阿拉伯芥AtMYBSs基因參與在糖訊息及離層酸訊息傳遞之研究

以作者查詢圖書館館藏

、以作者查詢臺灣博碩士

、以作者查詢全國書目

、勘誤回報

、線上人數：136

、訪客IP：3.135.183.89

姓名

吳珮綺(Pei-Chi Wu) 查詢紙本館藏

畢業系所

生命科學系

論文名稱

阿拉伯芥AtMYBSs基因參與在糖訊息及離層酸訊息傳遞之研究
(Functional analysis of AtMYBSs involved in Sugar- and ABA-signal pathway in Arabidopsis)

相關論文

★ 水稻CAF1基因之功能分析-水稻CAF1基因的選殖、定性及表現	★ 水稻OsDEADl-1基因的功能性探討
★ 利用水稻細胞之懸浮培養建立蛋白質高效率分泌系統	★ 水稻CCR4基因之功能分析- 水稻CCR4基因的選殖、定性及表現
★ 阿拉伯芥 AtMYBS 基因功能性探討	★ 水稻OsMYBS2基因的功能性分析
★ 水稻CCR4基因的功能分析- 繁衍大量表現和靜默表現的基因轉殖水稻	★ 水稻OsVALs基因的功能性分析- 水稻OsVALs基因的選殖、定性及表現
★ 分析水稻T-DNA插入突變株: M0022150, M0023563, M0023580, M0037352及M0032079	★ 以水稻懸浮培養細胞蛋白質生產系統生產mGMCSF
★ 建立表現耐熱澱粉普魯南糖酶基因之轉植甘藷	★ I. II.
★ 探討αAmy3、OsCIN1與Os33KD信號肽在水稻懸浮培養細胞中的功能及特性	★ 水稻CAF1基因在水稻懸浮培養細胞之研究
★ 探討阿拉伯芥兩個MYB-related轉錄因子在糖訊息傳遞中所扮演的角色	★ 水稻中五個DEAD-box RNA helicase - RH2、RH6、RH22、RH42和RH51基因之探討

檔案

[Endnote RIS 格式]

[Bibtex 格式]

[相關文章]

[文章引用]

[完整記錄]

[館藏目錄]

至系統瀏覽論文 ( 永不開放)

摘要(中)

水稻中OsMYBSs屬於R1/R2這類的MYB轉錄因子，已知會參與在糖訊息的途徑中，藉由結合在啟動子之TA-Box（TATCCA）序列，調控下游基因αAmy3的表現。利用水稻OsMYBSs的蛋白質氨基酸序列，與阿拉伯芥蛋白質資料庫進行比對，得到四個相似度極高且同時具有R1/R2家族高度保留區的序列，分別命名AtMYBS1、AtMYBS2、AtMYBS3與AtMYBS5。本論文主要利用reverse genetics的方式，以gain-of-function（大量表現AtMYBSs）以及loss-of -function（T-DNA插入之atmybs突變株）做為研究材料，來研究AtMYBSs在阿拉伯芥中的功能。同時考量研究轉錄因子時，常因為有基因redundancy的現象而增加研究的複雜度，我們利用Chimeric repressor silence technology（CRES-T）此項技術解決基因redundancy的問題。大量表現AtMYBS-SRDX的轉殖株也做為我們的研究材料，利用觀察這些植株的表徵，來推測AtMYBSs在阿拉伯芥中扮演的功能為何。在一般正常生長情況（16h light/8h dark）下，突變株atmybs2、atmybs3及tamybs5與wild-type植株外表性狀並未有明顯的差異。而且AtMYBS並未參與在澱粉代謝的途徑中。在種子萌芽時期，atmybs2的突變株對於高濃度糖與植物荷爾蒙ABA具有不敏感的外表型，atmys2突變株對於熱逆境及鹽害逆境並未有較高的抗性，顯示AtMYBS2在種子萌芽的階段可能參與在糖與ABA調控的途徑中。DIN3 (dark-induced 3)和DIN4 (dark-induced 4)基因為branched-chain ?-ketoacid dehydrogenase的酵素複合體的成員蛋白質，參與在支鏈胺基酸的代謝途徑中，其啟動子的位置上具有TA-box like的序列，在幼苗時期經由qRT-PCR分析，發現AtMYBS2負調控DIN3和DIN4基因的表現。CwINV4 (cell wall invertases)啟動子的位置上也有TA-box，也被AtMYBS2負調控著，綜合以上結果，推測AtMYBS2參與在糖訊息與植物荷爾蒙ABA傳遞途徑中，藉由結合在TA box的序列而調控下游基因的表現，而影響阿拉伯芥種子的萌芽。

