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研究生(外文):Ming-wei Li
論文名稱(外文):Cloning and sequence analysis of the peroxidase genes in High and Low rooting line of Cinnamomum kanehirae
指導教授(外文):Ching-Huang LiuChang-Hung Chou
外文關鍵詞:sugar repression responsivenessperoxidaselignificationadventitious root formationindole-3-butyric acidCinnamomum kanehirae
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生長素(auxin)能誘導植物發不定根,其中,人工合成的生長激素,引朵丁酸(indole-3-butyric acid, IBA)能有效地促進牛樟插穗不定根的誘發。推測IBA藉由抑制過氧化酶(peroxidase,POX)的活性誘導逆分化,在不定根發育時形成新的根源母細胞。因此,藉其他物種已發表的序列設計退化性引子,在牛樟(Cinnamomum kanehirae)幼根中篩選出POX cDNA,並利用5’和3’ RACE (Rapid Amplification of cDNA Ends)完成整段的cDNA序列。我們探討牛樟高發根(H)與低發根(L)品系POX基因的序列,發現兩個不同發根植株具有極相似的cDNA,因此針對此相似POX cDNA的上游調控區進行序列分析。調控區的序列包含auxin及木質素合成等相關element。高發根植株之牛樟幼根所含有的POX基因其調控區域有醣抑制反應(sugar repression responsiveness, SRS)的element。此element可導致POX基因表現受醣的抑制,推測可能導致高發根品系幼根與組織內含較少之POX有關。
Auxin can induce adventitious rooting. Synthetic auxin, indole-3-butyric acid (IBA), effectively promoted the rooting in Cinnamomum kanehirae. In Cinnamomum kanehirae, there are high (H) and low (L) rooting cultivar. The peroxidase (POX) activity significantly decreased in the IBA-treated tissues as compared with the control. The inhibition on POX activity may lead to the redifferentiation processes induced by IBA. In this investigation, we cloned POX cDNA from the young roots. Degenerate primers were designed from the conservative regions of other published POXs to amplify the expectant DNA fragment. We found that the H and L line have similar genes (>99%). The Full-length cDNA of the POX genes were cloned by the method of 5''and 3'' Rapid Amplification of cDNA Ends (RACE). The deduced amino acid were compared with the previously reported POX and showed between 40% and 70% identity with other plant POXs. Further studies on the promoter elements of POX in High-rooting cultivar and Low-rooting line show that some elements are related to auxin response, lignification, pathogen invasion and stress response.
The regulatory elements of the POX gene in High-rooting line contain sugar repression responsiveness (SRS) elements that might repress the expression of POX gene, causing the lower POX activity.
Chinese abstract ------------------------------------- I

English abstract ------------------------------------- II

Table of contents ---------------------------------- III

List of tables and figures ------------------------- IV

Introduction ------------------------------------------ 1

Materials and Methods ----------------------------- 5

Results ---------------------------------------------- 12

Discussion ------------------------------------------ 17

Reference ------------------------------------------- 23

Tables and figures --------------------------------- 28
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