沙蒿AQP基因结构预测及干旱胁迫下的时空表达模式研究
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引用本文:张 丹,胡晓莹,张 莉,马海龙,王 超,马玉花.沙蒿AQP基因结构预测及干旱胁迫下的时空表达模式研究[J].西北植物学报,2019,39(9):1543~1550
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张 丹,胡晓莹,张 莉,马海龙,王 超,马玉花* (青海大学 农牧学院西宁 810016) 
基金项目:国家自然科学基金(31660071);
中文摘要:为探究水通道蛋白(AQP)在沙蒿响应干旱胁迫中的作用机制,该研究以青海省柴达木盆地沙蒿为试验材料,采用RACE技术对其AQP基因进行扩增,获得沙蒿AQP全长克隆并对AQP蛋白进行结构预测和分析;采用qRT PCR对沙蒿AQP基因在不同程度干旱胁迫以及不同组织部位的表达模式进行分析。结果表明:(1)成功克隆获得沙蒿AQP基因长746 bp的片段1和长534 bp的片段2,经拼接后得到全长cDNA序列,沙蒿AQP基因总长为864 bp。(2)亚细胞定位表明沙蒿AQP基因定位于细胞膜上;同源比对显示沙蒿与向日葵、莴苣、橡胶树等植物的AQP基因具有较高的相似性;结构预测表明AQP蛋白含6个跨膜螺旋结构且亲水性较弱,α螺旋和无规则卷曲为AQP蛋白二级结构的主要构成元件。(3)qRT PCR分析表明,沙蒿AQP基因随着干旱胁迫的加重呈现有规律的变化,根、茎、叶中表达均上调,且叶中AQP基因表达量上调幅度最大。研究表明沙蒿AQP基因结构特征及其表达模式都是沙蒿对干旱胁迫的一种适应。
中文关键词:沙蒿  水道通蛋白  结构预测  干旱胁迫  表达模式
 
Prediction of AQP Gene Structure of Artemisia ordosica and Spatiotemporal Expression Pattern under Drought Stress
Abstract:In order to explore the mechanism of aquaporin (AQP) in response to drought stress in Artemisia ordosica, we amplified the AQP gene by RACE using A. ordosica in the Qaidam Basin of Qinghai Province. The full length gene of AQP was obtained and the structure of AQP protein was predicted and analyzed. At the same time, qRT PCR was used to analyze the expression patterns of AQP gene of A. ordosica in different degrees of drought stress and different tissues. The results showed that: (1) the 746 bp fragment 1 and the 534 bp fragment 2 of AQP gene were successfully cloned. The full length cDNA sequence was obtained after splicing. The total length of AQP gene of A. ordosica was 864 bp. (2) Subcellular localization indicated that the AQP gene of A. ordosica was located on the cell membrane. The homology alignment showed that the A. ordosica had high similarity with the AQP genes of plants such as Helianthus annuus, Lactuca sativa and Hevea brasiliensis. Structural prediction indicated that the AQP protein contained six transmembrane helix structures and was weak in hydrophilicity. In the protein secondary structure, α helix and random coil were the main constituent elements. (3) According to qRT PCR analysis, the result showed that AQP gene of A. ordosica showed regular changes with the increase of drought stress. The expression of AQP gene was up regulated in roots, stems and leaves, the highest was in leaves. So the above results indicated that the AQP gene had distinct structural characteristics and expression patterns to adapt to drought stress. This study provided the foundation for further characterizing the mechanism of A. ordosica drought resistance.
keywords:Artemisia ordosica  AQP  structure prediction  drought stress  expression pattern
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