利用cDNA微陣列技術探討蕃茄根部組織受過量銅誘導之基因; Functional Genomics Study in the Roots of Tomato (Lycopersicon escolentum) in Response to Excess of Copper Using cDNA Microarray Technique

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题名:	利用cDNA微陣列技術探討蕃茄根部組織受過量銅誘導之基因;Functional Genomics Study in the Roots of Tomato (Lycopersicon escolentum) in Response to Excess of Copper Using cDNA Microarray Technique
作者:	楊明華;Ming-Hua Yang
贡献者:	生命科學研究所
关键词:	蕃茄;重金屬;銅離子恆定機制;相減式cDNA雜交法;cDNA微陣列;功能性基因組;cDNA microarray;subtraction cDNA hybridization;copper homeostasis;heavy metal;tomato;functional genomics
日期:	2002-07-05
上传时间:	2009-09-22 10:16:52 (UTC+8)
出版者:	國立中央大學圖書館
摘要:	中文摘要對植物而言，銅是一必需的微量元素。在植物細胞內，銅可作為許多參與氧化反應、光合作用酵素的輔助因子。然而，過量銅存在會引起毒性反應如氧化性之壓迫，導致植物矮小以及死亡。另一方面，在缺乏銅的環境下，葉子會有彎曲變形以及提早老化的病徵。根是植物吸收土壤中金屬微量元素的重要組織器官，因此根部細胞可說是擔任控制銅離子恆定作用的第一道前線。然而目前根部細胞內銅離子恆定機制及生理功能並不清楚，所以本論文是採用功能性基因組之研究探討蕃茄根部組織在過量銅環境下誘導的特定基因，由這些基因已知的生理作用及相關途徑，來說明其在植物根部組織中銅離子恆定作用的機制。而我主要的實驗是利用差異式基因選殖與cDNA微陣列技術來大量收集受過量銅誘導的基因，且獲得其核苷酸序列並利用基因資料庫作序列的比對推斷其可能的身分。目前已從10 mM CuSO4處理四天之蕃茄根部細胞的相減式cDNA基因庫中，挑出48個受銅誘導兩倍以上的基因，而以基因的生理作用可分類為：（一）與抗氧化逆境相關之基因；（二）與抗病源體感染相關之基因；（三）與賀爾蒙的合成相關之基因；（四）與細胞的修復相關之基因；（五）其它基因群。 Abstract Copper is an essential trace element in plants. It is a cofactor for many cellular enzymes involving oxidation and photosynthesis. Excess amount of copper causes oxidative stress in cells and may lead stunted growth or death. On the other hand, the deficiency of copper causes several symptoms such as distortion and premature senescence of young leaves. Plants acquire trace elements such as copper through root system. Thus, root tissue stays in the front line and plays a key role in regulating copper homeostasis. However, the physiological mechanism of controlling copper concentration in root is still unclear. This thesis adopts functional genomic approach to unfold specific gene expression in various physiological processes under the treatment of excess copper to tomato root tissue. The up-regulated genes may help to illustrate the partial scheme of copper homeostasis in root system. The differential cloning method is employed to select copper-induced clones. Subsequently, the copper responsive genes were collected by cDNA microarray. The identities may be revealed by DNA sequencing and by the alignment of the gene sequences in Gene Bank. Forty-eight up-regulated clones of over two folds were identified from the cDNA subtraction library that was generated from the continuous treatment of 10 μM copper sulfate for 4 days in tomato roots. Based on their physiological implication, they can be grouped into five categories: (1) anti-oxidation (2) pathogen response (3) hormone biosynthesis (4) cellular organization (5) others.
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