Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum

Xianghai Tang; Rencheng Yu; Mingjiang Zhou; Zhigang Yu

doi:10.1007/s00343-012-1142-7

Your Location：

Home >

Browse articles >

Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum

OriginalPaper | Updated：2023-05-16

- Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum
- Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum
- 海洋湖沼学报（英文） 2012年30卷第2期页码：256-263
- Affiliations：
  
  1. Key Laboratory of Marine Chemistry Theory and Technology Ministry of Education Ocean University of China
  2. Key Laboratory of Marine Ecology and Environmental Sciences Institute of Oceanology Chinese Academy of Sciences
- Author bio：
- Funds：
  
  Supported by the National Natural Science Foundation of China (No. 41006082), the Postdoctoral Science Foundation (No. 20090461272), and the National Natural Science Foundation of China for Creative Research Groups (No. 40821004)
- DOI：10.1007/s00343-012-1142-7
  中图分类号：
- 网络首发：2012-03-01，
  
  纸质出版：2012
- Accepted：
Scan QR Code
Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum[J]. 海洋湖沼学报（英文）, 2012,30(2):256-263.

Tang, X., Yu, R., Zhou, M. et al. Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum, ., Chin. J. Ocean. Limnol. 30, 256–263 (2012). https://doi.org/10.1007/s00343-012-1142-7
Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum[J]. 海洋湖沼学报（英文）, 2012,30(2):256-263. DOI： 10.1007/s00343-012-1142-7.

Tang, X., Yu, R., Zhou, M. et al. Application of rRNA probes and fluorescence in situ hybridization for rapid detection of the toxic dinoflagellate Alexandrium minutum, ., Chin. J. Ocean. Limnol. 30, 256–263 (2012). https://doi.org/10.1007/s00343-012-1142-7 DOI：

摘要

Abstract

The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology

making identification difficult. In this study

species-specific rRNA probes were designed for whole-cell fluorescence in situ hybridization (FISH) to distinguish A. minutum from two phylogenetic clades. We acquired the complete SSU to LSU rDNA sequences (GenBank accession numbers JF906989-JF906999) of 11 Alexandrium strains and used these to design rRNA targeted oligonucleotide probes. Three ribotype-specific probes

M-GC-1

M-PC-2

and M-PC-3

were designed. The former is specific for the GC clade (“Global clade”) of A. minutum

the majority of which have been found non-toxic

and the latter two are specific for the PSP (paralytic shellfish poisoning)-producing PC clade (“Pacific clade”). The specificity of these three probes was confirmed by FISH. All cells in observed fields of view were fluorescently labeled when probes and target species were incubated under optimized FISH conditions. However

the accessibility of rRNA molecules in ribosomes varied among the probe binding positions. Thus

there was variation in the distribution of positive signals in labeled cells within nucleolus and cytosol (M-GC-1

M-PC-3)

or just nucleolus (M-PC-2). Our results provide a methodological basis for studying the biogeography and population dynamics of A. minutum

and providing an early warning of toxic HABs.

关键词

Keywords

references

Adachi M, Sako Y, Ishida Y. 1996a. Analysis of Alexandrium (Dinophyceae) species using sequence of the 5.8S ribosomal DNA and internal transcribed spacer regions. J. Phycol., 32: 424–432.

Adachi M, Sako Y, Ishida Y. 1996b. Identification of the toxic dinoflagellates Alexandrium catenella and A. tamarense (Dinophyceae) using DNA probes and whole-cell hybridization. J. Phycol., 32: 1 049–1 052.

Ahn S, Kulis D M, Erdner D L et al. 2006. Fiber-optic microarray for simultaneous detection of multiple harmful algal bloom species. Appl. Environ. Microbiol., 72(9): 5 742–5 749.

Anderson D M, Kulis D M, Keafer B A et al. 1999. Detection of the toxic dinoflagellate Alexandrium fundyense (Dinophyceae) with oligonucleotide and antibody probes: variability in labeling intensity with physiological condition. J. Phycol., 35(4): 870–883.

