Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (2): 302-310.doi: 10.3864/j.issn.0578-1752.2012.02.012

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Sequence Analysis of the Aquaporins Gene SlAQP in Tomato

 LI  Ren, WU  Xin-Xin, LI  Wei, YANG  Rong-Chao, ZHAO  Yong-Qin, WEN  Chang-Long, ZHAO  Bing, GUO  Yang-Dong   

  1. 1.中国农业大学农学与生物技术学院，北京 100193
  • Received:2011-07-12 Online:2012-01-15 Published:2011-11-22

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Abstract: 【Objective】 To provide basal data for mechanism of tomato’s drought resistance and cultivar development, the sequence characteristics of the aquaporin gene SlAQP in tomato were analyzed, the coded protein was intracellar located and the expression profiling of the Mirco Tom tomato was studied after treatment under drought stress. 【Method】 The rapid-amplification of cDNA ends (RACE) was used to amplify the full-length SlAQP gene, and the bioinformatics software was used to analyze the structures and function of the coded protein. In order to follow the intracellular localisation of the protein, the GFP sequence was fused downstream to the SlAQP coding region and the fusion gene SlAQP::GFP transferred into onion epidermal cells by biolistic method. The Mirco Tom tomato was used as material, the real time-PCR was adopted to study the expression profile of gene SlAQP.【Result】 The full-length cDNA of SlAQP gene (GenBank Accessin No. HQ433337) consists of 1 107 bp and contains a 852 bp open reading frame（ORF） encoding 283 amino acid proteins. Bioinformatics analysis demonstrated that SlAQP gene exhibited a typical structure with six membrane-spanning domains and an internal symmetry showing two highly conserved Asn-Pro-Ala (NPA) motifs, and possessing the MIP family signal consensus sequence. The SlAQP amino acids showed high identity with other 10 plant species PIP subfamily by NCBI homology comparison analysis. Phylogenetic analysis among 11 species indicated that SlAQP was clustered with the StAQP from Solanum tuberosum. The result of transient expression showed that SlAQP gene was located in the membrane. Southern blot analysis indicated that SlAQP was a single-copy gene. Real time-PCR analysis showed that the expression of SlAQP gene was down-regulated by drought stress.【Conclusion】The expression profiling of this gene indicates that the SlAQP gene may regulared by drought stress. This paper provides important information for the future study on the gene-expression regulation during drought stress.

Key words: tomato, aquaporins, sequence analysis, intracellular localization, Real time-PCR

[1]Kammerloher W, Fischer U, Piechottka G P, Schiiffner A R. Water channels in the plant plasma membrane cloned by immunoselection from an expression system. The Plant Journal, 1994, 6: 187-199.

[2]Holm L M, Jahn T P, Meller A B, Schjoerring J K, Ferri D, Klaerke D A, Zeuthen T. NH3 and NH4+permeability in aquaporin-expressing Xenopus oocytes. Pflugers Archive European Journal of Physiology, 2005, 450: 415-428.

[3]Robinson D C, Sieber H, Kammerloher W, Schaffner A R. PIP1 aquaporins are concentrated in plasma lemmasomes of Arabidopsis thaliana mesophyll. Plant Physiology, 1996, 111: 645-649.

[4]Johnson K D, Herman E M, Chrispeels M J. An abundant highly conserved tonoplast protein in seeds. Plant Physiology, 1989, 91: 1006-1013.

[5]Johansson I, Karlsson M, Johanson U, Larsson C, Kjellbom P. The role of aquaporins in cellular and whole plant water balance. Biochimica Biophysica Acta, 2000, 1465: 324-342.

[6]Kaldenhoff R, Kolling A, Meyers J, Karmann N U, Ruppel G, Richter G. The blue light-responsive AthH2 gene of Arabidopsis thaliana is primarily expressed in expanding as well as in differentiating cells and encodes a putative channel protein of the plasma membrane. Plant Journal, 1995, 7: 87-95.

[7]Barrieu F, Chaumont F, Chrispeels M J. High expression of the tonoplast aquaporin ZmTIP1 in epidermal and conducting tissues of maize. Plant Physiology, 1998, 117: 1153-1163.

[8]Tyerman S D, Bohnert H J, Maurel C, Steudle E, Smith J A C. Plant aquaproins: their molecular biology, biophysics and significance for plant water relations. Journal of Experimental Botany, 1998, 49: 1583-1587.

[9]Reisen D, Nathalie L C, Özalp C, Chaumont F, Marty F. Expression of a cauliflower tonoplast aquaporin tagged with GFP in tobacco suspension cells correlates with an increase in cell size. Plant Molecular Biology, 2003, 52: 387-400.

[10]O’Brien M, Bertrand C, Daniel P. Matton. Characterization of a fertilization-induced and developmentally regulated plasma- membrane aquaporin expressed in reproductive tissues, in the wild potato Solanum chacoense Bitt. Planta, 2002, 215: 485-493.

[11]Karlsson M, Johansson I, Bush M, McCann M C, Maurel C, Larsson C, Kjellbom P. An abundant TIP expressed in mature highly vacuolated cells. Plant Journal, 2000, 21: 83-90.

