Rapid detection and quantification of Alternaria solani in tomato
Rapid detection and quantification of Alternaria solani in tomato
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DOI：
10.1016/j.scienta.2012.12.026
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发表时间：
2013-02-28
影响因子：
4.3
通讯作者：
Srivastava, Alok Kumar
中科院分区：
文献类型：
--
作者：
Kumar, Sudheer;Singh, Ruchi;Srivastava, Alok Kumar
番茄早疫病的早期诊断是防治该病的关键。为了监测和量化这种真菌的发生，开发了一种基于实时PCR的诊断工具。根据β-微管蛋白基因设计特异性引物，用A.从不同地理来源获得的Solani分离物和相关物种。引物对A. Solani，因为从13个其它密切相关的分类群中没有观察到扩增信号。引物AS 1（5 ′-GCTCCCACTCCTTCCGCGC-3 ′）和AS 2（5 ′-GGAGGIGGAGTTACCGACAA-3 ′）从所有的A. Solani分离物。该方法的最低检测限为0.5 pg，并在人工感染的番茄幼苗上进行了验证，能够检测到A.接种后20天的Solani。据我们所知，这是第一个报告的定量真实的时间PCR检测方法的快速和特异性检测A。solani中，用从β-微管蛋白区域设计的引物组。(C)2013 Elsevier B. V.保留所有权利。
Early detection of infection is very crucial for efficient management of early blight of tomato caused by Alternaria solani. To monitor and quantify the occurrence of this fungus, a diagnostic tool based on real-time PCR was developed. Specific-primers were designed from beta-tubulin gene and specificity was checked with A. solani isolates and related species obtained from different geographical origins. The primers were highly specific for A. solani, as no amplification signal was observed from thirteen other closely related taxa. Primer set, AS1 (5'-GCTCCCACTCCTTCCGCGC-3') and AS2 (5'-GGAGGIGGAGTTACCGACAA-3') amplified a specific amplicon of 289 bp from all A. solani isolates. The lowest detection limit of the real-time PCR assay with designed primer set (AS1 and AS2) was 0.5 pg. The assay was also successfully validated on artificially infested tomato seedlings and able to detect A. solani up to 20 days post inoculation. To the best of our knowledge, this is the first report of a quantitative real time PCR detection method for rapid and specific detection of A. solani with a primer set designed from beta-tubulin region. (C) 2013 Elsevier B.V. All rights reserved.