Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/36137
標題: 鐵炮百合兩絨氈層基因LLA-1271和LLA-115之特性分析和表現
Characterization and expression of two tapetal genes LLA-1271 and LLA-115 in Lilium longiforum.
作者: 葉芳凌
Yeh, Fung-Ling
關鍵字: Lilium longiforum;鐵炮百合;tapetal;anther;花藥;絨氈層
出版社: 生物科技學研究所
引用: 曾稚鐙。2006鐵炮百合花藥發與早期基因之選殖與特性分析。國立中興大學碩士論文。 Bate N, Twell D (1998) Functional architecture of a late pollen promoter: pollen-specific transcription is developmentally regulated by multiple stage-specific and co-dependent activator elements. Plant Mol Biol 5: 859-869. Bianka SA, Jinxiang WA, Margret S (2006) Interactions between ethylene, gibberellin and abscisic acid regulate emergence and growth rate of adventitious roots in deepwater rice. Planta 223: 604–612. Bisht NC, Jagannath A, Burma PK, Pradhan AK, Pental D (2007) Retransformation of a male sterile barnase line with the barstar gene as an efficient alternative method to identify male sterile-restorer combinations for heterosis breeding. Plant Cell Rep 26: 727-733. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 284-282. Burnette WN (1981) “Western blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radioiodinated protein A. Anal Biochem 112: 195-203. Charbonnel CL, Lauga B, Combes D (2000) Isolation of a type 2 metallothionein-like gene preferentially expressed in the tapetum in Zea mays. Gene 254: 199–208. Chen PW, Chiang CM, Tseng TH, Yu SM (2006) Interaction between rice MYBGA and the gibberellin response element controls tissue-specific sugar sensitivity of alpha-amylase genes. Plant Cell 18: 2326-2340. Cigan AM, Unger E, Xu RJ, Kendall T, Fox TW (2001) Phenotypic complementation of ms45 maize requires tapetal expression of MS45. Sex Plant Reprod 14:135–142. Colcombet J, Boisson DA, Ros PR, Vera CE, Schroeder JI (2005) Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR KINASES 1 and 2 are essential for tapetum development and microspore maturation. Plant Cell 17: 3350-3361. Crossley SJ, Greenland AJ, Dickinsion HG (1995) The characterization of tapetum-specific cDNAs isolated from a Lilium henryi L. meiocyte subtractive cDNA library. Planta 196: 523-529. Ding ZJ, Wu XH, Wang T (2002) The rice tapetum-specific gene RA39 encodes a type I ribosome-inactivating protein. Sex Plant Reprod 15: 205–212. Edelín R, María DG, Philippe E, Michael W, Francisco M, José PB Luis AC (2007) The PsEND1 promoter: a novel tool to produce genetically engineered male-sterile plants by early anther ablation. Plant Cell Rep 26:313-325. Endo M, Matsubaraa H, Kokubuna T, Masukoa H, Takahataa Y, Tsuchiyab T, Fukudacid H, Demurad T, Watanabea M (2002) The advantages of cDNA microarray as an effective tool for identification of reproductive organ-specific genes in a model legume, Lotus japonicus. FEBS Lett 514: 229-237. Gupta V, Khurana R, Tyagi AK (2007) Promoters of two anther-specific genes confer organ-specific gene expression in a stage-specific manner in transgenic systems. Plant Cell Rep 26: 1919-1931. Hird DL, Worrall D, Hodge R, Smartt S, Paul W, Scott R (1993) The anther-specific protein encoded by the Brassica napus and Arabidopsis thaliana A6 gene displays similarity to beta-1,3-glucanases. Plant J 4: 1023-1033. Höfgen R, Willmitzer L (1988) Storage of competent cells for Agrobacterium trasformation. Nucleic