Multiplex Quantitative PCR for the Detection of Bacteria Associated with Huanglongbing ‘Candidatus Liberibacter asiaticus,’ ‘Ca. L. americanus,’ and 16Sr IX Group Phytoplasma

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MetadadosDescriçãoIdioma
Autor(es): dc.contributorFundo de Defesa da Citricultura – Fundecitrus-
Autor(es): dc.contributorUniversidade Estadual Paulista (UNESP)-
Autor(es): dc.creatorMartins, Elaine C.-
Autor(es): dc.creatorTeixeira, Diva C.-
Autor(es): dc.creatorColetti, Daniela A.B.-
Autor(es): dc.creatorWulff, Nelson A.-
Data de aceite: dc.date.accessioned2025-08-21T16:19:49Z-
Data de disponibilização: dc.date.available2025-08-21T16:19:49Z-
Data de envio: dc.date.issued2025-04-29-
Data de envio: dc.date.issued2025-03-01-
Fonte completa do material: dc.identifierhttp://dx.doi.org/10.1094/PDIS-05-24-0970-RE-
Fonte completa do material: dc.identifierhttps://hdl.handle.net/11449/299668-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/11449/299668-
Descrição: dc.descriptionThe occurrence of ‘Candidatus Liberibacter’ spp. and ‘Ca. Phytoplasma’ spp. associated with blotchy mottle symptoms poses challenges to huanglongbing (HLB) diagnosis using molecular techniques. The ability to detect multiple targets simultaneously and specifically is a key aspect met by quantitative PCR (qPCR). A set of primers and hydrolysis probes useful in either single or multiplex reactions for the detection and quantification of HLB-associated bacteria were developed. Sequences from conserved genes of the ribosomal proteins for Liberibacter and phytoplasma circumvent the lack of specificity and cross-reactivity problems related to 16Sr DNA gene amplification, allowing precise and specific detection of HLB-associated bacteria in citrus and in the Liberibacter vector, Diaphorina citri. The triplex reaction exhibited high quality and precision as a robust tool for quantifying ‘Ca. L. asiaticus’ (CLas), ‘Ca. L. americanus’ (CLam), and 16Sr IX phytoplasma. Triplex qPCR showed consistent results and comparable sensitivity to the ribonuclease reductase test, although quantification cycle (Cq) values were higher when compared with 16SrDNA qPCR. Detection tests using field samples indicate that the qPCR triplex can identify HLB-associated bacteria in samples with varying levels of symptoms, ranging from typical to asymptomatic. Assessment of field samples from growers indicated more than 78.6% had Cq lower than 35.0, below the cutoff established for qPCR reactions used in this work. qPCR triplex is a safe, specific, and sufficiently sensitive technique for detecting CLas, CLam, and 16Sr IX phytoplasma simultaneously, in both citrus and D. citri samples. Its application is of importance in assisting growers in making decisions for HLB management.-
Descrição: dc.descriptionFundo de Defesa da Citricultura-
Descrição: dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)-
Descrição: dc.descriptionDepartamento de Pesquisa & Desenvolvimento Fundo de Defesa da Citricultura – Fundecitrus-
Descrição: dc.descriptionInstituto de Química Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP-
Descrição: dc.descriptionInstituto de Química Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP-
Descrição: dc.descriptionFundo de Defesa da Citricultura: 04 3008-
Descrição: dc.descriptionFAPESP: 2007/ 55013-9-
Formato: dc.format623-632-
Idioma: dc.languageen-
Relação: dc.relationPlant Disease-
???dc.source???: dc.sourceScopus-
Palavras-chave: dc.subjectcitrus-
Palavras-chave: dc.subjectD. citri-
Palavras-chave: dc.subjectgreening-
Palavras-chave: dc.subjectHuanglongbing-
Palavras-chave: dc.subjectLiberibacter-
Palavras-chave: dc.subjectmultiplex-
Palavras-chave: dc.subjectPhytoplasma-
Palavras-chave: dc.subjectpsyllid-
Palavras-chave: dc.subjectqPCR-
Palavras-chave: dc.subjectquantification-
Título: dc.titleMultiplex Quantitative PCR for the Detection of Bacteria Associated with Huanglongbing ‘Candidatus Liberibacter asiaticus,’ ‘Ca. L. americanus,’ and 16Sr IX Group Phytoplasma-
Tipo de arquivo: dc.typelivro digital-
Aparece nas coleções:Repositório Institucional - Unesp

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