Cloning and Expression Analysis of VmNHX Gene in Vigna marina
Vigna marina(Burm.)Merr.is a perennial creeping or climbing herb of the Fabaceae,Vigna Savi genus,with strong salinity and drought resistance,and is a good candidate species for saline land reclaim.In order to ana-lyze the characteristics and functions of the VmNHX gene sequence of the Na+/H+transporter in Vigna marina,bioin-formatics,molecular biology and real-time PCR were used in this study.The results show that the cDNA of the VmNHX gene is 1 617 bp and encodes 538 amino acids.The molecular formula of the protein was C2729H4214N670O761S30.The molecular weight was 59.5 kD,and the theoretical isoelectric point was 5.89,which is speculated to be an acidic protein;this protein contains 1 b_cpal super family conserved domain and 11 transmembrane domains,which is a typical non-secretory,hydrophobic and transmembrane protein.The secondary structure is mostly a spi-ral structure,with a small number of chain structures.Prediction of VmNHX subcellular localization is localized in the plasma membrane and is consistent with tobacco subcellular localization results.Evolutionary tree and mul-ti-sequence alignment analysis showed that the VmNHX gene was compared to Glycine max(Linn.)Merr.I1KNP2_SOYBN,Arabidopsis thaliana(L.)Heynh At_NHX5 and At_NHX6 genes have high homology and belong to the CPA1 family.Real-time PCR analysis showed that under normal circumstances,the VmNHX gene was expressed in roots,stems and leaves,and the highest expression was expressed in stems.Under salt stress,expression in the roots and stems is significantly up-regulated.Analysis of Na+content showed that the accumulation of Na+in the root system of Vigna marina under salt treatment was significantly higher than that in the aboveground part.It is speculated that VmNHX plays an important role in the salt ion isolation of the root system of Vigna marina,which provides a reference for further analysis of the molecular mechanism of salt tolerance in Vigna marina,and the use of excellent salt tolerance gene of Vigna marinafor salt tolerant crop breeding.
Vigna marina(Burm.)Merr.Gene cloningNa+/H+antitransportersSalinity resistanceCPA1 family
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