[12]Shiota H, Sudoh T, Tanaka I. Expression analysis of genes encoding plasma membrane aquaporins during seed and fruit development in tomato. Plant Science, 2006, 171: 277-285.

[13]Alleva1 K, Marquez M, Villarreal N, Mut P, Bustamante C, Bellati J, Mart?´nez G, Civello M, Amodeo G. Cloning, functional characterization, and co-expression studies of a novel aquaporin (FaPIP2;1) of strawberry fruit. Journal of Experimental Botany, 2010, 61: 3935-3945.

[14]Henzler T, Steudle E. Reversible closing of water channels in chara internodes provides evidence for a composite transport model of the plasma membrane. Journal of Experimental Botany, 1995, 46: 199-209.

[15]Smart L B, Moskal W A, Cameron K D, Bennett A B. MIP genes are down-regulated under drought stress in Nicotiana glauca. Plant and Cell Physiology, 2001, 42(7): 686-693.

[16]马  亮, 梅晓宏, 许文涛, 罗云波, 周  忻, 唐小革, 黄昆仑. 玉米In5-2启动子功能区域在洋葱瞬时表达系统中的分析. 农业生物技术学报, 2010, 18(6): 1073-1078.

Ma L, Mei X H, Xu W T, Luo Y B, Zhou X, Tang X G, Huang K L. Analyses of functional region of maize In5-2 promoter in transient expression system of onion. Journal of Agricultural Biotechnology, 2010, 18(6): 1073-1078. (in Chinese)

[17]Maurel C. Aquaporins and water permeability of plant membranes. Annual Review of Plant Physiology and Plant Molecular Biology, 1997, 48: 399-429.

[18]PARK J H, SAIER M H Jr. Phylogenetic characterization of the MIP family of transmembrane channel proteins . Journal of Membrane Biology, 1996, 153( 3) : 171-180.

[19]王云霄, 张  颖, 江璐玎, 牛  蓓, 陈  放. 麻疯树水通道蛋白新基因J cP IP干旱胁迫下的功能分析. 热带亚热带植物学报, 2008, 16 (4) : 289-295.

Wang Y X, Zhang Y, Jiang L D, Niu B, Chen F. Cloning of a new aquaporin gene (JcPIP) from Jatropha curcas and analysis of its function under drought stress. Journal of Tropical and Subtropical Botany, 2008, 16 (4) : 289-295. (in Chinese)

[20]Chalfie M, Tu Y, Euskirchen G. Green fluorescent protein as a marker for gene expression. Science, 1994, 263: 802-805.

[21]Mahdieh M, Mostajeran A, Horie T, Katsuhara M. Drought stress alters water relations and expression of PIP-type aquaporin genes in Nicotiana tabacum plants. Plant and Cell Physiology, 2008, 49(5): 801-813.

[22]吴旺泽, 彭晓莉, 王晓明, 王  蒂. 马铃薯水通道蛋白基因cDNA 克隆、序列分析及表达. 农业生物技术学报, 2007, 15 (4): 677-683.

Wu W Z, Peng X L, Wang X M, Wang D. Cloning, sequence analysis and expression of cDNA encoding plasma aquaporins StPIP1 of Solanum tuberosum. Journal of Agricultural Biotechnology, 2007, 15(4): 677-683. (in Chinese)

[23]Jang J Y, Kim D G, Kim Y O, Kim J S, Kang H S. An expression analysis of a gene family encoding plasma membrane aquaporins in response to abiotic stresses in Arabidopsis thaliana. Plant Molecular Biology, 2004, 54: 713-725.

[24]Kaldenhoff R, Fischer M. Aquaporins in plants. Acta Physiology, 2006, 187: 169-176.

[25]Barrieu F, Marty-Mazars D, Thomas D, Chaumont F, Charbonnier M, Marty F. Desiccation and osmotic stress increase the abundance of mRNA of the tonoplast aquaporin BobTIP26-1 in cauliflower cells. Planta, 1999, 209: 77-86.

[26]Sarda X, Tousch D, Ferrare K, Legrand E, Dupuis J M, Casse-Delbart F, Lamaze T. Two TIP-like genes encoding aquaporins are expressed in sunflower guard cells. Plant Journal, 1997, 12: 1103-1111.

[27]陈儒钢，朱文超，巩振辉，李大伟，尹延旭，逯明辉. 辣椒水通道蛋白基因CaAQP 的克隆与序列分析. 中国农业科学, 2010, 43(20): 4323-4329.

Chen R G, Zhu W C, Gong Z H, Li D W, Yin Y X, Lu M H. Cloning and sequence analysis of the aquaporins gene CaAQP in pepper. Journal of Agricultural Biotechnology, 2010, 43(20): 4323-4329. (in Chinese)

[28]Jiang W, Fu F L, Jiang W, Mu Y, Yong T M, Li W C. Cloning and characterization of functional trehalose-6-phosphate synthase gene in maize. Journal of Plant Biology, 2010, 53:134-141.
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