大豆耐盐相关基因GmNcl1的序列单倍型及表达分析

doi:10.3864/j.issn.0578-1752.2014.03.001

Abstract

Abstract: 【Objective】The objective of the study is to confirm sequence polymorphisms of GmNcl1 gene and its differential pattern in stress. 【Method】Amino acid sequence of GmNCl1 was compared by BLASTP to find homologous genes. Fifty soybean varieties were used for the sequencing of GmNcl1 promoter and gene, and indicating of salt tolerance, and haplotype cluster and phylogenetic tree were analyzed by MAGE, looking for the correlation between salt-tolerance and sequence of GmNcl1. The expression pattern of GmNcl1 by real time PCR analysis in roots of salt-tolerant cultivar Tiefeng 8 and salt-sensitive cultivar 85-140 under a series of treatments including the protein inhibitors, pH, ABA treatments and salt, alkali and drought stress. 【Result】 GmNcl1 was homologous with Na +(K+)/H+ exchanger CHX. Sixteen single nucleotide polymorphism loci and two InDel loci were found in GmNcl1 gene sequence, including one G/A (salt-sensitive/salt-tolerant) base change in exon 3, which caused the nonsynonymous mutation from alanine to threonine. Eighteen polymorphism loci were composed into fourteen haplotypes, of which there were three salt-tolerant haplotypes and eleven salt-sensitive haplotypes. The haplotypes, GAGATATTC (salt-tolerant)/TTT ---- CT (salt-sensitive) composed of six loci were high-effectively distinguished between the salt tolerance and sensitivity. In the roots of soybean seedling, the expression of GmNcl1 gene was increased under the Na+/H+ antiporter inhibitor, amiloride, and unchanged under Ca2+-ATPase inhibitor, sodium vanadate. The expression of GmNcl1 was increased under alkaline pH, and showed fluctuations under acidic pH. The expression of GmNcl1 was also induced by ABA, and up-regulated by salt, alkali and drought stress. The greater stress, the more up-regulated expression of GmNcl1. Expression of GmNcl1 showed a similar trend in Tiefeng 8 and 85-140 under salt stress, but there was the greater increase in 85-140 than Tiefeng 8.【Conclusion】GmNcl1 was highly similar with Na+(K+)/H+ exchanger, its sequence polymorphism was related to salt-tolerance and sensitivity. GmNcl1 was inducted by ABA and GmNcl1 might be involved in the balance of the organism internal environment, and be a key factor in the stress response to salt, alkali and drought stress in soybean.

Key words: soybean , cation/proton exchanger , stress-tolerance , expression analysis

HE Wei-1, LIU Lin-2, GUAN Rong-Xia-2, QIU Li-Juan-2. Haplotypes and Expression Analysis of Salt-Tolerant Gene GmNcl1 in Soybean[J].Scientia Agricultura Sinica, 2014, 47(3): 411-421.

References

[1]FAO (Food, Agriculture Organization of the United Nations). FAO Production Yearbook. FAO, Rome, 2004.

[2]Chang A B, Lin R, Keith Studley W, Tran C V, Saier Jr M H. Phylogeny as a guide to structure and function of membrane transport proteins. Molecular Membrane Biology, 2004, 21: 171-181.

[3]Nagata T, Iizumi S, Satoh K, Kikuchi S. Comparative molecular biological analysis of membrane transport genes in organisms. Plant Molecular Biology, 2008, 66: 565-585.

[4]Chanroj S, Wang G, Venema K, Zhang M W, Delwiche C F, Sze H. Conserved and diversified gene families of monovalent cation/H+ antiporters from algae to flowering plants. Frontiers in Plant Science, 2012, 3: 25.

[5]Sze H, Padmanaban S, Cellier F, Honys D, Cheng N H, Bock K W, Conejero G, Li X, Twell D, Ward J M, Hirschi K D. Expression patterns of a novel AtCHX gene family highlight potential roles in osmotic adjustment and K+ homeostasis in pollen development. Plant Physiology, 2004, 136(1): 2532-2547.

[6]Chanroj S, Lu Y, Padmanaban S, Nanatani K, Uozumi N, Rao R, Sze H. Plant-speci?c cation/H+ exchanger 17 and its homologs are endomembrane K+ transporters with roles in protein sorting. The Journal of Biological Chemistry, 2011, 286: 33931-33941.