摘要(英)

MYB transcription factors are one of the largest gene families in plants. There are three types of MYB proteins, R1R2R3, R2R3, R1/2. Previously, three novel R1/2 type MYB proteins in rice, OsMYBS1, OsMYBS2, and OsMYBS3, which are reported involved in sugar- and hormone-regulated α-amylase genes expression during seed germination. In order to further investigate the biological roles of MYBSs in plants, Arabidopsis is applied in this study because of its powerful bioinformatics and comprehensive mutant lines. Here, both gain-of-function and loss-of-function strategies were applied by for studying the physiological roles of AtMYBSs. Gene redundancy interfere functional analyses of MYBSs transcription factors. Therefore, Chimeric REpressor Silencing Technology (CRES-T), fused the short transcriptional repressor domain (SRDX) to AtMYBSs, is also used.
There are no obvious different phenotypes and leaf starch content existing in gene knock-out of AtMYBSs in Arabidopsis. The atmybs2 presents glucose and abscisic acid (ABA)-insensitive phenotype during seed germination stage. The atmybs2 also presents ABA-insensitive in the seedling root growth. But the atmybs2 is not insensitive to heat stress or slat stress. The DIN3 (dark-induced 3) and DIN4 (dark-induced 4) which are subunits of a branched-chain ?-ketoacid dehydrogenase functions involved in amino acid degradation, both are downstream regulated genes of AtMYBS2. In addition to, one of cell wall invertases, CwINV4, is also a target gene of AtMYBS2. TA-box is identified on promoter region of these 3 AtMYBS2-responsive genes. According to these results, we conclude that AtMYBS2 may involve in seed germination pathway by regulating the TA-box containing gene(s) expression.

關鍵字(中)

★ ABA
★ 糖
★ MYB轉錄因子

關鍵字(英)

★ Myb transcription factor
★ sugar
★ ABA

論文目次

本文目錄
一、前人研究……………………………………………………….1
1-1植物糖訊息傳遞及基因調控研究………………………………1
1-2 MYB轉錄因子之介紹……………………………………………5
1-3利用CRES-T方法研究轉錄因子之功能…………………………8
二、研究動機與目的………………………………………………10
三、材料與方法……………………………………………………11
3-1使用植株、載體與植物品系…………………………………11
3-2轉殖株基因型的分析………………………………………….11
3-3基因表現分析…………………………………………………12
3-4 CRES-T基因構築…………………………………14
3-5植株分析方法………………………………………………………………………17
3-6短暫性基因表現分析………………………………………………17
四、實驗結果……………………………………………………………………….20
4-1阿拉伯芥突變株及轉殖株分析……………………………………20
4-2 阿拉伯芥參與在糖訊息調控的外觀分析…………………………22
4-3 阿拉伯芥參與在ABA訊息調控的外觀分析………………………23
4-4 AtMYBSs下游基因分析…………………………………………24
五、討論……………………………………………………………………27
5-1 AtMYBS2參與在糖訊息調控途徑中…………………………………………………27
5-2 AMYBS2 對下游基因表現的調控…………………………………………………….28
5-3 AtMYBS2在糖訊息及ABA訊息傳遞途徑中所扮演的角色……………30
六、未來研究與建議………………………………………………………………………32
七、參考文獻……………………………………………………………………68
附錄I…………………………………………………………………74
附錄II………………………………………………………………76
附錄III………………………………………………………………79

