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A conventional PCR technique to detect Septoria tritici in wheat seeds
V. F.
Consolo A C,
C. M.
Albani A,
C. M.
Berón A,
G. L.
Salerno A,
C. A.
Cordo B
A
Centro de Estudios de Biodiversidad y Biotecnología-Centro de Investigaciones Biológicas Fundación para Investigaciones Biológicas Aplicadas (CEBB-CIB FIBA), C.C. 1348, 7600 Mar del Plata, Argentina.
B
CIC (Comisión de Investigaciones Científicas, Provincia de Buenos Aires), CIDEFI, Facultad de Ciencias Agrarias y Forestales Universidad Nacional de La Plata, Calle 60 y 119, 1900, La Plata, Argentina.
C
Corresponding author. Email: faconsolo@fiba.org.ar
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Australasian Plant Pathology 38(3) 222–227 http://dx.doi.org/10.1071/AP08099
Submitted: 11 June 2008
Accepted: 18 November 2008
Published online: 3 April 2009
Abstract
A PCR assay was developed for detection of wheat seed naturally contaminated with Septoria tritici. S. tritici specific primers were derived from strict alignment of ITS and α-tubulin sequences of the pathogen. The specificity of four sets of synthesised oligonucleotide pairs (A, B, C and D) were tested using isolates from S. tritici, other selected fungi and wheat seeds. A single DNA fragment was amplified from S. tritici isolates with all primer pairs, whereas no product was generated from other DNA sources. S. tritici was also detected in wheat seed lots collected from plants with variable pycnidial coverage on the upper two leaves. PCR detection of as little as 0.5 pg of S. tritici genomic DNA was possible. This is the first report on the detection of S. tritici DNA in naturally infested wheat seeds. This PCR based assay is simple, rapid, specific, sensitive and suitable for routine detection of the wheat pathogen in infested wheat seeds.
Keywords:
Mycosphaerella graminicola, seedborne pathogen, Septoria leaf blotch, wheat.
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Australasian Plant Disease Notes
