Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/21733
標題: Rice functional genomics study with T-DNA insertion mutants--Characterization and gene expression analysis of fertility and development mutants, M0017031 and M0017091
利用T-DNA插入突變體進行水稻基因功能研究--稔實與發育異常突變體M0017031、M0017091之特性及基因表現分析
作者: Chen, Chi-Wen
陳季緯
關鍵字: 水稻
T-DNA
突變
稔實率
類黃酮
木質素
fetility
Flavonoids
lignins
出版社: 分子生物學研究所
引用: 張正賢 (1988) 稻作學。國立編譯館,台北市。 洪承裕 (2004) “美女櫻花瓣中 flavonoid 3’,5’-hydroxylase (F3’,5’H) 基因之選殖”國立中山大學分子生物學所碩士論文。 吳妤憶 (2003) 利用 RNA 干擾法探討水稻穀粒發育相關基因之功能。國立中興大學分子生物學研究所碩士論文。 吳采馨 (2005) 利用 T-DNA 插入突變體之水稻基因功能研究—生長發育異常突變體 M0019081 之特性及基因表現分析。國立中興大學分子生物學研究所碩士論文。 Abrahams, S., Tanner, G.J., Larkin, P.J., and Ashton, A.R. (2002) Identification and biochemical characterization of mutants in the proanthocyanidin pathway in Arabidopsis. Plant Physiol 130: 561-576. Alonso, J.M., and Stepanova, A.N. (2003) T-DNA mutagenesis in Arabidopsis. Methods Mol Biol 236: 177-188. Alonso, J.M., Stepanova, A.N., Leisse, T.J., Kim, C.J., Chen, H., Shinn, P., Stevenson, D.K., Zimmerman, J., Barajas, P., Cheuk, R., Gadrinab, C., Heller, C., Jeske, A., Koesema, E., Meyers, C.C., Parker, H., Prednis, L., Ansari, Y., Choy, N., Deen, H., Geralt, M., Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weigel, D., Carter, D.E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W.L., Berry, C.C., and Ecker, J.R. (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301: 653-657. Baltas, M., Lapeyre, C., Bedos-Belval, F., Maturano, M., Saint-Aguet, P., Roussel, L., Duran, H., and Grima-Pettenati, J. (2005) Kinetic and inhibition studies of cinnamoyl-CoA reductase 1 from Arabidopsis thaliana. Plant Physiol Biochem 43: 746-753. Bohm, H., Boeing, H., Hempe, J., Raab, B., and Kroke, A. (1998) Flavonols, flavone and anthocyanins as natural antioxidants of food and their possible role in the prevention of chronic diseases. Z Ernahrungswiss 37: 147-163. Britsch, L., and Grisebach, H. (1986) Purification and characterization of (2S)-flavanone 3-hydroxylase from Petunia hybrida. Eur J Biochem 156: 569-577. Burbulis, I.E., and Winkel-Shirley, B. (1999) Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway. Proc Natl Acad Sci U S A 96: 12929-12934. Chalker-Scott L, Scott JD. (2004) Elevated ultraviolet-B radiation induces cross-protection to cold in leaves of Rhododendron under field conditions. Photochem Photobiol. 79(2):199-204. Charrier B., Trinh H., Poirier S., Kondorosi A., and Ratet P., (1998) Flavanone 3-hydroxylase (FHT) Expression and Flavonoid Localization in Nodules of Three Legume Plants Reveal Distinct Tissue Specificities. Molecular Plant-Microbe Interactions. 11: 924-932. Coberly, L.C., and Rausher, M.D. (2003) Analysis of a chalcone synthase mutant in Ipomoea purpurea reveals a novel function for flavonoids: amelioration of heat stress. Mol Ecol 12: 1113-1124. Deboo, G.B., Albertsen, M.C., and Taylor, L.P. (1995) Flavanone 3-hydroxylase transcripts and flavonol accumulation are temporally coordinate in maize anthers. Plant J 7: 703-713. Feinbaum RL, Ausubel FM. (1988) Transcriptional regulation of the Arabidopsis thaliana chalcone synthase gene. Mol Cell Biol. 5:1985-92. Mol Cell Biol. 1988 May;8(5):1985-92. Giusti, M. M. and Wrolstad, R. E. (2003) Acylated anthocyanins from edible sources and their applications in food systems. Biochem. Eng. J. 14:217-225. Guo ZJ, Lamb C, Dixon RA (1998) Potentiation of the oxidative burst and isoflavonoid phytoalexin accumulation by serine protease inhibitors. Plant Physiol. 