Characterization of PyGUS gene silencing in the red macroalga, Pyropia yezoensis

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• 作者：Yoon Ju Shin ; Jong-Min Lim ; Ji Hyun Park ; Dong-Woog Choi…
• 关键词：Pyropia yezoensis ; Macroalgae ; Transgene silencing ; Argonaute ; Dicer
• 刊名：Plant Biotechnology Reports
• 出版年：2016
• 出版时间：November 2016
• 年：2016
• 卷：10
• 期：6
• 页码：359-367
• 全文大小：
• 刊物主题：Plant Sciences; Cell Biology; Plant Biochemistry; Biotechnology; Agriculture;
• 出版者：Springer Japan
• ISSN：1863-5474
• 卷排序：10

文摘

Transgene silencing is a common phenomenon in higher plants and microalgae. In the present study, we analyzed transgene silencing in the macroalga Pyropia yezoensis. This is the first report of transgene silencing in macroalgae. Transgenic lines were generated by transformation with the PyGUS gene and a hygromycin resistance gene (PtAph7-1). Histochemical GUS staining detected PyGUS expression in only three out of 12 lines. Southern and northern blotting, polymerase chain reaction (PCR), and reverse transcription (RT)-PCR analyses indicated that the PyGUS gene was silenced in the other 9 lines. Interestingly, PyGUS gene silencing in transgenic line 7 was repeatedly initiated at a later stage, in the developmental process from monospore to thallus through asexual reproduction. We identified genes encoding Argonaute and Dicer-like protein, the major components of the RNA-silencing pathway, in the genome and transcripts of P. yezoensis. Interestingly, we found the DDA motif in the PIWI domain of the AGO, suggesting that translational repression may be the major gene-silencing pathway in P. yezoensis. In this study, the observation of PyGUS gene silencing and identification of RNA-silencing components indicate that the gene (transgene) silencing machinery functions actively in the macroalga P. yezoensis. These results will be useful to study the regulation of gene expression and RNA-silencing mechanisms in Pyropia species and related macroalgae.