Screening by a 3-primer PCR of North American PVYN isolates for European-type members of the tuber necrosis-inducing PVYNTN subgroup

R. Singh, Munshi Singh, J. Mcdonald
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引用次数: 22

Abstract

Oligonucleotide primers for a 3-primer polymerase chain reaction (PCR) were optimized by adjusting their Tm values to identify European-type members of the potato tuber necrotic ringspot disease (PTNRD) inducing PVYNTN sub-group of PVYN. Fiftyseven North American isolates of PVYN were screened using this assay and one, originating from garden potato tubers in Ontario, was found to be positive. This isolate was confirmed to induce the diagnostic PTNRD, using a panel of 20 potato cultivars. AC Chaleur, AC Novachip, Century Russet, Cherokee, and Frontier Russet reacted severely and developed PTNRD either at the time of harvest or during storage. About 800 tubers collected from New Brunswick and Prince Edward Island fields, from plants exhibiting mosaic and leafroll symptoms were also screened for various strains of PVY using PCR primers. Results indicate that primer pairs developed are specific for PVY spp., PVYN, and PVYNTN.
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用3引物PCR方法筛选北美PVYN分离物中引起块茎坏死的PVYNTN亚群的欧洲型成员
通过调整Tm值对3引物聚合酶链反应(PCR)的寡核苷酸引物进行优化,以鉴定马铃薯块茎坏死环斑病(PTNRD)诱导PVYNTN亚群的欧洲型成员。用这种方法筛选了57株PVYN北美分离株,其中一株来自安大略省的花园马铃薯块茎,被发现呈阳性。通过20个马铃薯品种的鉴定,证实该分离物可诱导诊断性PTNRD。AC Chaleur, AC Novachip, Century Russet, Cherokee和Frontier Russet在收获或储存期间反应严重并发展PTNRD。从新不伦瑞克省和爱德华王子岛地区采集的约800块块茎,采用PCR引物对具有花叶病和叶卷病症状的植物进行了PVY菌株的筛选。结果表明,该引物对PVY spp.、PVYN和PVYNTN具有特异性。
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