118(4):1487-94 Halbwirth, H., Fischer, T.C., Roemmelt, S., Spinelli, F., Schlangen, K., Peterek, S., Sabatini, E., Messina, C., Speakman, J.B., Andreotti, C., Rademacher, W., Bazzi, C., Costa, G., Treutter, D., Forkmann, G., and Stich, K. (2003) Induction of antimicrobial 3-deoxyflavonoids in pome fruit trees controls fire blight. Z Naturforsch [C] 58: 765-770. Han, J.-J., and An, G., (2003) Flower-preferential poly(A) binding (PAB) protein gene from rice. Plant Sci. 165: 103-112. Harborne, J.B., and Williams, C.A. (2000) Advances in flavonoid research since 1992. Phytochemistry 55: 481-504. Heller W, Forkmann G (1993) Biosynthesis of flavonoids. In: Harborne J.B. The flavonoids, advances in research since 1986. Chapman & Hall, London, 399–425. Holton, T.A., Brugliera, F., Lester, D.R., Tanaka, Y., Hyland, C.D., Menting, J.G., Lu, C.Y., Farcy, E., Stevenson, T.W., and Cornish, E.C. (1993) Cloning and expression of cytochrome P450 genes controlling flower colour. Nature 366: 276-279. Holton, T.A., and Cornish, E.C. (1995) Genetics and Biochemistry of Anthocyanin Biosynthesis. Plant Cell 7: 1071-1083. Hrazdina, G., and Wagner, G.J. (1985) Metabolic pathways as enzyme complexes: evidence for the synthesis of phenylpropanoids and flavonoids on membrane associated enzyme complexes. Arch Biochem Biophys 237: 88-100. Inagaki Y, Johzuka-Hisatomi Y, Mori T, Takahashi S, Hayakawa Y, Peyachoknagul S, Ozeki Y, Iida S. (1999) Genomic organization of the genes encoding dihydroflavonol 4-reductase for flower pigmentation in the Japanese and common morning glories.Gene. 226(2):181-188. Itoh, Y., Higeta, D., Suzuki, A., Yoshida, H., and Ozeki, Y. (2002) Excision of transposable elements from the chalcone isomerase and dihydroflavonol 4-reductase genes may contribute to the variegation of the yellow-flowered carnation (Dianthus caryophyllus). Plant Cell Physiol 43: 578-585. IRGSP, (2005) The map-based sequence of the rice genome. Nature 463: 793-800. Jeon, J.S., Lee, S., Jung, K.H., Jun, S.H., Jeong, D.H., Lee, J., Kim, C., Jang, S., Yang, K., Nam, J., An, K., Han, M.J., Sung, R.J., Choi, H.S., Yu, J.H., Choi, J.H., Cho, S.Y., Cha, S.S., Kim, S.I., and An, G. (2000) T-DNA insertional mutagenesis for functional genomics in rice. Plant J 22: 561-570. Jin Z, Grotewold E, Qu W, Fu G, Zhao D. (2006) Cloning and characterization of a flavanone 3-hydroxylase gene from Saussurea medusa. DNA Seq. 6(2):121-9. Johzuka-Hisatomi, Y., Hoshino, A., Mori, T., Habu, Y., and Iida, S. (1999) Characterization of the chalcone synthase genes expressed in flowers of the common and Japanese morning glories. Genes Genet Syst 74: 141-147. Katsube, T., Ogawa, K., Hamaguchi, K., Shimao, K., Yamaguchi, K., Konno, S., Shimakawa, T., Naritaka, Y., Yagawa, H., and Aiba, M. (2002) Modification of endoscopic aspiration mucosectomy (EAM) for early gastric cancer: EAM with pre-cutting. Hepatogastroenterology 49: 1510-1513. Kawai, T., Wada, Y., Enmoto, T., Nakajima, S., Nishiyama, K., Kitaura, K., Sato, S., and Oka, T. (1995) Surgical palliation of cardiac malformations associated with right isomerism. Surg Today 25: 525-531. Kawasaki, T., Koita, H., Nakatsubo, T., Hasegawa, K., Wakabayashi, K., Takahashi, H., Umemura, K., Umezawa, T., and Shimamoto, K. (2006) Cinnamoyl-CoA reductase, a key enzyme in lignin biosynthesis, is an effector of small GTPase Rac in defense signaling in rice. Proc Natl Acad Sci U S A 103: 230-235. Koes, B.W., van Tulder, M.W., van der Windt, W.M., and Bouter, L.M. (1994) The efficacy of back schools: a review of randomized clinical trials. J Clin Epidemiol 47: 851-862. Koornneef, L. (1990) [Blow-out fractures of the orbit]. Ned Tijdschr Geneeskd 134: 552-553. Lange, E., Borresen, A.L., Chen, X., Chessa, L., Chiplunkar, S., Concannon, P., Dandekar, S., Gerken, S., Lange, K., Liang, T., and et al. (1995) Localization of an ataxia-telangiectasia