辽东湾毒害微藻潜在风险分布及环境关联
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摘要
以18S rDNA V4区作为目标基因,利用自行设计的真核浮游植物鉴定引物V4(F/R),结合高通量测序技术,对辽东湾2014年四季海水中真核藻类多样性进行了检测.结果发现了辽东湾毒害藻类共29种,曾引发赤潮或褐潮的种类有19种,可导致鱼类大规模死亡的有8种,主要为甲藻门.有18种疑似为辽东湾新记录种,其中9种在中国海域未见报道;发现麻痹性贝毒(PSP)产毒藻8种,腹泻性贝毒(DSP)产毒藻2种,产虾夷扇贝毒素(YTX)产毒藻3种,神经性贝毒(NSP)产毒藻2种,溶血毒素(hemolysin)产毒藻13种,另外还有1种褐潮产毒藻(EPS).辽东湾检出的毒害藻类中有8种自养型、1种异养型、20种混合营养型藻类.麻痹性贝毒、虾夷扇贝毒素、溶血毒素在夏季风险较高,神经性贝毒在秋季风险较高,腹泻性贝毒在春、夏季风险均较高,辽东湾东西两岸是贝毒风险高发区.辽东湾春季水温对YTX、NSP产毒藻影响显著负相关;夏季COD、Oil、DIN、Pb、Cd对各产毒藻类影响显著正相关;秋季各环境因子对各产毒藻类影响不显著;冬季Zn、As、Oil、DIN对各产毒藻类影响显著正相关,水温对各产毒藻类影响显著负相关,各产毒藻类与环境因子关联性有待于进一步研究.
Taking the V4 region of 18 S rDNA as the target gene, we designed the primer V4(F/R) and adopted high-throughput sequencing technology to evaluate the diversity and relative abundance of the eukaryotic phytoplankton community in different seasons of the year 2014 in Liaodong Bay. The results showed that there were 29 toxic algae in Liaodong Bay, including 8 kinds of paralytic shellfish poisoning(PSP) algae, 2 kinds of diarrhetic shellfish poisoning(DSP) algae, 3 kinds of yessotoxic(YTX) algae, 2 kinds of neurotoxic shellfish poisoning(NSP) algae, 13 kinds of hemolysin, and a kind of exopolysaccharide(EPS) algae. Of these, 19 toxic algae were reported to have caused either red or brown tides. Eight of the toxic algae, most of which belonged to Dinophyta, were capable of causing large-scale fish deaths. We found 18 algal species that had been previously undetected or unreported in Liaodong Bay, in which 9 algal species even had not previously appeared in other Chinese waters. The algal species had different trophic modes, including 8 kinds of autotrophs, 20 kinds of mixotrophs, and a kind of heterotroph. There were high risks from PSP, YTX, and NSP in summer;high risks from NSP in autumn; high risks from DSP in spring and summer; and the eastern and western waters of Liaodong Bay were prone to shellfish toxicity disasters. During spring, the water temperature was significantly and negatively correlated with the biomass of YTX and NSP algae. During summer, some environmental factors, such as chemical oxygen demand, oil, dissolved inorganic nitrogen(DIN), lead, and cadmium, were significantly and positively correlated with toxin-producing algae, but zinc, arsenic, oil, and DIN were positively correlated with toxic algae in the winter. The environmental factors were not significantly correlated with toxic algae in the autumn. The correlations between toxic algae and environmental factors should be further explored.
Taking the V4 region of 18 S rDNA as the target gene, we designed the primer V4(F/R) and adopted high-throughput sequencing technology to evaluate the diversity and relative abundance of the eukaryotic phytoplankton community in different seasons of the year 2014 in Liaodong Bay. The results showed that there were 29 toxic algae in Liaodong Bay, including 8 kinds of paralytic shellfish poisoning(PSP) algae, 2 kinds of diarrhetic shellfish poisoning(DSP) algae, 3 kinds of yessotoxic(YTX) algae, 2 kinds of neurotoxic shellfish poisoning(NSP) algae, 13 kinds of hemolysin, and a kind of exopolysaccharide(EPS) algae. Of these, 19 toxic algae were reported to have caused either red or brown tides. Eight of the toxic algae, most of which belonged to Dinophyta, were capable of causing large-scale fish deaths. We found 18 algal species that had been previously undetected or unreported in Liaodong Bay, in which 9 algal species even had not previously appeared in other Chinese waters. The algal species had different trophic modes, including 8 kinds of autotrophs, 20 kinds of mixotrophs, and a kind of heterotroph. There were high risks from PSP, YTX, and NSP in summer;high risks from NSP in autumn; high risks from DSP in spring and summer; and the eastern and western waters of Liaodong Bay were prone to shellfish toxicity disasters. During spring, the water temperature was significantly and negatively correlated with the biomass of YTX and NSP algae. During summer, some environmental factors, such as chemical oxygen demand, oil, dissolved inorganic nitrogen(DIN), lead, and cadmium, were significantly and positively correlated with toxin-producing algae, but zinc, arsenic, oil, and DIN were positively correlated with toxic algae in the winter. The environmental factors were not significantly correlated with toxic algae in the autumn. The correlations between toxic algae and environmental factors should be further explored.
