Construction of Plant Expression Vector with GmNHX1 and LcVP1 Dual-gene and Obtaining Transgenic Cotton

doi:10.7668/hbnxb.2014.04.010

Abstract

Abstract: By use of DNA recombination method,a vector,p1301-GmNHX1-LcVP1, harboring two open reading frames of soybean tonoplast Na⁺/H⁺ antiporter GmNHX1 and Lotus corniculatus H⁺-PPase LcVP1,was constructed through sequential restriction digests and ligations recombination.The two genes driven by CaMV 35S promoters were successfully transferred into cotton cultivars Coker 312 via Agrobacterium with hygromycin selection system.The molecular and physiological analysises of T₁ generation indicated that genes were integrated into the cotton genome and enhanced transgenic cotton salt tolerance.

Key words: Cotton, Salt tolerance, Soybean tonoplast Na⁺/H⁺ antiporter GmNHX1, Lotus corniculatus H⁺-PPase LcVP1, Genetic transformation

CLC Number:

Q78
S562.03

FENG Xue, JIA Yong-hong, GUO Hong-zhen, ZHANG Yi-ming, ZHAO Xue-liang, SUN Yan-xiang. Construction of Plant Expression Vector with GmNHX1 and LcVP1 Dual-gene and Obtaining Transgenic Cotton[J]. journal1, 2014, 29(4): 56-62. doi: 10.7668/hbnxb.2014.04.010.

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http://www.hbnxb.net/EN/Y2014/V29/I4/56

References

[1] Jiang L J,Duan L S,Tian X L,et al.NaCl salinity stress decreased Bacillus thuringiensis(Bt) protein content of transgenic Bt cotton (Gossypium hirsutum L.) seedlings[J].Environ Exp Bot,2006,55(3):315-320.
[2] Apse M P,Aharon G S,Snedden W A,et al.Salt tolerance conferred by overexpression of a vacuolar Na⁺/H⁺ antiport in Arabidopsis[J].Science,1999,285(5431):1256-1268.
[3] Gaxiola R A,Li J,Undurraga S,et al.Drought-and salt-tolerant plants result from overexpression of the AVP1 H⁺-pump[J].Proc Natl Acad Sci,2001,98(20):11444-11449.
[4] Zhao F Y,Zhang X J,Li P H,et al.Co-expression of the Suaeda salsa SsNHX1 and Arabidopsis AVP1 confer greater salt tolerance to transgenic rice than the single SsNHX1 [J].Mol Breeding,2006,17(4),341-353.
[5] He C,Yan J,Shen G,et al.Expression of an Arabidopsis vacuolar sodium/proton antiporter gene in cotton improves photosynthetic performance under salt conditions and increases fiber yield in the field[J].Plant Cell Physiol,2005,46(11):1848-1854.
[6] Pasapula V,Shen G,Kuppu S,et al.Expression of an Arabidopsis vacuolar H⁺-pyrophosphatase gene (AVP1) in cotton improves drought-and salt tolerance and increases fibre yield in the field conditions[J].Plant Biotechnol J,2011,9(1):88-99.
[7] Park S,Li J,Pittman J K,et al.Up-regulation of a H⁺-pyrophosphatase (H⁺-PPase) as a strategy to engineer drought-resistant crop plants[J].Proc Natl Acad Sci,2005,102(52):18830-18835.
[8] Leidi E O,Barragán V,Rubio L,et al.The AtNHX1 exchanger mediates potassium compartmentation in vacuoles of transgenic tomato[J],Plant J.2010,61(3):495-506.
[9] 孙艳香,王丹,白艳玲,等.大豆 GmNHX1 在百脉根中的过表达研究:体内Na⁺含量的降低是耐盐性提高的基础[J].科学通报,2006,51(8):928-936.
[10] 孙艳香,王勇.百脉根液泡膜H⁺-PPase基因的cDNA克隆与分析[J].植物生理通讯,2007,43(4):657-663.
[11] 孙艳香,冯雪,赵学良.百脉根液泡膜H⁺-PPase基因 LcVP1 植物表达载体构建与转基因毛白杨的获得[J].中国农学通报.2011,27(4):134-138.
[12] Wu S J,Wang H H,Li F F,et al.Enhanced Agrobacterium-mediated transformation of embryogenic calli of upland cotton via efficient selection and timely subculture of somatic embryos[J].Plant Mol Biol Rep,2008,26(3):174-185.
[13] Zhang T,Wu S J.Enhanced Agrobacterium-mediated transformation of embryogenic calli of upland cotton[J].Methods Mol Biol,2012,847:245-253.
[14] Paterson A H,Brubaker C L,Wendel J F.A rapid method for extraction of cotton (Gossypium spp.) Genomic DNA suitable for RFLP or PCR analysis[J].Plant Mol Biol Rep,1993,11(2):122-127.
[15] Sambrook J,Fritsch E F,Maniatist T.Molecular Cloning:A laboratory mannual[M].2 nd eds.New York:Cold Spring Harbor Laboratory Press,1989.
[16] 中国科学院上海植物生理研究所,上海市植物生理学会.现代植物生理学实验指南[M].北京:科学出版社,1999.
[17] Shi H,Quintero F J,Pardo J M,et al.The putative plasma membrane Na⁺/H⁺ antiporter SOS1 controls long-distance Na⁺ transport in plants[J].Plant Cell,2002,14(2):465-477.
[18] Qiu Q S,Guo Y,Quintero F J,et al.Regulation of vacuolar Na⁺/H⁺ exchange in Arabidopsis thaliana by the salt-overly-sensitive (SOS) pathway[J].J Biol Chem,2004,279(1):207-215.
[19] Mennen H,Jacoby B,Marschner H.Is sodium proton antiport ubiquitous in plant cells[J].J Plant Physiol,1990,137:180-183.
[20] 蒋玉蓉,吕有军,祝水金.棉花耐盐机理与盐害控制研究进展[J].棉花学报,2006,18(4):248-254.
[21] 吴霞,吴艾,上官小霞,等.转耐盐基因 GhNHX1 棉花株系耐盐性初步鉴定[J].山西农业科学,2012,40(1):16-17,4.
[22] 张国新,王秀萍,鲁雪林,等.自然盐分胁迫下棉花耐盐性评价[J].河南农业科学,2011,40(8):95-97.
[23] 李燕青,孙文彦,许建新,等.华北盐碱地耐盐经济作物筛选[J].华北农学报,2013,28(增刊):227-232.
[24] Hassidim M,Braun Y,Lerner H R,et al.Na/H and K/H antiport in root membrane vesicles isolated from the halophyte atriplex and the glycophyte cotton[J].Plant Physiol,1990,94(4):1795-1801.
[25] Lin H,Salus S S,Schumaker K S.Salt sensitivity and the activities of the H⁺-ATPases in cotton seedlings[J].Crop Sci,1997,37:190-197.
[26] Wu C A,Yang G D,Meng Q W,et al.The cotton GhNHX1 gene encoding a novel putative tonoplast Na(+)/H(+) antiporter plays an important role in salt stress[J].Plant Cell Physiol,2004,45(5):600-607.
[27] De Block M,Debrouwer D.Two T-DNA's co-transformed into Brassica napus by a double Agrobacterium tumefaciens infection are mainly integrated at the same locus[J].Theor Appl Genet,1991,82:257-263.
[28] 燕丽,崔继哲,张亮,等.共转化法及其在植物基因工程中的应用[J].生物技术通报,2008(1):91-94.
[29] 武维华.植物生理学[M].2版.北京:科学出版社,2008.

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