gene to an approximately 500-kb interval on chromosome 11q23.1: linkage analysis of 176 families by an international consortium. Am J Hum Genet 57: 112-119. Lee, S., Kim, J., Han, J.J., Han, M.J., and An, G. (2004) Functional analyses of the flowering time gene OsMADS50, the putative SUPPRESSOR OF OVEREXPRESSION OF CO 1/AGAMOUS-LIKE 20 (SOC1/AGL20) ortholog in rice. Plant J 38: 754-764. Lindsey K, Topping JF, Muskett PR, Wei W, Horne KL. (1998) Dissecting embryonic and seedling morphogenesis in Arabidopsis by promoter trap insertional mutagenesis. Symp Soc Exp Biol. 51:1-10. Review. Lindstedt S, Odelhog B, Rundgren M. (1977) Purification and some properties of 4-hydroxyphenylpyruvate dioxygenase from Pseudomonas sp. P. J. 874. Biochemistry.16(15):3369-3377. Liu, Y.L., Ho, D.K., Cassady, J.M., Cook, V.M., and Baird, W.M. (1992) Isolation of potential cancer chemopreventive agents from Eriodictyon californicum. J Nat Prod 55: 357-363. Lo Piero, A.R., Puglisi, I., and Petrone, G. (2006) Gene characterization, analysis of expression and in vitro synthesis of dihydroflavonol 4-reductase from [Citrus sinensis (L.) Osbeck]. Phytochemistry 67: 684-695. Lukacin, R., Urbanke, C., Groning, I., and Matern, U. (2000) The monomeric polypeptide comprises the functional flavanone 3beta-hydroxylase from Petunia hybrida. FEBS Lett 467: 353-358. Madhuri, D., Chand, P., and Sadana, J.R. (1999) Comparison of a dot immunobinding assay and the serum agglutination test for detecting serological responses in vaccinated and unvaccinated chickens following challenge with Salmonella gallinarum. Avian Pathol 28: 98-101. Martienssen, R.A. (1998) Functional genomics: probing plant gene function and expression with transposons. Proc Natl Acad Sci U S A 95: 2021-2026. Masclaux, F.G., Pont-Lezica, R., and Galaud, J.P. (2005) Relationship between allelic state of T-DNA and DNA methylation of chromosomal integration region in transformed Arabidopsis thaliana plants. Plant Mol Biol 58: 295-303. McElver, J., Tzafrir, I., Aux, G., Rogers, R., Ashby, C., Smith, K., Thomas, C., Schetter, A., Zhou, Q., Cushman, M.A., Tossberg, J., Nickle, T., Levin, J.Z., Law, M., Meinke, D., and Patton, D. (2001) Insertional mutagenesis of genes required for seed development in Arabidopsis thaliana. Genetics 159: 1751-1763. Meinke, D.W., and Sussex, I.M. (1979) Embryo-lethal mutants of Arabidopsis thaliana. A model system for genetic analysis of plant embryo development. Dev Biol 72: 50-61. Monties, J.R., Goudard, A., Blin, D., Mesana, T., Mouly-Bandini, A., Camillieri, J.F., Leroy, C., Pagbe, J.J., and Gulino, R. (1989) [Atypical heart transplantation. Technical considerations. Apropos of a case]. Ann Chir 43: 591-595. Pelletier, M.K., and Shirley, B.W. (1996) Analysis of flavanone 3-hydroxylase in Arabidopsis seedlings. Coordinate regulation with chalcone synthase and chalcone isomerase. Plant Physiol 111: 339-345. Pelletier MK, Murrell JR, Shirley BW (1997) Characterization of flavonol synthase and leucoanthocyanidin dioxygenase genes in Arabidopsis. Further evidence for differential regulation of "early" and "late" genes. Plant Physiol. 113(4):1437-45 Pelt, J.L., Downes, W.A., Schoborg, R.V., and McIntosh, C.A. (2003) Flavanone 3-hydroxylase expression in Citrus paradisi and Petunia hybrida seedlings. Phytochemistry 64: 435-444. Sarkanen, K.V. (1971). Precursors and their polymerization. In Lignins -Occurrence, Formation, Structure and Reactions. (Sarkanen, K.V. & Ludwig, C.H., eds) 95-l 63, Wiley Interscience, New York. Ramos, R. L. B., Tovar F. J., Junqueira,R. M., Lino F. B. and G. Sachetto-Martins (2001) Sugarcane expressed sequences tags (ESTs) encoding enzymes involved in lignin biosynthesis pathways Genetics and Molecular Biology, 24 (1-4), 235-241 Ryu C.