引文
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[6]宋伦,刘卫东,吴景,等.有害甲藻Stoeckeria algicida在辽东湾的时空分布[J].生态学报,2017,37(4):1339-1345.
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[15]Dong Shaojie,Bi Xiangdong,N.WANG,Song Lun,et al.Algicidal activities of Cladophora fracta on red tide-forming microalgae Heterosigma akashiwo and Gymnodinium breve.Allelopathy Journal,2016,37(2):231-240.
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[19]Quast C,Pruesse E,Yilmaz P,et al.The SILVA ribosomal RNA gene database project:improved data processing and web-based tools.Nucleic Acids Research,2013,41:590-596.
[20]Rao Fu,Jun Gong.Single cell analysis linking ribosomal(r)DNA and r RNA copy numbers to cell size and growth rate provides insights into molecular protistan ecology[J].Journal of Eukaryotic Microbiology2017,64:885–896.
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[22]赵冬至.中国典型海域赤潮灾害发生规律[M].北京:海洋出版社,2010:1-389.
[23]宋伦,宋广军.辽东湾浮游植物生态特征研究[M].沈阳:辽宁科学技术出版社,2016:437-440.
[24]Xu Xin,Yu Zhiming,Cheng Fangjin,et al.Molecular diversity and ecological characteristics of the eukaryotic phytoplankton community in the coastal waters of the Bohai sea,China[J].Harmful Algae,2017,61:13-22.
[25]Jeong H J,Kim J S,Park J Y,et al.Stoeckeria algicida n.gen.,n.sp.(Dinophyceae)from the Coastal Waters off Southern Korea:Morphology and Small Subunit Ribosomal DNA Gene Sequence[J].The Journal of Eukaryotic Microbiology,2005,52(4):382-390.
[26]Jeong H J,Kim J S,Kim J H,et al.Feeding and grazing impact of the newly described heterotrophic dinoflagellate Stoeckeria algicida on the harmful alga Heterosigma akashiwo[J].Marine Ecology Progress Series,2005,295:69-78.
[27]Xing Xiaoli,Lin Xuyin,Chen Changping,et al.Observations of several cryptomonad flagellates from China Sea by scanning electron microscopy[J].Journal of Systematics Evolution,2008,46(2):205-212.
[28]Sarno D,Wiebe H C F K,Linda K M,et al.Diversity in the genus Skeletonema(Bacillariophyceae).II.An assesment of the taxonomy of S.costatum-like species with the description of four new species[J].Journal of Phycology,2005,41(1):151-176.
[29]王雪芹,王光华,乔飞,等.高通量测序及其在食物网解析中的应用进展[J].生态学报,2017,37(8):2530-2539.
[30]宋伦,宋广军,王年斌,等.辽东湾网采浮游植物粒级结构的胁迫响应[J].中国环境科学,2015,35(9):2764-2771.
[31]宋伦,宋广军,王年斌,等.辽东湾浮游生物粒径结构稳定性分析[J].中国环境科学,2015,35(10):3117-3126.
[32]Bruland K W,Orians K J,Cowen J P.Reactive trace metals in the stratified central North Pacific[J].Geochimica et Cosmochimica Acta,1994,58(15):3171-3182.
[33]张玉凤,田金,杨爽,等.大连湾海域营养盐时空分布、结构特征及其生态响应[J].中国环境科学,2015,1:236-243.
[34]李清雪,陶建华.应用浮游植物群落结构指数评价海域富营养化[J].中国环境科学,1999,19(6):548-551.
[35]邓春梅,于志刚.东海、南黄海浮游植物粒级结构及环境影响因素分析[J].中国海洋大学学报,2008,38(5):791-798.
[2]Pearse M C,H?kan E,Philipp H.Freeze-drying for the stabilisation of shellfish toxinsin mussel tissue(Mytilus edulis)reference materials[J].Analytical and Bioanalytical Chemistry,2007,387:2475–2486.
[3]Suzuki T,Quilliam M A.LC-MS/MS analysis of diarrhetic shellfish poisoning(DSP)toxins,okadaic acid and dinophysistoxin analogues,and other lipophilic toxins[J].Analytical Sciences,2011,27(6):571-584.
[4]Armi Z,Turki S,Trabelsi E,et al.Occurrence of diarrhetic shellfish poisoning(DSP)toxins in clams(Ruditapes decussatus)from Tunis north lagoon[J].Environ Monit Assess,2012,184(8):5085-5095.
[5]宋伦,宋广军,王年斌,等.黄渤海产毒藻类及贝毒研究现状[J].水产科学,2013,32(7):430-436.
[6]宋伦,刘卫东,吴景,等.有害甲藻Stoeckeria algicida在辽东湾的时空分布[J].生态学报,2017,37(4):1339-1345.
[7]田华.麻痹性贝毒的累积、转化、排出过程及预警诊断指标研究[D].青岛:中国海洋大学,2009:1-93.