Acids Res 16 9877 Honys D, Twell D (2004) Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biol 5: R85. Hsieh K, Huang AH (2007) Tapetosomes in Brassica tapetum accumulate endoplasmic reticulum-derived flavonoids and alkanes for delivery to the pollen surface. Plant Cell 19: 582-596. Hsu YF, Tzeng JD, Liu MC, Yei FL, Chung MC, Wang CS (2008) Identification of anther-specific/predominant genes regulated by gibberellin during development of lily anthers. J Plant Physiol, in press. Hsu YF, Wang CS, Raja R (2007) Gene expression pattern at desiccation in the anther of Lilium longiflorum. Planta 226: 311-322. Hurkman WJ, Tanaka CK (1986) Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis. Plant Physiol 81: 802-806. Jeon JS, Chung YY, Lee S, Yi GH, Oh BG, An G (1999) Isolation and characterization of an anther-specific gene, RA8, from rice (Oryza sativa L.). Plant Mol Biol 391:35-44. Judith LGP, Diego MP, Ingo D, Jorge EM, Bernd MR (2007) Genome-wide analysis of ABA-responsive elements ABRE and CE3 reveals divergent patterns in Arabidopsis and rice. BMC Genomics 8: 260. Jung KH, Han MJ, Lee YS, Kim YW, Hwang I, Kim MJ, Kim YK, Nahm B H, An G (2005) Rice Undeveloped Tapetum1 is a major regulator of early tapetum development. Plant Cell 17: 2705-2722. Kaneko M, Itoh H, Inukai Y, Sakamoto T, Ueguchi TM, Ashikari M, Matsuoka M (2003) Where do gibberellin biosynthesis and gibberellin signaling occur in rice plants? Plant J 35: 104-115. Kapoor S, Kobayashi A, Takatsuji H (2002) Silencing of the tapetum-specific zinc finger gene TAZ1 causes premature degeneration of tapetum and pollen abortion in petunia. Plant Cell 14: 2353-2367. Kiss HG, Koning RE (1989) Endogenous levels and transport of 1-Aminocyclopropane-1-Carboxylic acid in stamens of ipomoea nil (convolvulaceae). Plant Physiol 90: 157-161. Koltunow AM, Truettner J, Cox KH, Wallroth M, Goldberg RB (1990) Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201-1224. Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157: 105-132. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685. Lauga B, Charbonnel CL, Combes D(2000) Characterization of MZm3-3, a Zea mays tapetum-specific transcript. Plant Sci 157: 65-75. Lee YH, Chung KH, Kim HU, Jin YM, Kim HI, Park BS (2003) Induction of male sterile cabbage using a tapetum-specific promoter from Brassica campestris L. ssp. pekinensis. 22: 268-273. Li N, Zhang DS, Liu HS, Yin CS, Li XX, Liang WQ, Yuan Z, Xu B, Chu HW, Wang J, Wen TQ, Huang H, Luo D, Ma H, Zhang DB (2006) The rice tapetum degeneration retardation gene is required for tapetum degradation and anther development. Plant Cell 18: 2999-3014. Liesbeth DG, Wim HV, Sophie B, Andy P, Ana MV, Peter H Dominique VDS (2007) Reciprocal influence of ethylene and gibberellins on response-gene expression in Arabidopsis thaliana. Planta 226: 485-498. Luo H, Lee JY, Hu Q, Nelson VK, Eitas T K, Lickwar C, Kausch AP, Chandlee JM, Hodges TK (2006) RTS, a rice anther-specific gene is required for male fertility and its promoter sequence directs tissue-specific gene expression in different plant species. Plant Mol Biol 62: 397-408. Ma H (2005) Molecular genetic analyses of microsporogenesis and microgametogenesis in flowering plants. Annu Rev Plant Biol 56: 393-434. Maliga P, Klessig DF, Cashmore