Artemisia annua, a traditional Chinese medicinal plant, remains the only plant source for artemisinin production, yet few genes have been identified to be involved in both the response to biotic stresses, such as pathogens, and artemisinin biosynthesis. Here, we isolated and identified the WRKY transcription factor (TF) AaWRKY17, which could significantly increase the artemisinin content and resistance to Pseudomonas syringae in A. annua. Yeast one-hybrid (Y1H), dual-luciferase (dual-LUC), and electrophoretic mobility shift assay (EMSA) results showed that AaWRKY17 directly bound to the W-box motifs in the promoter region of the artemisinin biosynthetic pathway gene amorpha-4,11-diene synthase (ADS) and promoted its expression. Real-time quantitative PCR (RT-qPCR) analysis revealed that the transcript levels of two defense marker genes, Pathogenesis-Related 5 (PR5) and NDR1/HIN1-LIKE 10 (NHL10), were greatly increased in AaWRKY17-overexpressing transgenic A. annua plants. Additionally, overexpression of AaWRKY17 in A. annua resulted in decreased susceptibility to P. syringae. These results indicated that AaWRKY17 acted as a positive regulator in response to P. syringae infection. Together, our findings demonstrated that the novel WRKY transcription factor AaWRKY17 could potentially be used in transgenic breeding to improve the content of artemisinin and pathogen tolerance in A. annua.

中文翻译：

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AaWRKY17是青蒿素生物合成的正调节因子，参与青蒿对丁香假单胞菌的抗性


青蒿是一种传统的中药植物，仍然是生产青蒿素的唯一植物来源，但很少有基因被发现参与对生物胁迫（如病原体）的反应和青蒿素生物合成。在这里，我们分离并鉴定了WRKY转录因子（TF）AaWRKY17，它可以显着提高青蒿中的青蒿素含量和对丁香假单胞菌的抵抗力。酵母单杂交（Y1H）、双荧光素酶（dual-LUC）和电泳迁移率变动分析（EMSA）结果表明，AaWRKY17直接与青蒿素生物合成途径基因amorpha-4启动子区的W-box基序结合,11-二烯合酶（ADS）并促进其表达。实时定量 PCR (RT-qPCR) 分析显示，在过表达 AaWRKY17 的转基因 A.青蒿植物。此外，AaWRKY17 在青蒿中的过度表达导致对丁香假单胞菌的敏感性降低。这些结果表明 AaWRKY17 作为响应丁香假单胞菌感染的正调节因子。总之，我们的研究结果表明，新型 WRKY 转录因子 AaWRKY17 有可能用于转基因育种，以提高青蒿中青蒿素的含量和病原体耐受性。