| 摘要: |
| 【目的】构建NaCl胁迫下柽柳根部cDNA文库，在盐胁迫下检测柽柳水通道蛋白（Aquaporins，AQPs）基因的表达，探讨AQPs与柽柳耐盐性的关系。【方法】以0.4 mol/L NaCl胁迫处理的刚毛柽柳（Tamarix hispida）根部组织为试材，分别构建未经盐胁迫及胁迫24，48 h的cDNA文库，并测定文库滴度。对从文库中获得的EST序列进行拼接，获得刚毛柽柳AQP（ThAQP）的单一序列，对其进行生物信息学分析。采用实时荧光定量PCR方法，研究ThAQP基因在盐胁迫后的刚毛柽柳根和叶中的表达情况。【结果】构建的NaCl胁迫0，24，48 h 3个文库的初始滴度分别为1.0×106，1.4×106和1.2×106 pfu/mL，插入片段的平均长度分别为0.8，0.9 和0.85 kb，平均重组率为98.2%。文库克隆随机测序获得了刚毛柽柳的7条ThAQP基因单一序列,这7条ThAQP基因分别与水通道蛋白的4个亚家族成员基因具有很高的同源性。在根和叶中，这7条ThAQP基因表达对盐胁迫的响应不同，在根部ThAQP1～ThAQP4相对表达量增多，而其他基因的相对表达量变化不明显；在叶部，ThAQP7在胁迫24 h时相对表达量出现小幅度上升，而其他6条ThAQP的相对表达量则明显下降。【结论】ThAQP基因可能与柽柳抵御盐胁迫有关。 |
| 关键词: 柽柳 cDNA文库 NaCl胁迫 水通道蛋白 |
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| 基金项目:国家自然科学基金项目（30972386）；中央高校基本科研业务费专项资金项目（DL09BA22） |
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| Construction of cDNA library of Taramix hispida and expression of ThAQP genes |
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| Abstract: |
| 【Objective】The study was to construct three cDNA libraries of root tissues of Taramix hispida,to observe the expression of aquaporins under NaCl stress,and to study mechanisms of salt tolerance of aquaporins from T.hispida.【Method】Three cDNA libraries were constructed from the root tissues of T.hispida of well watered and exposed to NaCl (0.4 mol/L) for 24 and 48 h,respectively (named as control,24 and 48 h library,respectively).And the titers of unamplified libraries were performed;EST sequences from three libraries were assembled,then seven AQP unigenes were obtained and analyzed using bioinformatics,and the expression patterns of seven AQPs under NaCl treatment were researched using real-time PCR.【Result】Average primary titer of the three libraries was 1.0×106,1.4×106 and 1.2×106 pfu/mL,respectively,with a rate of recombination about 98.2%.PCR detection revealed that their average insert-size was respectively 0.8,0.9 and 0.85 kb.The seven ThAQP genes were obtained through random selection of sequencing process,and sequence analysis showed that the seven genes were highly similar to the members of four subfamilies of AQPs respectively.The seven ThAQP genes had different expression patterns in roots and leaves under salt treatment.In root tissue,the expression of ThAQP1-ThAQP4 was up-regulated,but those of the others remained basically stable under salt treatment.In contrast,in leaf tissue,in addition to ThAQP7 gene whose expression was slightly up-regulated at 24 h of stress,the expression of other six ThAQP genes showed obviously down regulation.【Conclusion】The ThAQP may be involved in the adaptation of plant salt tolerance. |
| Key words: Taramix sp. cDNA library NaCl treatment aquaporin |