[7]Lu Y, Chanroj S, Zulkifli L, Johnson M A, Uozumi N, Cheung A, Sze H. Pollen tubes lacking a pair of K+ transporters fail to target ovules in Arabidopsis. The Plant Cell, 2011, 23(1): 81-93.

[8]Cellier F, Conejero G, Ricaud L, Luu D T, Lepetit M, Gosti F, Casse F. Characterization of AtCHX17 a member of the cation/H+ exchangers CHX family from Arabidopsis thaliana suggests a role in K+ homeostasis. The Plant Journal, 2004, 39(6): 834-846.

[9]Zhao J, Cheng N H, Motes C M, Blancaflor E B, Moore M, Gonzales N, Padmanaban S, Sze H, Ward J M, Hirschi K D. AtCHX13 is a plasma membrane K+ transporter. Plant Physiology, 2008, 148(2): 796-807.

[10]Kreps J A, Wu Y, Chang H S, Zhu T, Wang X, Harper J F. Transcriptome changes for Arabidopsis in response to salt, osmotic, and cold stress. Plant Physiology, 2002, 130(4): 2129-2141.

[11]Maathuis F J, Filatov V, Herzyk P, Krijger G C, Axelsen K B, Chen S, Green B J, Li Y, Madagan K L, Sanchez-Fernandez R, Forde B G, Palmgren M G, Rea P A, Williams L E, Sanders D, Amtmann A. Transcriptome analysis of root transporters reveals participation of multiple gene families in the response to cation stress. The Plant Journal, 2003, 35(6): 675-692.

[12]Hall D, Evans A R, Newbury H J, Pritchard J. Functional analysis of CHX21: A putative sodium transporter in Arabidopsis. Journal of Experimental Botany, 2006, 57(5): 1201-1210.

[13]Song C P, Guo Y, Qiu Q, Lambert G, Galbraith D W, Jagendorf A, Zhu J K. A probable Na+(K+)/H+ exchanger on the chloroplast envelope functions in pH homeostasis and chloroplast development in Arabidopsis thaliana. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(27): 10211-10216.

[14]Padmanaban S, Chanroj S, Kwak J M, Li X, Ward J M, Sze H. Participation of endomembrane cation/H+ exchanger AtCHX20 in osmoregulation of guard cells. Plant Physiology, 2007, 144: 82-93.

[15]姜静涵, 郭勇, 常汝镇, 关荣霞, 邱丽娟. 大豆苗期耐盐性的简便鉴定方法. 作物学报, 2013, 39(7): 1248-1256.

Jiang J H, Guo Y, Chang R Z, Guan R X, Qiu L J. Simple evaluation method of tolerance to salt at seedling stage in soybean. Acta Agronomica Sinica, 2013, 39(7): 1248-1256. (in Chinese)

[16]Tester M, Davenport R. Na+ tolerance and Na+ transport in higher plants. Annals of Botany, 2003, 91: 503-527.

[17]Chen L H, Zhang B, Xu Z Q. Salt tolerance conferred by overexpression of Arabidopsis vacuolar Na+/H+ antiporter gene AtNHX1 in common buckwheat (Fagopyrum esculentum). Transgenic Research, 2008, 17(1): 121-32.

[18]Galvez F J, Baghour M, Hao G, Cagnac O, Rodriguez-Rosales M P, Venema K. Expression of LeNHX isoforms in response to salt stress in salt sensitive and salt tolerant tomato species. Plant Physiology and Biochemistry, 2012, 51: 109-115.

[19]Jiang X, Leidi E O, Pardo J M. How do vacuolar NHX exchangers function in plant salt tolerance? Plant Signal Behave, 2010, 5(7): 792-795.

[20]Zorb C, Noll A, Karl S, Leib K, Yan F, Schubert S. Molecular characterization of Na+/H+ antiporters (ZmNHX) of maize (Zea mays L.) and their expression under salt stress. Journal of Plant Physiology, 2005, 162(1): 55-66.

[21]邵桂花, 常汝镇, 陈一舞, 闫淑荣. 大豆耐盐性遗传的研究. 作物学报, 1994, 20(6): 721-726.

