Cloning and Function Analysis of Aquaporin Protein Gene sikPIP1 from Saussurea involucrata Kar. et Kir

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.013

Abstract

Abstract: Saussurea involucrata aquaporin protein gene sikPIP₁ was cloned from established cDNA library by PCR technique. Then a plant expression vector of pBI121-sikPIP₁ was constructed and transgenic tobacco plants were obtained by Agrobacterium-mediated method. The analysis by PCR and RT-PCR indicated that the aquaporin protein gene sikPIP₁ had been integrated and transcribed into the tobacco genome. The drought and cold resistance by water stress and low-temperature stress for the positive plants from above PCR and RT-PCR examination were analyzed. The results showed that：(1)The plasma membrane aquaporin gene sikPIP₁ was cloned with the length of 880 bp and complete ORF of 840 bp. (2)Under the water stress, the growth phenotype of transgenic tobacco was evidently better than wild-type after 7 days’ water-break, the results of physiological index showed that relative conductivity and MDA content of transgenic tobacco were lower than those of wild-type. However, the relative water content of transgenic tobacco was higher than that of wild-type. (3)The growth phenotype of transgenic tobacco was significantly better than that of wild-type under the different temperature stress. Especially below the 0 degree, the wild-type ones were wilting seriously, however, the transgenic tobacco had less damages. The results of physiological index showed that relative conductivity and MDA content of transgenic tobacco were lower than that of wild-type. The testing results showed that sikPIP₁ gene improved tobacco's tolerance of drought and cold.

Key words: Sasussured involucrate, aquaporin protein, sikPIP₁ gene, tobacco, drought tolerance, cold tolerance

Li Yushun, Liu Yiling, Liu Bucang, Mu Jianqiang, Zhu Jianbo. Cloning and Function Analysis of Aquaporin Protein Gene sikPIP₁ from Saussurea involucrata Kar. et Kir[J]. Biotechnology Bulletin, 2015, 31(9): 97-105.

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] Kaldenhoff R, Ribas-Carbo M, Sans JF, et al. . Aquaporins and plant water balance[J]. Plant Cell Environ, 2008, 31(5)：658-666.
[3] Chaumont F, Moshelion M, Daniels MJ. Regulation of plant aquaporin activity[J]. Biol Cell, 2005, 97(10)：749-764.
[4] 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.
[5] Hachez C, Besserer A, Chevalier AS, et al. Insights into plant plasma membrane aquaporin trafficking[J]. Plant Sci, 2013, 18(6)：344-352.
[6] 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.
[7] Izumi CM, Jiye R, Mineo S, et al. CO₂ transport by PIP2 aquaporins of barley[J]. Plant Cell Physiol, 2014, 55(2)：251-257.
[8] 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.
[9] 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.
[10] 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.
[11] 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.
[12] 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.
[13] 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.
[14] 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.
[15] Danielson JAH, Johanson U. Unexpected complexity of the aquaporin gene family in the moss Physcomitrella patens[J]. BMC Plant Biol, 2008, 8：45.
[16] 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.
[17] 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.
[18] 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.
[19] 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.
[20] 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.
[21] 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.
[22] 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.
[23] 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.
[24] 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.
[25] 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.
[26] 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.
[27] 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.
[28] 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.
[29] 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.
[30] Park JH, Saier MH. Phylogenetic characterization of the MIP family of tranmembrare channel protein[J]. Membr Biol, 1996(153)：171-180.
[31] 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.
[32] 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.
[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.

Cloning and Function Analysis of Aquaporin Protein Gene sikPIP₁ from Saussurea involucrata Kar. et Kir

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