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標題: 水稻OSE705蛋白特性及功能研究
Characterization and functional study of rice OSE705 protein
作者: 邱思嘉
Chiou, Sz-Jia
關鍵字: rice
seed maturation and germination

出版社: 分子生物學研究所
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摘要: A rice gene, Ose705 has been characterized as one of the rice embryo-specific gene; however its functional role during embryo development has not been well characterized. The previous results of Ose705 mRNA expressed from 3 to 40 days after pollination (DAP) and the OSE705 protein can only be detected between 11 to 20 DAP were further confirmed in this study. In addition, this study discovered that Ose705 mRNA expressed with very high level on the first 3 days after seed imbibitions. These observations suggested that Ose705 may play important role during seed germination and seed development. To better understand the functional role of this gene, RNA interference (RNAi) approach to knock down/ out the expression of Ose705 was performed RNAi transgenic line with 35S or Ose705 promoter constructs (35S::705i and 705::705i) were available from previous study. The morphological phenotype of these two RNAi lines revealed no difference from those of their host variety TNG67 during vegetative growth, however, the 35S::705i showed much lower fertility (30.07%±11.97%) than that of TNG67. But, the relationship between the low fertility and suppression of Ose705 expression was not consistent. No futher study in this part was conducted. While no significant phenotype was observed in the 705::705i line, I futher focus on the development of 10-day-old embryos. I initially observed the size of 10-day-old embryos were bigger than those of TNG67 raised in the second growth season of 2008, however, the results were reverted in the first growth season of 2009. Tissue section of these 10-day-old embryos were conducted to check their structure and measure their exact size. The results showed the structures of all the embryos were normal except the size of 705::705i embryos were small. These inconsistent results were discussed in detail in the text. As result showed that Ose705 mRNA expressed with very high level on the early stage of seed germination, I examined the expression of target gene driven by 1.7 Kb Ose705 promoter on transgenic lines (705::LAA and 705::LNCA) available in our laboratory. The results indicated that these target genes could be deteceted at the early stage of seed germination, however the expression patterns were different from those of the native Ose705 gene.
根據本實驗室已有資訊,推測Ose705可能是水稻胚發育特有表現的基因, 然而該基因功能尚不清楚。本研究發現Ose705 mRNA在授粉後3到40天種子表現,而該蛋白則表現於授粉後的11到20天種子。另外,本研究新發現於萌芽後的前3天胚芽,Ose705 mRNA表現強而之後逐漸減少。綜合上述的研究成果,推測該基因參與種子發育及發芽過程。本研究以RNA干擾法抑制Ose705基因,探討該基因功能。本實驗室已獲得35S::705i及705::705i構築之RNAi同質品系轉殖株,而且於種子發育及萌芽時期Ose705基因均受抑制。觀察RNAi轉殖株生長過程外觀性狀,只發現35S::705i轉殖株有低稔實性狀,然而低稔實與RNAi抑制Ose705基因表現,較無直接的關係。進一步觀察不同期作RNAi轉殖株及TNG67種子發育過程,發現第二期作授粉後10天705::705i 轉殖種子發育階段較快,而第一期作無此現象,分析此時期之穀粒,發現含有RNAi構築之轉殖種子Ose705 mRNA及其蛋白均被抑制。進一步以組織切片方式量測和觀察授粉後10天胚,發現該轉殖胚較小,然而其結構及形態無差異。另外萌芽試驗中,發現種子萌芽率沒有差異。本研究發現種子萌芽初期Ose705 mRNA即大量表現,之後可以運用該啟動子組織特異表現的特性。先使用本實驗室已有以1.7Kb Ose705 啟動子表現LNCA及LAA之轉殖株對水稻萌芽後不同天數胚芽進行分析,發現萌芽時期可測得LNCA及LAA mRNA和各自蛋白表現,確認該啟動子於種子萌芽時期可以表現外源基因,然而外源基因表現之情況與內生性Ose705基因是不同。
其他識別: U0005-1208200918015800
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