First report on the detection and quantification of Verticillium dahliae from Estonian strawberry fields using quantitative real-time PCR

Abstract

The main aim of this investigation was to develop a SYBRGreen real-time polymerase chain reaction (PCR) assay for detection and quantification of Verticillium dahliae directly from affected strawberry roots and soils. The proposed assay utilizes a specifically designed primer pair on the basis of an internal transcribed spacer (ITS) sequence. During 2014-2015, plant and soil samples were randomly collected from different areas including Vasula, Rohu, Unipiha, Utsu and Marjamaa in Estonia and analyzed for V dahliae. Real-time PCR technique using primers designed to the rDNA ITS1 was highly sensitive and accurate and so allowed reliable quantification of the pathogen DNA at low inoculation in soils (3 x 10(-1) pg mu l(-1)) and even in root of symptomless plants (5 x 10(-1) pg mu l(-1)). This is the first study of using this technique to quantify the population of V dahliae in strawberry fields from Estonia.