-H., You J.-H., Kang H.-G., Hur J., Kim Y.-H., Han M.-J., An K., Chung B.-C., Lee C.-H. and An G.. (2004) Generation of T-DNA tagging lines with a bidirectional gene trap vector and the establishment of an insertion-site database. Plant Molecular Biology. 54: 489–502. Sarkanen S, Razal RA, Piccariello T, Yamamoto E, Lewis NG. (1991) Lignin peroxidase: toward a clarification of its role in vivo. J Biol Chem. 266(6):3636-43. Schinella, G.R., Giner, R.M., Recio, M.C., Mordujovich de Buschiazzo, P., Rios, J.L., and Manez, S. (1998) Anti-inflammatory effects of South American Tanacetum vulgare. J Pharm Pharmacol 50: 1069-1074. Sedgley M., (1979) Structural changes in the pollinated and unpollinated avocado stigma and style. J Cell Sci. 38: 49-60. Sheng, J., and Citovsky, V. (1996) Agrobacterium-plant cell DNA transport: have virulence proteins, will travel. Plant Cell 8: 1699-1710. Shirley BW, Hanley S, Goodman HM. (1992) Effects of ionizing radiation on a plant genome: analysis of two Arabidopsis transparent testa mutations. Plant Cell. 4(3):333-47. Shirley, B.W., Kubasek, W.L., Storz, G., Bruggemann, E., Koornneef, M., Ausubel, F.M., and Goodman, H.M. (1995) Analysis of Arabidopsis mutants deficient in flavonoid biosynthesis. Plant J 8: 659-671. Stafford HA. (1991) Flavonoid Evolution: An Enzymic Approach. Plant Physiol. 96(3):680-685. Tissier, A.F., Marillonnet, S., Klimyuk, V., Patel, K., Torres, M.A., Murphy, G., and Jones, J.D. (1999) Multiple independent defective suppressor-mutator transposon insertions in Arabidopsis: a tool for functional genomics. Plant Cell 11: 1841-1852. Tokunaga N, Sakakibara N, Umezawa T, Ito Y, Fukuda H, Sato Y. (2005) Involvement of extracellular dilignols in lignification during tracheary element differentiation of isolated Zinnia mesophyll cells. Plant Cell Physiol. 46(1):224-32. van der Rest, B., Danoun, S., Boudet, A.M., and Rochange, S.F. (2006) Down-regulation of cinnamoyl-CoA reductase in tomato (Solanum lycopersicum L.) induces dramatic changes in soluble phenolic pools. J Exp Bot 57: 1399-1411. Vogt T, Taylor LP. (1995) Flavonol 3-O-glycosyltransferases associated with petunia pollen produce gametophyte-specific flavonol diglycosides. Plant Physiol. 108(3):903-911 Wang, H., Cao, G., and Prior, R L. (1997) Oxygen radical absorbing capacity of anthocyanins. J. Agric. Food Chem. 45 (2):1612-1615. Weisshaar, B., and Jenkins, G.I. (1998) Phenylpropanoid biosynthesis and its regulation. Curr Opin Plant Biol 1: 251-257. Winkel-Shirley, B. (2001) It takes a garden. How work on diverse plant species has contributed to an understanding of flavonoid metabolism. Plant Physiol 127: 1399-1404. Winkel-Shirley, B. (2001) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126: 485-493. Winkel-Shirley, B. (2002) Biosynthesis of flavonoids and effects of stress. Curr Opin Plant Biol 5: 218-223. Woerdenbag, H.J., Merfort, I., Passreiter, C.M., Schmidt, T.J., Willuhn, G., van Uden, W., Pras, N., Kampinga, H.H., and Konings, A.W. (1994) Cytotoxicity of flavonoids and sesquiterpene lactones from Arnica species against the GLC4 and the COLO 320 cell lines. Planta Med 60: 434-437. Ylstra, B., Touraev, A., Moreno, R.M., Stoger, E., van Tunen, A.J., Vicente, O., Mol, J.N., and Heberle-Bors, E. (1992) Flavonols Stimulate Development, Germination, and Tube Growth of Tobacco Pollen. Plant Physiol 100: 902-907. Zupan, J.R., and Zambryski, P. (1995) Transfer of T-DNA from Agrobacterium to the plant cell. Plant Physiol 107: 1041-1047. Zipfel C. and Felix G. (2005) Plants and animals: a different taste for microbes? Curr Opin Plant Biol.8(4):353-360.
