Comparative genomics provide a rapid detection of Fusarium oxysporum f. sp. conglutinans

doi:10.1016/S2095-3119(15)61237-0

Journal of Integrative Agriculture ›› 2016, Vol. 15 ›› Issue (4): 822-831.DOI: 10.1016/S2095-3119(15)61237-0

Comparative genomics provide a rapid detection of Fusarium oxysporum f. sp. conglutinans

LING Jian*, ZHANG Ji-xiang*, ZENG Feng, CAO Yue-xia, XIE Bing-yan, YANG Yu-hong

Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China

收稿日期:2015-06-05 出版日期:2016-04-01 发布日期:2016-04-07
通讯作者: YANG Yu-hong, Tel: +86-10-82109545, E-mail: yangyuhong@caas.cn
作者简介:LING Jian, E-mail: lingjian2005@126.com
基金资助:
This research was supported in part by the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, P.R.China, the National Natural Science Foundation of China (31571962, 31272003), the National Key Technology R&D Program of China (2012BAD19B06), the Special Fund for Agro-Scientific Research in the Public Interest, China (200903049-04), the National Staple Vegetable Industry Technology System Construction Project, China (CARS-25-B-01), and the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences.

Comparative genomics provide a rapid detection of Fusarium oxysporum f. sp. conglutinans

LING Jian*, ZHANG Ji-xiang*, ZENG Feng, CAO Yue-xia, XIE Bing-yan, YANG Yu-hong

Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China

Received:2015-06-05 Online:2016-04-01 Published:2016-04-07
Contact: YANG Yu-hong, Tel: +86-10-82109545, E-mail: yangyuhong@caas.cn
About author:LING Jian, E-mail: lingjian2005@126.com
Supported by:
This research was supported in part by the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, P.R.China, the National Natural Science Foundation of China (31571962, 31272003), the National Key Technology R&D Program of China (2012BAD19B06), the Special Fund for Agro-Scientific Research in the Public Interest, China (200903049-04), the National Staple Vegetable Industry Technology System Construction Project, China (CARS-25-B-01), and the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences.

摘要/Abstract

摘要： Fusarium oxysporum f. sp. conglutinans (Foc) is the causal agent of Fusarium wilt disease of Brassica oleracea. A rapid, accurate, and reliable method to detect and identify plant pathogens is vitally important to integrated disease management. In this study, using a comparative genome analysis among Fusarium oxysporum (Fo), we developed a Foc-specific primer set (Focs-1/Focs-2) and established a multiplex-PCR assay. In the assay, the Focs-1/Focs-2 and universal primers for Fusarium species (W106R/F106S) could be used to detect Foc isolates in a single PCR reaction. With the optimized PCR parameters, the multiplex-PCR assay showed a high specificity for detecting Foc and was very sensitive to detect as little as 100 pg of pure Foc genomic DNA or 1 000 spores in 1 g of twice-autoclaved soil. We also demonstrated that Foc isolates were easily detected from infected plant tissues, as well as from natural field soils, using the multiplex-PCR assay. To our knowledge, this is a first report on detection Fo by comparative genomic method.

关键词: Fusarium oxysporum , wilt disease , cabbage , multiplex-PCR , molecular detection , comparative genomics

Abstract: Fusarium oxysporum f. sp. conglutinans (Foc) is the causal agent of Fusarium wilt disease of Brassica oleracea. A rapid, accurate, and reliable method to detect and identify plant pathogens is vitally important to integrated disease management. In this study, using a comparative genome analysis among Fusarium oxysporum (Fo), we developed a Foc-specific primer set (Focs-1/Focs-2) and established a multiplex-PCR assay. In the assay, the Focs-1/Focs-2 and universal primers for Fusarium species (W106R/F106S) could be used to detect Foc isolates in a single PCR reaction. With the optimized PCR parameters, the multiplex-PCR assay showed a high specificity for detecting Foc and was very sensitive to detect as little as 100 pg of pure Foc genomic DNA or 1 000 spores in 1 g of twice-autoclaved soil. We also demonstrated that Foc isolates were easily detected from infected plant tissues, as well as from natural field soils, using the multiplex-PCR assay. To our knowledge, this is a first report on detection Fo by comparative genomic method.

Key words: Fusarium oxysporum , wilt disease , cabbage , multiplex-PCR , molecular detection , comparative genomics

LING Jian, ZHANG Ji-xiang, ZENG Feng, CAO Yue-xia, XIE Bing-yan, YANG Yu-hong. Comparative genomics provide a rapid detection of Fusarium oxysporum f. sp. conglutinans[J]. Journal of Integrative Agriculture, 2016, 15(4): 822-831.

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Comparative genomics provide a rapid detection of Fusarium oxysporum f. sp. conglutinans

Comparative genomics provide a rapid detection of Fusarium oxysporum f. sp. conglutinans

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