Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/21925
標題: Functional analysis of an abnormal growth mutant M0039314
利用T-DNA插入突變體探討水稻基因之功能-穀粒發育異常突變體M0039314之功能分析
作者: Chen, Po-Ju
陳柏儒
關鍵字: T-DNA
水稻T-DNA突變體
OsSPL16
OsMADS45
出版社: 分子生物學研究所
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摘要: T-DNA突變株M0039314具有稻稈外擴、穗型直立、花梗及穗長較短、花藥發育異常、稔實率低及種子發育不完全等特異的農藝性狀。經由南方墨點法分析顯示此M0039314突變株為由單一個T-DNA插入在第8對染色體中編號PAC P0702E04的48,951 bp處。T-DNA構築上之35S強化子可以活化T-DNA插入點兩側基因OsSPL16和OsMADS45。M0039314經過數代自交後,子代產生分離現象,其異質品系 (heterozygous,TW),可依稔實率及株高分成TWo和TWx兩型,其中TWo之株型較高、稔實率佳,與野生型 (TNG67) 相似,而TWx則呈現半矮化及低稔實率之性狀,與此突變株之同質品系較相似。進一步分別將OsSPL16及OsMADS45大量表現於TNG67野生型中,以探討影響M0039314外表性狀之主要基因,分別獲得大量表現7株35S::SPL16和8株ubi::MADS45的轉殖株。並經由轉殖基因片段確認、RT-PCR、南方墨點法和西方墨點法等,確認其DNA、RNA蛋白質表現及插入基因拷貝數等,進而觀察ubi::MADS45轉殖株之T0和T1後代,結果顯示轉殖株株高較矮、分糵數較多、稔實率較低、劍葉外擴、抽穗時間較早及葉色較深。這些現象與M0039314突變株的株高、稔實率及葉色相近。而35S::SPL16 T0轉殖株可呈現與M0039314相同之株高較矮、分糵數較少、葉鞘及劍葉筆直及葉色較深等,此外35S::SPL16亦表現出根較粗且根數較少還有晚抽穗等M0039314所缺乏的性狀。OsSPL16和OsMADS45都屬於轉錄調控因子,而水稻中 OsMADS14、OsMADS15、OsMADS18和OsMADS20均屬SQUAMOA-like 基因推測可能會受到OsSPL16調控。利用RT-PCR偵測,發現OsMADS14和OsMADS15基因可以在大量表現OsSPL16轉殖株中被活化。M0039314 T-DNA突變體與35S::SPL16及ubi::MADS45轉殖株,在株高生長、葉片形態、稔實率或抽穗時間等廣泛的外表性狀,是否由於受到OsSPL16及OsMADS45轉錄調節因子調控所導致,因此本研究亦進一步利用蛋白質體學方式,偵測M0039314及TNG67的未成熟幼穗蛋白,結果顯示有差異點的產生,未來將進一步分析定序之,以探討OsSPL16及OsMADS45之下游基因功能及生理之調節角色。
URI: http://hdl.handle.net/11455/21925
其他識別: U0005-1208200811461100
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1208200811461100
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