Development of a quantitative real-time PCR assay using SYBR Green for early detection and quantification of Austropuccinia psidii in Eucalyptus grandis

Autor(es)

Bini, Andressa Peres ^[1] ; Quecine, Maria Carolina ^[1] ; da Silva, Thalita Moraes ^[1] ; Silva, Luciana Duque ^[2] ; Labate, Carlos Alberto ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Escola Super Agr Luiz de Queiroz, Dept Genet, Ave Padua Dias 11, BR-13418900 Piracicaba, SP - Brazil

^[2] Univ Sao Paulo, Escola Super Agr Luiz de Queiroz, Dept Ciencias Florestais, Ave Padua Dias 11, BR-13418900 Piracicaba, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: European Journal of Plant Pathology; v. 150, n. 3, p. 735-746, MAR 2018.
• Citações Web of Science: 4

Resumo

Commercial areas containing Eucalyptus plantations have expanded in recent years due to increased demands for pulp, paper and bioenergy. One of the threats that can reduce Eucalyptus production is the eucalyptus rust disease caused by Austropuccinia psidii, a biotrophic fungus that affects a broad range of Myrtaceae. An accurate diagnosis tool for the early detection of rust disease could be useful in breeding programs for selection of resistant plants against rust, in phytosanitary purposes or in rust epidemics studies. The aim of the present work was to develop a SYBR Green-based quantitative real-time PCR (qPCR) assay for the early detection and quantification of A. psidii in Eucalyptus grandis leaves. Three sets of primers based on the A. psidii ribosomal DNA intergenic space region (IGS), beta-tubulin and elongation factor genes were designed and evaluated. The assays using the IGS primer set resulted in the highest detection efficiency, detecting a lower limit of 0.5 pg of A. psidii DNA. Under artificial inoculation in plants, A. psidii was detected immediately after pathogen inoculation until 240 h post-inoculation using qPCR. In field validation of the method, A. psidii was detected using qPCR in naturally infected leaves with or without rust symptoms. This easy and fast method can be used for an efficient detection of A. psidii in E. grandis leaves. The implications of this tool for rust studies are discussed below. (AU)

• Processo FAPESP: 08/50361-1 - Genômica funcional aplicada à descoberta de genes de resistência à ferrugem do eucalipto
• Beneficiário: Carlos Alberto Labate
• Linha de fomento: Auxílio à Pesquisa - Parceria para Inovação Tecnológica - PITE
• Processo FAPESP: 14/16804-4 - Aplicação das ômicas no entendimento da interação Puccinia psidii x Eucaliptus grandis
• Beneficiário: Maria Carolina Quecine Verdi
• Linha de fomento: Auxílio à Pesquisa - Apoio a Jovens Pesquisadores
• Processo FAPESP: 10/50445-0 - Estado da variabilidade genética de Puccinia psidiiwinter agente causal de ferrugem em Eucalyptus spp
• Beneficiário: Maria Carolina Quecine Verdi
• Linha de fomento: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/07596-6 - Estudo molecular do fitopatógeno Puccinia psidii Winter, agente causal da ferrugem, in vitro e durante sua interação com Eucalyptus spp
• Beneficiário: Andressa Peres Bini
• Linha de fomento: Bolsas no Brasil - Doutorado Direto