PCR-Based Detection for the Quarantine Fungus <i>Colletotrichum kahawae</i>, a Biosecurity Threat to the Coffee (<i>Coffea arabica</i>) Industry Worldwide.

Ferrucho, Rosa Lilia; Marín-Ramírez, Gustavo Adolfo; Ochoa-Corona, Francisco; Ángel C, Carlos Ariel

PCR-Based Detection for the Quarantine Fungus Colletotrichum kahawae, a Biosecurity Threat to the Coffee (Coffea arabica) Industry Worldwide.

Ferrucho, Rosa Lilia; Marín-Ramírez, Gustavo Adolfo; Ochoa-Corona, Francisco; Ángel C, Carlos Ariel.

Afiliação

Ferrucho RL; Discipline of Plant Pathology, Colombian National Coffee Research Center (Cenicafé) - Colombian Coffee Growers Federation (CCGF), Manizales, Caldas 170009, Colombia.
Marín-Ramírez GA; Discipline of Plant Pathology, Colombian National Coffee Research Center (Cenicafé) - Colombian Coffee Growers Federation (CCGF), Manizales, Caldas 170009, Colombia.
Ochoa-Corona F; Institute for Biosecurity and Microbial Forensics, Oklahoma State University, Stillwater, OK, U.S.A.
Ángel C CA; Discipline of Plant Pathology, Colombian National Coffee Research Center (Cenicafé) - Colombian Coffee Growers Federation (CCGF), Manizales, Caldas 170009, Colombia.

Plant Dis ; 108(9): 2615-2624, 2024 Sep.

Article em En | MEDLINE | ID: mdl-38616394

ABSTRACT

ABSTRACT

Coffee berry disease is caused by Colletotrichum kahawae, a quarantine fungus still absent from most coffee-producing countries. Given the potential adverse effects on coffee berry production, it is a severe worldwide threat to farmers and industry. Current biosecurity management focuses on exclusion by applying quarantine measures, including the certification of coffee plants and their products. However, methods for detecting C. kahawae by National Plant Protection Organization (NPPO) laboratories still need approval. This research aims to functionally demonstrate, standardize, and validate a method for detecting and discriminating C. kahawae from other Colletotrichum species that may be present in coffee plant samples. The method proposes to use an end-point PCR marker for the mating type gene (MAT1-2-1) and a confirmatory test with a real-time quantitative PCR (qPCR) marker developed on the glutamine synthetase gene. The C. kahawae amplicons for the Cen-CkM10 qPCR marker exhibited specific melting temperature values and high-resolution melt profiles that could be readily differentiated from other tested species, including their relatives. Given the fungus's quarantine status, specificity was tested using artificial mixtures of DNA of C. kahawae with other Colletotrichum species and coffee plant DNA. The described method will enable NPPOs in coffee-producing and exporting countries, especially Colombia, to prevent this pathogen's entry, establishment, and spread.[Formula see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Assuntos

Coffea; Colletotrichum; Doenças das Plantas; Colletotrichum/genética; Colletotrichum/isolamento & purificação; Doenças das Plantas/microbiologia; Doenças das Plantas/prevenção & controle; Coffea/microbiologia; Reação em Cadeia da Polimerase em Tempo Real/métodos; Reação em Cadeia da Polimerase/métodos

Palavras-chave

CBD (coffee berry disease); Colletotrichum kahawae; PCR; biosecurity; certification; etiology; pathogen detection; quarantine

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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Doenças das Plantas / Colletotrichum / Coffea Idioma: En Revista: Plant Dis Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Colômbia País de publicação: Estados Unidos

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