Title page for 972204003


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Student Number 972204003
Author Ying-yu Kuo(郭盈妤)
Author's Email Address jennykuo0101@yahoo.com.tw
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Department Life Science
Year 2010
Semester 2
Degree Master
Type of Document Master's Thesis
Language zh-TW.Big5 Chinese
Title Physiological function assay of a heat- and abscisic acid-regulated gene, AtRZFP33
Date of Defense 2011-05-20
Page Count 69
Keyword
  • ABA signaling pathway
  • heat shock response
  • stomatal aperture
  • Abstract Plants respond to evaluated temperature by inducing the heat shock response. The heat shock response (HSR) protects plants from cellular damage not only by accumulation of heat shock protein, but also by accelerated cooling down of biological system mediated by stomatal opening. The plant hormone abscisic acid (ABA) normally accumulates in response to drought, cold and salt stresses. Previous study indicated that rice OsRZFP34 was identified as a heat- and ABA- responsive gene according to analyses of microarray expression profile and reverse transcriptase polymerase chain reaction (RT-PCR). The complex cascades of gene expression in various stresses can contribute to stress response, which further enhances the stress tolerance. To identify the crosslink between high temperature and ABA-related response in Arabidopsis, we characterized the Arabidopsis At5g22920, also named AtRZFP33, which is a homolog of rice OsRZFP34. AtRZFP33 belongs to zinc finger subfamily, containing a deduced RING zinc finger signature. Using quantitative RT-PCR to analyze tissue specificity and expression pattern of AtRZFP33, we found that AtRZFP33 was significantly reduced by heat and ABA treatments. Although zinc finger protein is usually characterized as a transcription factor, subcellular localization of AtRZFP33-GFP in onion epidermal cells showed that AtRZFP33 was localized not only in nucleus but in the cytosol. To study the role of AtRZFP33 in resistance to stresses, we examined the loss of function of AtRZFP33 in a T-DNA insertion mutant, SALK_017562. Thermotolerance was decreased in atrzfp33 mutant line in seed stage; in seedling, we found atrzfp33 mutant line was resistant to drought and chilling stresses, resulting from its stomatal closure. The stomatal aperture was smaller in atrzfp33 mutant line, but 20% higher in transgenic plants overexpressing AtRZFP33 relative to wild type. It reflects that AtRZFP33 may play a negative role in stomatal closure controlling mechanism. Using quantitative RT-PCR to analyze the expression of ion channels related to stomata opening, we found that KAT1, OST2 and MRP5,were upregulated in atrzfp33 mutant, and downregulated in overexpression plants. Moreover, we transformed 35S::AtRZFP33 construct into nced3, abi1 and abi2 mutant lines. Stomatal aperture of nced3/AtRZFP33OX and abi2/AtRZFP33OX were higher than single mutants, but there was no difference between abi1/AtRZFP33OX and its single mutant. Collectively, these results indicated that AtRZFP33 may functions in heat and ABA stress response by regulating the stomatal mechanism.
    Table of Content 摘要 ........ III
    縮寫對照表 .......................... V
    Abstract....... VI
    壹、緒論............ 1
    植物與環境逆境.........1
    熱休克(Heat shockHeat) 反應................ 2
    離層酸(Abscisic acid)反應 .... 2
    氣孔的調節............... 4
    RING(Ring Interesting New Gene)鋅手指蛋白質....... 5
    OsRZFP34 .... 6
    研究起源與目的....................... 7
    貳、材料與方法............................ 9
    一、基因表現分析 ............... 9
    二、AtRZFP33蛋白質在細胞中的表現位置 ........ 12
    三、阿拉伯芥農桿菌轉殖...................... 17
    四、植株生理功能分析........................ 19
    叁、結果................................ 22
    AtRZFP33在阿拉伯芥中的表現部位 ............... 22
    AtRZFP33突變株的鑑定 ............. 23
    突變株之逆境抗性分析........................ 24
    AtRZFP33過量表現轉殖株之鑑定 .................. 26
    轉殖株之氣孔開闔分析....................... 27
    基因表現量之分析 .......................... 30
    肆、討論 .................................... 32
    AtRZFP33在阿拉伯芥的地上部表現且受高溫與ABA抑制表現..... 32
    AtRZFP33-GFP結合蛋白質表現在細胞質與細胞核中.................... 33
    AtRZFP33之生理功能分析 ..................... 33
    水稻OsRZFP34與AtRZFP33之生理功能相似...................... 36
    AtRZFP33參與的訊息傳遞路徑 ................. 37
    未來研究方向.............................. 37
    伍、參考文獻 ........................... 39
    Reference Abe, H., Urao, T., Ito, T., Seki, M., Shinozaki, K., Yamaguchi-Shinozaki, K. (2003) Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant Cell 15: 63-78
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    Fujita, Y., Fujita, M., Satoh, R., Maruyama, K., Parvez, M.M., Seki, M., Hiratsu, K., Ohme-Takagi, M., Shinozaki, K. and Yamaguchi-Shinozaki, K. (2005) AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis. Plant Cell 17: 3470-3488
    Gong, M, Li, Y.J. and Chen, S.Z. (1998). Abscisic acid induced thermotolerance in maize seedlings is mediated by Ca+2 and associated with antioxidant systems. J. Plant Physiol. 153:488–496.
    Advisor
  • Ching-hui Yeh(葉靖輝)
  • Files
  • 972204003.pdf
  • disapprove authorization
    Date of Submission 2011-06-22

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