Reference genes for gene expression studies by RT-qPCR in Brevipalpus yothersi (Acari: Tenuipalpidae), the mite vector of citrus leprosis virus

Autor(es)

Rogerio, Luana Aparecida ^[1] ; Galdeano, Diogo Manzano ^[1] ; Arena, Gabriella Dias ^[1] ; Nunes, Maria Andreia ^[1] ; Machado, Marcos Antonio ^[1] ; Novelli, Valdenice Moreira ^[1]

Afiliação do(s) autor(es)

^[1] Sylvio Moreira Citrus Ctr, Agron Inst IAC, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 9, APR 25 2019.
• Citações Web of Science: 0

Resumo

Quantitative reverse transcription PCR (RT-qPCR) is a high-throughput method to analyze the transcriptional expression of genes. Currently, no reference genes have been described for evaluating gene expression in Brevipalpus yothersi, the false spider mite, a polyphagous that act as vector of the citrus leprosis virus C (CiLV-C), an important citrus disease. This study aimed to identify the most stable reference genes in B. yothersi. The RT-qPCR expression data for selected genes were evaluated from three conditions: different developmental stages, plant hosts and acquisition of CiLV-C. To analyze the stability of the candidate reference genes we used Delta Cq method, GeNorm, NormFinder, BestKeeper and RefFinder. Ubiq and GAPDH are best suited for normalizing gene expression data in viruliferous and non-viruliferous mites. Ubiq, EF1 alpha and GAPDH are the most stable for different developmental stages. RPL13 and RPL32 are the best reference genes for approaches to B. yothersi in different host plants. Considering all the experimental conditions, Ubiq, EF1 alpha, and GAPDH were the most stable genes. Here we developed an accurate and comprehensive RT-qPCR strategy for use in B. yothersi gene expression analysis. These results will improve the understanding of the biology of the false spider mites and their role as virus vectors. (AU)

• Processo FAPESP: 16/21749-8 - Leprose dos citros e seu vetor: aspectos moleculares e biológicos
• Beneficiário: Valdenice Moreira Novelli
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/50880-0 - INCT 2014: de genômica comparativa e funcional e melhoramento assistido de citros
• Beneficiário: Marcos Antonio Machado
• Modalidade de apoio: Auxílio à Pesquisa - Temático