2010.充气和搅动对球形棕囊藻生长及囊体形成的影响.生态学报,30(12):3368-3374
王艳,齐雨藻,李韶山,2007.球形棕囊藻生长的营养需求研究.水生生物学报,31(1):24-29
王艳,齐雨藻,沈萍萍等,2003.温度和盐度对球形棕囊藻细胞DMSP产量的影响.水生生物学报,27(4):367-371
王超,李新辉,赖子尼等,2010.珠江口球形棕囊藻(Phaeocystis globosa)赤潮后期的浮游植物群落结构特征研究.生态科学,29(2):140-146
王小冬,王艳,2012.高摄食压力下球形棕囊藻凝聚体的形成.生态学报,32(2):414-420
刘玉芳,赵玲,尹平河等,2011.季磷盐改性蒙脱土去除球形棕囊藻的实验研究.中国环境科学,31(8):1295-1299
齐雨藻,沈萍萍,王艳,2001.棕囊藻属(Phaeocystis)的分类与生活史(综述).热带亚热带植物学报,9(2):174-184
江涛,江天久,李鹏等,2006.湛江球形棕囊藻赤潮除藻试验.热带亚热带植物学报,14(1):14-18
曲凌云,吕颂辉,高春蕾等,2008.棕囊藻渤海株核糖体18SrDNA和ITS基因结构序列分析.海洋科学进展,26(2):200-206
杨和福,2004.棕囊藻的生物学概述I.形态分类和生理生态学.东海海洋,22(1):49-63
沈萍萍,王艳,齐雨藻等,2000.球形棕囊藻的生长特性及生活史研究.水生生物学报,24(6):635-643
陈月琴,王宁,周惠等,2002.棕囊藻赤潮原因种的分子鉴定和起源分析.海洋学报,24(6):99-103
陈丽芬,章群,骆育敏等,2003.18S rDNA序列分析鉴定棕囊藻香港株P2为球形棕囊藻.生态科学,22(4):349-350
陈菊芳,徐宁,江天久等,1999.中国赤潮新记录种-球形棕囊藻(Phaeocystis globosa).暨南大学学报(自然科学版),20(3):124-129
晏荣军,尹平河,潘剑宇等,2006.棕囊藻囊泡的培养与去除研究.热带海洋学报,25(3):69-71
唐赢中,胡章喜,邓蕴彦,2016.休眠孢囊作为甲藻有害藻华年际频发和地理扩散一种关键机制的研究进展.海洋科学集刊,51:132-154
曹西华,俞志明,邱丽霞,2017.改性黏土法消除球形棕囊藻赤潮的现场实验与效果评估.海洋与湖沼,48(4):753-759
覃仙玲,赖俊翔,陈波等,2016.棕囊藻北部湾株的18SrDNA分子鉴定.热带亚热带植物学报,24(2):176-181
Andersen R A,Bailey J C,Decelle J et al,2015.Phaeocystis rex sp.nov.(Phaeocystales,Prymnesiophyceae):a new solitary species that produces a multilayered scale cell covering.European Journal of Phycology,50(2):207-222
Antajan E,Chrétiennot-Dinet M J,Leblanc C et al,2004.19′-Hexanoyloxyfucoxanthin may not be the appropriate pigment to trace occurrence and fate of Phaeocystis:the case of P.globosa in Belgian coastal waters.Journal of Sea Research,52(3):165-177
Brussaard C P D,Kuipers B,Veldhuis M J W,2005.A mesocosm study of Phaeocystis globosa population dynamics:I.Regulatory role of viruses in bloom control.Harmful Algae,4(5):859-874
Cadée G C,1996.Accumulation and sedimentation of Phaeocystis globosa in the Dutch Wadden Sea.Journal of Sea Research,36(3-4):321-327
Darriba D,Taboada G L,Doallo R et al,2012.jModelTest 2:more models,new heuristics and parallel computing.Nature Methods,9(8):772
Daugbjerg N,Hansen G,Larsen J et al,2000.Phylogeny of some of the major genera of dinoflagellates based on ultrastructure and partial LSU rDNA sequence data,including the erection of three new genera of unarmoured dinoflagellates.Phycologia,39(4):302-317
Edvardsen B,Eikrem W,Green J C et al,2000.Phylogenetic reconstructions of the Haptophyta inferred from 18s ribosomal DNA sequences and available morphological data.Phycologia,39(1):19-35
Guillard R R L,1975.Culture of phytoplankton for feeding marine invertebrates.In:Smith W L,Chanley M H eds.Culture of Marine Invertebrate Animals.New York,USA:Plenum Press,26-60
Guindon S,2010.Bayesian estimation of divergence times from large sequence alignments.Molecular Biology and Evolution,27(8):1768-1781
Guindon S,Gascuel O,2003.A simple,fast,and accurate algorithm to estimate large phylogenies by maximum likelihood.Systematic Biology,52(5):696-704
Guiselin N,Courcot L,Artigas L F et al,2009.An optimised protocol to prepare Phaeocystis globosa morphotypes for scanning electron microscopy observation.Journal of Microbiological Methods,77(1):119-123
Hall T A,1999.BioEdit:a user-friendly biological sequence alignment editor and analysis program for Windows95/98/NT.Nucleic Acids Symposium Series,41:95-98
Hallegraeff G M,1983.Scale-bearing and loricate nanoplankton from the East Australian Current.Botanica Marina,26(11):493-516
Hamm C E,2000.Architecture,ecology and biogeochemistry of Phaeocystis colonies.Journal of Sea Research,43(3-4):307-315