參考文獻

馬翎甄。2006。阿拉伯芥 AtMYBS 基因功能性探討阿拉伯芥 AtMYBS 基因功能性探討。國立中央大學生命科學研究所碩士論文。台灣，桃園。
Arenas-Huertero, F., Arroyo, A., Zhou, L., Sheen, J. and Leon, P. (2000) Analysis of arabidopsis glucose insensitive mutants, gin5 and gin6, reveals a central role of the plant hormone ABA in the regulation of plant vegetative development by sugar. Genes Dev., 14(16), 2085-2096.
Baena-Gonzalez, E. and Sheen, J. (2008) Convergent energy and stress signaling. Trends Plant Sci., 13(9), 474-482.
Baena-Gonzalez, E., Rolland, F., Thevelein, J. M. and Sheen, J. (2007) A central integrator of transcription networks in plant stress and energy signalling. Nature, 448(7156), 938-942.
Bajguz, A. (2009) Brassinosteroid enhanced the level of abscisic acid in chlorella vulgaris subjected to short-term heat stress. J. Plant Physiol., 166(8), 882-886.
Carre, I.A. and Kim, J. Y. (2002) MYB transcription factors in the arabidopsis circadian clock. J. Exp. Bot., 53(374), 1551-1557.
Cheng, W.H., Endo, A., Zhou, L., Penney, J., Chen, H. C., Arroyo, A., Leon, P., Nambara, E., Asami, T., Seo, M., Koshiba, T. and Sheen, J. (2002) A unique short-chain dehydrogenase/reductase in arabidopsis glucose signaling and abscisic acid biosynthesis and functions. Plant Cell, 14(11), 2723-2743.
Churin, Y., Adam, E., Kozma-Bognar, L., Nagy, F. and Borner, T. (2003) Characterization of two myb-like transcription factors binding to CAB promoters in wheat and barley. Plant Mol. Biol., 52(2), 447-462.
Dubos, C., Le Gourrierec, J., Baudry, A., Huep, G., Lanet, E., Debeaujon, I., Routaboul, J. M., Alboresi, A., Weisshaar, B. and Lepiniec, L. (2008) MYBL2 is a new regulator of flavonoid biosynthesis in arabidopsis thaliana. Plant J., 55(6), 940-953.
Finkelstein, R.R. and Lynch, T. J. (2000) The arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor. Plant Cell, 12(4), 599-609.
Frampton, J., Leutz, A., Gibson, T. and Graf, T. (1989) DNA-binding domain ancestry. Nature, 342(6246), 134.
Fujiki, Y., Sato, T., Ito, M. and Watanabe, A. (2000) Isolation and characterization of cDNA clones for the e1beta and E2 subunits of the branched-chain alpha-ketoacid dehydrogenase complex in arabidopsis. J. Biol. Chem., 275(8), 6007-6013.
Fujiki, Y., Yoshikawa, Y., Sato, T., Inada, N., Ito, M., Nishida, I. and Watanabe, A. (2001) Dark-inducible genes from arabidopsis thaliana are associated with leaf senescence and repressed by sugars. Physiol. Plant., 111(3), 345-352.
Fujita, Y., Fujita, M., Satoh, R., Maruyama, K., Parvez, M. M., Seki, M., Hiratsu, K., Ohme-Takagi, M., Shinozaki, K. and Yamaguchi-Shinozaki, K. (2005) AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in arabidopsis. Plant Cell, 17(12), 3470-3488.
Graham, I.A. and Eastmond, P. J. (2002) Pathways of straight and branched chain fatty acid catabolism in higher plants. Prog. Lipid Res., 41(2), 156-181.
Hiratsu, K., Mitsuda, N., Matsui, K. and Ohme-Takagi, M. (2004) Identification of the minimal repression domain of SUPERMAN shows that the DLELRL hexapeptide is both necessary and sufficient for repression of transcription in arabidopsis. Biochem. Biophys. Res. Commun., 321(1), 172-178.
Hiratsu, K., Matsui, K., Koyama, T. and Ohme-Takagi, M. (2003) Dominant repression of target genes by chimeric repressors that include the EAR motif, a repression domain, in arabidopsis. Plant J., 34(5), 733-739.
Hiratsu, K., Ohta, M., Matsui, K. and Ohme-Takagi, M. (2002) The SUPERMAN protein is an active repressor whose carboxy-terminal repression domain is required for the development of normal flowers. FEBS Lett., 514(2-3), 351-354.
Jin, H. and Martin, C. (1999) Multifunctionality and diversity within the plant MYB-gene family. Plant Mol. Biol., 41(5), 577-585.
