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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8907完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 洪傳揚 | |
| dc.contributor.author | Yi-Chieh Lin | en |
| dc.contributor.author | 林依潔 | zh_TW |
| dc.date.accessioned | 2021-05-20T20:03:51Z | - |
| dc.date.available | 2019-07-03 | |
| dc.date.available | 2021-05-20T20:03:51Z | - |
| dc.date.copyright | 2009-08-21 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-08-18 | |
| dc.identifier.citation | 戶刈義次 (1963). 作物學試驗法. 東京農業技術學會印行 第159-176頁
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Mutations of the multi-drug resistance-associated protein ABC transporter gene 5 result in reduction of phytic acid in rice seeds. Theor Appl Genet 119, 75-83. Yamaguchi-Shinozaki, K., and Shinozaki, K. (2005). Organization of cis-acting regulatory elements in osmotic- and cold-stress-responsive promoters. Trends Plant Sci 10, 88-94. Yamaguchi, H., Nishizawa, N.K., Nakanishi, H., and Mori, S. (2002). 1D17, a new iron-regulated ABC transporter from barley roots, localizes to the tonoplast. J Exp Bot 53, 727-735. Yasuno, N., Takamure, I., Kidou, S., Tokuji, Y., Ureshi, A.N., Funabiki, A., Ashikaga, K., Yamanouchi, U., Yano, M., and Kato, K. (2009). Rice shoot branching requires an ATP-binding cassette subfamily G protein. New Phytol 182, 91-101. Yazaki, K. (2005). Transporters of secondary metabolites. Curr Opin Plant Biol 8, 301-307. Yazaki, K., Shitan, N., Takamatsu, H., Ueda, K., and Sato, F. (2001). A novel Coptis japonica multidrug-resistant protein preferentially expressed in the alkaloid-accumulating rhizome. J Exp Bot 52, 877-879. Yew, W.W., and Leung, C.C. (2008). Management of multidrug-resistant tuberculosis: Update 2007. Respirology 13, 2 1-46. Yuan, H., Li, X., Wu, J., Li, J., Qu, X., Xu, W., and Tang, W. (2008). Strategies to overcome or circumvent P-glycoprotein mediated multidrug resistance. Curr Med Chem 15, 470-476. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/8907 | - |
| dc.description.abstract | ABC轉運蛋白(ATP-binding cassette transporter)廣泛存在各種物種之間，對各種不同基質的轉運扮演重要的角色。目前植物的ABC轉運蛋白功能以阿拉伯芥研究較多，水稻中ABC轉運蛋白功能性分析的相關研究非常少。因此本試驗擬針對水稻ABC轉運蛋白MDR次基因家族成員OsABCB10基因進行功能性分析。胺基酸比對結果顯示，OsABCB10為全分子轉運蛋白，具有ABC轉運蛋白的保守性功能區域Walker A、ABC signature以及Walker B。即時定量聚合酶連鎖反應(Q-RT- PCR)分析OsABCB10組織專一性基因表現，結果顯示孕穗期水稻的葉身、葉鞘及花序中都有相當高OsABCB10基因的表現，未成熟種子中表現量更強。以植物荷爾蒙處理兩週大水稻幼苗，結果顯示NAA、JA及SA都會誘導OsABCB10基因在地上部或根部表現。高鹽、ABA以及低溫(4℃)等非生物逆境都會誘導地上部OsABCB10的表現，而根部則受高鹽、乾旱及低溫誘導表現，其中低溫處理可強烈誘導根部OsABCB10的表現超過1000倍以上。分析POsABCB10-GUS轉殖水稻，結果發現GUS主要累積在葉片以及未成熟花序中，尤其集中於內穎、外穎、小穗軸及雌蕊部分。以低溫處理POsABCB10-GUS轉殖株2天，結果顯示GUS受誘導表現累積於根部。將POsABCB10-GUS種子浸潤後，結果顯示浸潤後2小時啟動子活性最高，隨浸潤時間增加而活性降低。分析OsABCB10:GFP融合蛋白於細胞中表現的位置，結果顯示OsABCB10是一個表現於細胞膜上的蛋白質。進一步分析基因剔除突變株abcb10同型合子，外表性狀與野生型無明顯差異，但是溫室或是田間試驗的結果顯示，abcb10稔實率較野生型大幅降低60%以上，同時abcb10花粉也有近40%表現出發育不良的情形。反之，OsABCB10基因大量表現的轉殖系，則與野生型無明顯差異。這些結果顯示，OsABCB10可能是維持水稻穀粒稔實率的重要基因。另外，試驗中亦觀察到abcb10種子有發芽遲緩及發芽率降低的現象，顯示OsABCB10可能也參與了種子發芽過程的調節。此試驗的結果說明了低溫誘導表現的OsABCB10可能是參與調節水稻穀粒充實率或種子發芽的ABC轉運蛋白。 | zh_TW |