Anderson D M, Kulis D, Keafer B A et al. 2005. Identification and enumeration of Alexandrium spp. from the Gulf of Maine using molecular probes. Deep-Sea Res. II, 52: 2 467–2 490.

Balech E. 1995. The Genus Alexandrium Halim (Dinoflagellata). Sherkin Island Marine Station, Cork, Ireland.

Cembella A D. 1998. Ecophysiology and metabolism of paralytic shellfish toxins in marine microalgae. In: Anderson D M, Cembella A D, Hallegraeff G M eds. Physiological Ecology of Harmful Algal Blooms. NATO ASI Series. Springer, Berlin. p.381–403.

Dyhrman S T, Erdner D, Du J L et al. 2006. Molecular quantification of toxic Alexandrium fundyense in the Gulf of Maine using real-time PCR. Harmful Algae, 5(3): 242–250.

Guilllard R R L, Ryther J H. 1962. Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt and Detonula confervacea (Cleve) Gran. Can. J. Microl., 8: 229–239.

Hallegraeff G M. 1993. A review of harmful algal blooms and their apparent global increase. Phycologia, 32: 79–99.

Hansen G, Daugbjerg N, Franco J M. 2003. Morphology, toxin composition and LSU rDNA phylogeny of Alexandrium minutum (Dinophyceae) from Denmark, with some morphological observations on other European strains. Harmful Algae, 2(4): 317–335.

Higman W A, Stone D M, Lewis J M. 2001. Sequence comparison of toxic and non-toxic Alexandrium tamarense (Dinophyceae) isolates from UK waters. Phycologia, 40: 256–262.

Hosoi-Tanabe S, Sako Y. 2005a. Rapid detection of natural cells of Alexandrium tamarense and A. catenella (Dinophyceae) by fluorescence in situ hybridization. Harmful Algae, 4(2): 319–328.

Hosoi-Tanabe S, Sako Y. 2005b. Species-specific detection and quantification of toxic marine dinoflagellates Alexandrium tamarense and A. catenella by Real-Time PCR assay. Mar. Biotechnol., 7(5): 506–514.

John U, Cembella A, Hummert C et al. 2003. Discrimination of the toxigenic dinoflagellates Alexandrium tamarense and A. ostenfeldii in co-occurring natural populations from Scottish coastal waters. Eur. J. Phycol., 38(1): 25–40.

John U, Medlin L K, Groben R. 2005. Development of specific rRNA probes to distinguish between geographic clades of the Alexandrium tamarense species complex. J. Plankton Res., 27(2): 199–204.

Ki J S, Han M S. 2005. Molecular analysis of complete SSU to LSU rDNA sequence in the harmful dinoflagellate Alexandrium tamarense (Korean isolate, HY970328M). Ocean Sci. J., 40: 155–166.

Ki J S, Han M S. 2007. Informative characteristics of 12 divergent domains in complete large subunit rDNA sequences from the harmful dinoflagellate genus, Alexandrium (Dinophyceae). J. Eukaryot. Microbiol., 54(2): 210–219.

Kim C J, Kim C H, Sako Y. 2005. Development of molecular identification method for genus Alexandrium (Dinophyceae) using whole-cell FISH. Mar. Biotechnol., 7(3): 215–222.

Lilly E L, Halanych K M, Anderson D M. 2005. Phylogeny, biogeography, and species boundaries within the Alexandrium minutum group. Harmful Algae, 4(6): 1 004–1 020.

McCauley L A R, Erdner D L, Nagai S et al. 2009. Biogeographic analysis of the globally distributed harmful algal bloom species Alexandrium minutum (dinophyceae) based on rRNA gene sequences and microsatellite markers. J. Phycol., 45(2): 454–463.

Pitcher G C, Cembella A D, Joyce L B et al. 2007. The dinoflagellate Alexandrium minutum in Cape Town harbour (South Africa): Bloom characteristics, phylogenetic analysis and toxin composition. Harmful Algae, 6(6): 823–836.

