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福建石兜水库产毒拉氏尖头藻(Raphidiopsis raciborskii)遗传多样性

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谭凤娇, 肖鹏, 左俊, 金磊, 程晨, 雷腊梅, 谢丽强, 杨军. 福建石兜水库产毒拉氏尖头藻(Raphidiopsis raciborskii)遗传多样性[J]. 生态毒理学报, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
引用本文: 谭凤娇, 肖鹏, 左俊, 金磊, 程晨, 雷腊梅, 谢丽强, 杨军. 福建石兜水库产毒拉氏尖头藻(Raphidiopsis raciborskii)遗传多样性[J]. 生态毒理学报, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
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Tan Fengjiao, Xiao Peng, Zuo Jun, Jin Lei, Cheng Chen, Lei Lamei, Xie Liqiang, Yang Jun. Genetic Diversity of Toxin-Producing Raphidiopsis raciborskii (Cyanobacterium) Isolated from Shidou Reservoir in Fujian Province[J]. Asian journal of ecotoxicology, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
Citation: Tan Fengjiao, Xiao Peng, Zuo Jun, Jin Lei, Cheng Chen, Lei Lamei, Xie Liqiang, Yang Jun. Genetic Diversity of Toxin-Producing Raphidiopsis raciborskii (Cyanobacterium) Isolated from Shidou Reservoir in Fujian Province[J]. Asian journal of ecotoxicology, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
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福建石兜水库产毒拉氏尖头藻(Raphidiopsis raciborskii)遗传多样性

  • 1. 中国科学院城市环境研究所, 福建省流域生态学重点实验室, 城市环境与健康重点实验室, 水生态健康研究组, 厦门 361021;

  • 2. 中国科学院南京地理与湖泊研究所, 湖泊与环境国家重点实验室, 南京 210008;

  • 3. 中国科学院大学, 北京 100049;

  • 4. 暨南大学生命科学技术学院生态学系, 广州 510632

    • 作者简介: 谭凤娇(1991—),女,博士研究生,研究方向为环境科学、有害蓝藻监测,E-mail:fjtan@iue.ac.cn
    通讯作者: 杨军, E-mail: jyang@iue.ac.cn
  • 基金项目:

    国家自然科学基金资助项目（91851104）；福建省自然科学基金资助项目（2019J02016，2020J01119）；福建省环保科技计划项目（2021R009）

  • 中图分类号: X171.5

Genetic Diversity of Toxin-Producing Raphidiopsis raciborskii (Cyanobacterium) Isolated from Shidou Reservoir in Fujian Province

  • 1. Aquatic EcoHealth Group, Fujian Key Laboratory of Watershed Ecology, Key Laboratory of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen 361021, China;

  • 2. State Key Laboratory of Lake Science and Environment, Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008, China;

  • 3. University of Chinese Academy of Sciences, Beijing 100049, China;

  • 4. Department of Ecology, College of Life Science and Technology, Jinan University, Guangzhou 510632, China

    • Corresponding author: Yang Jun, jyang@iue.ac.cn
  • Fund Project:

