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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林彥蓉 | |
| dc.contributor.author | Meng-Ying Lin | en |
| dc.contributor.author | 林孟穎 | zh_TW |
| dc.date.accessioned | 2021-06-08T00:13:46Z | - |
| dc.date.copyright | 2013-08-14 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-02 | |
| dc.identifier.citation | 朱昌蘭, 沈文飆, 翟虎渠, 萬建民 (2004) 水稻低直鏈澱粉含量基因育種利用的研究進展。中國農業科學 37: 157-162
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17449 | - |
| dc.description.abstract | 水稻穀粒呈不透明的突變體，包含粉質、糯性及半糯胚乳，為食品和工業利用上之重要性狀，不僅可作為研究充實期養分儲藏代謝過程的遺傳材料，也能促進了解澱粉生合成相關基因的調控網絡。本研究所使用的粉質胚乳材料為臺稉 8 號經 EMS 誘變且經多代自交固定而得之突變品系 WY14，其直鏈澱粉含量 (14.20 ± 0.05 %) 顯著低於臺稉 8 號 (18.15 ± 0.58 %)，第一期作穀粒充實較慢，最終累積乾重較輕，但在第二期作則無顯著差異。WY14 與臺稉 8 號之蛋白質種類及含量無顯著差異，而胚乳內脂肪酸含量和組成不同，WY14 總脂肪酸含量 (0.38 ± 0.03 %) 顯著高於臺稉 8 號 (0.09 ± 0.01 %)，棕櫚酸及花生酸比例顯著較高，且額外偵測到 0.16 % 長鏈飽和脂肪酸山萮酸。粉質突變基因經定位選殖的結果為 FLO7 的第一個外顯子有一核苷酸序列突變造成非同義置換，經基因及功能比對顯示 FLO7 為新穎的粉質基因，預測為 ABC 轉運蛋白，推測在水稻穀粒充實期澱粉生合成路徑中扮演運送物質的角色。以 RNA 干擾擾技術弱化 FLO7 之基因表現，22 個 RNAi 系之 T1 種子中，11 個轉殖系的榖粒透明與粉質分離，再度證明 FLO7 可造成水稻粉質胚乳。利用 3’ RACE 進行四種預測 FLO7 轉錄物之確認，FLO7 在胚乳內主要轉錄物為 FLO7-2a。以即時定量 PCR 反應進行 FLO7 及其他澱粉生合成相關基因於 101 年第一及第二期作水稻三葉齡組織或未成熟穎果的基因表現分析，FLO7 於三葉齡組織及未成熟穎果皆有表現，無明顯組織專一性，WY14 及臺稉 8 號於授粉後第 15 天表現量差異最大。以 WY14 相較臺稉 8 號之基因表現結果觀察，與澱粉生合成相關之上游路徑中，在細胞質作用的基因表現量大致呈現上升趨勢，如 UGP、OsAGPL2 及 OsAPGS2b 等；而下游路徑中，在澱粉體作用的基因表現量則下降，如OsGBSSI、OsSSIIIa 及 OsBEI 等。WY14 之 PPDKA 及 cyPPDKB 表現顯著高於臺稉 8 號，可能為脂肪酸含量較高之原因。推測 FLO7 可能參與在澱粉生合成之相關路徑中，影響特定物質轉運至澱粉體並導致澱粉排列改變。本研究結果對水稻胚乳內澱粉生合成相關基因調控機制及遺傳網絡的研究有所貢獻，日後亦可利用分子標幟輔助選種之方式將粉質胚乳性狀導入現行品種，增加稻米的多元應用。 | zh_TW |
| dc.description.abstract | Opaque rice endosperm including floury, glutinous, and dull grains is important for food and industrial applications. The corresponding mutants not only provide valuable genetic materials for elucidating metabolic processes related to nutrient storage during grain filling but also facilitate the understanding the gene network of starch synthesis related genes. A floury mutant, WY14, is one of the mutants from TK8 induced by EMS which possessed lower amylose content (14.20 ± 0.05%) compared to that of wild type TK8 (18.15 ± 0.58 %). WY14 proceeded worse grain filling and accumulated lower dry weight of grains in the first crop season of 2012 than TK8 did, but not in the second crop season of 2012. Profiles of total storage proteins in WY14 and TK8 revealed no difference. However, WY14 had higher amount of fatty acid and proportion of palmitic acid and arachidic acid than TK8, and WY14 possessed additional 0.16 % of long-chain saturated fatty acid, behenic acid. The mutated gene, conferring floury endosperm of WY14, was isolated by positional cloning, which was named FLO7 and annotated as ABC transporter.
