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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王俊能(Chun-Neng Wang) | |
| dc.contributor.author | Yun-Yu Chen | en |
| dc.contributor.author | 陳運佑 | zh_TW |
| dc.date.accessioned | 2021-06-16T05:15:29Z | - |
| dc.date.available | 2019-08-21 | |
| dc.date.copyright | 2014-08-21 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-18 | |
| dc.identifier.citation | Abramoff MD, Magalhaes PJ, Ram SJ (2004) Image processing with ImageJ. Biophotonics International 11: 36-42
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56101 | - |
| dc.description.abstract | 苦苣苔科的堇蘭(Streptocarpus rexii)不具有頂端分生組織，其細胞分裂活力位於葉子上，使葉子取代頂端分生組織的功能而具有生長新器官與維持植株發育的能力。過去研究顯示堇蘭這種異位的分生組織會受到外加細胞分裂素(cytokinin)的促進，而這和模式植物中細胞分裂素負責分生組織活力的維持，具有相似的性質。故我們嘗試解開細胞分裂素是如何參與堇蘭異位分生組織活力的調控。本研究自堇蘭分離出細胞分裂素生合成異戊烯基轉移酶基因(ISOPENTENYLTRANSFERASE, IPT) ，並檢測其在堇蘭幼苗的分生組織中的表現模式及是否受到外加植物荷爾蒙的調控。我們自堇蘭分離出五個IPT同源基因(SrIPT1, SrIPT2, SrIPT3, SrIPT5, SrIPT9)，其中SrIPT1與SrIPT3集中表現在花或根部，SrIPT5和SrIPT9則在各個器官都有表現，而SrIPT2具有提前終止密碼子，可能是一個假基因(pseudogene)。我們更進一步發現SrIPT1, SrIPT5與SrIPT9均大量表現在堇蘭的異位分生組織內(groove meristem和basal meristem)，而SrIPT5在分生組織處(葉子基部)的表現量比不具有分生組織活力分佈的葉尖端的表現量還高。這些結果支持異位分生組織的活力維持需要高濃度細胞分裂素的假設。最後，我們發現外加的荷爾蒙如生長素與吉貝素會抑制SrIPT3, SrIPT5, SrIPT9表現，而細胞分裂素會促進SrIPT1, SrIPT3與SrIPT9表現，暗示植物荷爾蒙調節SrIPT的表現量可能與堇蘭異位分生組織的維持有關。我們的研究證實IPT基因在堇蘭的分生組織內的表現，顯示其可能參與維持分生組織的活力。這也進一步暗示細胞分裂素生合成可能和堇蘭的分生組織自莖頂異位至葉子後的活力維持有關。 | zh_TW |
| dc.description.abstract | Streptocarpus rexii (Gesneriaceae) lacks the conventional shoot apical meristem (SAM). On the other hand, unorthodox meristems form on the leaves of S. rexii, and are capable of producing lateral organs and maintaining plant’s growth. Previous researches indicated that the exogenous cytokinin (CK) treatment affected the activities of the unorthodox meristems of S. rexii. We therefore aimed to understand how CK is involved in maintaining the unorthodox meristem activity of S. rexii. In this study, I isolated the CK biosynthesis IPT gene (ISOPENTENYLTRANSFERASE) and examined their expression patterns in relation to meristem activity in the seedling of S. rexii. I also tested whether the expression levels of SrIPT genes can be altered by exogenous hormone treatments in order to understand whether SrIPTs can be regulated by major phytohormones. Five IPT homologous genes were isolated from S. rexii (SrIPTs): SrIPT1, SrIPT2, SrIPT3, SrIPT5, and SrIPT9, while SrIPT2 is perhaps a pseudogene because of the formation of a premature stop codon. Real-time PCR of the other four SrIPTs revealed that SrIPT1 mainly expressed in floral organs and SrIPT3 mainly in roots, and SrIPT5 and SrIPT9 expressed in all organs examined. Furthermore, I found the strong expression signals of SrIPT1, SrIPT5 and SrIPT9 within the unorthodox meristems (basal and groove meristems) via RNA in situ hybridization. I also discovered that SrIPT5 strongly expressed in meristem-located (proximal end) rather than no-meristem-located (distal end) lamina tissue of older leaves. These results supported the hypothesis that meristem-localized CK level is required for the maintenance of these unorthodox meristems. Exogenous hormone treatments such as auxin and gibberellin can inhibit certain SrIPTs expression but CK increase their expression instead, implying that hormone regulated SrIPTs expression may be important for meristem maintenance. Overall, our results indicated that certain CK biosynthesis IPT genes expressed in unorthodox meristems, suggesting they have a role on meristem maintenance. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T05:15:29Z (GMT). No. of bitstreams: 1 ntu-103-R01b44006-1.pdf: 4486535 bytes, checksum: 5acfd3c914933a60f15323ce2ad0610b (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 中文摘要……………………………………………………………………………… I
