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新疆雪莲水孔蛋白sikPIP1基因的克隆与功能分析

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.09.013, PP. 97-105

Keywords: 新疆雪莲,水孔蛋白,sikPIP1基因,烟草,抗寒性,抗旱性

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Abstract:

利用PCR技术从已建立的新疆雪莲cDNA文库中克隆雪莲水孔蛋白基因sikPIP1,构建植物表达载体pBI121-sikPIP1,利用农杆菌介导法获得转基因烟草,PCR和RT-PCR检测证明该基因成功导入并得以转录,并对检测结果均为阳性的植株通过水分胁迫和温度胁迫进行抗旱性和抗寒性分析。结果显示:(1)克隆得到长为880bp,具有水孔蛋白特性的sikPIP1基因,完整ORF为840bp。(2)水分胁迫中,断水7d后转基因烟草生长表型明显优于野生型烟草,生理指标测定结果显示,转基因烟草的相对电导率和丙二醛含量均低于野生型烟草,相对含水量高于野生型烟草。(3)不同温度胁迫处理,转基因烟草表型显著优于野生型,特别是0℃以下低温胁迫,野生型烟草出现严重的萎蔫,而转基因烟草受伤害程度较轻;生理指标结果显示转基因烟草相对电导率、丙二醛含量低于野生型烟草。结果表明,转sikPIP1基因提高了烟草抗旱能力和抗寒能力。

