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Semi-quantitative RT-PCR analysis of photoregulated gene expression in marine diatoms.

TitleSemi-quantitative RT-PCR analysis of photoregulated gene expression in marine diatoms.
Publication TypeJournal Article
Year of Publication1999
AuthorsLeblanc, C, Falciatore, A, Watanabe, M, Bowler, C
JournalPlant Mol Biol
Volume40
Issue6
Pagination1031-44
Date Published1999 Aug
ISSN0167-4412
KeywordsActins, Algal Proteins, Amino Acid Sequence, Diatoms, DNA, Gene Expression Regulation, Genes, Light, Light-Harvesting Protein Complexes, Molecular Sequence Data, Photosynthetic Reaction Center Complex Proteins, Reverse Transcriptase Polymerase Chain Reaction, RNA, Messenger, Sequence Alignment, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Spectrophotometry
Abstract

The low cell densities of diatoms and other phytoplankton in culture has precluded the use of classical RNA analysis techniques for routine studies of gene expression in large numbers of samples. This has seriously hampered studies of the basic biology of such organisms. To circumvent this problem, we have developed a high-throughput semi-quantitative RT-PCR-based protocol and used it to monitor expression of a gene encoding a fucoxanthin, chlorophyll a/c-binding protein (FCP) in the centric planktonic diatom Thalassiosira weissflogii. Analysis of FCP gene expression in dark-adapted diatom cultures revealed that mRNA levels increase 5- to 6-fold in response to white light irradiation and peak around 6 to 8 h. To determine the photoreceptors involved in this response action spectra of FCP gene expression were determined using the Okazaki large spectrograph. Responses consistent with the presence of cryptochrome-, rhodopsin- and phytochrome-type receptors could be detected. The apparent presence of phytochrome-mediated responses is of particular interest given the low fluences of red and far-red light wavelengths in the marine environment.

Alternate JournalPlant Mol. Biol.
PubMed ID10527427