小麦质膜蛋白基因TaPM19-1的克隆及其对非生物胁迫的响应

doi:10.3864/j.issn.0578-1752.2012.12.020

摘要/Abstract

摘要： 【目的】分析小麦质膜蛋白基因TaPM19-1的特征及其对非生物胁迫的响应，并探讨其在小麦抗逆调控过程中的生物学功能。【方法】利用RACE技术克隆了该基因cDNA全长，采用生物信息学方法分析克隆基因编码蛋白的特性，并通过半定量RT-PCR分析该基因在非生物胁迫条件下的表达特性。【结果】TaPM19-1的cDNA全长1 090 bp，无内含子，编码蛋白包含182个氨基酸，分子量为19.02 kD，包含有典型的AWPM19保守域；依据氨基酸序列将来源于不同植物的16个AWPM19类蛋白分为3组，TaPM19-1与2个来源于大麦及1个二穗短柄草的AWPM19类蛋白亲源关系最近，同属于第三组。高级结构分析显示，TaPM19-1可形成4个由α-螺旋组成的跨膜域，N端位于膜内，包含由27个氨基酸组成的信号肽，C端近40个氨基酸位于膜内。表达分析显示，TaPM19-1除在发育后期的种子中有较高水平表达外，其它组织中未检测到表达；在所检测的2个小麦品种根系中，TaPM19-1的表达受ABA诱导，但在叶片中的表达量极低；水分胁迫条件下，该基因在根系和叶片中均呈现较强的诱导表达特性；在高盐和高温胁迫条件下，该基因在洛旱2号小麦中均可诱导表达，而在中国春小麦中未检测到该基因表达；在低温胁迫条件下，未检测到TaPM19-1的表达。【结论】获得了小麦质膜蛋白基因TaPM19-1的cDNA全长，其编码蛋白可形成典型的跨膜结构特征；该基因受植物激素ABA的诱导，在洛旱2号中也可被干旱、高盐和高温胁迫诱导，但在中国春中对高盐和高温胁迫无响应。推测TaPM19-1在洛旱2号和中国春中存在不同的转录调控机制。

关键词: 小麦, 质膜蛋白, 基因克隆, 非生物胁迫, 表达

Abstract: 【Objective】 The characteristics of a plasma membrane protein gene TaPM19-1 and its response to abiotic stresses in wheat were studied, which will provide an insight into its biological roles involved in the molecular mechanisms of abiotic-sterss response in wheat. 【Method】 RACE technology was used in full-length cDNA cloning and proper bioinformatics software was applied for characterizing the cloned gene and its deduced protein. Semi-quantitative RT-PCR was performed to study the expression patterns of TaPM19-1 under abiotic stresses. 【Result】 The full-length cDNA of TaPM19-1 is 1 090 bp and no introns were included. TaPM19-1 consists of 182 amino acids with a molecular weight of 19.02 kD, and an typical AWPM19 conserved domain was found in it. Based on the amino acid sequences, 16 AWPM19-like proteins of different plant species were sorted into 3 categories, TaPM19-1 and 3 others, 2 AWMP19-like proteins of Hordeum vulgare and 1 of Brachypodium distachyon, were highly similar and included in the third category. The structural prediction of TaPM19-1 revealed that 4 transmembrane regions with alpha helixes could be formed, the N-terminal (including a signal peptide consists of 27 amino acids) was located inside the membrane, while the C-terminus (about 40 amino acids) located inside. The RT-PCR analysis showed that the expression of TaPM19-1 was not detected in all tissues except in later period of developing seeds. The expression of TaPM19-1 was greatly induced by ABA in roots of two detected wheat cultivars, while its expression in leaves was very low. Under the water stress, the gene was immediately and highly induced both in roots and leaves. The expression of TaPM19-1 can be induced by salt- and heat-stress in Luohan 2, while not induced in Chinese Spring. Under lower temperature treatment, no expression of TaPM19-1 was detected in both wheat varieties. 【Conclusion】 The full-length cDNA of TaPM19-1 was obtained and some typical transmembrane structural features were found in its deduced protein. The expression of TaPM19-1 can be induced by plant hormone ABA. TaPM19-1 can also be induced by drought-, salt- and heat-stress in Luohan 2, while not response to salt- and heat-stress in Chinese Spring. The results suggested that TaPM10-1 might have different transcriptional regulating mechanisms in Luohan 2 and Chinese Spring.

