Sample Preparation of the Macro Alga Pyropia yezoensis for the Determination of Messenger RNA

ABSTRACT

Few nutrients that limit the growth of photoautotrophs in marine environments have been identified. Observing phytoplankton growth after nutrient addition is an effective but time-consuming way of identifying key nutrients. One possible and potentially more efficient approach is to characterize the nutrients that photoautotrophs preferentially absorb. Quantification of transcript levels is suitable for this purpose and may be performed by reverse transcription followed by the polymerase chain reaction. However, since this analysis cannot be performed on-site, a suitable sample preparation method was developed using the marine macroalga Pyropia yezoensis. This study showed that specimen transport, even at temperatures below 10°C, may cause changes in transcript abundance. This was especially true for transcripts associated with the metabolism of macronutrients such as nitrogen and phosphorus. To prevent transport-induced changes in transcript levels and erroneous conclusions, on-site freezing of specimens is recommended when sampling.

KEYWORDS:

• Macro alga
• nutritional status
• Pyropia yezoensis
• reverse transcription quantitative polymerase chain reaction