SBIR Phase I: Bioluminescent Reporter Phage for the Rapid and Specific Detection of the Causative Agent of Bacterial Blight

这个小企业创新研究第一阶段项目将产生一个“光标记”的报告噬菌体，可以赋予植物病原体假单胞菌pv.泽泻黑腐病菌泽泻是细菌性枯萎病的病原体，细菌性枯萎病是一种危害十字花科蔬菜的传染性和破坏性疾病。由于疾病的严重性使作物无法销售，因此必须能够正确识别无症状和患病植物以及接种库中的药剂。第一阶段的目标是开发一种生物发光的报告噬菌体，用于检测P.泽泻目的是产生特异性的P. Alisalensis光标记的报道噬菌体，然后进行可行性研究以证明光标记的报道基因可以用作P. alisalingae pv.泽泻检测系统该研究预计将产生一个可行的重组报告噬菌体，可以快速，灵敏，特异性地赋予生物发光信号响应，从而检测P.该项目的更广泛的影响/商业潜力是P. alisalensis诊断可能有助于减少对美国各地商业种植的蔬菜作物的损害。症状首先出现在较低的叶子上，表现为小的水浸泡的绒毛。这些软组织或病变扩大并被亮黄色边界包围，最终合并形成大面积坏死区。细菌性枯萎病感染可导致60%的商业田地的作物受损。黑腐病菌泽泻具有广泛的宿主范围，危害单子叶植物（加州雀麦、燕麦）、十字花科植物（花椰菜、花椰菜、花椰菜拉布、布鲁塞尔子甘蓝、卷心菜、萝卜、芝麻菜）和番茄。在美国，仅甘蓝类蔬菜（例如花椰菜、花椰菜、卷心菜和布鲁塞尔子甘蓝）的价值估计为每年13亿美元。因此，由于细菌性枯萎病造成的经济损失可能是严重的。然而，迄今为止，没有市售的或批准的P.泽泻检测方法。本研究的长期目标是开发一种新的诊断试剂盒，用于检测这种农业上重要的植物病原体。该研究也为其他农业重要细菌病原菌生物传感器的开发提供了基础技术。

This Small Business Innovation Research Phase I project will generate a "light-tagged" reporter phage that can confer a bioluminescent signal to the phytopathogen Pseudomonas syringae pv. alisalensis. P. syringae pv. alisalensis is the causative agent of bacterial blight, a contagious and damaging disease afflicting cruciferous vegetables. Since the severity of the disease renders the crop unmarketable, it is essential to be able to correctly identify the agent on asymptomatic and diseased plants, and from inoculum reservoirs. The Phase I goal is to develop a bioluminescent reporter phage for the detection of P. syringae pv. alisalensis. The objective is to generate a specific P. syringae pv. alisalensis light-tagged reporter phage and then perform feasibility studies to demonstrate that the light-tagged reporter can be used as a P. syringae pv. alisalensis detection system. The research is expected to generate a viable recombinant reporter phage that can rapidly, sensitively, and specifically confer a bioluminescent signal response, and hence detect P. syringae pv. alisalensis.The broader impact/commercial potential of this project is that the P. syringae pv. alisalensis diagnostic may help to reduce damage to commercially grown vegetable crops throughout the U.S. Symptoms first appear as small water-soaked flecks on the lower foliage. These flecks or lesions expand and become surrounded by bright yellow borders that eventually coalesce to form large necrotic areas. Bacterial blight infestations can result in crop damage to 60% of the commercial field. P. syringae pv. alisalensis has a broad host range afflicting monocots (California brome, oats), crucifers (cauliflower, broccoli, broccoli raab, brussels sprouts, cabbage, radish, arugula) and tomato. The value of Brassica oleracea vegetables alone (e.g. broccoli, cauliflower, cabbage and brussels sprouts) is estimated at $1.3 billion annually in the U.S. Consequently, the economic losses due to bacterial blight are potentially severe. However, to date, there are no commercially available or approved P. syringae pv. alisalensis detection methodologies. The long-term goal of this research is to develop a novel diagnostic kit for the detection of this agriculturally important phytopathogen. The research also may provide the foundation technology for the development of biosensors for other agriculturally important bacterial pathogens.