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研究生:邱祿棠
研究生(外文):Lu-Tung Chou
論文名稱:植物中NAC-like基因之選殖及特性分析
論文名稱(外文):Molecular Cloning and Function Analysis of NAC-like Genes from Various Plant Species
指導教授:楊長賢楊長賢引用關係
指導教授(外文):Chang-Hsien Yang
學位類別:碩士
校院名稱:國立中興大學
系所名稱:農業生物科技學研究所
學門:農業科學學門
學類:農業技術學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:94
中文關鍵詞:NAC-like 基因分子選殖
外文關鍵詞:NACGENE
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植物頂芽分生組織的細胞分裂主要是靠一群NAC-like基因的調控。此類NAC-like gene有一特性-在其靠近N端之amino acid序列中會包含一個大約150 amino acid的NAC domain。為了研究不同植物種類中細胞分裂受NAC-like基因調控的情形,我們從木本植物洋紫荊中分離出BpNACL1及一年生植物阿拉伯芥中分離出AtNACL1、AtNACL2,此三基因都是屬於NAC-like基因。BpNACL1會轉譯出229個胺基酸序列,此序列跟水稻中的一個NAC-like基因OsNAC4,有68%的一致性跟79%的相似度。在BpNACL1及OSNAC4的NAC domain中,則有92%的一致性。BpNACL1之mRNA會在植物發育初期的根、子葉、植物頂芽分生組織及營養葉中偵測到。異位大量表現BpNACL1的轉基因阿拉伯芥植株其花序與根產生變異。與野生種比較起來轉基因植株花序多了許多分歧點而且有欒生性狀,另方面,根部有短根和叢生性狀。此結果顯示BpNACL1應與莖及根分生組織的發育皆有關。進一步分析發現,在轉基因植株中與莖分生組織發育相關的FASCIATA基因及與根分生組織發育相關的DBP基因表現量有上升的情形。在本研究中,二個在序列上與BpNACL1具相當高相似度的NAC-like基因亦由阿拉伯芥中選殖出。AtNACL1會轉譯出269個胺基酸而AtNACL2則會轉譯出317個胺基酸,它們與BpNACL1有53%及42%的一致性。在NAC domain中,與BpNACL1則有75% (AtNACL1)及58% (AtNACL2)的一致性。與BpNACL1相似,AtNACL1 mRNA亦在根及頂芽分生組織中偵測到,而在營養葉中則有顯著的下降。相對的,AtNACL2 mRNA會在頂芽分生組織偵測到,在營養葉中亦有下降的趨勢但在根的部位卻偵測不到表現量。此外,這二基因都會在花苞中表現。當大量表現正向AtNACL1於野生種阿拉伯芥中時發現有分生組織增生的性狀,而大量表現反向AtNACL1於野生種阿拉伯芥中後則使得此類轉基因植物有分生組織停止發育的趨勢。

The proper cell division in apical meristem of plants was mainly regulated by a group of NAC-like genes that contained a conserved 150 amino acid NAC domain at their N-terminus of proteins. To investigate the cell division in various plant species, three NAC-like genes were isolated and characterized from woody plant Bauhinia purpurea (BpNACL1) and weed Arabidopsis (AtNACL1 and AtNACL2). BpNACL1, encodes a 229 amino acid protein that showed 68% identity and 79% similarity to rice NAC-like gene OsNAC4. In NAC domain, 92% amino acids are identical between BpNACL1 and OsNAC4. BpNACL1 mRNA was detected in root, cotyledon and apical meristem and was absent in vegetative leaves of young seedlings. Transgenic Arabidopsis plants ectopically expressed BpNACL1 produced shoot with more branches and root with more lateral roots than wild-type plants. This result indicated that BpNACL1 should involve in the regulation of both shoot and root apical meristematic activity. Further analysis indicated that the expression of genes involved in meristematic development such as FASCIATA and DBP was up-regulated in 35S::BpNACL1 plants. In Arabidopsis, AtNACL1 encodes a 269 amino acid protein whereas AtNACL2 encodes a 317 amino acid protein that showed 53% and 42% identity to BpNACL1 respectively. In NAC domain, 75% (AtNACL1) and 58% (AtNACL2) amino acids are identical to BpNACL1. Similar to BpNACL1, AtNACL1 mRNA was also detected in root, apical meristem and its expression was significantly decreased in vegetative leaves. By contrast, AtNACL2 mRNA was detected in apical meristem, decreased in vegetative leaves and absent in root. Both AtNACL1 and AtNACL2 were expressed in floral buds. Transgenic Arabidopsis plants ectopically expressed AtNACL1 produced more meristematic structures whereas Arabidopsis plants ectopically expressed antisense AtNACL1 severe altered the meristem formation.

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壹、前言------------------------------------------------------------------- 5
貳、前人研究------------------------------------------------------------- 7
參、材料與方法---------------------------------------------------------- 13
肆、結果------------------------------------------------------------------- 39
伍、討論、---------------------------------------------------------------- 48
陸、參考文獻------------------------------------------------------------- 53
柒、圖---------------------------------------------------------------------- 62
捌、表---------------------------------------------------------------------- 81
玖、附圖------------------------------------------------------------------- 82

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