| dc.description.abstract | ATP binding cassette (ABC) transporters ubiquitously existed in all living organisms, and play important roles in transporting various substances across membrane systems. The functions of ABC transporters have been investigated extensively in Arabidopsis, however, very few studies are reported in rice. To this end, this study focuses on the functional characterization of the rice OsABCB10 ABC transporter. Amino acid alignment analysis showed that the OsABCB10 is a full molecule MDR-type ABC transporter containing typical Walker A、ABC signature and Walker B domains. Quantitative RT-PCR analysis indicated that the OsABCB10 was highly expressed in blade, sheath and inflorescence at booting stage, and strongly expressed in the immature seeds. Treatment of two-week-old rice seedlings with different plant hormones revealed that the OsABCB10 was induced by NAA, JA and SA both in shoot and root. Furthermore, the OsABCB10 was slightly induced by salt, ABA and cold (4℃) in shoot tissue, while it was slightly induced by salt and drought in root tissue. Surprisingly, cold treatment dramatically increased more than 1000 folds of OsABCB10 transcript accumulated in root tissue. Transgenic rice harboring POsABCB10-GUS construct showed that GUS protein slightly accumulated in blade and inflorescence, and strongly expressed in palea, lemma, rachilla and pistil. Upon cold treatment, GUS protein was induced in root tissue of POsABCB10-GUS plants as compared with control plants. Moreover, OsABCB10 promoter activity was enhanced after 2 hours imbibition, and then gradually declined with imbibition time. Subcellular localization assay showed that OsABCB10 is localized at plasma membrane. The OsABCB10 knock-out mutant abcb10 showed a striking seed fertility reduced phenotype either cultivated in the greenhouse or in the field. Meanwhile, up to 40% non-fertile pollen was observed in abcb10. In contrast, rice overexpressing OsABCB10 had no effect on seed fertility. These results demonstrated that OsABCB10 is required for seed development, and may be a crucial gene to maintain seed fertility in rice. In addition, seeds of abcb10 showed a germination delayed phenotype and possessed lower germination rate as compared with WT, indicating that OsABCB10 plays a role in modulating seed germination. Taken together, our results provide evidences to support that cold treatment highly induced MDR-type ABC transporter OsABCB10 may be involved in mediating seed fertility or seed germination of rice. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T20:03:51Z (GMT). No. of bitstreams: 1 ntu-98-R96623004-1.pdf: 3974924 bytes, checksum: dbbcc856d337b22aa577a6ecfc4011f4 (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 中文摘要……………………………………………………………IV
