抗性稗草（Echinochloa crus-galli）种群发展将严重威胁我国水稻的安全生产，前期研究发现乙烯应答转录因子ERF基因及其蛋白表达量在稗草多抗性生物型与敏感型间存在显著性差异，初步表明ERF在稗草抵抗除草剂胁迫中发挥着重要的功能。申请者拟以稗草多抗性（兼抗二氯喹啉酸和五氟磺草胺）、单抗性（二氯喹啉酸或五氟磺草胺）和敏感生物型为研究材料，首先克隆稗草的ERF基因，并确定其蛋白功能域、系统发生树及其所属的亚家族类。在此基础上，利用实时荧光定量PCR技术分析供试稗草ERF基因在供试药剂胁迫下的时空表达特性差异；并采用原核表达、亚细胞定位和转录激活研究方法探讨不同来源ERF蛋白的生物学功能差异性；通过转基因技术验证ERF在转基因烟草中的表达功能，并捕获ERF参与的信号转导相关蛋白，最终揭示ERF转录因子调控杂草产生抗药性的新机制，为挖掘农田杂草绿色治理的新途径提供理论依据。

The evolution of barnyardgrass (Echinochloa crus-galli) resistant to multiple herbicides will seriously threat rice production in China. In our previous work, we found that gene and protein expressions of ethylene-responsive factor (ERF) had significant differences between multiple-resistant and susceptible biotypes of E. crus-galli and preliminary showed that ERF played an important function in herbicide resistance. Therefore, using multi-herbicide resistant (quinclorac and penoxsulam), single herbicide resistant (quinclorac or penoxsulam) and susceptible barnyardgrass biotypes as materials, the objectives of this study will be to (1) identify ERF genes from E. crus-galli and determine gene function domain and subfamily class using phylogenetic tree analysis; (2) compare temporal and spatial variation in ERF gene expression under herbicide stress in E. crus-galli using quantitative real-time PCR method; (3) determine ERF biological functions through prokaryotic expression, subcellular localization and transcriptional activation analysis; (4) demonstrate ERF gene functions in transgenic tobacco and unravel the responsive genes related to hormone signaling pathways under the control of ERF transcription factor. In general, these above results will explore the new mechanism of barnyardgrass resistant to multiple-herbicide and will provide a theoretical basis for making a new strategy to control weeds safety and ensure crops high quality in farmland systems.