摘要: In this study, seven T-DNA insertion mutant lines, M0017031, M0017091, M0019675, M0019845, M0023854, M0023858 and M0030289, were selected by their distinct grain development phenotypes. DNA sequences flanked to T-DNA insertion sites for each mutant were obtained by plasmid rescue and TAIL-PCR. Confirmation of the T-DNA insertion events and their co-segregation with the phenotype were performed through progeny genotyping and phenotype observation. Among them, only mutants M0017031 and M0017091 had well co-segregation between genotype and phenotype. They were then selected for further study. In mutant M0017031, a T-DNA was inserted in the first intron of a putative flavanone 3-hydroxylase (designated as 031-10) in the BAC clone of OSJNBa0084K01 located in the rice chromosome number 4. This mutant has distinct phenotypes such as short plant height, narrow leaves and low fertility. The expression of gene 031-10 was detected through all life cycle of TNG67, but no in mutant lines. Gene 031-10, encoding a putative flavanone 3-hydroxylase, is an important enzyme in the flavonoids biosynthesis pathway and has been suggested to involve in regulation of male activities and embryo developments. Thus, knock-out of gene 031-10 by T-DNA insertion may result in the phenotype of M0017031. In mutant M0017091, the T-DNA was inserted in the intron 4 of a putative cinnamoyl-CoA reductase (CCR, designated as 091-3) and upstream of another CCR (091-4). Mutant M0017091 revealed distinct phenotypes of short plant height, wide and dark-green leaves, small grain size, short panicle length and spikelets closely-packed on panicles. While the expression of 091-3 was not detected in both the mutant and TNG67, the expression of 091-4 was enhanced in M0017091. CCR is a co-enzyme in lignin biosynthesis pathway and lignin is responsible for the growth of the secondary cell wall. The abnormal expression of CCR may result abnormal lignin biosynthesis and therefore result in the distinct phenotypes observed in M0017091. To further understand the functions of the above mentioned genes, use transgenic approach to create gain-of-function or loss-of-function transgenic rice plants with these target genes will be necessary.
本研究以水稻台農67號 (TNG67) 之T-DNA 突變庫為材料,篩選具有特異性狀的七株突變株:M0017031、M0017091、M0019675、M0019845、M0023854、M0023858和M0030289 進行基因功能的研究。利用質體救援和 TAIL-PCR 取得T-DNA 插入點的兩側序列,再比對 NCBI資料庫取得 T-DNA 插入位置,並於RiceGAAS 資料庫預測插入點的基因分佈。以南方墨點法分析基因型與外表性狀分佈吻合的突變株,得知M0017031、M0017091的 T/T 基因型分別有2個以及1 個 T-DNA 插入事件。其中,突變株M0017031的T-DNA插入在 flavanone 3-hydroxylase (FHT, 031-10) 第一個 intron 上。已知FHT是一種 2OG dihydroxylase,為植物類黃酮素 (flavonoids) 生合成路徑重要的酵素,除影響色素形成功能外,與調控雄蕊生成和胚之發育有關。基因型 T/T的株形矮小、葉片較細,低稔實率,還有授粉失敗及種子發育異常的穀粒比率上升。經過分析,得知FHT 在水稻營養生長期生及殖生長期都有表現,尤其在孕穗部位具有較高表現量,並確認突變株 T/T 缺乏FHT 基因表現,推測FHT 基因與植株生長和花器發育有關。另外,突變株M0017091 的T-DNA插入在cinnamoyl-CoA reductase (CCR, 091-4) 基因表現方向的上游。已知CCR在木質素生合成途徑扮演重要角色,而木質素是植物次級細胞壁的主要成份,具生長和抵抗病原體入侵的功能。基因型 T/T 和 T/W株型矮小,葉片寬且呈深綠色,稻穗短小與穀粒排列密集。分析得知突變株可能因為被 T-DNA 插入而增加CCR (091-4) 的表現量,推測突變株的木質素合成路徑被改變後,造成突變株細胞壁組成異常,而產生矮株等外表性狀。未來將再深入探討 RNA 和蛋白質的表現時期、位置,與突變株外表性狀之相關性特性的關係,並利用基因轉殖策略,瞭解基因的功能。
URI: http://hdl.handle.net/11455/21733
其他識別: U0005-1008200615415400
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1008200615415400
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