苦荞麦FtNHX1基因的克隆及表达分析

发布时间：2018-06-05 15:38

本文选题：苦荞麦 + 盐胁迫　；参考：《华北农学报》2017年04期

【摘要】：为深入研究液泡膜Na~+/H~+逆向转运蛋白在植物耐盐中的作用,以耐盐苦荞麦品种川荞1号为材料,利用同源克隆方法得到NHX基因,命名为FtNHX1,并在Gen Bank中注册,登录号为KY438929;序列分析表明,该基因开放阅读框1 662 bp,共编码553个氨基酸,预测蛋白分子量61.24 kDa,等电点5.15。系统进化树分析表明,FtNHX1与拟南芥(AtNHX1)、水稻(OsNHX1)和小麦(TaNHX1)的亲缘关系较近,氨基酸同源率分别为60.22%,58.95%和57.30%;荧光定量PCR分析表明,随着Na Cl胁迫浓度的增加,FtNHX1基因在苦荞麦根部、茎基部和叶片的相对表达量极显著增加,150 mmol/L NaCl胁迫下增加幅度最大,分别比对照增加了254.10%,311.35%和256.18%;Na Cl胁迫下FtNHX1基因在苦荞麦根部、茎基部和叶片的平均表达量分别比对照增加了109.46%,145.67%和155.94%,茎基部和叶片的平均表达量较高,说明苦荞麦FtNHX1基因的表达明显受盐胁迫诱导和调节,FtNHX1基因与苦荞麦的耐盐性有密切联系。
[Abstract]:In order to study the role of vacuolar membrane Na ~ / H ~ antiporter in salt tolerance of plants, a salt tolerant buckwheat variety Chuanzao 1 was used to obtain NHX gene named FtNHX1 by homology cloning method, and it was registered in GenBank. The accession number was KY438929, and the sequence analysis showed that the open reading frame 1662 BP encoded 553 amino acids, the predicted molecular weight of protein was 61.24 kDaand the isoelectric point was 5.15. Phylogenetic tree analysis showed that FtNHX1 was closely related to Arabidopsis thaliana AtNHX1, rice OsNHX1) and wheat TaNHX1), the amino acid homology was 60.2222% and 57.300.The fluorescence quantitative PCR analysis showed that FtNHX1 gene was located in the root of buckwheat with the increase of NaCl stress concentration. The relative expression of FtNHX1 in stem base and leaf was significantly increased under 150 mmol / L NaCl stress, and the FtNHX1 gene was increased by 254.1010 ~ 311.35% and 256.1818% respectively in the root of Tartary buckwheat under NaCl stress. The average expression of FtNHX1 in stem base and leaf increased by 145.67% and 155.94%, respectively, and the average expression of FtNHX1 gene in stem base and leaf was higher than that in control, which indicated that the expression of FtNHX1 gene in Tartary buckwheat was induced and regulated by salt stress, and there was a close relationship between FtNHX1 gene and salt-tolerance of Tartary buckwheat.
【作者单位】：青岛农业大学生命科学学院山东省高校植物生物技术重点实验室;
【基金】：国家自然科学基金项目(31371552) 山东省自然科学基金项目(ZR2010CL019)
【分类号】：Q943.2;S517

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