AR, Gruissem W, Varner JE (1995) Method in Plant Molecular Biology, a Laboratory Course Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. p 357-359. McCormick S (1993) Male gametophyte development. Plant Cell 10: 1265–1275. Millar AA, Gubler F (2005) The Arabidopsis GAMYB-like genes, MYB33 and MYB65, are microRNA-regulated genes that redundantly facilitate anther development. Plant Cell 17: 705-721. Muschietti J, Dircks L, Vancanneyt G, McCormick S (1994) LAT52 protein is essential for tomato pollen development: pollen expressing antisense LAT52 RNA hydrates and germinates abnormally and cannot achieve fertilization. Plant J 6: 321-338. Paul W, Hodge R, Smartt S, Draper J, Scott R (1992) The isolation and characterisation of the tapetum-specific Arabidopsis thaliana A9 gene. Plant Mol Biol 19: 611-622. Pina C, Pinto F, Feijo JA, Becker JD (2005) Gene family analysis of the Arabidopsis pollen transcriptome reveals biological implications for cell growth, division control, and gene expression regulation. Plant Physiol 138: 744-756. Rabilloud T, Carpentier G, Tarroux P (1988) Improvement and simplification of low-background silver staining of proteins by using sodium sithonite. Electrophoresis 9: 288-291 Rombauts S, Florquin K, Lescot M, Marchal K, Rouzé P, Van de Peer Y (2003) Computational approaches to identify promoters and cis-regulatory elements in plant genomes. Plant Physiol 132: 1162-1176. Roque E, Gómez MD, Ellul P, Wallbraun M, Madueño F, Beltrán JP, Cañas LA (2007) The PsEND1 promoter: a novel tool to produce genetically engineered male-sterile plants by early anther ablation. Plant Cell Rep 26: 313-325. Rubinelli P, Hu Y, Ma H (1998) Identification, sequence analysis and expression studies of novel anther-specific genes of Arabidopsis thaliana. Plant Mol Biol 37: 607-619. Sambrook JE, Fritsch ET, Maniatis R (1989) Molecular cloning: A laboratory manual, 2rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. Semarjit S, Ravindra K, Sipra GM (2006) Isolation of pollen early genes and analysis of expression pattern during the development of male gametophyte. Plant Science 170: 417-425. Shen JB, Hsu FC (1992) Brassica anther-specific genes: characterization and in situ localization of expression. Mol Gen Genet 234: 379-379. Smith DE, Fisher PA (1984) Identification, developmental regulation and response to heat shock of two antigenically related forms of a major nuclear enveloped method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots. J Cell Biol 99: 20-28. Solano R, Nieto C, Avila J, Canas L, Diaz I, Paz-Ares J (1995) Dual DNA binding specificity of a petal epidermis-specific MYB transcription factor (MYB.Ph3) from Petunia hybrida. EMBO J 14: 1773-1784. Suen DF, Wu SS, Chang HC, Dhugga KS, Huang AH (2003) Cell wall reactive proteins in the coat and wall of maize pollen: potential role in pollen tube growth on the stigma and through the style. J Biol Chem 44: 43672-43681 Swain SM, Singh DP, Helliwell CA, Poole AT (2005) Plants with increased expression of ent-kaurene oxidase are resistant to chemical inhibitors of this gibberellin biosynthesis enzyme. Plant Cell Physiol 46: 284-291. Takada K, Ishimaru K, Kamada H, Ezura H. (2006). Anther-specific expression of mutated melon ethylene receptor gene Cm-ERS1/H70A affected