| 关丽梅1,2，马伟华1，林拥军1，陈浩1*.一个水稻褐飞虱为害诱导型启动子的克隆[J].植物保护,2016,42(1):19-25. |
| 一个水稻褐飞虱为害诱导型启动子的克隆 |
| Cloning of a brown planthopper (Nilaparavata lugens) infection inducible promoter in rice |
| 投稿时间：2014-11-23 修订日期：2015-01-07 |
| DOI： |
| 中文关键词: 水稻 诱导性启动子 褐飞虱 抗虫 基因工程 |
| 英文关键词:Oryza sativa inducible promoter Nilaparavata lugens insect-resistance genetic engineering |
| 基金项目:转基因生物新品种培育重大专项(2014ZX0810-002) |
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| 中文摘要: |
| 目前组成型启动子在基因工程中的应用最为广泛, 然而驱动外源基因在各组织中持续恒定地表达可能引起植物发育迟缓等问题。诱导型启动子能够在特定条件下实现目的基因定时优势表达, 最大限度减少由于非必需蛋白在转基因植物内的积累对其造成的伤害。依据基因芯片数据并通过定量RT-PCR验证, 找到了一个受水稻褐飞虱(Nilaparavata lugens)为害诱导表达基因(LOC_Os01g73940)。用PCR技术从籼稻品种‘TN1’的基因组中获得Os01g73940上游1 953 bp的启动子片段, 命名为BPHIP。将BPHIP连接到带有β-glucuronidase (GUS) 报告基因的植物表达载体上, 通过农杆菌介导转化水稻品种‘中花11’。通过GUS组织化学染色法及定量RT-PCR检测证明, BPHIP是一个受褐飞虱为害和茉莉酸处理诱导上调的启动子。利用此类启动子在基因工程中可以减少对水稻生理方面产生的副作用, 对抗虫转基因水稻具有良好的应用价值。 |
| 英文摘要: |
| The constitutive promoters are widely applied in plant genetic engineering. However, it may cause growth retardation and other adverse effects due to the constant expression of exogenous genes in transgenic plants. The inducible promoters can control the expression of target genes more economically and minimize the possible harms to transgenic plants due to the accumulation of non-essential proteins. Based on the analysis of DNA microarray and quantitative RT-PCR, we found the expression of a rice gene (LOC_Os01g73940) could be induced by brown planthopper infection. The 1 953 bp upstream region of Os01g73940 gene was isolated from the indica rice variety ‘TN1’ by PCR, which was referred to as brown planthopper induced promoter (BPHIP). BPHIP was ligated with a β-glucuronidase (GUS) report gene to construct a fusion gene, and then transformed into a japonica rice variety ‘Zhonghua 11’. The results of GUS histochemical staining and quantitative RT-PCR confirmed that BPHIP is a promoter induced by brown planthopper infection and jasmonic acid treatment. This inducible promoter may mitigate the possible adverse effects of constitutive promoters in genetic engineering and has enormous application potential in insect-resistant transgenic breeding. |
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