  • 摘要: 近几十年以来，全球内陆深水水体拉氏尖头藻（Raphidiopsis raciborskii）的检出率和水华事件日益增多，这种新型水华蓝藻具有产毒和无毒株系，其中能合成拟柱胞藻毒素（cylindrospermopsins，CYNs）的株系主要分布在澳大利亚和东南亚，严重威胁水生态系统和人类健康。本研究于2018年从福建省厦门市石兜水库分离、培养了4株拉氏尖头藻（XM1～XM4），对其形态、遗传多样性和产毒特征进行了研究。显微观察结果表明，4株拉氏尖头藻形态特征基本一致，细胞平均长度为3.79～4.26 μm，藻丝体呈直线型、平均长度为122～199 μm，在环境样品中可观察到异形胞（固氮细胞），使用富含氮的BG11培养基培养后异形胞会消失。酶联免疫吸附（ELISA）试剂盒测定显示，4株拉氏尖头藻均为产毒藻株；液相色谱串联质谱（LC-MS/MS）分析表明，4个藻株能同时合成CYN和脱氧拟柱胞藻毒素（deoxy-CYN），其中XM1、XM2和XM3藻株以CYN为主，在对数生长后期平均浓度为0.71～1.02 pg·cell-1，XM4则以deoxy-CYN为主，平均浓度为0.42 pg·cell-1。16S rRNA基因系统发育分析表明，XM2藻株与非洲藻株同源相似性较高，其他3株（XM1、XM3和XM4）与亚洲、欧洲的藻株同源相似性较高，表明石兜水库拉氏尖头藻种群遗传多样性较高。序列分析发现，XM2藻株具有完整的产CYN基因簇，但是部分关键基因发生了重排变异，cyrB和cyrI基因分别被分割成2部分和3部分，XM2藻株与浙江杭州藻株产毒基因簇最为相似。在气候变化和人类活动多重影响下，我国水源地水库产毒拉氏尖头藻的来源、传播扩散、遗传分化和生态效应有待进一步研究。
    Abstract: In recent decades, the detection of the cyanobacterium Raphidiopsis raciborskii in deep freshwaters has increased, and its blooms were influenced by global climate change and human activity. R. raphidiopsis can synthesize cylindrospermopsins (CYNs), a potent hepatotoxin, which seriously threatens aquatic ecosystems and human health. However, populations of R. raciborskii include both toxic and non-toxic strains in aquatic ecosystems, making risk assessment difficult and precise knowledge of local populations important. The CYNs-producing strains are currently only distributed in tropical and subtropical regions of Australia and Southeast Asia. Recently, toxic stains have been sporadically reported in South China, causing concern for aquatic health. In this study, four new R. raciborskii strains (XM1~XM4) from Shidou Reservoir, Xiamen City, Fujian Province, Southeast China were isolated and cultured in 2018. Their morphological, phylogenetic and toxin-production characteristics were investigated and compared. Our results showed that the morphological characteristics of the four strains of R. raciborskii were the same with straight trichome. The average vegetative cell length of the four strains was 3.79~4.26 μm, and the average trichome length ranged from 122 to 199 μm. Heterocysts (specialized nitrogen fixing cells) were observed in the field samples, but they gradually disappeared after culturing in nitrogen-rich medium (i.e. BG11). The enzyme linked immunosorbent assay (ELISA) indicated that the four strains could produce CYNs. LC-MS/MS analysis revealed that the four strains co-produced CYN and deoxy-cylindrospermopsin (deoxy-CYN). However, XM1, XM2 and XM3 strains mainly produced CYN with mean concentration ranged from 0.71 to 1.02 pg·cell-1, while XM4 strain mainly produced deoxy-CYN with mean concentration of 0.42 pg·cell-1. Phylogenetic analysis, using 16S rRNA sequence, showed that XM2 strain exhibited a higher homology with strains from Africa, while the other three strains (XM1, XM3 and XM4) had higher homology with Asian and European strains. The results suggest that the genetic diversity of the R. raciborskii population in Shidou Reservoir is high. Based on DNA sequencing, we found XM2 strain has the complete cyr gene cluster but gene rearrangement with the cyrB and cyrI genes spilt into two and three parts, respectively. The cyr gene cluster of XM2 strain exhibited a higher similarity with Hangzhou strains (China) than Australian strains. Further research is needed to clarify the strains' source and geographic dispersal, its genomic differentiation and evolution, and the ecological consequences of this bloom-forming R. raciborskii in China.
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  • 收稿日期:  2021-10-12
谭凤娇, 肖鹏, 左俊, 金磊, 程晨, 雷腊梅, 谢丽强, 杨军. 福建石兜水库产毒拉氏尖头藻(Raphidiopsis raciborskii)遗传多样性[J]. 生态毒理学报, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
引用本文: 谭凤娇, 肖鹏, 左俊, 金磊, 程晨, 雷腊梅, 谢丽强, 杨军. 福建石兜水库产毒拉氏尖头藻(Raphidiopsis raciborskii)遗传多样性[J]. 生态毒理学报, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
shu
Tan Fengjiao, Xiao Peng, Zuo Jun, Jin Lei, Cheng Chen, Lei Lamei, Xie Liqiang, Yang Jun. Genetic Diversity of Toxin-Producing Raphidiopsis raciborskii (Cyanobacterium) Isolated from Shidou Reservoir in Fujian Province[J]. Asian journal of ecotoxicology, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
Citation: Tan Fengjiao, Xiao Peng, Zuo Jun, Jin Lei, Cheng Chen, Lei Lamei, Xie Liqiang, Yang Jun. Genetic Diversity of Toxin-Producing Raphidiopsis raciborskii (Cyanobacterium) Isolated from Shidou Reservoir in Fujian Province[J]. Asian journal of ecotoxicology, 2022, 17(3): 56-67. doi: 10.7524/AJE.1673-5897.20211012004
shu

