Cloning and Expression under Drought of NHX6 in Hippophae rhamnoides
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摘要:
目的 克隆中国沙棘HrNHX6基因，分析其在干旱胁迫下的表达，以期为HrNHX6基因的抗旱功能研究提供参考依据。 方法 对中国沙棘HrNHX6基因进行扩增，并利用生物软件分析HrNHX6基因序列，预测其蛋白结构，在此基础上对干旱胁迫下HrNHX6基因的表达进行分析。 结果 获得的HrNHX6基因属于NHX家族NHX6基因，其编码蛋白质定位于液泡膜，无信号肽，含有13个跨膜螺旋区，具有N-糖基化位点、蛋白激酶磷酸化位点、豆蔻酰化位点、酰胺化位点、糖转运蛋白signature 位点等多种类型的修饰位点。qRT-PCR分析结果表明，干旱胁迫下不同组织之间HrNHX6基因的表达水平具显著差异，而不同干旱程度胁迫处理之间HrNHX6基因的表达水平呈极显著差异。 结论 HrNHX6蛋白的结构特性及HrNHX6基因在干旱胁迫下的表达模式表明其与中国沙棘抗旱关系密切且受干旱胁迫诱导。 Abstract:Objective NHX6 of Hippophae rhamnoides was cloned and expression under drought determined to study the stress resistance mechanism. Method HrNHX6 was cloned, amplified, and sequenced for analysis by biological software. Expression of the gene under drought stress was determined. Result Located on the tonoplast, HrNHX6 had no signal peptide and contained 13 transmembrane helical regions with numerous types of modification sites, such as N-glycosylation, protein kinase phosphorylation lation, myristoylation, amidation, and sugar transporter signature. The rusults of qRT-PCR analysis showed that the expression level of HrNHX6 gene was significantly different between different tissues under draught stress, and the expression level of HrNHX6 gene was significantly different between different drought stress treatment. Conclusion The drought resistance of H. rhamnoides was closely related to HrNHX6 as shown by the structure and expression under drought stress of the gene. The important role indicated a significant venue for improving the draught resistance of the medicinal plant. -
Key words:
- Hippophae rhamnoides /
- HrNHX6 /
- sequence analysis /
- protein structure /
- gene expression
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图 2 NHX6蛋白质系统发育树和保守基序
a：HrNHX6的系统发育树；b：HrNHX6的保守基序。
Figure 2. Phylogenetic tree and conserved motifs of NHX6 protein
a: Phylogenetic tree of HrNHX; b: Conserved motifs of HrNHX6.
图 3 HrNHX6与其他植物NHX6蛋白序列比对分析
A：氨氯吡嗪脒结合位点，B：Na+/H+交换泵。黑色背景表示相同氨基酸，红色、蓝色阴影分别表示75%、50%以上的保守性。
Figure 3. Alignment on amino acid residues of HrNHX6 and genes of other plants
A: Binding sites for ammonia chloride pyrazine microphones; B: Na+/H+ exchange pump; black background represents same amino acid; red and blue shadows, more than 75% and 50% conservatism, respectively.
图 6 干旱胁迫下HrNHX6基因表达模式
不同大写字母表示不同胁迫处理时间下差异极显著（P＜0.01），不同小写字母表示不同组织之间差异显著（P＜0.05）。
Figure 6. Expression of HrNHX6 under drought stress
Different capital letters indicate that the difference is extremely significant under different stress treatment time at P＜0.01; those with different lowercase letters, significant difference between different tissues at P＜0.05.
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