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Agronomy 2019, 9(2), 45; https://doi.org/10.3390/agronomy9020045

Detection and Quantification of Fusarium spp. (F. oxysporum, F. verticillioides, F. graminearum) and Magnaporthiopsis maydis in Maize Using Real-Time PCR Targeting the ITS Region

1
ICAAM-Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Instituto de Investigação e Formação Avançada, Universidade de Évora, Polo da Mitra, Ap. 94, 7006-554 Évora, Portugal
2
Departamento de Biologia, ICAAM-Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Escola de Ciências e Tecnologia, Universidade de Évora, Polo da Mitra, Ap. 94, 7006-554 Évora, Portugal
3
Departamento de Fitotecnia, ICAAM-Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Escola de Ciências e Tecnologia, Universidade de Évora, Polo da Mitra, Ap. 94, 7006-554 Évora, Portugal
*
Author to whom correspondence should be addressed.
Received: 12 December 2018 / Revised: 10 January 2019 / Accepted: 18 January 2019 / Published: 22 January 2019
(This article belongs to the Special Issue Etiology and Control of Crop Diseases)
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Abstract

Fusarium spp. and Magnaporthiopsis maydis are soil-inhabiting fungi and respectively the causal agents of fusarium ear rot and late wilt, two important diseases that can affect maize, one of the most important cereal crops worldwide. Here, we present two sensitive real-time PCR TaqMan MGB (Minor Groove Binder) assays that detect and discriminate several Fusarium spp. (F. oxysporum, F. verticillioides, and F. graminearum) from M. maydis. The method is based on selective real-time qPCR amplification of the internal transcribed spacer (ITS) region and allows the quantification of the fungi. The applicability of this newly developed TaqMan methodology was demonstrated in a field experiment through the screening of potentially infected maize roots, revealing a high specificity and proving to be a suitable tool to ascertain Fusarium spp. and M. maydis infection in maize. Its high sensitivity makes it very efficient for the early diagnosis of the diseases and also for certification purposes. Thus, qPCR through the use of TaqMan probes is here proposed as a promising tool for specific identification and quantification of these soil-borne fungal pathogens known to cause disease on a large number of crops. View Full-Text
Keywords: TaqMan probes; fungi detection; ITS region; fusarium ear rot; late wilt; Zea mays L. TaqMan probes; fungi detection; ITS region; fusarium ear rot; late wilt; Zea mays L.
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Campos, M.D.; Patanita, M.; Campos, C.; Materatski, P.; Varanda, C.M.R.; Brito, I.; Félix, M.R. Detection and Quantification of Fusarium spp. (F. oxysporum, F. verticillioides, F. graminearum) and Magnaporthiopsis maydis in Maize Using Real-Time PCR Targeting the ITS Region. Agronomy 2019, 9, 45.

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