福建石兜水库产毒拉氏尖头藻(Raphidiopsis raciborskii)遗传多样性

    通讯作者: 杨军, E-mail: jyang@iue.ac.cn
    作者简介: 谭凤娇(1991—),女,博士研究生,研究方向为环境科学、有害蓝藻监测,E-mail:fjtan@iue.ac.cn
  • 1. 中国科学院城市环境研究所, 福建省流域生态学重点实验室, 城市环境与健康重点实验室, 水生态健康研究组, 厦门 361021;
  • 2. 中国科学院南京地理与湖泊研究所, 湖泊与环境国家重点实验室, 南京 210008;
  • 3. 中国科学院大学, 北京 100049;
  • 4. 暨南大学生命科学技术学院生态学系, 广州 510632
  • 收稿日期:  2021-10-12
基金项目:

国家自然科学基金资助项目（91851104）；福建省自然科学基金资助项目（2019J02016，2020J01119）；福建省环保科技计划项目（2021R009）

摘要: 近几十年以来，全球内陆深水水体拉氏尖头藻（Raphidiopsis raciborskii）的检出率和水华事件日益增多，这种新型水华蓝藻具有产毒和无毒株系，其中能合成拟柱胞藻毒素（cylindrospermopsins，CYNs）的株系主要分布在澳大利亚和东南亚，严重威胁水生态系统和人类健康。本研究于2018年从福建省厦门市石兜水库分离、培养了4株拉氏尖头藻（XM1～XM4），对其形态、遗传多样性和产毒特征进行了研究。显微观察结果表明，4株拉氏尖头藻形态特征基本一致，细胞平均长度为3.79～4.26 μm，藻丝体呈直线型、平均长度为122～199 μm，在环境样品中可观察到异形胞（固氮细胞），使用富含氮的BG11培养基培养后异形胞会消失。酶联免疫吸附（ELISA）试剂盒测定显示，4株拉氏尖头藻均为产毒藻株；液相色谱串联质谱（LC-MS/MS）分析表明，4个藻株能同时合成CYN和脱氧拟柱胞藻毒素（deoxy-CYN），其中XM1、XM2和XM3藻株以CYN为主，在对数生长后期平均浓度为0.71～1.02 pg·cell-1，XM4则以deoxy-CYN为主，平均浓度为0.42 pg·cell-1。16S rRNA基因系统发育分析表明，XM2藻株与非洲藻株同源相似性较高，其他3株（XM1、XM3和XM4）与亚洲、欧洲的藻株同源相似性较高，表明石兜水库拉氏尖头藻种群遗传多样性较高。序列分析发现，XM2藻株具有完整的产CYN基因簇，但是部分关键基因发生了重排变异，cyrB和cyrI基因分别被分割成2部分和3部分，XM2藻株与浙江杭州藻株产毒基因簇最为相似。在气候变化和人类活动多重影响下，我国水源地水库产毒拉氏尖头藻的来源、传播扩散、遗传分化和生态效应有待进一步研究。

English Abstract

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