碩士 / 國立臺灣海洋大學 / 生命科學暨生物科技學系 / 104 / Polyunsaturated aldehydes (PUAs) derived from wounded-diatoms are considered as secondary metabolites belonging to the oxylipins known to act mainly as teratogenic compounds in different marine organism. Moreover, PUAs can also be toxic to diatoms themselves, and were shown to induce programmed cell death (PCD), which is proposed as an important cellular mechanism responsible for massive cell loss during phytoplankton bloom succession. Previous studies have showed that environmental stresses (e.g. cell aging or light limitation) trigger DSPs expression to lead to PCD via NO signaling. However, information for the signal transduction and regulatory mechanisms eliciting PCD in PUA-stressed diatoms is still in its infancy. In this study, a marine diatom, (Skeletonema tropicum, Kaohsiung strain), was applied as the model for this purpose. The mRNA levels of DSP1 and DSP2 in S. tropicum cells were measured after the treatments of the PUAs, 2,4-heptadienal (HEPTA) and 2,4-octadienal (OCTA) and 2,4-Decadienal (DECA). Our results indicated that only HEPTA and OCTA, but not DECA, were able to induce DSP1 and DSP2 expression. In addition, cells pre-treated with ROS scavenger, Tempol, or NO scavenger, c-PTIO, for 30 min prior to HEPTA addition could obviously reduce the DSP1 and DSP2 expression. These results implied that ROS and NO were the secondary messengers to upregulate DSP1 and DSP2 expression via PUA signaling. Furthermore, cells under different treatments of PUAs (i.e. HEPTA and OCTA), H2O2 (ROS donor) and DEANO (NO donor) were harvested for RNA-seq libraries construction. The analysis have been performed using pair-end sequencing approach by Illumina NextSeq 500. A total of 117 Giga bases (150 bases per read length) with a coverage more than 50 times was obtained. As expected, all of the DSP1 and DSP2 expressions level in the PUAs, H2O2, and DEANO treatments were significantly greater than the no-treatment control. Therefore, these RNA-seq libraries are confident for further elucidating the molecular regulatory mechanisms in S. tropicum cells under the stresses mediated by PUA signaling.
| Identifer | oai:union.ndltd.org:TW/104NTOU5613001 |
| Date | January 2016 |
| Creators | Lee, Siao-Sian, 李筱仙 |
| Contributors | Chen, Shiow-Yi, Chung, Chih-Ching, 陳秀儀, 鍾至青 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | en_US |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 42 |
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