Xiao Jiaming, Wu Jin, Liu Dongdong, Li Xiawei, Liu Junlong, Su Xunwen, Wang Yonglin
School of Technology, Beijing Forestry University, Beijing 100083, China.
State Key Laboratory of Efficient Production of Forest Resources, Beijing Key Laboratory for Forest Pest Control, College of Forestry, Beijing Forestry University, Beijing 100083, China.
Plant Dis. 2025 Apr;109(4):862-874. doi: 10.1094/PDIS-06-24-1221-RE. Epub 2025 Apr 23.
Pine wilt disease caused by the pine wood nematode, , has profound implications for global forestry ecology. Conventional PCR methods need long operating time and are complicated to perform. The need for rapid and effective detection methodologies to curtail its dissemination and reduce pine felling has become more apparent. This study initially proposed the use of fluorescence recognition for the detection of pine wood nematode disease, accompanied by the development of a dedicated fluorescence detection system based on deep learning. This system possesses the capability to perform excitation, detection, as well as data analysis and transmission of test samples. In exploring fluorescence recognition methodologies, the efficacy of five conventional machine learning algorithms was juxtaposed with that of You Only Look Once version 5 and You Only Look Once version 10, both in the pre- and post-image processing stages. Moreover, enhancements were introduced to the You Only Look Once version 5 model. The network's aptitude for discerning features across varied scales and resolutions was bolstered through the integration of Res2Net. Meanwhile, a SimAM attention mechanism was incorporated into the backbone network, and the original PANet structure was replaced by the Bi-FPN within the Head network to amplify feature fusion capabilities. The enhanced YOLOv5 model demonstrates significant improvements, particularly in the recognition of large-size images, achieving an accuracy improvement of 39.98%. The research presents a novel detection system for pine nematode detection, capable of detecting samples with DNA concentrations as low as 1 fg/μl within 20 min. This system integrates detection instruments, laptops, cloud computing, and smartphones, holding tremendous potential for field application.
由松材线虫引起的松树萎蔫病对全球森林生态具有深远影响。传统的聚合酶链式反应(PCR)方法操作时间长且执行复杂。为遏制其传播并减少松树砍伐,对快速有效的检测方法的需求变得更加明显。本研究最初提出利用荧光识别来检测松材线虫病,并开发了基于深度学习的专用荧光检测系统。该系统具备对测试样品进行激发、检测以及数据分析和传输的能力。在探索荧光识别方法时,将五种传统机器学习算法的效果与You Only Look Once版本5(YOLOv5)和You Only Look Once版本10(YOLOv10)在图像预处理和后处理阶段的效果进行了并列比较。此外,对YOLOv5模型进行了改进。通过集成Res2Net增强了网络在不同尺度和分辨率下辨别特征的能力。同时,在主干网络中引入了SimAM注意力机制,并在头部网络中将原始的路径聚合网络(PANet)结构替换为双向特征金字塔网络(Bi-FPN)以增强特征融合能力。改进后的YOLOv5模型展示出显著提升,尤其是在大尺寸图像识别方面,准确率提高了39.98%。该研究提出了一种用于松材线虫检测的新型检测系统,能够在20分钟内检测出DNA浓度低至1 fg/μl的样品。该系统集成了检测仪器、笔记本电脑、云计算和智能手机,具有巨大的现场应用潜力。