An efficient procedure to extract RNA from a single Aphis gossypii and Aphis spiraecola for detecting Citrus tristeza virus by nested RT-PCR

Dandlen, Susana Anahi; Russo, Guilherme; Horta, Soraia; Duarte, Amilcar; Paiva, Paulo; Tomás Marques, Natália

http://hdl.handle.net/10400.1/27083

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Authors

Dandlen, Susana Anahi

Tomás Marques, Natália

Abstract(s)

A simple protocol to extract RNA from a single wingless aphid, Aphis gossypii and Aphis spiraecola, using liquid nitrogen and Tri-Reagent is described. A nested reverse-transcription polymerase chain reaction with degenerate primers enabled Citrus tristeza virus (CTV) detection in these aphid species collected from an infected tangor ‘Ortanique’ (Citrus × sinensis (L.) Osbeck × Citrus reticulata Blanco) in the field. The procedure enabled a CTV detection rate of 75% in the A. gossypii and A. spiraecola evaluated.