Department of Plant Pathology, University of California, Davis, CA, USA.
Plant Biotechnol J. 2011 Feb;9(2):205-15. doi: 10.1111/j.1467-7652.2010.00544.x.
Non-expresser of pathogenesis-related genes 1 (NPR1) is the master regulator of salicylic acid-mediated systemic acquired resistance. Over-expression of Arabidopsis NPR1 and rice NH1 (NPR1 homolog1)/OsNPR1 in rice results in enhanced resistance. While there are four rice NPR1 paralogs in the rice genome, none have been demonstrated to function in disease resistance. To study rice NPR1 paralog 3, we introduced constructs into rice and tested for effects on resistance to infection by Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial blight. While over-expression of NH3 using the maize ubiquitin-1 promoter failed to enhance resistance, introduction of an extra copy of NH3 driven by its own promoter (nNT-NH3) resulted in clear, enhanced resistance. Progeny analysis confirms that the enhanced resistance phenotype, measured by Xoo-induced lesion length, is associated with the NH3 transgene. Bacterial growth curve analysis indicates that bacterial population levels are reduced 10-fold in nNT-NH3 lines compared to control rice lines. The transgenic plants exhibit higher sensitivity to benzothiadiazole (BTH) and 2,6-dichloroisonicotinic acid (INA) treatment as measured by increased cell death. Expression analysis of pathogenesis-related (PR) genes showed that nNT-NH3 plants display greatly enhanced induction of PR genes only after treatment with BTH. Our study demonstrates an alternative method to employ a regulatory protein to enhance plant defence. This approach avoids using undesirable constitutive, high-level expression and may prove to be more practical for engineering resistance.
非表达致病性相关基因 1(NPR1)是水杨酸介导的系统获得性抗性的主要调节因子。拟南芥 NPR1 和水稻 NH1(NPR1 同源物 1)/OsNPR1 在水稻中的过表达导致抗性增强。虽然水稻基因组中有四个 NPR1 同源物,但没有一个被证明在抗病性中起作用。为了研究水稻 NPR1 同源物 3,我们将构建体引入水稻中,并测试其对感染稻黄单胞菌 pv.oryzae(Xoo)的抗性的影响,Xoo 是细菌性条斑病的病原体。虽然使用玉米泛素-1 启动子过表达 NH3 未能增强抗性,但引入由其自身启动子驱动的 NH3 的额外拷贝(nNT-NH3)导致明显的增强抗性。后代分析证实,用 Xoo 诱导的病变长度衡量的增强抗性表型与 NH3 转基因有关。细菌生长曲线分析表明,与对照水稻系相比,nNT-NH3 系中的细菌种群水平降低了 10 倍。与对照水稻系相比,转基因植物对苯并噻二唑(BTH)和 2,6-二氯异烟酸(INA)处理的敏感性更高,表现为细胞死亡增加。PR 基因表达分析表明,只有在用 BTH 处理后,nNT-NH3 植物才会显示出 PR 基因的极大增强诱导。我们的研究证明了一种利用调节蛋白增强植物防御的替代方法。这种方法避免了使用不理想的组成型、高水平表达,并可能被证明在工程抗性方面更实用。