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Cloning and Expression Characteristic of CSEBF1 Gene in Cucumber (Cucumis stavius L.) |
JIN Xiao-xia1, DONG Yan-long2, QIN Zhi-wei3, YU Li-jie1, WU Shu-ju1, SONG Jian1 |
1. College of Life Science and Technology, Harbin Normal University, Harbin 150025, China; 2. Horticulture branch, Heilongjiang academy of agricultural sciences, Harbin 150060, China; 3. College of horticulture, Northeast Agriculture University, Harbin 150030, China |
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Abstract Based on selecting SSH of stem tips treated by ethrel, the cDNA sequence of cucumber EIN3-Binding F box protein 1 gene, named CSEBF1(GenBank: KF366911) was obtained by RT-PCR and in silico cloning. The cDNA sequence of the CSEBF1 gene was 1 964bp, contained a predicted protein sequence of 640 amino acids, the F-box sequences and essential structure for ubiquitin-mediated proteolysis—LRRs (leucine-rich repeats). The predicted amino acids sequence of cucumber CSEBF1 shared 60.47% identity with Arabidopsis. The mRNA expressions of CSEBF1 in different tissues treated by ethrel were detected through real-time quantitative RT-PCR, and results revealed that highest level of CSEBF1 mRNA was increased by ethrel at 8h in leaf and root, and at 16h in stem. Meanwhile, the expressions of genes involved in ethylene signal transduction were detected through RT-PCR, revealed that CSEIN3 expression was induced in stem and leaf at 4 h; the mRNA accumulation of CSCTR1 was detected in stem at 8 h and leaf at 16 h; the expression of CSACS2 was induced in leaf at 2 h, and increased subsequently. The mRNA of CSACS1G was increased in leaf at 4 h, and then continued to lower levels.
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Received: 20 December 2013
Published: 25 March 2014
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[1] Frugis G, Chua N H. Ubiquitin-mediated proteolysis in plant hormone signal transduction. Trends Cell Biol, 2002, 12: 308-311. [2] Potuschak T, Lechner E, Parmentier Y, et al. EIN3-Dependent regulation of plant ethylene hormone signaling by two Arabidopsis F box proteins:EBF1 and EBF2.Cell, 2003, 115(6): 679-689. [3] Guo H, Ecker J R. Plant responses to ethylene gas are mediated by SCF EBF1/EBF2-dependent proteolysis of EIN3 transcription factor. Cell, 2003, 115(6): 667-677. [4] Kepinski S, Leyser O. SCF-mediated proteolysis and negative regulation in ethylene signaling. Cell, 2003, 115(6): 647-648. [5] 吴玉, 杨迎伍, 邓伟, 等. 番茄EBF2基因的克隆、亚细胞定位与遗传转化. 核农学报, 2010, 24(3):490-494. Wu Y, Yang Y W, Deng W, et al. Cloneing, subcellular localization and transformation of EBF2 gene in tomato. Journal of Nuclear Agricultural Sciences, 2010, 24(3):490-494. [6] Guo H, Ecker J R. Plant responses to ethylene gas are mediated by SCF EBF1/EBF2-dependent proteolysis of EIN3 transcription factor. Cell, 2003, 115(6): 667-677. [7] Gagne J M, Downes B P, Shiu S H, et al. The F-box subunit of the SCF E3 complex is encoded by a diverse superfamily of genes in Arabidopsis. Proc Natl Acad Sci, 2002, 99: 11519-11524. [8] Glickman M H, Ciechanover A. The ubiquitin proteasome proteolytic pathway destruction for the sake of construction. Physiol. Rev, 2002, 82: 373-428. [9] Ni W M, Xie D X, Hobbie L, et al. Regulation of flower development in Arabidopsis by SCF complexes. Plant Physiology, 2004, 134: 1574-1585. [10] Gagne J M, Smalle J, Gingerich D J, et al. Arabidopsis EIN3-binding F-box 1 and 2 form ubiquitin-protein ligases that repress ethylene action and promote growth by directing EIN3 degradation. Proc Natl Acad Sci. USA, 2004, 101: 6803-6808. [11] Alonso J M, Stepanova A N, Solano R, et al. Five components of the ethylene-response pathway identified in a screen for weak ethyleneinsensitive mutants in Arabidopsis. Proc