Katoh K,Misawa K,Kuma K I et al,2002.MAFFT:a novel method for rapid multiple sequence alignment based on fast Fourier transform.Nucleic Acids Research,30(14):3059-3066
Lange M,Chen Y Q,Medlin L K,2002.Molecular genetic delineation of Phaeocystis species(Prymnesiophyceae)using coding and non-coding regions of nuclear and plastid genomes.European Journal of Phycology,37(1):77-92
Medlin L,Zingone A,2007.A taxonomic review of the genus Phaeocystis.Biogeochemistry,83(1-3):3-18
Moestrup?,1979.Identification by electron microscopy of marine nanoplankton from New Zealand,including the description of four new species.New Zealand Journal of Botany,17(1):61-95
Parke M,Green J C,Manton I,1971.Observations on the fine structure of zoids of the genus Phaeocystis[Haptophyceae].Journal of the Marine Biological Association of the United Kingdom,51(4):927-941
Qi Y Z,Chen J F,Wang Z H et al,2004.Some observations on harmful algal bloom(HAB)events along the coast of Guangdong,southern China in 1998.Hydrobiologia,512(1-3):209-214
Riegman R,Noordeloos A A M,Cadée G C,1992.Phaeocystis blooms and eutrophication of the continental coastal zones of the North Sea.Marine Biology,112(3):479-484
Ronquist F,Huelsenbeck J P,2003.MrBayes 3:Bayesian phylogenetic inference under mixed models.Bioinformatics,19:1572-1574
Rousseau V,Chrétiennot-Dinet M J,Jacobsen A et al,2007.The life cycle of Phaeocystis:state of knowledge and presumptive role in ecology.Biogeochemistry,83(1-3):29-47
Rousseau V,Lantoine F,Rodriguez F et al,2013.Characterization of Phaeocystis globosa(Prymnesiophyceae),the blooming species in the Southern North Sea.Journal of Sea Research,76:105-113
Rousseau V,Vaulot D,Casotti R et al,1994.The life cycle of Phaeocystis(Prymnesiophycaea):evidence and hypotheses.Journal of Marine Systems,5(1):23-39
Schoemann V,Becquevort S,Stefels J et al,2005.Phaeocystis blooms in the global ocean and their controlling mechanisms:a review.Journal of Sea Research,53(1-2):43-66
Scholin C A,Herzog M,Sogin M et al,1994.Identification of group-and strain-specific genetic markers for globally distributed Alexandrium(Dinophyceae).II.Sequence analysis of a fragment of the LSU rRNA gene.Journal of Phycology,30(6):999-1011
Shen P P,Qi Y Z,Wang Y et al,2011.Phaeocystis globosa Scherffel,a harmful microalga,and its production of dimethylsulfoniopropionate.Chinese Journal of Oceanology and Limnology,29(4):869-873
Smith Jr W O,Liu X,Tang K W et al,2014.Giantism and its role in the harmful algal bloom species Phaeocystis globosa.Deep Sea Research Part II:Topical Studies in Oceanography,101:95-106
Spurr A R,1969.A low-viscosity epoxy resin embedding medium for electron microscopy.Journal of Ultrastructure Research,26(1-2):31-43
Vaulot D,Birrien J L,Marie D et al,1994.Morphology,ploidy,pigment composition,and genome size of cultured strains of Phaeocystis(Prymnesiophyceae).Journal of Phycology,30(6):1022-1035
Veldhuis M J W,Brussaard C P D,Noordeloos A A M,2005.Living in a Phaeocystis colony:a way to be a successful algal species.Harmful Algae,4(5):841-858
Verity P G,Brussaard C P,Nejstgaard J C et al,2007.Current understanding of Phaeocystis ecology and biogeochemistry,and perspectives for future research.Biogeochemistry,83(1-3):311-330
Wang X D,Wang Y,Ou L J et al,2015.Allocation costs associated with induced defense in Phaeocystis globosa(Prymnesiophyceae):the effects of nutrient availability.Scientific Reports,5:10850