Shao G H, Chang R Z, Chen Y W, Yan S R. Study on inheritance of salt tolerance in soybean. Acta Agronomica Sinica, 1994, 20(6): 721-726. (in Chinese)

[22]刘光宇, 关荣霞, 常汝镇, 邱丽娟. 大豆不同器官Na+含量与苗期耐盐性的相关分析. 作物学报, 2011, 37(7): 1266-1273.

Liu G Y, Guan R X, Chang R Z, Qiu L J. Correlation between Na+ contents in different organs of soybean and salt tolerance at the seedling stage. Acta Agronomica Sinica, 2011, 37(7): 1266-1273. (in Chinese)

[23]Counillon L, Franchi A, Pouyssegur J. A point mutation of the Na+/H+ exchanger gene (NHE1) and amplification of the mutated allele confer amiloride resistance upon chronic acidosis. Proceedings of the National Academy of Sciences of the United States of America, 1993, 90(10): 4508-4512.

[24]Darley C P, van Wuytswinkel O C, van der Woude K, Mager W H, de Boer A H. Arabidopsis thaliana and Saccharomyces cerevisiae NHX1 genes encode amiloride sensitive electroneutral Na+/H+ exchangers. The Biochemical Journal, 2000, 351(1): 241-249.

[25]Nelson M T, Blaustein M P. Effects of ATP and vanadate on calcium efflux from barnacle muscle fibres. Nature, 1981, 289(5795): 314-316.

[26]Froman D P, Feltmann A J. Fowl (Gallus domesticus) sperm motility depends upon mitochondrial calcium cycling driven by extracellular sodium. Biology of Reproduction, 2005, 72(1): 97-101.

[27]Maresova L, Sychrova H. Arabidopsis thaliana CHX17 gene complements the kha1 deletion phenotypes in Saccharomyces cerevisiae. Yeast, 2006, 123: 167-171.

[28]Rodriguez-Rosales M P, Jiang X, Galvez F J, Aranda M N, Cubero B, Venema K. Overexpression of the tomato K+/H+ antiporter LeNHX2 confers salt tolerance by improving potassium compartmentalization. The New Phytologist, 2008, 179: 366-377.

[29]Leidi E O, Barragan V, Rubio L, El-Hamdaoui A, Ruiz M T, Cubero B, Fernandez J A, Bressan R A, Hasegawa P M, Quintero F J, Pardo J M. The AtNHX1 exchanger mediates potassium compartmentation in vacuoles of transgenic tomato. The Plant Journal, 2010, 61: 495-506.

[30]Bassil E, Ohto M A, Esumi T, Tajima H, Zhu Z, Cagnac O, Belmonte M, Peleg Z, Yamaguchi T, Blumwald E. The Arabidopsis intracellular Na+/H+ antiporters NHX5 and NHX6 are endosome associated and necessary for plant growth and development. The Plant Cell, 2011, 23: 224-239.

[31]Quintero F J, Martinez-Atienza J, Villalta I, Jiang X, Kim W Y, Ali Z, Fujii H, Mendoza I, Yun D J, Zhu J K, Pardo J M. Activation of the plasma membrane Na/H antiporter Salt-Overly-Sensitive 1 (SOS1) by phosphorylation of an autoinhibitory C-terminal domain. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108: 2611-2616.

[32]Venema K, Belver A, Marin-Manzano M C, Rodriguez-Rosales M P, Donaire J P. A novel intracellular K+/H+ antiporter related to Na+/H+ antiporters is important for K+ ion homeostasis in plants. The Journal of Biological Chemistry, 2003, 278(25): 22453-22459.

[33]Shi H, Zhu J K. Regulation of expression of the vacuolar Na+/H+ antiporter gene AtNHX1 by salt stress and abscisic acid. Plant Molecular Biology, 2002, 50(3): 543-550.

[34]Shi H, Ishitani M, Kim C, Zhu J K. The Arabidopsis thaliana salt tolerance gene SOS1 encodes a putative Na+/H+ antiporter. Proceedings of the National Academy of Sciences of United States of America, 2000, 97(12): 6896-901.

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Haplotypes and Expression Analysis of Salt-Tolerant Gene GmNcl1 in Soybean

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