Key words: wheat, plasma membrane protein, gene cloning, abiotic stress, expression

李永春, 张春艳, 张宁, 孟凡荣, 任江萍, 牛洪斌, 王翔, 尹钧. 小麦质膜蛋白基因TaPM19-1的克隆及其对非生物胁迫的响应[J]. 中国农业科学, 2012, 45(12): 2502-2509.

LI Yong-Chun, ZHANG Chun-Yan, ZHANG Ning, MENG Fan-Rong, REN Jiang-Ping, NIU Hong-Bin, WANG Xiang, YIN Jun. Cloning of a Plasma Membrane Protein Gene TaPM19-1 and Its Response to Abiotic Stresses in Wheat[J]. Scientia Agricultura Sinica, 2012, 45(12): 2502-2509.

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参考文献

[1]Tuteja N, Tiburcio A F, Fortes A M, Bartels D. Plant abiotic stress. Introduction to PSB special issue. Plant Signaling Behavior, 2011, 6: 173-174.

[2]张瑞越, 徐兆师, 李连城, 陈明, 马有志. 一个新的小麦非生物胁迫诱导基因的克隆及表达特性. 中国农业科学, 2007, 40: 875-881.

Zhang R Y, Xu Z S, Li L C, Chen M, Ma Y Z. Cloning and expression analysis of a novel abiotic stress-induced wheat genes. Scientia Agricultura Sinica, 2007,40: 875-881. (in Chinese)

[3]Qin F, Shinozaki K, Yamaguchi-Shinozaki K. Achievements and challenges in understanding plant abiotic stress responses and tolerance. Plant Cell Physiology, 2011, 52: 1569-1582.

[4]Hirayama T, Shinozaki K. Research on plant abiotic stress responses in the post-genome era: past, present and future. The Plant Journal, 2010, 61: 1041-1052.

[5]刘诗航, 王彩香, 毛新国, 刘惠民, 李昂, 景蕊莲. 小麦蛋白磷酸酶2A 调节亚基基因TaBβ-1 的克隆及其在非生物胁迫下的表达特性. 中国农业科学, 2010, 43(11): 2197-2208.

Liu S H, Wang C X, Mao X G, Liu H M, Li A, Jing R L. Cloning of protein phosphatase 2A regulatory subunit gene TaBβ-1 and its expression patterns under abiotic stresses in wheat. Scientia Agricultura Sinica, 2010, 43(11): 2197-2208. (in Chinese)

[6]Sanchez A, Shin J, Davis S J. Abiotic stress and the plant circadian clock. Plant Signaling Behavior, 2011, 6: 223-231.

[7]Xiong L, Zhu J K. Molecular and genetic aspects of plant responses to osmotic stress. Plant Cell Environment, 2002, 25: 131-139.

[8]Hennig L. Plant gene regulation in response to abiotic stress. Biochimica et Biophysica Acta, 2012, 1819: 85.

[9]Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K. AP2/ERF family transcription factors in plant abiotic stress responses. Biochimica et Biophysica Acta, 2012, 1819: 86-96.

[10]Alexandersson E, Saalbach G, Larsson C, Kjellbom P. Arabidopsis plasma membrane proteomics identifies components of transport, signal transduction and membrane trafficking. Plant Cell Physiology, 2004, 45: 1543-1556.

[11]Hossain Z, Nouri M Z, Komatsu S. Plant cell organelle proteomics in response to abiotic stress. Journal of Proteome Research, 2012, 11: 37-48.