tapetum degeneration and pollen grain production in transgenic tobacco plants. Plant Cell Rep 25: 936-941. Turgut K, Barsby T, Craze M, Freeman J, Hodge R, Paul W, Scott R (1994) The highly expressed tapetum-specific A9 gene is not required for male fertility in Brassica napus. Plant Mol Biol 24: 97-104. Varnier AL, Mazeyrat GF, Sangwan RS, Clément CJ (2005) Programmed cell death progressively models the development of anther sporophytic tissues from the tapetum and is triggered in pollen grains during maturation. J Struct Biol 2: 118-128. Verwoerd TC, Dekker BM, Hoekema A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 17: 2362. Wang S, Tiwari SB, Hagen G, Guilfoyle TJ (2005) AUXIN RESPONSE FACTOR7 restores the expression of auxin-responsive genes in mutant Arabidopsis leaf mesophyll protoplasts. Plant Cell 17: 1979-1993. Wang CS, Walling LL, Eckard KJ, Lord, EM (1992a) Patterns of protein accumulation in developing anther of Lilium longiflorum correlate with histological events. Amer J Bot 79: 118-127. Wang CS, Walling LL, Eckard KJ, Lord, EM (1992b) Immunological characterization of a tapetal protein in developing anthers of Lilium longiflorum. Plant Physiol 99: 822-829. Watanabe N, Lam E (2006) Arabidopsis Bax inhibitor-1 functions as an attenuator of biotic and abiotic types of cell death. Plant J 45: 884-894. Xu SX, Liu GS, Chen RD (2006) Characterization of an anther- and tapetum-specific gene and its highly specific promoter isolated from tomato. Plant Cell Rep 25: 231-240. Yang SL, Xie LF, Mao HZ, Puah CS, Yang WC, Jiang L, Sundaresan V, Ye D (2003) Tapetum Determinant 1 is required for cell specialization in the Arabidopsis anther. Plant Cell 15: 2792-2804. Zhang ZB, Zhu J, Gao JF, Wang C, Li H, Li H, Zhang HQ, Zhang S, Wang DM, Wang QX, Huang H, Xia HJ, Yang ZN (2007) Transcription factor AtMYB103 is required for anther development by regulating tapetum development, callose dissolution and exine formation in Arabidopsis. Plant J 23: 528-538. Zhang X, Feng B, Zhang Q, Zhang D, Altman N, Ma H (2005) Genome-wide expression profiling and identification of gene activities during early flower development in Arabidopsis. Plant Mol Biol 58: 401-419.
摘要: 
利用扣除抑制雜合 (suppression subtraction hybridization)的方法從鐵炮百合(Lilium longiflorum)花藥的小孢子發育期cDNA集合庫選殖出LLA-1271和LLA-115。LLA-1271 cDNA利用5´-RACE得到全長為970 bp,可轉譯出225個胺基酸,扣除N端26個胺基酸的訊息胜肽,估計成熟蛋白質的分子量為21 kDa。基因庫比對不到任何已知的蛋白質,為一未知嶄新的蛋白質。特別是LLA-1271蛋白質除了N端和C端外,整個蛋白為八個重複的保留序列所涵蓋。LLA-115全長為545 bp,可轉譯出112個胺基酸,扣除N端25個胺基酸的訊息胜肽,估計成熟蛋白質之分子量為9 kDa,基因庫比對後與擬南芥、油菜的細胞壁蛋白相似。利用北方墨漬法得知LLA-1271和LLA-115 mRNA均表現在小孢子發育時期的花藥中。進一步由原位雜合的實驗證實LLA-1271 和LLA-115 mRNAs在絨氈層中均有顯著的表現。利用激勃素抑制劑uniconazol和乙烯抑制劑2,5-norbornadien處理,顯示兩個基因均受激勃素的誘導,而誘導的路徑接受乙烯的抑制調控。利用合成LLA-1271蛋白小段胜肽片段,對兔子進行免疫注射製備抗體。LLA-1271-胜肽2抗血清,經親和純化(Affinity-purified)後發現在小孢子發育時期偵測到32 kDa大小的蛋白,且只在花藥中有訊號。利用二維電泳分離蛋白質與西方墨漬分析,得知LLA-1271蛋白質具多型性,至少有四個isofroms。花藥分層分析顯示LLA-1271蛋白在花藥壁、小孢子壁和小孢子分層中累積,顯示LLA-1271蛋白沈積在小孢子壁上。LLA-115抗血清則利用大腸桿菌大量表現融合蛋白後製備的。LLA-115基因不含插入子。利用TAIL-PCR找到LLA-115基因622 bp的啟動子區域,其中包含了兩個花粉專一AGAAA調控區和其他荷爾蒙有關的調控區。將LLA-115基因的啟動子分別構築含有β-glucuronidase (GUS)、barnase、barstar基因,轉殖到擬南芥中,以便進一步探討其花藥專一性,並測試花粉的稔性。
URI: http://hdl.handle.net/11455/36137
其他識別: U0005-0402200819144600
Appears in Collections:生物科技學研究所

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