Sako Y, Hosoi-Tanabe S, Uchida A. 2004. Fluorescence in situ hybridization using rRNA-targeted probes for simple and rapid identification of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella. J. Phycol., 40(3): 598–605.

Scholin C A, Herzog M, Sogin M et al. 1994. Identification of group- and strain-specific genetic markers for globally distributed Alexandrium (Dinophyceae) species. II. Sequence analysis of a fragment of the large-subunit ribosomal RNA gene. J. Phycol., 30: 999–1 011.

Scorzetti G, Brand L E, Hitchcock G L et al. 2009. Multiple simultaneous detection of Harmful Algal Blooms (HABs) through a high throughput bead array technology, with potential use in phytoplankton community analysis. Harmful Algae, 8(2): 196–211.

Tang X H, Yu R C, Yan T et al. 2006. Sequence analysis and molecular phylogenetics of ribosomal RNA gene of Alexandrium (dinophyceae) strains in china. Oceanol. Limnol. Sin., 37(6): 530–535. (in Chinese with English abstract)

Taylor F J R, Fukuyo Y, Larsen J. 1995. Taxonomy of harmful dinoflagellates. In: Hallegraeff G M, Anderson D M, Cembella A D eds. Manual on Harmful Marine Microalgae. IOC Manuals and Guides No.33. IOC of UNESCO. p.283–317.

Touzet N, Davidson K, Pete R et al. 2010. Co-occurrence of the west European (Gr.III) and north American (Gr.I) ribotypes of Alexandrium tamarense (Dinophyceae) in Shetland, Scotland. Protist, 161(3): 370–384.

Touzet N, Raine R. 2007. Discrimination of Alexandrium andersoni and A. minutum (Dinophyceae) using LSU rRNA-targeted oligonucleotide probes and fluorescent whole-cell hybridisation. Phycologia, 46(2): 168–177.

Turner J T, Doucette G J, Powell C L et al. 2000. Accumulation of red tide toxins in larger size fractions of zooplankton assemblages from Massachusetts Bay, USA. Mar. Ecol. Prog. Ser., 203: 95–107.

Wang D Z, Jiang T J, Hsieh D P H. 2005. Toxin composition variations in cultures of Alexandrium species isolated from the coastal waters of southern China. HarmfuI Algae, 4(1): l09–121.

Wang Z H, Matsuoka K, Qi Y Z et al. 2004. Dinoflagellate cysts in recent sediments from Chinese coastal waters. Mar. Ecol., 25(4): 289–311.

Yu R C, Hummert C, Luekas B et al. 1998. Analysis of PSP toxins in algae and shellfish samples from China using a modified HPLC method. Chromatographia, 48(9): 67l–677.

Yu R C, Tang X H, Zhang Q C et al. 2006. Application of fluorescence in-situ hybridization (FISH) method to detect “tamarense/catenella species complex” (“Temperate Asian” ribotype) in Genus Alexandrium along Chinese coast. Acta Sci. Circumst., 26(4): 646–651. (in Chinese with English abstract)

Zhen Y, Mi T Z, Yu Z G. 2009. Detection of several harmful algal species by sandwich hybridization integrated with a nuclease protection assay. Harmful Algae, 8(5): 651–657.

Zhou M J, Li J, Luekas B et al. 1999. A recent shellfish toxin investigation in China. Mar. Pollut. Bull., 39(1): 33l–334.

Zhou M J, Zhu M Y, Zhang J. 2001. Status of harmful algal blooms and related research activities in China. Chin. Bull. Life Sci., 13(2): 54–59. (in Chinese with English abstract)

浏览量

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

工具集

关联资源

Chromosomal mapping of 5S and 18S-5.8S-25S rRNA genes in Saccharina japonica (Phaeophyceae) as visualized by dual-color fluorescence in situ hybridization

基于NSGA II算法分布度改进的ATREX发动机性能优化方法研究

测试布局对涡轮效率的影响研究

基于模态频率的V型缺口梁裂纹扩展速率模型

压电阻抗法表征固体推进剂老化的研究