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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 葉開溫 | |
dc.contributor.author | Chung-Yi Chiou | en |
dc.contributor.author | 邱崇益 | zh_TW |
dc.date.accessioned | 2021-06-15T04:59:02Z | - |
dc.date.available | 2010-07-29 | |
dc.date.copyright | 2010-07-29 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-07-29 | |
dc.identifier.citation | Adams, K.L., Percifield, R., Wendel, J.F. (2004). Organ-specific silencing of
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46229 | - |
dc.description.abstract | 第一部分
文心蘭為台灣重要的經濟花卉,在切花市場上極具經濟價值,但其色素形成機制之研究卻很少。本研究目的在選殖及分析與文心蘭花朵色素生合成相關之基因,進一步探討南西文心蘭(Oncidium Gower Ramsey)花朵中花青素分布特異性之調控機制。首先利用高效液相層析儀鑑定南西文心蘭花朵中花青素組成種類分別為malvidin-3-O-galactoside、peonidin-3-O-glucoside、delphinidin-3-O-glucoside 和cyanidin-3-O-glucoside;反之,在花朵的唇辦中偵測不到花青素的累積。分析四個參與花青素生合成基因的表達,發現chalcone synthase (OgCHS)、chalcone isomerase (OgCHI)、dihydroflavonol 4-reductase (OgDFR) 和 anthocyanidin synthase (OgANS)會隨著花朵發育成熟而增加表達量,而OgCHI與OgDFR卻在花朵的唇辦中不表達。利用基因槍將花苞專一啟動子(Pchrc)分別驅動OgCHI與OgDFR的基因載體送入唇辦組織中進行短暫性表現分析,發現花青素因OgCHI與OgDFR的活化表現而再度累積。而花青素生合成的調控因子如:OgMYB1、OgbHLH與OgWD40,也會伴隨花朵發育成熟而增加表達量,而OgMYB1在花朵的唇辦中也不表達。利用基因槍將花苞專一啟動子(Pchrc)驅動OgMYB1的基因載體送入唇辦組織中進行短暫性表現分析,發現花青素會因OgMYB1調控OgCHI與OgDFR的轉錄表現而再度累積。由此證明花青素在南西文心蘭花朵中分布的特異性是由OgMYB1的差異表達所造成。而OgMYB1差異轉錄層次的調控是由其DNA甲基化所影響。 第二部分 香吉士(Oncidium Gower Ramsey ‘Sunkist’)與白玉(Oncidium Gower Ramsey’White Jade’ ) 為南西文心蘭(Oncidium Gower Ramsey)的突變種,本研究目的在分析及研究這三者文心蘭中花色差異與類胡蘿蔔素形成機制之相關性。首先利用高效液相層析儀鑑定香吉士文心蘭花朵中類胡蘿蔔素組成的種類與南西文心蘭幾乎相同(lutein、violaxathin、9-cis-violaxathin和neoxanthin),但額外多出β-carotene;反之,在白玉文心蘭的花朵中偵測不到類胡蘿蔔素的累積。分析九個參與類胡蘿蔔素生合成與代謝基因在花苞成熟時期的表達,發現香吉士文心蘭中β-hydroxylase (OgHyb)與zeaxathin epoxidase (OgZep)的表現量比南西文心蘭低,此結果可能造成香吉士文心蘭花苞中β-carotene的累積;而白玉文心蘭中與類胡蘿蔔素生合成相關基因的表達與南西文心蘭並無差異,但參與類胡蘿蔔素代謝的基因carotenoid cleavage dioxygenase 1 (OgCCD1)在白玉文心蘭中卻大量表現,此結果可能造成白玉文心蘭花苞中類胡蘿蔔素無法累積。利用基因槍將花苞專一啟動子(Pchrc)驅動OgCCD1的基因載體送入南西唇辦組織中進行短暫性表現分析,發現類胡蘿蔔素會因OgCCD1的活化表現而分解類胡蘿蔔素。進一步釣取OgCCD1啟動子進行甲基化特異PCR方法分析(MSP),發現OgCCD1啟動子在南西文心蘭中有被甲基化的現象,而白玉文心蘭卻無此現象發生,由此證明OgCCD1參與白玉文心蘭白色花朵的形成。 第三部分 南西文心蘭是台灣目前最大宗的外銷切花作物,但由於人工雜交育種不易,故現在仍以此黃花品系為主,利用遺傳工程技術進行分子育種是改良品種的另一條出路。為了轉殖花色基因必須選殖一個花苞專一啟動子。首先分析了OgCHRC基因之表達圖譜(expression profile)發現此基因在根、葉不表達,但會隨著花朵發育成熟而增加表達量。因此,其5’端DNA片段應為是理想的花瓣專一表達啟動子,將其啟動子片段分離出來之後,結合GUS報導基因,藉由測試報導基因短暫性表現分析,顯示此啟動子具有花瓣組織細胞表達專一性,且特別於單子葉植物表達較佳。本研究也利用此啟動子驅動文心蘭重要花色基因,如OgCHS, OgMYB1, OgPds,進行文心蘭花色改良,以期未來有新花色品種文心蘭的產出。 | zh_TW |
dc.description.provenance | Made available in DSpace on 2021-06-15T04:59:02Z (GMT). No. of bitstreams: 1 ntu-99-D94b42004-1.pdf: 4675378 bytes, checksum: b3eef617da2aa8cb8ccdce05d68b3328 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | TABLE OF CONTENTS
