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.

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

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