芹菜品种‘津南实芹’AgICE1基因的克隆及其对非生物胁迫的响应
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摘要
从芹菜(Apium graveolens Linn.)品种‘津南实芹’(‘Jinnan Shiqin’)叶片RNA中克隆获得1个转录因子基因,命名为AgICE1。序列分析结果显示:AgICE1基因含有1个长度为1 455 bp的开放阅读框(ORF),编码484个氨基酸。AgICE1转录因子氨基酸序列的理论相对分子质量为53 140,理论等电点为pI 5.09,为亲水性蛋白。AgICE1转录因子的氨基酸组成中脂肪族氨基酸的比例最高,酸性氨基酸的比例次之,碱性氨基酸的比例较低,芳香族氨基酸的比例最低。序列比对与系统进化分析结果表明:ICE1转录因子在不同植物中高度保守。实时荧光定量PCR检测结果表明:AgICE1基因在叶片中的相对表达水平显著高于在根和叶柄中的相对表达水平;在高温(38℃)、低温(4℃)、干旱(质量体积分数20%PEG6000)和高盐(0.2 mol·L~(-1) NaCl)处理下,AgICE1基因的相对表达水平总体上升高。研究结果表明:AgICE1转录因子在芹菜非生物胁迫调控中起重要作用。
A transcription factor gene was cloned from RNA in leaf blade of cultivar ‘Jinnan Shiqin' of Apium graveolens Linn., which was named AgICE1. Sequence analysis result shows that there is an open reading frame(ORF) in AgICE1 gene with length of 1 455 bp, which encodes 484 amino acids. Theoretical relative molecular mass of amino acid sequence of transcription factor AgICE1 is 53 140, theoretical isoelectric point is pI 5.09, and amino acid sequence of transcription factor AgICE1 is a hydrophilic protein. The percentage of aliphatic amino acid is the highest in amino acid composition of transcription factor AgICE1, that of acidic amino acid comes second, that of basic amino acid is relatively low, and that of aromatic amino acid is the lowest. Sequence alignment and phylogenetic analysis results show that transcription factor ICE1 is highly conserved in different species. Real-time fluorescence quantitative PCR determination result shows that relative expression level of AgICE1 gene in leaf blade is significantly higher than that in root and petiole, and its relative expression level increases in general in treatments of high temperature(38 ℃), low temperature(4 ℃), drought(mass volume fraction of 20% PEG6000), and high salt(0.2 mol·L~(-1) NaCl). It is suggested that transcription factor AgICE1 plays an important role in regulation of abiotic stresses of A. graveolens.
A transcription factor gene was cloned from RNA in leaf blade of cultivar ‘Jinnan Shiqin' of Apium graveolens Linn., which was named AgICE1. Sequence analysis result shows that there is an open reading frame(ORF) in AgICE1 gene with length of 1 455 bp, which encodes 484 amino acids. Theoretical relative molecular mass of amino acid sequence of transcription factor AgICE1 is 53 140, theoretical isoelectric point is pI 5.09, and amino acid sequence of transcription factor AgICE1 is a hydrophilic protein. The percentage of aliphatic amino acid is the highest in amino acid composition of transcription factor AgICE1, that of acidic amino acid comes second, that of basic amino acid is relatively low, and that of aromatic amino acid is the lowest. Sequence alignment and phylogenetic analysis results show that transcription factor ICE1 is highly conserved in different species. Real-time fluorescence quantitative PCR determination result shows that relative expression level of AgICE1 gene in leaf blade is significantly higher than that in root and petiole, and its relative expression level increases in general in treatments of high temperature(38 ℃), low temperature(4 ℃), drought(mass volume fraction of 20% PEG6000), and high salt(0.2 mol·L~(-1) NaCl). It is suggested that transcription factor AgICE1 plays an important role in regulation of abiotic stresses of A. graveolens.
引文
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[15] 却枫, 黄莹, 王枫, 等. 胡萝卜中DcDofD1转录因子的克隆及其对非生物逆境胁迫的响应分析[J]. 植物遗传资源学报, 2015, 16(5): 1073-1079.
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[22] 邵天恒, 孙曙光, 许海峰, 等. 芹菜中转录因子基因AgDREB1的分离与表达研究[J]. 南京农业大学学报, 2014, 37(6): 19-27.
[23] 李瑞梅, 惠杜娟, 刘姣, 等. 植物抗寒转录因子CBF和ICE研究进展[J]. 广东农业科学, 2012, 39(23): 132-135, 138.
[24] 甄伟, 陈溪, 孙思洋, 等. 冷诱导基因的转录因子CBF1转化油菜和烟草及抗寒性鉴定[J]. 自然科学进展, 2000, 10(12): 1104-1108.
[25] 王建华, 刘鸿先, 徐同. 超氧物歧化酶(SOD)在植物逆境和衰老生理中的作用[J]. 植物生理学通讯, 1989(1): 1-7.
[26] MITTLER R. Oxidative stress, antioxidants and stress tolerance[J]. Trends in Plant Science, 2002, 7(9): 405-410.
[27] CHAPARZADEH N, D’AMICO M L, KHAVARI-NEJAD R-A, et al. Antioxidative responses of Calendula officinalis under salinity conditions[J]. Plant Physiology and Biochemistry, 2004, 42(9): 695-701.
[28] 邵常荣, 张旸, 解莉楠, 等. 植物对非生物逆境响应的转录调控和代谢谱分析的研究进展[J]. 植物生理学报, 2011, 47(5): 443-451.