[8]刘卫东,宋伦,吴景.环境样本中微型和微微型浮游植物高通量测序的引物优化[J].生态学报,2017,37(12):4208-4216.
[9]Amaral Z L,Artigas L F,Baross J,et al.A global census of marine microbes.life in the world′s oceans:diversity,distribution and abundance[M].Oxford:Blackwell Publishing Ltd.,2010:223-245.
[10]于杰.浮游生物多样性高效检测技术的建立及其在渤海褐潮研究中的应用[D].青岛:中国海洋大学,2014.
[11]宋伦,吴景,宋永刚,等.褐潮致灾种抑食金球藻在辽东湾的分布[J].环境科学研究,2017,30(4):537-544.
[12]宋伦,毕相东.渤海海洋生态灾害及应急处置[M].沈阳:辽宁科学技术出版社,2015:175-210.
[13]Song Lun,Yang Guojun,Wang Nianbin,et al.Relationship between environmental factors and plankton in the Bayuquan Port,Liaodong Bay,China:a five-year study[J].Chinese Journal of Oceanology and Limnology.2016,34(4):654-671.
[14]DB21/T 2254-2014海洋贝毒预警监测技术规程[S].
[15]Dong Shaojie,Bi Xiangdong,N.WANG,Song Lun,et al.Algicidal activities of Cladophora fracta on red tide-forming microalgae Heterosigma akashiwo and Gymnodinium breve.Allelopathy Journal,2016,37(2):231-240.
[16]GB/T 12763-2007海洋调查规范[S].
[17]GB17378-2007海洋监测规范[S].
[18]宋伦,吴景,刘卫东,等.渤海长兴岛海域微型和微微型浮游植物多样性[J].环境科学研究,2016,29(11):1635-1642.
[19]Quast C,Pruesse E,Yilmaz P,et al.The SILVA ribosomal RNA gene database project:improved data processing and web-based tools.Nucleic Acids Research,2013,41:590-596.
[20]Rao Fu,Jun Gong.Single cell analysis linking ribosomal(r)DNA and r RNA copy numbers to cell size and growth rate provides insights into molecular protistan ecology[J].Journal of Eukaryotic Microbiology2017,64:885–896.
[21]刘瑞玉.中国海洋生物名录[M].北京:科学出版社,2008:301-870.
[22]赵冬至.中国典型海域赤潮灾害发生规律[M].北京:海洋出版社,2010:1-389.
[23]宋伦,宋广军.辽东湾浮游植物生态特征研究[M].沈阳:辽宁科学技术出版社,2016:437-440.
[24]Xu Xin,Yu Zhiming,Cheng Fangjin,et al.Molecular diversity and ecological characteristics of the eukaryotic phytoplankton community in the coastal waters of the Bohai sea,China[J].Harmful Algae,2017,61:13-22.
[25]Jeong H J,Kim J S,Park J Y,et al.Stoeckeria algicida n.gen.,n.sp.(Dinophyceae)from the Coastal Waters off Southern Korea:Morphology and Small Subunit Ribosomal DNA Gene Sequence[J].The Journal of Eukaryotic Microbiology,2005,52(4):382-390.
[26]Jeong H J,Kim J S,Kim J H,et al.Feeding and grazing impact of the newly described heterotrophic dinoflagellate Stoeckeria algicida on the harmful alga Heterosigma akashiwo[J].Marine Ecology Progress Series,2005,295:69-78.
[27]Xing Xiaoli,Lin Xuyin,Chen Changping,et al.Observations of several cryptomonad flagellates from China Sea by scanning electron microscopy[J].Journal of Systematics Evolution,2008,46(2):205-212.
[28]Sarno D,Wiebe H C F K,Linda K M,et al.Diversity in the genus Skeletonema(Bacillariophyceae).II.An assesment of the taxonomy of S.costatum-like species with the description of four new species[J].Journal of Phycology,2005,41(1):151-176.
[29]王雪芹,王光华,乔飞,等.高通量测序及其在食物网解析中的应用进展[J].生态学报,2017,37(8):2530-2539.
[30]宋伦,宋广军,王年斌,等.辽东湾网采浮游植物粒级结构的胁迫响应[J].中国环境科学,2015,35(9):2764-2771.
[31]宋伦,宋广军,王年斌,等.辽东湾浮游生物粒径结构稳定性分析[J].中国环境科学,2015,35(10):3117-3126.
[32]Bruland K W,Orians K J,Cowen J P.Reactive trace metals in the stratified central North Pacific[J].Geochimica et Cosmochimica Acta,1994,58(15):3171-3182.
[33]张玉凤,田金,杨爽,等.大连湾海域营养盐时空分布、结构特征及其生态响应[J].中国环境科学,2015,1:236-243.
[34]李清雪,陶建华.应用浮游植物群落结构指数评价海域富营养化[J].中国环境科学,1999,19(6):548-551.
[35]邓春梅,于志刚.东海、南黄海浮游植物粒级结构及环境影响因素分析[J].中国海洋大学学报,2008,38(5):791-798.
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