103(1)_str15

 

ISSN 1392-3196 / e-ISSN 2335-8947
Zemdirbyste-Agriculture, vol. 103, No. 1 (2016), p. 115–118
DOI 10.13080/z-a.2016.103.015

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

Seyed Mahyar MIRMAJLESSI, Inmaculada LARENA, Marika MÄND, Evelin LOIT

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 × 10-1 pg µl-1) and even in root of symptomless plants (5 × 10-1 pg µl-1). This is the first study of using this technique to quantify the population of V. dahliae in strawberry fields from Estonia.

Key words: quantitative detection, real-time PCR.

Full text: 103_1_str15.pdf