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

作　　者：关丽梅[1,2] 马伟华[1] 林拥军[1] 陈浩[1] 

机构地区：[1]华中农业大学作物遗传改良国家重点实验室,武汉430070 [2]江西省科学院,南昌330029

出　　处：《植物保护》2016年第1期19-25,共7页Plant Protection

基　　金：转基因生物新品种培育重大专项(2014ZX0810-002)

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 953bp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。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 mieroarray 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 OsOlg73940 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 fl-glucuronidase （GUS） report gene to construct a fusion gene, and then transformed in- to 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.

关 键 词：水稻 诱导型启动子 褐飞虱 抗虫 基因工程 

分 类 号：S435.112.3[农业科学—农业昆虫与害虫防治]