Kawamura, M., Ito, S., Nakamichi, N., Yamashino, T. and Mizuno, T. (2008) The function of the clock-associated transcriptional regulator CCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) in arabidopsis thaliana. Biosci. Biotechnol. Biochem., 72(5), 1307-1316.
Klempnauer, K.H., Gonda, T. J. and Bishop, J. M. (1982) Nucleotide sequence of the retroviral leukemia gene v-myb and its cellular progenitor c-myb: The architecture of a transduced oncogene. Cell, 31(2 Pt 1), 453-463.
Koch, K.E., Nolte, K. D., Duke, E. R., McCarty, D. R. and Avigne, W. T. (1992) Sugar levels modulate differential expression of maize sucrose synthase genes. Plant Cell, 4(1), 59-69.
Laby, R.J., Kincaid, M. S., Kim, D. and Gibson, S. I. (2000) The arabidopsis sugar-insensitive mutants sis4 and sis5 are defective in abscisic acid synthesis and response. Plant J., 23(5), 587-596.
Leon, P. and Sheen, J. (2003) Sugar and hormone connections. Trends Plant Sci., 8(3), 110-116.
Lin, P.C., Hwang, S. G., Endo, A., Okamoto, M., Koshiba, T. and Cheng, W. H. (2007) Ectopic expression of ABSCISIC ACID 2/GLUCOSE INSENSITIVE 1 in arabidopsis promotes seed dormancy and stress tolerance. Plant Physiol., 143(2), 745-758.
Lu, C.A., Lim, E. K. and Yu, S. M. (1998) Sugar response sequence in the promoter of a rice alpha-amylase gene serves as a transcriptional enhancer. J. Biol. Chem., 273(17), 10120-10131.
Lu, C.A., Ho, T. H., Ho, S. L. and Yu, S. M. (2002) Three novel MYB proteins with one DNA binding repeat mediate sugar and hormone regulation of alpha-amylase gene expression. Plant Cell, 14(8), 1963-1980.
Lu, C.A., Lin, C. C., Lee, K. W., Chen, J. L., Huang, L. F., Ho, S. L., Liu, H. J., Hsing, Y. I. and Yu, S. M. (2007) The SnRK1A protein kinase plays a key role in sugar signaling during germination and seedling growth of rice. Plant Cell, 19(8), 2484-2499.
Matsui, K., Umemura, Y. and Ohme-Takagi, M. (2008) AtMYBL2, a protein with a single MYB domain, acts as a negative regulator of anthocyanin biosynthesis in arabidopsis. Plant J., 55(6), 954-967.
Matsui, K., Hiratsu, K., Koyama, T., Tanaka, H. and Ohme-Takagi, M. (2005) A chimeric AtMYB23 repressor induces hairy roots, elongation of leaves and stems, and inhibition of the deposition of mucilage on seed coats in arabidopsis. Plant Cell Physiol., 46(1), 147-155.
Mitsuda, N., Hiratsu, K., Todaka, D., Nakashima, K., Yamaguchi-Shinozaki, K. and Ohme-Takagi, M. (2006) Efficient production of male and female sterile plants by expression of a chimeric repressor in arabidopsis and rice. Plant. Biotechnol. J., 4(3), 325-332.
Ohta, M., Matsui, K., Hiratsu, K., Shinshi, H. and Ohme-Takagi, M. (2001) Repression domains of class II ERF transcriptional repressors share an essential motif for active repression. Plant Cell, 13(8), 1959-1968.
Pandey, G.K., Grant, J. J., Cheong, Y. H., Kim, B. G., Li, L. and Luan, S. (2005) ABR1, an APETALA2-domain transcription factor that functions as a repressor of ABA response in arabidopsis. Plant Physiol., 139(3), 1185-1193.
Penfield, S., Li, Y., Gilday, A. D., Graham, S. and Graham, I. A. (2006) Arabidopsis ABA INSENSITIVE4 regulates lipid mobilization in the embryo and reveals repression of seed germination by the endosperm. Plant Cell, 18(8), 1887-1899.
Pinfield-Wells, H., Rylott, E. L., Gilday, A. D., Graham, S., Job, K., Larson, T. R. and Graham, I. A. (2005) Sucrose rescues seedling establishment but not germination of arabidopsis mutants disrupted in peroxisomal fatty acid catabolism. Plant J., 43(6), 861-872.
Rolland, F. and Sheen, J. (2005) Sugar sensing and signalling networks in plants. Biochem. Soc. Trans., 33(Pt 1), 269-271.
Rolland, F., Baena-Gonzalez, E. and Sheen, J. (2006) Sugar sensing and signaling in plants: Conserved and novel mechanisms. Annu. Rev. Plant. Biol., 57, 675-709.
