Real-time PCR for detection and quantification of fungal and oomycete tomato pathogens in plant and soil samples

• Published in: Plant science (Limerick) Vol. 171; no. 1; pp. 155 - 165
• Main Authors: Lievens, Bart, Brouwer, Margreet, Vanachter, Alfons C.R.C., Cammue, Bruno P.A., Thomma, Bart P.H.J.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 01.07.2006; Elsevier Science
• Subjects: Biological and medical sciences; diagnostics; dna; Fundamental and applied biological sciences. Psychology; Fungal plant pathogens; Fusarium solani; identification; Internal transcribed spacer (ITS); internal transcribed spacers; Lycopersicon esculentum; microsclerotia; molecular sequence data; nucleotide sequences; Oomycetes; pathogen identification; Pathology, epidemiology, host-fungus relationships. Damages, economic importance; Phytopathology. Animal pests. Plant and forest protection; phytophthora; plant pathogenic fungi; polymerase chain reaction; pythium; Pythium ultimum; quantitative analysis; quantitative pcr; Rhizoctonia solani; ribosomal DNA; roots; soil fungi; SYBR Green I; Thanatephorus cucumeris; Verticillium; Verticillium albo-atrum; Verticillium dahliae; Verticillium tricorpus; Internal transcribed spacer (ITS); Fusarium solani; Rhizoctonia solani; Pythium ultimum; SYBR Green I; Verticillium; Plant pathogen; Phycomycetes; Biomass; Fungi; Dicotyledones; Angiospermae; Lycopersicon esculentum; Spermatophyta; Fungi Imperfecti; Solanaceae; Thallophyta
• ISSN: 0168-9452; 1873-2259
• DOI: 10.1016/j.plantsci.2006.03.009

Although new, rapid detection and identification technologies are becoming available more and more for various plant pathogens, pathogen quantification remains one of the main challenges in the disease management of many crops. Currently, real-time polymerase chain reaction (PCR) is the most straightforward technique to quantify pathogen presence. This manuscript describes the use of real-time PCR to quantitatively assess the presence of a number of economically important fungal and oomycete tomato pathogens in biological samples. We demonstrate that pathogen DNA can be accurately quantified over at least four orders of magnitude. Additionally, we demonstrate the feasibility of the technique to quantify pathogen biomass in complex biological samples.