Natl Acad Sci. USA, 2003, 100: 2992-2997. [12] Ecker J R. The ethylene signal transduction pathway in plants. Science, 1995, 268: 667-675. [13] Chao Q, Rothenberg M, Solano R, et al. Activation of the ethylene gas tespoonse pathway in Arabidopsis by the nuclear protein ETHYLENE-INSENSITIVE3 and related proteins. Cell, 1997, 89: 1133-1144. [14] Potuschak T, Lechner E, Parmentier Y, et al. EIN3-dependent regulation of plant ethylene hormone signaling by two Arabidopsis F box proteins: EBF1 and EBF2. Cell, 2003, 115: 679-689. [15] Yang Z, Tian L, Latoszek-Green M, et al. Arabidopsis ERF4 is a transcriptional repressor capable of modulating ethylene and abscisic acid responses. Plant Mol. Biol, 2005: 585-596. [16] Linda R, Anna S, Jose A. Molecular Mechanisms of Ethylene–Auxin Interaction, Molecular Plant, 2013, 6(6): 1734–1737. [17] He W, Brumos J, Li H, Ji Y, Ke M, Gong X, Zeng Q, Li W, Zhang X, An F, et al. A small-molecule screen identifies L-kynurenine as a competitive inhibitor of TAA1/TAR activity in ethylene-directed auxin biosynthesis and root growth in Arabidopsis. Plant Cell, 2011, 23: 3944–3960. [18] Yin T J, Quinn J A. Tests of a mechanistic model of one hormone regulating both sexes in Cucumis sativus (Cucurbitaceae). American Journal of Botany, 1995, 82: 1537-1546. [19] Yamasaki S, FujⅡ N, Takahashi H. The ethylene-regulated expression of CS-ETR2 and CS-ERS genes in cucumber plants and their possible involvement with sex expression in flowers. Plant and Cell Physiology, 2000, 41: 608-616. [20] Yamasaki S, FujⅡ N, Matsuura S, et al. The M locus and ethylene-controlled sex determination in andromonoecious cucumber plants. Plant and Cell Physiology, 2001, 42: 608-619. [21] Yamasaki S, FujⅡ N, Takahashi H. Characterization of ethylene effects on sex determination in cucumber plants. Sexual Plant Reproduction, 2003, 16: 103-111. [22] Trebitsh T, Staub J E, O'Neill S D. Identification of a 1-Aminocyclopropane-1-Carboxylic acid synthase gene linked to the Female (F) locus that enhances female sex expression in Cucumber. Plant Physiology, 1997, 113: 987-995. [23] Mathooko F M, Mwaniki M W, Nakatsuka A, et al. Expression characteristics of CS-ACS1, CS-ACS2 and CS-ACS3, three members of the 1-aminocyclopropane-1-carboxylate synthase gene family in cucumber (Cucumis sativus L.) fruit under carbon dioxide stress. Plant and Cell Physiology, 1999, 40: 164-172. [24] Kamachi S, Sekimoto H, Kondo N, et al. Cloning of a cDNA for a 1-aminocyclopropane-1-carboxylate synthase that is expressed during development of female flowers at the apices of Cucumis sativus L. Plant and Cell Physiology, 1997, 38: 1197-1206. [25] Kamachi S, Mizusawa H, Mazuura S, et al. Expression of two 1-aminocyclopropane-1-carboxylate synthase genes, CS-ACS1 and CS-ACS2, correlated with sex phenotypes in cucumis plants (Cucumis sativus L.). Plant Biotechnology, 2000, 17: 69-74. [26] Saito S, FujⅡ N, Miyazawa Y, et al. Correlation between development of female flower buds and expression of the CS-ACS2 gene in cucumber plants. Journal of Experimental Botany, 2007, 58: 2897-2907. [27] Shiber A, Gaur R K, Rimon-Knopf R, et al. The origin and mode of function of the Female locus in cucumber. Cucurbitaceae, 2008: 263-270. [28] 程立宝, 秦智伟, 刘宏宇, 等. 黄瓜cs-acs1g基因克隆及不同时空表达的研究. 园艺学报, 2005, 32: 840-843. Cheng L B, Qin Z W, Liu H Y, et al. Cloning of cs-acs1g gene and research on expression of different periods and sites in cucumber. Acta Horticulturae Sinica, 2005, 32: 840-843. [29] 程立宝, 秦智伟, 李淑艳, 等. 与黄瓜雌性性状连锁的cs-acs1g基因特异片段克隆与鉴定. 中国农业科学, 2006, 39: 2545-2550. Cheng L B, Qin Z W, Li Y S, et al. Cloning and identification of special cs-acs1g gene-linked to gynoecious in cucumber. Scientia Agricultura Sinica, 2006, 39: 2545-2550. |
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