title: Use of two-dimensional gel electrophoresis to differentiate morphospecies of Alexandrium minutum, a paralytic shellfish poisoning toxin-producing dinoflagellate of harmful algal blooms.
abstract: Contamination of shellfish with paralytic shellfish poisoning toxins (PST) produced by toxic harmful algal blooms (HABs) have been negatively affecting the shellfish and aquaculture industries worldwide. Therefore, accurate and early identification of toxic phytoplankton species is crucial in HABs surveillance programs that allow fish-farmers to take appropriate preventive measures in shellfish harvesting and other aquaculture activities to overcome the negative impacts of HABs on human health. The identification of toxic dinoflagellates present in the water is currently a time-consuming operation since it requires skillful taxonomists and toxicologists equipped with optical and scanning electron microscopes as well as sophisticated equipment, for example, high-performance liquid chromotography-fluorescence detection. In this paper, a two-dimensional gel electrophoresis (2-DE)-based proteomic approach was applied to discriminate between toxic and nontoxic strains of Alexandrium minutum. Variation in morphological features between toxic and nontoxic strains was minimal and not significant. Also, variation in 2-DE protein patterns within either toxic or nontoxic strains was low, but pronounced differences were detected between toxic and nontoxic strains. The most notable differences between these strains were several abundant proteins with pIs ranging from 4.8 to 5.3 and apparent molecular masses between 17.5 and 21.5 kDa. Groups of proteins, namely NT1, NT2, NT3, and NT4, were consistently found in all nontoxic strains, while T1 and T2 were prominent in the toxic strains. These specific protein spots characteristic for toxic and nontoxic strains remained clearly distinguishable irrespective of the various growth conditions tested. Therefore, they have the potential to serve as "taxonomic markers" to distinguish toxic and nontoxic strains within A. minutum. Initial studies revealed that the expression pattern of T1 was tightly correlated to toxin biosynthesis in the examined alga and may be used to serve as a potential toxin indicator.
authors.affiliation: Department of Ecology and Biodiversity, The University of Hong Kong, Hong Kong.
authors.forename: Leo Lai
authors.initials: LL
authors.lastname: Chan
authors.forename: Ivor John
authors.initials: IJ
authors.lastname: Hodgkiss
authors.forename: Paul Kwan-Sing
authors.initials: PK
authors.lastname: Lam
authors.forename: Jennifer Man-Fan
authors.initials: JM
authors.lastname: Wan
authors.forename: Hong-Nong
authors.initials: HN
authors.lastname: Chou
authors.forename: John Hon-Kei
authors.initials: JH
authors.lastname: Lum
authors.forename: Maria Gar-Yee
authors.initials: MG
authors.lastname: Lo
authors.forename: Abby Sin-Chi
authors.initials: AS
authors.lastname: Mak
authors.forename: Wai-Hung
authors.initials: WH
authors.lastname: Sit
authors.forename: Samuel Chun-Lap
authors.initials: SC
authors.lastname: Lo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004141:Dinoflagellida / Q000145:classification / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D008387:Marine Toxins / Q000378:metabolism* / Q000506:poisoning; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D020543:Proteome / Q000378:metabolism*; D012758:Shellfish / Q000469:parasitology*; D057096:Shellfish Poisoning; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D008387:Marine Toxins; D020543:Proteome