Xu N,Huang B Z,Hu Z X et al,2017.Effects of temperature,salinity,and irradiance on the growth of harmful algal bloom species Phaeocystis globosa Scherffel(Prymnesiophyceae)isolated from the South China Sea.Chinese Journal of Oceanology and Limnology,35(3):557-565
Zingone A,Chrétiennot-Dinet M J,Lange M et al,1999.Morphological and genetic characterization of Phaeocystis cordata and P.jahnii(Prymnesiophyceae),two new species from the Mediterranean Sea.Journal of Phycology,35(6):1322-1337
王艳,王小冬,李韶山,2010.充气和搅动对球形棕囊藻生长及囊体形成的影响.生态学报,30(12):3368-3374
王艳,齐雨藻,李韶山,2007.球形棕囊藻生长的营养需求研究.水生生物学报,31(1):24-29
王艳,齐雨藻,沈萍萍等,2003.温度和盐度对球形棕囊藻细胞DMSP产量的影响.水生生物学报,27(4):367-371
王超,李新辉,赖子尼等,2010.珠江口球形棕囊藻(Phaeocystis globosa)赤潮后期的浮游植物群落结构特征研究.生态科学,29(2):140-146
王小冬,王艳,2012.高摄食压力下球形棕囊藻凝聚体的形成.生态学报,32(2):414-420
刘玉芳,赵玲,尹平河等,2011.季磷盐改性蒙脱土去除球形棕囊藻的实验研究.中国环境科学,31(8):1295-1299
齐雨藻,沈萍萍,王艳,2001.棕囊藻属(Phaeocystis)的分类与生活史(综述).热带亚热带植物学报,9(2):174-184
江涛,江天久,李鹏等,2006.湛江球形棕囊藻赤潮除藻试验.热带亚热带植物学报,14(1):14-18
曲凌云,吕颂辉,高春蕾等,2008.棕囊藻渤海株核糖体18SrDNA和ITS基因结构序列分析.海洋科学进展,26(2):200-206
杨和福,2004.棕囊藻的生物学概述I.形态分类和生理生态学.东海海洋,22(1):49-63
沈萍萍,王艳,齐雨藻等,2000.球形棕囊藻的生长特性及生活史研究.水生生物学报,24(6):635-643
陈月琴,王宁,周惠等,2002.棕囊藻赤潮原因种的分子鉴定和起源分析.海洋学报,24(6):99-103
陈丽芬,章群,骆育敏等,2003.18S rDNA序列分析鉴定棕囊藻香港株P2为球形棕囊藻.生态科学,22(4):349-350
陈菊芳,徐宁,江天久等,1999.中国赤潮新记录种-球形棕囊藻(Phaeocystis globosa).暨南大学学报(自然科学版),20(3):124-129
晏荣军,尹平河,潘剑宇等,2006.棕囊藻囊泡的培养与去除研究.热带海洋学报,25(3):69-71
唐赢中,胡章喜,邓蕴彦,2016.休眠孢囊作为甲藻有害藻华年际频发和地理扩散一种关键机制的研究进展.海洋科学集刊,51:132-154
曹西华,俞志明,邱丽霞,2017.改性黏土法消除球形棕囊藻赤潮的现场实验与效果评估.海洋与湖沼,48(4):753-759
覃仙玲,赖俊翔,陈波等,2016.棕囊藻北部湾株的18SrDNA分子鉴定.热带亚热带植物学报,24(2):176-181
Andersen R A,Bailey J C,Decelle J et al,2015.Phaeocystis rex sp.nov.(Phaeocystales,Prymnesiophyceae):a new solitary species that produces a multilayered scale cell covering.European Journal of Phycology,50(2):207-222
Antajan E,Chrétiennot-Dinet M J,Leblanc C et al,2004.19′-Hexanoyloxyfucoxanthin may not be the appropriate pigment to trace occurrence and fate of Phaeocystis:the case of P.globosa in Belgian coastal waters.Journal of Sea Research,52(3):165-177
Brussaard C P D,Kuipers B,Veldhuis M J W,2005.A mesocosm study of Phaeocystis globosa population dynamics:I.Regulatory role of viruses in bloom control.Harmful Algae,4(5):859-874
Cadée G C,1996.Accumulation and sedimentation of Phaeocystis globosa in the Dutch Wadden Sea.Journal of Sea Research,36(3-4):321-327
Darriba D,Taboada G L,Doallo R et al,2012.jModelTest 2:more models,new heuristics and parallel computing.Nature Methods,9(8):772
Daugbjerg N,Hansen G,Larsen J et al,2000.Phylogeny of some of the major genera of dinoflagellates based on ultrastructure and partial LSU rDNA sequence data,including the erection of three new genera of unarmoured dinoflagellates.Phycologia,39(4):302-317
Edvardsen B,Eikrem W,Green J C et al,2000.Phylogenetic reconstructions of the Haptophyta inferred from 18s ribosomal DNA sequences and available morphological data.Phycologia,39(1):19-35
Guillard R R L,1975.Culture of phytoplankton for feeding marine invertebrates.In:Smith W L,Chanley M H eds.Culture of Marine Invertebrate Animals.New York,USA:Plenum Press,26-60
Guindon S,2010.Bayesian estimation of divergence times from large sequence alignments.Molecular Biology and Evolution,27(8):1768-1781
Guindon S,Gascuel O,2003.A simple,fast,and accurate algorithm to estimate large phylogenies by maximum likelihood.Systematic Biology,52(5):696-704