Chinese abstract---------------------------------iii 1.Chapter I General introduction 1.1 The Oncidium plant----------------------------10 1.2 Introduction to flower color------------------10 1.3 Orchid transformation technology--------------12 1.4 Outline of the dissertation-------------------12 2.Chapter II Regulatory mechanism of anthocyanin pigmentation in Oncidium Gower Ramsey ------ Differential expression of MYB gene (OgMYB1) determines color patterning floral tissue of Oncidium Gower Ramsey ~ Plant Mol Biol. 2008 66 (4):379-88 2.1 ABSTRACT 2.1.1 Abstract in Chinese------------------------15 2.1.2 Abstract in English------------------------16 2.1.3 Abbreviation-------------------------------17 2.2 INTRODUCTION 2.2.1 Plant anthocyanin biosynthesis-------------18 2.2.2 Transcriptional factors regulate anthocyanin biosynthesis in Plant----------------------19 2.2.3 Goals of current research project----------21 2.3 RESULTS 2.3.1 HPLC analysis of anthocyanin in Oncidium floral tissues-------------------------------------22 2.3.2 Cloning and expression patterns of the four anthocyanin biosynthesis genes, chalcone synthase (OgCHS), chalcone isomerase (OgCHI), dihydroflavonol reductase (OgDFR), and anthocyanidin synthase (OgANS)------------------------------------------22 2.3.3 Production of red pigmentation in yellow lip tissues by transient expression of both OgCHI and OgDFR genes-------------------------------------23 2.3.4 Isolation and characterization of MYB- related genes from flower bud of Oncidium Gower Ramsey--24 2.3.5 OgMYB1 , OgWD40 and OgbHLH expression analysis, and regulation of anthocyanin biosynthetic genes by OgMYB1-------------------------------------------25 2.4 DISCUSSIONS 2.4.1 Absence of OgCHI and OgDFR cause disproportionate distribution of red pigments in floral tissue------26 2.4.2 Specific functions of OgMYB1 in mediating direct activation of OgCHI and OgDFR----------------------27 3.Chapter III Regulatory mechanism of carotenoid pigmentation in three Oncidium cultivars ------Differential expression of carotenoid-related genes determines diversifiedcarotenoid fingerprints in floral tissues of Oncidium cultivars ~ Planta (in press) 3.1 ABSTRACT 3.1.1 Abstract in Chinese-------------------------------38 3.1.2 Abstract in English-------------------------------39 3.1.3 Abbreviation--------------------------------------40 3.2 INTRODUCTION 3.2.1 Plant carotenoids biosynthesis------------------- 41 3.2.2 Plant carotenoids metabolism--------------------- 43 3.2.3 The epigenetic regulation by DNA methylation in Plant---------------------------------------------44 3.2.4 Goals of current research project-----------------45 3.3 RESULTS 3.3.1 Analysis of carotenoid composition in floral tissues of three Oncidium cultivars –Oncidium Gower Ramsey (GR), Oncidium Sunkist (Sun) and Oncidium White Jade (WJ)-----------------------------------46 3.3.2 Cloning and characterization of carotenoid-related genes---------------------------------------------47 3.3.3 Transient expression of OgCCD1 gene in floral lip tissues of Oncidium Gower Ramsey demonstrated the function of carotenoid degradation----------------49 3.3.4 Methylation effect on the OgCCD1 upstream region causes gene silence in GR cultivar----------------50 3.4 DISCUSSIONS------------------------------------------50 4. Chapter