Título:	Sensitive detection, quantifcation, and monitoring of Erwinia psidii colonization of guava plants using intercalating dye‑based real‑time PCR
Autor(es):	Hermenegildo, Pollyane da Silva Freitas, Rodrigo Galvão de Cascardo, Renan de Souza Guimarães, Lúcio Mauro Silva Pacheco, Jorge Luis Badel Alfenas‑Zerbini, Poliane Marques, Abi Soares dos Anjos Alfenas, Acelino Couto Ferreira, Marisa Alvares da Silva Velloso
ORCID:	https://orcid.org/0000-0002-4283-122X https://orcid.org/0000-0002-1298-9149 https://orcid.org/0000-0003-1397-8086 https://orcid.org/0000-0001-5021-6684 https://orcid.org/0000-0002-8203-5244 https://orcid.org/0009-0008-4751-1170 https://orcid.org/0000-0001-7776-3362 https://orcid.org/0000-0002-0677-861X
Afiliação do autor:	Universidade de Brasília, Departamento de Fitopatologia Universidade Federal de Viçosa, Departamento de Fitopatologia Universidade Federal de Viçosa, Departamento de Fitopatologia Universidade Federal de Viçosa, Departamento de Fitopatologia Universidade Federal de Viçosa, Departamento de Fitopatologia Universidade Federal de Viçosa, Departamento de Microbiologia Embrapa Recursos Genéticos e Biotecnologia, Gestão da Estação Quarentenária de Germoplasma Vegetal Universidade Federal de Viçosa, Departamento de Fitopatologia Universidade de Brasília, Departamento de Fitopatologia
Assunto:	Goiaba - doenças e pragas Goiaba - bactérias Diagnóstico bacteriológico
Data de publicação:	22-Nov-2022
Editora:	Springer Nature
Referência:	HERMENEGILDO, Pollyane da Silva et al. Sensitive detection, quantifcation, and monitoring of Erwinia psidii colonization of guava plants using intercalating dye‑based real‑time PCR. Tropical Plant Pathology, [S. l.], v. 48, p. 375–383, 2023. DOI: https://doi.org/10.1007/s40858-022-00542-9
Abstract:	Bacterial blight caused by Erwinia psidii is considered an important disease of the guava crop in Brazil. The disease has been disseminated to diferent geographic areas due to the movement of infected but asymptomatic propagating plant material. Consequently, methods showing high specifcity and sensitivity for early detection of latent infections are required to aid in the establishment of the use of pathogen-free seedlings and propagating material. In this study, an intercalating dye-based real-time PCR (qPCR) method using newly designed species-specifc primers targeting the recA gene sequence was developed. Primer specifcity was frst confrmed in silico and then by PCR amplifcation using DNA from strains of a collection of E. psidii and other plant-associated bacterial species. DNA from strains of other bacterial species obtained from uninfected guava and eucalypt leaves or from other plant species were not amplifed. When bacterial suspensions and purifed DNA were used in qPCR, detection limits were 103 CFU mL−1 and 102 genomic units µL−1, respectively. Using qPCR, E. psidii was detected in 100% of samples from symptomatic and in 57.1% of samples from asymptomatic trees collected in four guava orchards. The qPCR method allowed quantifcation of E. psidii populations in infected tissue of varieties Pedro Sato and Sassaoka as well as confrmation of the previously reported E. psidii acropetal and basipetal movement inside the plant. This new detection method with improved sensitivity has great potential not only for implementing disease diagnosis in guava and eucalypt nurseries and orchards but also for investigating relevant aspects of E. psidii life cycle and epidemiology.
Unidade Acadêmica:	Instituto de Ciências Biológicas (IB) Departamento de Fitopatologia (IB FIT)
Programa de pós-graduação:	Programa de Pós-Graduação em Fitopatologia
DOI:	https://doi.org/10.1007/s40858-022-00542-9
Versão da editora:	https://link.springer.com/article/10.1007/s40858-022-00542-9
Aparece nas coleções:	Artigos publicados em periódicos e afins

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