Guiselin N,Courcot L,Artigas L F et al,2009.An optimised protocol to prepare Phaeocystis globosa morphotypes for scanning electron microscopy observation.Journal of Microbiological Methods,77(1):119-123
Hall T A,1999.BioEdit:a user-friendly biological sequence alignment editor and analysis program for Windows95/98/NT.Nucleic Acids Symposium Series,41:95-98
Hallegraeff G M,1983.Scale-bearing and loricate nanoplankton from the East Australian Current.Botanica Marina,26(11):493-516
Hamm C E,2000.Architecture,ecology and biogeochemistry of Phaeocystis colonies.Journal of Sea Research,43(3-4):307-315
Katoh K,Misawa K,Kuma K I et al,2002.MAFFT:a novel method for rapid multiple sequence alignment based on fast Fourier transform.Nucleic Acids Research,30(14):3059-3066
Lange M,Chen Y Q,Medlin L K,2002.Molecular genetic delineation of Phaeocystis species(Prymnesiophyceae)using coding and non-coding regions of nuclear and plastid genomes.European Journal of Phycology,37(1):77-92
Medlin L,Zingone A,2007.A taxonomic review of the genus Phaeocystis.Biogeochemistry,83(1-3):3-18
Moestrup?,1979.Identification by electron microscopy of marine nanoplankton from New Zealand,including the description of four new species.New Zealand Journal of Botany,17(1):61-95
Parke M,Green J C,Manton I,1971.Observations on the fine structure of zoids of the genus Phaeocystis[Haptophyceae].Journal of the Marine Biological Association of the United Kingdom,51(4):927-941
Qi Y Z,Chen J F,Wang Z H et al,2004.Some observations on harmful algal bloom(HAB)events along the coast of Guangdong,southern China in 1998.Hydrobiologia,512(1-3):209-214
Riegman R,Noordeloos A A M,Cadée G C,1992.Phaeocystis blooms and eutrophication of the continental coastal zones of the North Sea.Marine Biology,112(3):479-484
Ronquist F,Huelsenbeck J P,2003.MrBayes 3:Bayesian phylogenetic inference under mixed models.Bioinformatics,19:1572-1574
Rousseau V,Chrétiennot-Dinet M J,Jacobsen A et al,2007.The life cycle of Phaeocystis:state of knowledge and presumptive role in ecology.Biogeochemistry,83(1-3):29-47
Rousseau V,Lantoine F,Rodriguez F et al,2013.Characterization of Phaeocystis globosa(Prymnesiophyceae),the blooming species in the Southern North Sea.Journal of Sea Research,76:105-113
Rousseau V,Vaulot D,Casotti R et al,1994.The life cycle of Phaeocystis(Prymnesiophycaea):evidence and hypotheses.Journal of Marine Systems,5(1):23-39
Schoemann V,Becquevort S,Stefels J et al,2005.Phaeocystis blooms in the global ocean and their controlling mechanisms:a review.Journal of Sea Research,53(1-2):43-66
Scholin C A,Herzog M,Sogin M et al,1994.Identification of group-and strain-specific genetic markers for globally distributed Alexandrium(Dinophyceae).II.Sequence analysis of a fragment of the LSU rRNA gene.Journal of Phycology,30(6):999-1011
Shen P P,Qi Y Z,Wang Y et al,2011.Phaeocystis globosa Scherffel,a harmful microalga,and its production of dimethylsulfoniopropionate.Chinese Journal of Oceanology and Limnology,29(4):869-873
Smith Jr W O,Liu X,Tang K W et al,2014.Giantism and its role in the harmful algal bloom species Phaeocystis globosa.Deep Sea Research Part II:Topical Studies in Oceanography,101:95-106
Spurr A R,1969.A low-viscosity epoxy resin embedding medium for electron microscopy.Journal of Ultrastructure Research,26(1-2):31-43
Vaulot D,Birrien J L,Marie D et al,1994.Morphology,ploidy,pigment composition,and genome size of cultured strains of Phaeocystis(Prymnesiophyceae).Journal of Phycology,30(6):1022-1035
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