Rook, F., Corke, F., Card, R., Munz, G., Smith, C. and Bevan, M. W. (2001) Impaired sucrose-induction mutants reveal the modulation of sugar-induced starch biosynthetic gene expression by abscisic acid signalling. Plant J., 26(4), 421-433.
Rubio-Somoza, I., Martinez, M., Diaz, I. and Carbonero, P. (2006) HvMCB1, a R1MYB transcription factor from barley with antagonistic regulatory functions during seed development and germination. Plant J., 45(1), 17-30.
Rubio-Somoza, I., Martinez, M., Abraham, Z., Diaz, I. and Carbonero, P. (2006) Ternary complex formation between HvMYBS3 and other factors involved in transcriptional control in barley seeds. Plant J., 47(2), 269-281.
Rylott, E.L., Rogers, C. A., Gilday, A. D., Edgell, T., Larson, T. R. and Graham, I. A. (2003) Arabidopsis mutants in short- and medium-chain acyl-CoA oxidase activities accumulate acyl-CoAs and reveal that fatty acid beta-oxidation is essential for embryo development. J. Biol. Chem., 278(24), 21370-21377.
Schellmann, S., Schnittger, A., Kirik, V., Wada, T., Okada, K., Beermann, A., Thumfahrt, J., Jurgens, G. and Hulskamp, M. (2002) TRIPTYCHON and CAPRICE mediate lateral inhibition during trichome and root hair patterning in arabidopsis. EMBO J., 21(19), 5036-5046.
Sheen, J., Zhou, L. and Jang, J. C. (1999) Sugars as signaling molecules. Curr. Opin. Plant Biol., 2(5), 410-418.
Sherson, S.M., Alford, H. L., Forbes, S. M., Wallace, G. and Smith, S. M. (2003) Roles of cell-wall invertases and monosaccharide transporters in the growth and development of arabidopsis. J. Exp. Bot., 54(382), 525-531.
Smith, A.M., Zeeman, S. C. and Smith, S. M. (2005) Starch degradation. Annu. Rev. Plant. Biol., 56, 73-98.
van Doorn, W.G. (2004) Is petal senescence due to sugar starvation? Plant Physiol., 134(1), 35-42.
Wang, S., Kwak, S. H., Zeng, Q., Ellis, B. E., Chen, X. Y., Schiefelbein, J. and Chen, J. G. (2007) TRICHOMELESS1 regulates trichome patterning by suppressing GLABRA1 in arabidopsis. Development, 134(21), 3873-3882.
Wang, Z.Y., Kenigsbuch, D., Sun, L., Harel, E., Ong, M. S. and Tobin, E. M. (1997) A myb-related transcription factor is involved in the phytochrome regulation of an arabidopsis lhcb gene. Plant Cell, 9(4), 491-507.
Yanhui, C., Xiaoyuan, Y., Kun, H., Meihua, L., Jigang, L., Zhaofeng, G., Zhiqiang, L., Yunfei, Z., Xiaoxiao, W., Xiaoming, Q., Yunping, S., Li, Z., Xiaohui, D., Jingchu, L., Xing-Wang, D., Zhangliang, C., Hongya, G. and Li-Jia, Q. (2006) The MYB transcription factor superfamily of arabidopsis: Expression analysis and phylogenetic comparison with the rice MYB family. Plant Mol. Biol., 60(1), 107-124.
Yu, S.M. (1999) Cellular and genetic responses of plants to sugar starvation. Plant Physiol., 121(3), 687-693.
Yu, S.M., Kuo, Y. H., Sheu, G., Sheu, Y. J. and Liu, L. F. (1991) Metabolic derepression of alpha-amylase gene expression in suspension-cultured cells of rice. J. Biol. Chem., 266(31), 21131-21137.
Yu, T.S., Zeeman, S. C., Thorneycroft, D., Fulton, D. C., Dunstan, H., Lue, W. L., Hegemann, B., Tung, S. Y., Umemoto, T., Chapple, A., Tsai, D. L., Wang, S. M., Smith, A. M., Chen, J. and Smith, S. M. (2005) Alpha-amylase is not required for breakdown of transitory starch in arabidopsis leaves. J. Biol. Chem., 280(11), 9773-9779.
Zhu, G., Ye, N. and Zhang, J. (2009) Glucose-induced delay of seed germination in rice is mediated by the suppression of ABA catabolism rather than an enhancement of ABA biosynthesis. Plant Cell Physiol., 50(3), 644-651.
Zou, J., Liu, A., Chen, X., Zhou, X., Gao, G., Wang, W. and Zhang, X. (2009) Expression analysis of nine rice heat shock protein genes under abiotic stresses and ABA treatment. J. Plant Physiol., 166(8), 851-861.
Zou, M., Guan, Y., Ren, H., Zhang, F. and Chen, F. (2008) A bZIP transcription factor, OsABI5, is involved in rice fertility and stress tolerance. Plant Mol. Biol., 66(6), 675-683.

指導教授

陸重安(Chung-An Lu)

審核日期

2009-8-12

推文