Disease Resistant Crops Research Unit, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
Plant Biotechnol J. 2015 Aug;13(6):753-65. doi: 10.1111/pbi.12303. Epub 2014 Dec 9.
The rice transcription factor WRKY45 plays a central role in the salicylic acid signalling pathway and mediates chemical-induced resistance to multiple pathogens, including Magnaporthe oryzae and Xanthomonas oryzae pv. oryzae. Previously, we reported that rice transformants overexpressing WRKY45 driven by the maize ubiquitin promoter were strongly resistant to both pathogens; however, their growth and yield were negatively affected because of the trade-off between the two conflicting traits. Also, some unknown environmental factor(s) exacerbated this problem. Here, we report the development of transgenic rice lines resistant to both pathogens and with agronomic traits almost comparable to those of wild-type rice. This was achieved by optimizing the promoter driving WRKY45 expression. We isolated 16 constitutive promoters from rice genomic DNA and tested their ability to drive WRKY45 expression. Comparisons among different transformant lines showed that, overall, the strength of WRKY45 expression was positively correlated with disease resistance and negatively correlated with agronomic traits. We conducted field trials to evaluate the growth of transgenic and control lines. The agronomic traits of two lines expressing WRKY45 driven by the OsUbi7 promoter (PO sUbi7 lines) were nearly comparable to those of untransformed rice, and both lines were pathogen resistant. Interestingly, excessive WRKY45 expression rendered rice plants sensitive to low temperature and salinity, and stress sensitivity was correlated with the induction of defence genes by these stresses. These negative effects were barely observed in the PO sUbi7 lines. Moreover, their patterns of defence gene expression were similar to those in plants primed by chemical defence inducers.
水稻转录因子 WRKY45 在水杨酸信号通路中发挥核心作用,并介导对多种病原体(包括稻瘟病菌和稻黄单胞菌)的化学诱导抗性。此前,我们报道了过表达 WRKY45 的水稻转化体,其驱动子为玉米泛素启动子,对这两种病原体均具有很强的抗性;然而,由于两种相互冲突的性状之间的权衡,它们的生长和产量受到了负面影响。此外,一些未知的环境因素加剧了这个问题。在这里,我们报告了开发出对这两种病原体具有抗性且农艺性状几乎与野生型水稻相当的转基因水稻品系。这是通过优化驱动 WRKY45 表达的启动子来实现的。我们从水稻基因组 DNA 中分离出 16 个组成型启动子,并测试了它们驱动 WRKY45 表达的能力。不同转化体系之间的比较表明,总体而言,WRKY45 表达的强度与疾病抗性呈正相关,与农艺性状呈负相关。我们进行了田间试验来评估转基因和对照系的生长情况。表达 WRKY45 的 OsUbi7 启动子(PO sUbi7 系)驱动的两个系的农艺性状与未转化的水稻几乎相当,且这两个系均对病原体具有抗性。有趣的是,过量的 WRKY45 表达使水稻植株对低温和盐度敏感,而对这些胁迫的敏感性与这些胁迫诱导防御基因的表达有关。在 PO sUbi7 系中几乎没有观察到这些负面影响。此外,它们的防御基因表达模式与化学防御诱导剂引发的植物相似。