Abstract.…………………………………………………………………………….… II Content………………………………………………………………………………III Index of figures and table……………………………………………………………V Abbreviation…………………………………………………………………………VI Overall goal……………………………………………………………………………1 Introduction…………………………………………………………………………….2 The uniqueness of Streptocarpus rexii……………………………………………….2 Cytokinin promotes meristem activity in Streptocarpus species…………………5 Similarities in regulating meristem activity shared by model plants and S. rexii…..8 The CK biosynthesis gene ISOPENTENYLTRANSFERASE (IPT) ………………10 Aims of this study…………………………………………………………………14 Materials and Methods…………………………………………………………16 Plant material………………………………………………………………16 Nucliec acid extraction, regular PCR and cloning protocol…………………16 Isolation of SrIPT genes…………………………………………………………20 3’RACE………………………………………………………………20 Inverse PCR (iPCR) ……………………………………………………23 TAIL PCR………………………………………………………………26 Isolation of SrIPT2 and SrIPT9 intron sequence………………………29 Homology assessment of SrIPTs…………………………………………………29 Sequence identity and similarity comparison…………………………29 Reconstruction of phylogenetic tree of IPT genes……………………30 Reverse transcription PCR and real-time PCR………………………………31 RT-PCR with plasmid control for relative expression analysis………31 Extraction of organs and phyllomorph RNA for expression analysis…37 Plant hormone treatment………………………………………………38 Gene expression analysis via real-time PCR…………………………39 RNA in situ hybridization……………………………………………………41 Preparation of plant tissue sample……………………………………41 Preparation of DIG-labeled RNA probe………………………………44 In situ hybridization……………………………………………………47 Results……………………………………………………………………………55 Isolation of IPT homologs in S. rexii……………………………………………55 Homology assessment of SrIPTs…………………………………………………55 Sequence identity and similarity………………………………………55 Reconstruction of phylogenetic tree of IPT genes……………………57 Verification of conserved domain shared by SrIPTs and model species IPT……58 The spatial expression analysis of SrIPTs…………………………………………62 Comparison of expression level among SrIPTs in seedlings…………62 Expression patterns of SrIPTs in different organs……………………63 SrIPTs expression level between proximal and distal phyllomorph…67 In situ expression patterns of SrIPTs in basal and groove meristem…70 Effect of hormone treatments on SrIPTs expression level……………75 Discussion…………………………………………………………………………77 SrIPTs are homologs to other model species IPT…………………………………77 SrIPT2 is likely to be a pseudogene………………………………………………77 Expression of SrIPTs associated with basal and groove meristem of S. rexii……78 SrIPTs expression overlapped with reported KNOX and GA related genes………81 Differentiated expression patterns of SrIPTs share similarities with related IPTs...82 SrIPTs may contribute to purposes other than meristem maintenance……………86 Phytohormones may involve in regulation of CK biosynthesis in S. rexii………87 Conclusion and Future Prospects………………………………………………90 Appendices………………………………………………………………………92 The biosynthesis pathway of cytokinin……………………………………………92 Summary of expression patterns of IPT homologs………………………………93 Summary of hormonal response of model plants IPTs……………………………96 cDNA sequences isolated in this study……………………………………………97 Primers used in this study………………………………………………………101 Calculating gene expression level using plasmid control………………………102 RNA in situ hybridization probe region…………………………………………103 Identity and similarity analysis of SrIPTs and AtIPTs…………………………105 Alignment of IPT homologs used in phylogenetic analysis……………………106 Premature stop codon in SrIPT2 and SlIPT5……………………………………107 References………………………………………………………………………108 | |
| dc.language.iso | en | |
| dc.subject | 苦苣苔科 | zh_TW |
| dc.subject | 堇蘭 | zh_TW |
| dc.subject | 細胞分裂素 | zh_TW |
| dc.subject | 細胞分裂素生合成 | zh_TW |
| dc.subject | 異位分生組織 | zh_TW |
| dc.subject | 異戊烯基轉移? | zh_TW |
| dc.subject | 分生組織活力 | zh_TW |
| dc.subject | cytokinin | en |
| dc.subject | meristem activity | en |
| dc.subject | Gesneriaceae | en |
| dc.subject | Streptocarpus rexii | en |
| dc.subject | isopentenyltransferase | en |
| dc.subject | unorthodox meristem | en |
| dc.subject | cytokinin biosynthesis | en |
| dc.title | 細胞分裂素生合成基因對子葉不等大堇蘭分生組織異位的維持角色 | zh_TW |
| dc.title | The role of cytokinin biosynthesis ISOPENTENYLTRANSFERASE gene in ectopic meristem maintenance of an unorthodox anisocotylous plants, Streptocarpus rexii (Gesneriaceae) | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳虹樺(Hong-Hwa Chen),蔡育彰(Yu-Chang Tsai),陳仁治(Jen-Chih Chen) | |
| dc.subject.keyword | 苦苣苔科,堇蘭,細胞分裂素,細胞分裂素生合成,異位分生組織,異戊烯基轉移?,分生組織活力, | zh_TW |
| dc.subject.keyword | Gesneriaceae,Streptocarpus rexii,cytokinin,cytokinin biosynthesis,unorthodox meristem,isopentenyltransferase,meristem activity, | en |
| dc.relation.page | 116 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-08-18 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生態學與演化生物學研究所 | zh_TW |
| 顯示於系所單位： | 生態學與演化生物學研究所 | |
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