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JPR Advance Access originally published online on February 27, 2006
Journal of Plankton Research 2006 28(6):613-620; doi:10.1093/plankt/fbi145
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© The Author 2006. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Nitric oxide synthase-like enzyme mediated nitric oxide generation by harmful red tide phytoplankton, Chattonella marina

Daekyung Kim, Kenichi Yamaguchi and Tatsuya Oda*

Division of Biochemistry, Faculty of Fisheries, Nagasaki University, Bunkyo-Machi 1-14, Nagasaki 852-8521, Japan

* Corresponding Author: t-oda{at}net.nagasaki-u.ac.jp

Received December 19, 2005; accepted in principle February 1, 2006; accepted for publication February 23, 2006; published online February 27, 2006
Communicating editor: K.J. Flynn

The unicellular marine phytoplankton Chattonella marina is known to exhibit potent fish-killing activity. Previous studies have demonstrated that C. marina produces reactive oxygen species (ROS), and ROS-mediated ichthyotoxic mechanism has been postulated. However, the exact toxic mechanism is still controversial. In this study, we obtained evidence that C. marina produces nitric oxide (NO) under normal growth conditions. We utilized chemiluminescence (CL) reaction between NO and luminol–H2O2 to detect NO in C. marina cell suspensions. In this assay, significant CL was observed in C. marina in a cell-number-dependent manner, and this was diminished by the addition of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO), a specific NO scavenger. The NO generation by C. marina was also confirmed by a spectrophotometric assay based on the measurement of the diazo-reaction-positive substances (NOx) and by fluorometric assay using highly specific fluorescent indicator of NO. The NO level in C. marina was significantly decreased by NG-nitro-L-arginine methyl ester (L-NAME), a specific NO synthase (NOS) inhibitor. The addition of L-arginine resulted in the increased NO level, whereas NaNO2 had no effect. These results suggest that a NOS-like enzyme is mainly responsible for NO generation in C. marina.


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