[12]Nouri M Z, Komatsu S. Comparative analysis of soybean plasma membrane proteins under osmotic stress using gel-based and LC MS/ MS-based proteomics approaches. Proteomics, 2010, 10: 1930-1945.

[13]Hashimoto M, Toorchi M, Matsushita K, Iwasaki Y, Komatsu S. Proteome analysis of rice root plasma membrane and detection of cold stress responsive proteins. Protein Peptide Letters, 2009, 16: 685-697.

[14]Nohzadeh M S, Habibi R M, Heidari M, Salekdeh G H. Proteomics reveals new salt responsive proteins associated with rice plasma membrane. Bioscience Biotechnology Biochemistry, 2007, 71: 2144-2154.

[15]Koike M, Sutoh K, Kawakami A, Torada A, Oono K, Imai R. Molecular characterization of a cold-induced plasma membrane protein gene from wheat. Molecular Genetics and Genomics, 2005, 274: 445-453.

[16]Gao Z, He X, Zhao B, Zhou C, Liang Y, Ge R, Shen Y, Huang Z. Overexpressing a putative aquaporin gene from wheat, TaNIP, enhances salt tolerance in transgenic Arabidopsis. Plant Cell Physiology, 2010, 51: 767-775.

[17]Ayadi M, Cavez D, Miled N, Chaumont F, Masmoudi K. Identification and characterization of two plasma membrane aquaporins in durum wheat (Triticum turgidum L. subsp. durum) and their role in abiotic stress tolerance. Plant Physiology and Biochemestry, 2011, 49: 1029-1039.

[18]李永春, 孟凡荣, 王潇, 陈雷, 任江萍, 牛洪斌, 李磊, 尹钧. 水分胁迫条件下“洛旱2号”小麦根系的基因表达谱. 作物学报, 2008, 34(12): 2126-2133.

Li Y C, Meng F R, Wang X, Chen L, Ren J P, Niu H B, Li L, Yin J. Gene expression profiling in roots of wheat cultivar “Luohan 2” under water stress. Acta Agronomica Sinica, 2008, 34(12): 2126-2133. (in Chinese)

[19]Meng F, Ni Z, Wu L, Sun Q. Identification of 17 differentially expressed cDNAs between wheat reciprocal cross-fertilized kernels and their parents. South African Journal of Botany, 2007, 73: 522-529.

[20]Arnold K, Bordoli L, Kopp J, Schwede T. The SWISS-MODEL workspace: A web-based environment for protein structure homology modelling. Bioinformatics, 2006, 22: 195-201.

[21]Rost B, Yachdav G, Liu J. The Predictprotein server. Nucleic Acids Research, 2004, 32: 321-326.

[22]Zhu J K. Salt and drought stress signal transduction in plants. Annual Review of Plant Biology, 2002, 53: 247-273.

[23]Nakashima K, Takasaki H, Mizoi J, Shinozaki K, Yamaguchi- Shinozaki K. NAC transcription factors in plant abiotic stress responses. Biochimica et Biophysica Acta, 2012, 1819: 97-103.

[24]Shinozaki K, Yamaguchi-Shinozaki K. Gene networks involved in drought stress response and tolerance. Journal of Experimental Botany, 2007, 58: 221-227.

[25]Ranford J C, Bryce J H, Morris P C. PM19, a barley (Hordeum vulgare L.) gene encoding a putative plasma membrane protein, is expressed during embryo development and dormancy. Journal of Experimental Botany, 2002, 53: 147-148.

[26]王静英, 李永春, 王潇, 孟凡荣, 任江萍, 牛洪斌, 尹钧. 小麦TaLEA4基因的克隆和表达. 麦类作物学报, 2008, 28(2): 183-186.

Wang J Y, Li Y C, Wang X, Meng F R, Ren J P, Niu H B, Yin J. Cloning and characterizing of TaLEA4 gene in wheat. Journal of Triticeae Crops, 2008, 28(2): 183-186. (in Chinese)