| 摘要: |
| 【目的】根据前期基因家族鉴定分析得到OfWRKY120，对该基因盐胁迫响应的功能进行分析，为桂花盐胁迫下的调控机制研究奠定基础。【方法】利用生物信息学方法对OfWRKY120保守结构域、系统进化进行分析，并采用qRT-PCR技术研究盐胁迫下OfWRKY120在桂花叶片中的相对表达量。构建OfWRKY120超表达载体，使用亚细胞定位、酵母自激活检测等探究其特性，将该基因瞬时转化本氏烟草后，对烟草进行盐处理，利用DAB、NBT染色和相关生理指标的测定，解析瞬时转化OfWRKY120对本氏烟草耐盐性的影响。【结果】OfWRKY120基因全长为1 422 bp，编码474个氨基酸，存在WRKY superfamily保守结构域。盐胁迫能够激活桂花叶片中OfWRKY120的表达，在72 h时相对表达量最高，与对照有显著差异。亚细胞定位结果表明，OfWRKY120定位于细胞核。酵母自激活结果显示OfWRKY120无自激活活性。盐胁迫后，OfWRKY120瞬时过表达植株的超氧阴离子(O-2·)和脯氨酸含量显著高于空载对照，且DAB和NBT染色表现出更深的颜色。本氏烟草中的qRT-PCR结果显示，OfWRKY120显著降低了NbCAT的表达，而显著提高了NbP5CS1、NbP5CS2和NbP5CR的表达。【结论】OfWRKY120过表达增加了盐胁迫下本氏烟草中活性氧（ROS）的含量，导致植物对盐胁迫的敏感性提高。 |
| 关键词: 桂花 OfWRKY120基因 盐胁迫 抗逆育种 |
| DOI： |
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| 基金项目:国家自然科学基金面上项目（32071828）；江苏高校优势学科建设工程资助项目 |
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| Identification and salt stress response of OfWRKY120 gene in Osmanthus fragrans |
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ZAI Zhouying,WANG Liu,DING Huifen,et al
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| Abstract: |
| 【Objective】According to previous gene family identification analysis,a WRKY gene of Osmanthus fragrans named OfWRKY120 was obtained and the response to salt stress was analyzed to provide basis for further analysis of the regulatory mechanism under salt stress.【Method】The conserved domain and phylogenetics of OfWRKY120 were analyzed by bioinformatics methods,and qRT-PCR was used to analyze the relative expression of OfWRKY120 in O. fragrans leaves under salt stress.A super-expression vector of OfWRKY120 was constructed and subcellular localization and autoactivation detection were carried out to explore the characteristics of the gene.The effect of OfWRKY120 on salt tolerance was analyzed by treating Nicotiana benthamiana with salt,determining relevant physiological indexes,and performing DAB & NBT staining.【Result】The OfWRKY120 gene was 1 422 bp in length,encoding 474 amino acids with a conserved WRKY superfamily.Salt stress activated the expression of OfWRKY120 in O. fragrans,and the relative expression was the highest at 72 h with significant difference from the control. Subcellular localization results showed that OfWRKY120 was located in the nucleus.Yeast autoactivation results showed no autoactivation activity of OfWRKY120.After salt treatment,the contents of superoxide anion (O-2·) and proline in transgenic leaves with OfWRKY120 overexpression were significantly higher than those of empty vector,and DAB and NBT staining showed darker colors.The qRT-PCR results of N.benthamiana showed that OfWRKY120 overexpression significantly reduced NbCAT expression,while significantly increased NbP5CS1,NbP5CS2 and NbP5CR expression.【Conclusion】OfWRKY120 increased contents of reactive oxygen species（ROS）in N.benthamiana under salt stress and improved sensitivity of plants to salt stress. |
| Key words: Osmanthus fragrans OfWRKY120 gene salt stress breeding for stress tolerance |
