Tegli Stefania, Biancalani Carola, Ignatov Aleksandr N, Osdaghi Ebrahim
Laboratorio di Patologia Vegetale Molecolare, Dipartimento di Scienze e Tecnologie Agrarie, Alimentari Ambientali e Forestali (DAGRI), Università degli Studi di Firenze, Via della Lastruccia 10, 50019 Sesto Fiorentino (Firenze), Italy.
Agrarian and Technological Institute (ATI), Peoples' Friendship University of Russia, Miklukho-Maklaya str.8, 117198 Moscow, Russia.
Microorganisms. 2020 Oct 31;8(11):1705. doi: 10.3390/microorganisms8111705.
pv. () is a Gram-positive phytopathogenic bacterium attacking leguminous crops and causing systemic diseases such as the bacterial wilt of beans and bacterial spot of soybeans. Since the early 20th century, is reported to be present in North America, where it still causes high economic losses. Currently, is an emerging plant pathogen, rapidly spreading worldwide and occurring in many bean-producing countries. Infected seeds are the main dissemination pathway for , both over short and long distances. remains viable in the seeds for long times, even in field conditions. According to the most recent EU legislation, is included among the quarantine pests not known to occur in the Union territory, and for which the phytosanitary inspection consists mainly of the visual examination of imported bean seeds. The seedborne nature of combined with the globalization of trades urgently call for the implementation of a highly specific diagnostic test for , to be routinely and easily used at the official ports of entry and into the fields. This paper reports the development of a LAMP (Loop-Mediated Isothermal Amplification) specific for , that allows the detection of in infected seeds, both by fluorescence and visual monitoring, after 30 min of reaction and with a detection limit at around 4 fg/μL of pure genomic DNA.
菜豆晕疫病菌()是一种革兰氏阳性植物病原菌,侵袭豆科作物并引发系统性病害,如菜豆细菌性萎蔫病和大豆细菌性斑点病。自20世纪初以来,据报道它存在于北美,在那里它仍然造成高额经济损失。目前,它是一种新兴的植物病原菌,在全球迅速传播,在许多豆类生产国均有发生。受感染的种子是菜豆晕疫病菌短距离和长距离传播的主要途径。即使在田间条件下,菜豆晕疫病菌在种子中也能长时间存活。根据欧盟最新法规,菜豆晕疫病菌被列入欧盟境内未知的检疫性有害生物名单,对其植物检疫检查主要包括对进口豆类种子的目视检查。菜豆晕疫病菌的种传特性以及贸易全球化迫切要求实施一种针对菜豆晕疫病菌的高度特异性诊断测试,以便在官方入境口岸和田间常规且简便地使用。本文报道了一种针对菜豆晕疫病菌的环介导等温扩增(LAMP)方法的开发,该方法能够在反应30分钟后,通过荧光和目视监测检测受感染种子中的菜豆晕疫病菌,检测限约为4 fg/μL纯病菌基因组DNA。