英文摘要………………………………………………………V 縮寫字對照表………….……………………………………VII 壹、前言 一、ABC轉運蛋白的發現…………………………………………………1 二、ABC轉運蛋白的基本特性……………………………………………2 三、ABC轉運蛋白的分類與命名…………………………………………4 四、植物的ABC轉運蛋白……………………………………………...7 五、植物ABC轉運蛋白的功能……………………………8 六、水稻ABC轉運蛋白基因的研究..………………………17 貳、研究目的………………………………………………………21 参、材料與方法 一、質粒的構築…………………………………………22 二、水稻基因轉殖……………………………………………………26 三、基因表現分析材料與處理…………………………………………28 四、基因表現分析………………………………………………………31 五、基因型分析(genotyping)………………………………………………35 六、啟動子特性分析…………………………………………………………35 七、蛋白質次細胞位置分析.……………………………………36 八、花粉活性鑑定………….……………………………………………37 九、種子發芽率分析……………………………………………37 肆、結果 一、水稻MDR基因家族分子特性分析.…………………………………38 二、OsABCB10基因表現特性分析.………………………………………39 三、OsABCB10啟動子特性分析…………………………………………41 四、OsABCB10次細胞定位分析.………………………………………43 五、OsABCB10基因活化標記(activation-tagged)轉殖株品系M0029907的分析.………………43 六、OsABCB10基因剔除突變株(NF2043)的篩選及外表性狀觀察.…………44 伍、討論.………………….………..…47 陸、參考文獻....................52 圖1、水稻MDR基因家族胺基酸保守性序列分析………………….……….………63 圖2、ABC轉運蛋白MDR(ABCB)次基因家族成員的親源演化樹分析……….……65 圖3、以生物資訊學方式進行OsABCB10胺基酸序列分析…………………………66 圖4、OsABCB10組織專一性之基因表現分析……………………………………..…67 圖5、OsABCB10於孕穗期穎花內部各花器之基因表現分析…...…………………...68 圖6、水稻幼苗於不同植物荷爾蒙處理下OsABCB10基因表現分析……………….69 圖7、水稻幼苗於不同重金屬處理下OsABCB10基因表現分析.……..…………..…70 圖8、水稻幼苗於不同非生物逆境處理下OsABCB10基因表現分析………….……71 圖9、OsABCB10啟動子cis-acting element區域分析………………………….…..…72 圖10、POsABCB10-GUS轉殖水稻的組織化學染色分析………….………….…….……73 圖11~13、POsABCB10-GUS轉殖水稻幼花序的組織化學染色分析…….……….....74~76 圖14、POsABCB10-GUS轉殖水稻穎花內部各構造的組織化學染色分析……….……..77 圖15、POsABCB10-GUS轉殖水稻於低溫逆境處理後組織化學染色分析………….…..78 圖16、POsABCB10-GUS轉殖水稻種子浸潤(imbibition)後化學組織染色分析….……..79 圖17、OsABCB10蛋白的次細胞定位(subcellular localization)………...….....……..80 圖18、OsABCB10基因活化標記(activation-tagged)轉殖株M0029907之分析.……..81 圖19、基因活化標記轉殖株M0029907中對OsABCB10兩翼基因之影響…...……..82 圖20、OsABCB10基因剔除(knock-out)突變株品系NF2043之篩選……….....……..83 圖21、野生型及基因剔除突變株NF2043開花期與孕穗期外表性狀差異......……..84 圖22、OsABCB10基因缺失使種子稔實率(Seed Fertility)大幅降低……….....……..85 圖23、OsABCB10基因的活化或缺失突變株種子外觀及千粒重分析………..……..86 圖24、OsABCB10基因缺失突變株顯著降低活性花粉數……………………..……..87 圖25、野生型(TNG67)及OsABCB10基因活化轉殖株之種子發芽率分析……..…..88 圖26、野生型(Nipponbare)及OsABCB10基因剔除突變株之種子發芽分析………..89 | |
| dc.language.iso | zh-TW | |
| dc.title | 水稻ABC類型ATP-Binding Cassette(ABC)轉運蛋白OsABCB10的功能分析 | zh_TW |
| dc.title | Functional Analysis of Rice MDR-type ATP-Binding Cassette (ABC) Transporter OsABCB10 | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 余淑美,盧虎生,陸重安,陳凱儀 | |
| dc.subject.keyword | 功能性分析,ABC轉運蛋白, | zh_TW |
| dc.subject.keyword | Functional analysis,ABC transporter, | en |
| dc.relation.page | 115 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2009-08-18 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 農業化學研究所 | zh_TW |
| 顯示於系所單位： | 農業化學系 | |
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