The mutation of FLO7 responsible for floury endosperm was confirmed by RNAi. T1 seeds of RNAi lines showed segregation of both translucent and floury endosperm. Identification of 4 FLO7 transcripts by 3’ RACE suggested that the main FLO7 transcript in endosperm is FLO7-2a. FLO7 expressed not only in vegetative tissues but also in immature caryopsis, and FLO7 expressed differently with significance between WY14 and TK8 at 15 DAP. Gene expression of most starch biosynthesis related enzymes acting in cytosol had higher level, but not those in amyloplast which were down-regulated in WY14 as compared to in TK8. Futhermore, higher expression of PPDKA and cyPPDKB may account for higher total amount of fatty acid in WY14 than in TK8. This research contributes to the relevant regulatory mechanism and genetic network related to starch synthesis in rice endosperm. The flo7 of WY14 may be applied to breed new floury varieties by marker assisted selection for multipurpose usage of rice grains. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T00:13:46Z (GMT). No. of bitstreams: 1 ntu-102-R00621111-1.pdf: 3363384 bytes, checksum: c117ede2d251d729d17d61b67641a7ca (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 目錄
致謝 I 中文摘要 II Abstract III 目錄 V 圖目錄 VII 縮寫表 VIII 壹、前言 1 貳、前人研究 4 一、水稻胚乳發育 4 二、水稻胚乳內蛋白質與脂質 5 三、水稻澱粉結構及其生合成 7 四、食味品質相關基因及 QTLs 10 五、水稻胚乳突變體 12 六、ABC 運輸蛋白 (ATP-binding cassette transporter) 14 七、FLO7 基因表現預測 15 參、材料和方法 17 一、試驗材料 17 二、穀粒成分分析 19 (一) 穀粒充實過程之穎果乾重 19 (二) 直鏈澱粉之測定 19 (三) 穎果蛋白質萃取 20 (四) 胚乳脂肪酸萃取 21 三、FLO7 缺失造成粉質胚乳之確認 22 (一) 大腸桿菌轉型及 FLO7 之 RNAi 構築 24 (二) 農桿菌轉型 (Agrobacterium-mediated transformation) 26 (三) 農桿菌轉殖 26 四、FLO7 和其他澱粉生合成相關基因之轉錄分析 27 (一) Total-RNA 用於 3’ RACE (rapid amplification of cDNA ends) 以確認 FLO7 在穀粒充實期表現之轉錄物 (transcript)，FLO7 於不同組織之表現及其他澱粉生合成相關基因之轉錄分析 27 (二) FLO7 之 3’-RACE 分析 28 (三) FLO7 和其他澱粉生合成相關基因之轉錄分析 30 肆、結果 32 一、穀粒成分分析 32 二、FLO7 於水稻胚乳內之功能及基因表現分析 39 三、胚乳內澱粉生合成相關基因表現分析 45 伍、討論 55 一、FLO7 對水稻穀粒成分的影響 55 二、FLO7 基因之轉錄分析 58 三、FLO7 之突變擾動澱粉生合成路徑 60 四、未來展望 64 陸、參考文獻 66 柒、附錄 76 | |
| dc.language.iso | zh-TW | |
| dc.title | 水稻粉質基因 FLO7 對穀粒成分及澱粉生合成相關基因之影響 | zh_TW |
| dc.title | The Effect of a Floury Gene Floury7 (FLO7) in Rice (Oryza sativa L.) on Grain Composition and Starch Synthesis Related Genes | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 邢禹依,洪傳揚,張孟基 | |
| dc.subject.keyword | 水稻,粉質胚乳,ABC轉運蛋白, | zh_TW |
| dc.subject.keyword | Oryza sativa,floury endosperm,ABC transporter, | en |
| dc.relation.page | 96 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2013-08-02 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 農藝學研究所 | zh_TW |
| 顯示於系所單位： | 農藝學系 | |
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