Journal
HARMFUL ALGAE
Volume 39, Issue -, Pages 26-39Publisher
ELSEVIER
DOI: 10.1016/j.hal.2014.06.005
Keywords
Alexandrium catenella; Cell cycle; Phosphate limitation; Suppression subtractive hybridization; Toxin
Categories
Funding
- Natural Science Foundation of China [41129001, 41176091, 41330959, 41130855]
- Chinese Academy of Sciences [XDA11020203-3]
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Molecular mechanisms regulating responses to phosphate (P) limitation in dinoflagellates are poorly understood. Here we investigated gene differential expression Alexandrium catenella strain ATHK using suppression subtractive hybridization (SSH) and reverse-transcription quantitative PCR (qRT-PCR) techniques. SSH revealed a score of genes responded to P-limitation, and comparison with EST sequencing results indicated superior efficiency of SSH in identifying differentially expressed genes. Among the P-limitation responsive genes were the up-regulated stress-related genes, the down-regulated phosphorus-transferring genes, and a number of other genes previously undocumented for dinoflagellates. Most notably, the previously undocumented cell cycle inhibitory regulator fizzy/cell division cycle 20-related protein gene was up-regulated while putatively cell cycle promoting genes (e.g. calcium-dependent protein kinase) down-regulated, consistent with the decrease in growth rate and cell cycle arrest in G1 phase under P-limitation observed microscopically and flow cytometrically respectively. Besides, both SSH and qRT-PCR showed that saxitoxin related genes, menaquinone biosynthesis methyltransferase, an unknown gene and some other genes were also significantly induced by P-limitation. Our results demonstrate the utility of SSH in gene regulation research and provide a set of promising genes for further studies to understand the regulation of cell division (and hence population growth) and toxin production by P-limitation in A. catenella and likely other dinoflagellates. (C) 2014 Elsevier B.V. All rights reserved.
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