References

[1]  Denker BM, Smith BL, Kuhajda FP, et al. Identification, purification, and partial characterization of a novel Mr 28, 000 integral membrane protein from erythrocytes and renal tubules[J]. J Biol Chem, 1988, 263(30):15634-15642
[2]  Chaumont F, Moshelion M, Daniels MJ. Regulation of plant aquaporin activity[J]. Biol Cell, 2005, 97(10):749-764.
[3]  Bienert GP, Heinen RB, Berny MC, et al. Maize plasma membrane aquaporin ZmPIP2;5, but not ZmPIP1;2, facilitates transmembrane diffusion of hydrogen peroxide[J]. Biochimica et Biophysica Acta, 2014, 1838:216-222.
[4]  Schuurmans JA, van Dongen JT, Rutjens BPW, et al. Members of the aquaporin family in the developing pea seed coat include representatives of the PIP, TIP, and NIP subfamilies[J]. Plant Mol Biol, 2003, 53(5):655-667.
[5]  Holm LM, Jahn TP, Moller AL, et al. NH3 and NH4+ permeability in aquaporin-expressing Xenopus oocytes[J]. Pflugers Arch, 2005, 450(6):415-428.
[6]  李红梅, 丁岳炼, 黄新敏, 等. 香石竹切花水孔蛋白基因DcPIP2的克隆及特征分析[J]. 园艺学报, 2011, 38(11):2199-2208.
[7]  Sakurai J, Ishikawa F, Yamaguchi T, et al. Identification of 33 rice aquaporin genes and analysis of their expression and function[J]. Plant Cell Physiol, 2005, 46(9):1568-1577.
[8]  Wudick MM, Luu DT, Maurel C. A look inside:localization patterns and functions of intracellular plant aquaporins[J]. New Phytol, 2009, 184(2):289-302.
[9]  Lee SH, Chung GC, Jang, JY, et al. Overexpression of PIP2;5 aquaporin alleviates effects of low root temperature on cell hydraulic conductivity and growth in Arabidopsis[J]. Plant Physiology, 2012, 159(1):479-488.
[10]  Liu CW, Fukumoto T, Matsumoto T, et al . Aquaporin OsPIP1;1 promotes rice salt resistance and seed germination[J]. Plant Physiol Biochem, 2013, 63:151-158.
[11]  李嵘, 牛向丽, 苗雁文, 等. 水通道蛋白基因OsPIP2;6的功能分析[J]. 中国农业科学, 2013, 46(15):3079-3086.
[12]  陈发菊. 我国雪莲植物的种类、生境分布及化学成分的研究进展[J]. 植物学通报, 1999, 16(5):561-566.
[13]  焦天奇, 孙辉, 祝建波, 等. 转天山雪莲sikPIP3基因烟草的获得及抗逆性鉴定[J]. 西北植物学报, 2012, 32(3):431-438.
[14]  程晨, 郭新勇, 王爱英, 祝建波. 转新疆雪莲去饱和酶基因sikSAD重组酵母低温和酒精耐受性分析[J]. 微生物学报, 2011, 38(11):1647-1656.
[15]  李合生. 植物生理生化实验原理和技术[M]. 北京:高等教育出版社, 2003:164-263.
[16]  Aharon R, Shahak Y, Wininger S, et al. Overexpression of a plasma membrane aquaporin in transgenic tobacco improves plant vigor under favorable growth conditions but not under drought or salt stress[J]. Plant Cell, 2003, 15(2):439-447.
[17]  Lin W, Peng Y, Li G, et al. Isolation and functional characterization of PgTIP1, a hormone-autotrophic cells-specific tonoplast aquaporin in ginseng[J]. J Experimental Botany, 2007, 58(5):947-956.
[18]  Kaldenhoff R, Ribas-Carbo M, Sans JF, et al. . Aquaporins and plant water balance[J]. Plant Cell Environ, 2008, 31(5):658-666.
[19]  Hachez C, Besserer A, Chevalier AS, et al. Insights into plant plasma membrane aquaporin trafficking[J]. Plant Sci, 2013, 18(6):344-352.
[20]  Izumi CM, Jiye R, Mineo S, et al. CO2 transport by PIP2 aquaporins of barley[J]. Plant Cell Physiol, 2014, 55(2):251-257.
[21]  Forrest KL, Bhave M. Major intrinsic proteins(MIPs)in plants:a complex gene family with major impacts on plant phenotype, Funt. Integr[J]. Genomics, 2007, 7(4):263-289.
[22]  Aroca R, Ferrante A, Vernieri P, et al. Drought, abscisic acid and transpiration rate effects on the regulation of PIP aquaporin gene expression and abundance in Phaseolus vulgaris plants[J]. Ann Bot, 2006, 98(6):1301-1310.
[23]  Heckwolf M, Pater D, Hanson DT, et al. The Arabidopsis thaliana aquaporin AtPIP1;2 is a physiologically relevant CO2 trasport facilitator[J]. Plant J, 2011, 67(5):795-804.
[24]  Johanson U, Karlsson M, Johansson I, et al. The complete set of genes encoding major intrinsic proteins in Arabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants[J]. Plant Physiol, 2001, 126(4):1358-1369.
[25]  Danielson JAH, Johanson U. Unexpected complexity of the aquaporin gene family in the moss Physcomitrella patens[J]. BMC Plant Biol, 2008, 8:45.
[26]  Jang JY, Lee SH, Rhee JY, et al. Transgenic arabidopsis and tobacco plants overexpressing an aquaporin respond differently to various abiotic stresses[J]. Plant Mol Biol, 2007, 64:621-632.
[27]  Aroca R, Porcel R, Ruiz-Lozano JM. Regulation of root water uptake under abiotic stress conditions[J]. J Exp Bot, 2012, 63(1):43-57.
[28]  Zhang J, Li DD, Zou D, et al. A cotton gene encoding a plasma membrane aquaporin is involved in seedling development and in response to drought stress[J]. Acta Biochim Biophys Sin, 2013, 45:104-114.
[29]  Matsumoto T, Lian HL, Su WA, et al. Role of the aquaporin PIP1 subfamily in the chilling tolerance of rice[J]. Plant Cell Physiol, 2009, 50(2):216-229.
[30]  庄丽, 李卫红, 孟丽红. 新疆雪莲资源的利用、研发与保护[J]. 干旱区资源与环境, 2006, 20(2):195-202.
[31]  郭新勇, 程晨, 祝建波, 等. 天山雪莲冷调节蛋白基因siCOR转化烟草植株的抗旱性分析[J]. 植物学报, 2012, 47(2):111-119.
[32]  Park JH, Saier MH. Phylogenetic characterization of the MIP family of tranmembrare channel protein[J]. Membr Biol, 1996(153):171-180.
[33]  Fetter K, Van Wilder V, Moshelion M, et al. Interactions between plasma membrane aquaporins modulate their water channel activity[J]. Plant Cell, 2004, 16(1):215-228.

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