稻属水通道蛋白基因家族之分子演化研究.doc

稻屬水通道蛋白基因家族之分子演化研究 Molecular Evolution of Aquaporin Gene Family in Oryza 研究生：吳佳蒂 Wu, Chia-Ti 指導教授：江友中 Chiang, Yu-Chung 廖培鈞 Liao, Pei-Chun 【摘要】 亞洲栽培稻 (Oryza sativa)為目前全球主要糧食作物之一，源自於亞洲野生稻 (O. rufipogon)馴化形成，依亞種系統分成印度亞種(subsp. indica)和日本亞種(subsp. japonica)，其中印度亞種又稱為秈稻，而日本亞種又可區分為溫帶梗稻和熱帶梗稻(subsp. javanica)；另外，亞洲栽培稻依生長習性又可分為旱稻 (upland rice)和水稻 (lowland rice)。隨著近年來全球暖化的影響，乾旱情形日趨嚴重，造成糧食收成短缺，而研究稻米的抗旱機制就更顯重要。根據先前研究，水通道蛋白 (Aquaporin or water channel protein)位於植物的細胞膜和液胞膜上，可選擇性調節水分子的運輸，且稻屬的水通道蛋白基因家族 (Rice water channel gene family, RWC)為抗旱的候選基因家族之一，所以本研究目的透過分子演化了解在其它稻屬植物、亞洲野生稻、亞洲栽培稻以及亞洲栽培稻再細分的各分群的水通道蛋白基因之核酸變異情形、變異是否受到天擇的影響和在天擇影響下的適應性。藉由PCR、選殖和定序技術得到第七號染色體和第九號染色體的水通道蛋白基因序列，透過遺傳歧異度分析 (genetic diversity analysis)，進一步重建親緣關係樹(reconstructing phylogenetic trees)，並進行中性假說測定 (neutrality test)和PAML (phylogenetic analysis by maximum likelihood)。本研究的結果呈現，在157個樣本中，栽培稻的遺傳歧異度較野生稻低，推測栽培稻在人類馴化過程受到瓶頸效應影響而使遺傳多樣性降低。在第七號染色體A基因座的當地稻分群可能近期有受到正向天擇(positive selection)作用，B基因座為偽基因 (pseudogene)，C基因座可能受到負向天擇(negative selection)和寬鬆性天擇(ralexation of selection)的作用，而第九號染色體A基因座符合中性演化。未來可針對RWC在這四個基因座核酸變異情形探討是否在抗旱適應性上有所差異。 關鍵字：稻屬、野生稻、栽培稻、旱稻、水通道蛋白、天擇模式 【Abstract】 Rice (Oryza sativa) is one of the major crops in the world. O. sativa, a common used cultivated rice domesticated from O. rufipogon, can be divided into spp. indica, spp. japonica, and spp. javanica according to the different origin of domesticated. Additionally, O. sativa also can be divided into upland and lowland rice according to the different growing habitats. Affecting by global warming, the extreme drought was decreased the food productivity which causes food deficiency. Therefore, studies on the mechanisms of drought resistance (or tolerance) in the crops is more and more important. The water channel protein is a kind of membrane protein to play a key role in water transport and iron pump controlling. The different expression rate of water channel protein 3(RWC3) between upland and lowland rice is leading to different what of drought resis