IV Cloning of floral specific promoter from Oncidium Gower Ramsey -----Characterization and promoter activity of chromoplast specific carotenoid associated gene (CHRC) from Oncidium Gower Ramsey Biotechnol Lett 2008 30:1861-1866 4.1 ABSTRACT 4.1.1 Abstract in Chinese------------------------------63 4.1.2 Abstract in English------------------------------64 4.2 INTRODUCTION 4.2.1 The biological signification of chromoplast specific carotenoidassociated gene (CHRC) in Plant--------65 4.2.2 Goals of current research project----------------66 4.3 RESULTS 4.3.1 Molecular characterization of a chromoplast specific carotenoid-associated protein gene, OgCHRC-------67 4.3.2 Isolation and sequence analysis of OgCHRC promoter (Pchrc) in silico--------------------------------67 4.3.3 Transient expression analysis of the OgCHRC promoter using particle bombardment------------------------68 4.3.4 Analysis of OgCHRC promoter activity in floral tissues of various Plants-------------------------68 4.3.5 Application of Pchrc promoter driving flower color genes in Oncidium transformation-----------------69 4.4 DISCUSSIONS-----------------------------------------69 5. CONCLUSION---------------------------------------- 71 6. FUTURE PROSPECTUS------------------------------------72 7. TABLE------------------------------------------------79 8. MATERIALS AND METHODS--------------------------------82 8.1 Plant materials-------------------------------------82 8.2 RNA extraction -------------------------------------82 8.3 Rapid amplification of cDNA end; RACE --------------84 8.4 RT PCR for gene expression (One-step RT PCR)--------90 8.5 Anthocyanins analysis-------------------------------91 8.6 Carotenoid analysis---------------------------------92 8.7 Northern blot analysis------------------------------93 8.8 Radioactive DNA probes------------------------------93 8.9 Methylation analysis using the bisulfite sequence method-------------------------------------94 8.10 Constructs for transient expression using particle bombardment-------------------------------94 8.11 Phylogenetic analysis 8.11.1 A phylogenetic analysis of the deduced amino acid sequences of MYBs Related to anthocyanin regulation or other physiological processes------95 8.11.2 Protein Sequence alignment of OgCCD1 with other plant CCDs, and OgNCED with other plant NCEDs involved in ABA biosynthesis--------------------96 9. REFERENCES-------------------------------------------98 | |
dc.language.iso | en | |
dc.title | 調控文心蘭花色形成機制之研究 | zh_TW |
dc.title | Regulatory mechanism of floral coloration in Oncidium cultivars | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 林秋榮,黃麗春,陳虹樺,謝旭亮,張耀乾,楊文彬 | |
dc.subject.keyword | 文心蘭,花青素,胡蘿蔔素, | zh_TW |
dc.subject.keyword | Oncidiun,anthocyanin,carotenoid, | en |
dc.relation.page | 117 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-07-29 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 植物科學研究所 | zh_TW |
顯示於系所單位: | 植物科學研究所 |
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