基于dna的无症状进口种薯中茄枯菌检测方法

A. Perera, O. Weerasena, P. Dasanayaka, M. Subhashini
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摘要

在斯里兰卡种植的蔬菜中,马铃薯是一种重要的经济作物,主要依赖于健康的马铃薯种子。每年约40%的种子马铃薯需求是通过从荷兰、美国、德国和法国进口种子马铃薯来满足的。种子马铃薯的进口使植物致病病虫害有可能进入斯里兰卡。青枯病是马铃薯最具破坏性的病害之一。引起马铃薯青枯病的Ralstonia solanacearum被认为是斯里兰卡重要的检疫意义植物病原体。现有的常规方法如培养法、生化法等检测进口种薯中的茄红病菌耗时长、费力且灵敏度低。虽然免疫诊断方法快速,但灵敏度不足以检测无症状或潜伏感染的种子马铃薯中的细菌。本研究采用基于dna的检测方法对斯里兰卡进口种子马铃薯进行筛选,30份检测样品中有5份(17%)呈番茄红病菌阳性。对进口种薯样品进行进一步研究,发现在进口种薯样品中检测到亚洲种型I和美洲种型II。在从美国和法国进口的种子马铃薯中检测到种型II(种3/生物变种2),在从荷兰进口的种子马铃薯中检测到种型I和种型II(种3/生物变种2),荷兰的大部分种子马铃薯进口到该国。应严格采取检疫措施,以避免II种型3种/ 2生物变种菌株在国内传播和建立,因为它是极具破坏性的马铃薯病原体,在斯里兰卡高海拔地区的分布有限。此外,基于dna的方法可用于鉴定病原菌,以避免番茄红霉的引进或进入该国,以改善斯里兰卡的马铃薯种植。关键词:种薯,青枯病,番茄枯萎病,检疫害虫,rsol_fllic
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Detection of Ralstonia solanacearum in Asymptomatic Imported Seed Potato using a DNA-based Method
Potato is an economically important crop among vegetables grown in Sri Lanka that mainly relies on healthy seed potatoes. About 40% of the annual seed potato requirement is fulfilled by the import of seed potatoes from Netherland, USA, Germany and France. Import of seed potatoes makes possibilities to enter plant pathogenic pests and diseases to Sri Lanka. Bacterial wilt is one of the most destructive diseases of potato. Ralstonia solanacearum, which causes bacterial wilt of potato, is considered as an important quarantine significant plant pathogen in Sri Lanka. The currently available conventional methods such as culture methods, biochemical methods are time consuming, very laborious and not sensitive for the detection of R. solanacearum in imported seed potatoes. Although immunodiagnostic methods are rapid, the sensitivity is not enough to detect the bacterium in asymptomatic or latently infected seed potatoes. In this study, a DNA-based detection method was applied to screen seed potatoes imported into Sri Lanka and 5 out of 30 tested samples (17%) were positive for R. solanacearum. The seed potato samples detected as infected with R. solanacearum were further studied and it revealed that the Asian phylotype I and the American phylotype II were detected from seed potato samples imported to the country. Phylotype II (Race 3/biovar 2) was detected in seed potatoes imported from USA and France and both phylotype I and phylotype II (Race 3/biovar 2) were detected in seed potatoes imported from Netherland from where majority of seed potatoes are imported into the country. The quarantine measures should be strictly followed to avoid the spread and establishment of phylotype II, Race 3/biovar 2 strains within the country, as it is the extremely destructive potato pathogen which have a restricted distribution in higher elevations of Sri Lanka. Further, the DNA-based method can be used to identify the pathogen to avoid the introduction or entry of R. solanacearum into the country for the betterment of potato cultivation in Sri Lanka. KEYWORDS: Seed potato, Bacterial wilt, Ralstonia solanacearum, Quarantine pest, Rsol_fliC
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