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一种基于聚合酶链式反应(PCR)的方法,用于检测银皮病感染的块茎组织和土壤中的马铃薯病原菌——茄丝核菌。

A PCR-based method for detection of potato pathogen, Helminthosporium solani, in silver scurf infected tuber tissue and soils.

作者信息

Errampalli D, Saunders J, Cullen D

机构信息

Crops and Livestock Research Centre, Agriculture and Agri-Food Canada, P.O. Box 6000, 4902 Victoria Ave. North, Vineland Station, ON, Canada L0R 2E0.

出版信息

J Microbiol Methods. 2001 Feb 1;44(1):59-68. doi: 10.1016/s0167-7012(00)00240-2.

Abstract

Silver scurf caused by Helminthosporium solani causes significant economic losses in table stock, seed and processing potatoes. Specific polymerase chain reaction (PCR) primers, Hs1F1/Hs2R1, from H. solani were used for the amplification of a 447-bp product from 20 tissue samples and 54 single spore H. solani isolates, from eastern Canada (27 isolates), western Canada (13 isolates) and North Dakota in USA (14 isolates), but not from other potato fungal pathogens. In addition to PCR analysis, all 54 isolates were studied using conventional detection methods, visual disease symptoms and/or colony morphology and microscopic examination of the morphology of conidiophores and conidia. The PCR assay successfully detected H. solani and the PCR results correlated well with assessments based on conventional techniques. The detection of H. solani by PCR (1 day) is rapid and offers an alternative to the time consuming conventional diagnostic techniques (4-5 weeks). Nested PCR assay was necessary for the detection of H. solani in soils and thus can provide a sensitive technique to study the epidemiology of silver scurf in soils.

摘要

由茄丝核菌引起的银皮病会给鲜食马铃薯、种薯和加工型马铃薯造成重大经济损失。来自茄丝核菌的特异性聚合酶链反应(PCR)引物Hs1F1/Hs2R1,用于从20个组织样本和54个单孢茄丝核菌分离株中扩增出一个447 bp的产物,这些分离株来自加拿大东部(27个分离株)、加拿大西部(13个分离株)和美国北达科他州(14个分离株),但未从其他马铃薯真菌病原体中扩增出该产物。除了PCR分析外,还使用传统检测方法、肉眼观察病害症状和/或菌落形态以及对分生孢子梗和分生孢子形态进行显微镜检查,对所有54个分离株进行了研究。PCR检测成功检测到了茄丝核菌,且PCR结果与基于传统技术的评估结果高度相关。通过PCR检测茄丝核菌(1天)速度很快,为耗时的传统诊断技术(4 - 5周)提供了一种替代方法。巢式PCR检测对于检测土壤中的茄丝核菌是必要的,因此可以提供一种敏感技术来研究土壤中银皮病的流行病学。

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