Characterization of the DAF interacting proteins in regulating anther dehiscence in Arabidopsis thaliana
| Autor: | Chiu-Ling Chen, 陳秋伶 |
|---|---|
| Rok vydání: | 2017 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 105 DAF 【DEFECTIVE IN ANTHER DEHISCENCE1 (DAD1)-Activating Factor】, a RING-finger E3 ligase protein, regulates anther dehiscence by activating the expression of DAD1 in the jasmonic acid (JA) biosynthesis pathway. To identify the putative targets which will be degraded by DAF, yeast two-hybrid analysis was performed and several candidate target proteins such as CSD190, CSD760 and CSD220 were identified. In this study, we further investigated the function of these candidate proteins. The ectopic expression of either CSD190, CSD760 or CSD220 in Arabidopsis showed similar phenotype of anther indehiscence and male sterility throughout flower development. The expression of DAD1 was also down-regulated in the over-expressing transgenic plants. This result are similar to that of 35S::DAF antisense/RNAi flowers. The result of Alexander’s staining indicated that pollen viability of the 35S::CSD190, 35S::CSD760 and 35S::CSD220 transgenic plants is similar to the wild-type pollen. When manually pollinating the transgenic pollen to its stigma, normal elongation of the siliques was observed. This indicates that the function of the pistil for the transgenic plants is normal. Especially, 35S::CSD190 plants showed earlier germination of pollen tube. In pCSD190::GUS plants, the GUS activity was detected in late stages of pollen development but the GUS activity of pDAF::GUS was not detected in this stage. We suggested that CSD190 may play a role in pollen development with other E3 ligase proteins to regulate pollen development since CSD190 is an ubiquitin-like protein and 35S::CSD190 plants showed male-sterility. The expression pattern of CSD190 overlapped with the expression of DAF in stamens after the flower development stage 12. However, CSD190 seemed not to be degrade by DAF, thus CSD190 and DAF may has auxiliary relationship. CSD760 and CSD220 still have the potential to be the substrate of DAF since the expression pattern of CSD760 and CSD220 is similar to the pattern of DAF. 35S::CSD760 and 35S::CSD220 plants also showed similar phenotype to 35S::DAF antisense/RNAi plants. Furthermore, on the part of the pistil, DAFL1 is a protein similar to DAF, acting as E3 ligase. DAFL1 is shown on the pistil alone. According to the performance location, inference CSD190 in the pistil is also playing to help DAFL1 role. CSD760 is located in the pistil position and DAF is the same, suggesting that may belong to the DAFL1 substrate. These results provided new information for understanding the mechanism of DAF and its interacting proteins in regulating anther dehiscence in plants. Further experiments will be performed for investigating the correlation. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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