[2] 李梦瑶, 王枫, 侯喜林, 等. 芹菜过敏原蛋白Api g 1基因的克隆与表达分析[J]. 南京农业大学学报, 2013, 36(2): 13-19.
[3] 杨献光, 梁卫红, 齐志广, 等. 植物非生物胁迫应答的分子机制[J]. 麦类作物学报, 2006, 26(6): 158-161.
[4] 尹盈, 张玥, 胡靖妍, 等. 茶树低温胁迫转录因子CsICE的亚细胞定位及表达分析[J]. 园艺学报, 2013, 40(10): 1961-1968.
[5] GUY C L, NIEMI K J, BRAMBL R. Altered gene expression during cold acclimation of spinach[J]. Proceedings of the National Academy of Sciences of the United States of America, 1985, 82(11): 3673-3677.
[6] 张丽丽, 李景富, 王傲雪. 转录激活因子CBF基因在植物抗冷分子机制中的作用[J]. 园艺学报, 2008, 35(5): 765-771.
[7] CHINNUSAMY V, OHTA M, KANRAR S, et al. ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis[J]. Genes and Development, 2003, 17(8): 1043-1054.
[8] ZARKA D G, VOGEL J T, COOK D, et al. Cold induction of Arabidopsis CBF genes involves multiple ICE (inducer of CBF expression) promoter elements and a cold-regulatory circuit that is desensitized by low temperature[J]. Plant Physiology, 2003, 133(2): 910-918.
[9] CHEN Y, JIANG J, SONG A, et al. Ambient temperature enhanced freezing tolerance of Chrysanthemum dichrum CdICE1 Arabidopsis via miR398[J]. BMC Biology, 2013, 11(1): 121.
[10] 向殿军, 殷奎德, 满丽莉, 等. 萝卜低温胁迫转录因子的克隆及遗传转化[J]. 西北植物学报, 2011, 31(10): 1933-1941.
[11] FENG X M, ZHAO Q, ZHAO L L, et al. The cold-induced basic helix-loop-helix transcription factor gene MdCIbHLH1 encodes an ICE-like protein in apple[J]. BMC Plant Biology, 2012, 12(1): 22.
[12] 黄莹, 徐志胜, 王枫, 等. 胡萝卜低温胁迫转录因子DcICE1基因克隆与非生物逆境响应分析[J]. 西北植物学报, 2015, 35(1): 30-36.
[13] 郑银英, 崔百明, 常明进, 等. 转拟南芥ICE1基因增强烟草抗寒性的研究[J]. 西北植物学报, 2009, 29(1): 75-79.
[14] 李海燕. 芹菜品种——津南实芹[J]. 天津农林科技, 2004(4): 21.
[15] 却枫, 黄莹, 王枫, 等. 胡萝卜中DcDofD1转录因子的克隆及其对非生物逆境胁迫的响应分析[J]. 植物遗传资源学报, 2015, 16(5): 1073-1079.
[16] FENG K, HOU X L, LI M Y, et al. CeleryDB: a genomic database for celery[J]. Database: The Journal of Biological Databases and Curation, 2018: bay070.
[17] GASTEIGER E, GATTIKER A, HOOGLAND C, et al. ExPASy: the proteomics server for in-depth protein knowledge and analysis[J]. Nucleic Acids Research, 2003, 31(13): 3784-3788.
[18] TAMURA K, PETERSON D, PETERSON N, et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Molecular Biology and Evolution, 2011, 28(10): 2731-2739.
[19] 谭国飞, 王枫, 王广龙, 等. 鸭儿芹肉桂酸4-羟化酶基因的克隆与不同温度下的表达分析[J]. 西北植物学报, 2014, 34(7): 1298-1304.
[20] 李合生. 现代植物生理学[M]. 2版. 北京: 高等教育出版社, 2006: 342.
[21] SAKUMA Y, LIU Q, DUBOUZET J G, et al. DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs, transcription factors involved in dehydration- and cold-inducible gene expression[J]. Biochemical and Biophysical Research Communications, 2002, 290(3): 998-1009.
[22] 邵天恒, 孙曙光, 许海峰, 等. 芹菜中转录因子基因AgDREB1的分离与表达研究[J]. 南京农业大学学报, 2014, 37(6): 19-27.
[23] 李瑞梅, 惠杜娟, 刘姣, 等. 植物抗寒转录因子CBF和ICE研究进展[J]. 广东农业科学, 2012, 39(23): 132-135, 138.
[24] 甄伟, 陈溪, 孙思洋, 等. 冷诱导基因的转录因子CBF1转化油菜和烟草及抗寒性鉴定[J]. 自然科学进展, 2000, 10(12): 1104-1108.
[25] 王建华, 刘鸿先, 徐同. 超氧物歧化酶(SOD)在植物逆境和衰老生理中的作用[J]. 植物生理学通讯, 1989(1): 1-7.
[26] MITTLER R. Oxidative stress, antioxidants and stress tolerance[J]. Trends in Plant Science, 2002, 7(9): 405-410.
[27] CHAPARZADEH N, D’AMICO M L, KHAVARI-NEJAD R-A, et al. Antioxidative responses of Calendula officinalis under salinity conditions[J]. Plant Physiology and Biochemistry, 2004, 42(9): 695-701.
[28] 邵常荣, 张旸, 解莉楠, 等. 植物对非生物逆境响应的转录调控和代谢谱分析的研究进展[J]. 植物生理学报, 2011, 47(5): 443-451.