PCR detection of Fusarium oxysporum fsp gladioli race 1, causal agent of Gladiolus yellows disease, from infected corms

• Published in: Plant pathology Vol. 49; no. 1; pp. 89 - 100
• Main Authors: Haan, L.A.M. de, Numansen, A, Roebroeck, E.J.A, Doorn, J. van
• Format: Journal Article
• Language: English
• Published: Oxford, U.K. and Cambridge, USA Blackwell Science Ltd 01.01.2000; Blackwell; Wiley Subscription Services, Inc
• Subjects: ADN; bioassay; bioassays; Biological and medical sciences; CORMO; CORMS; CORMUS; DETECTION; disease diagnosis; DNA; ENFERMEDADES DE LAS PLANTAS; ENFERMEDADES FUNGOSAS; epiphytes; etiology; Fundamental and applied biological sciences. Psychology; FUNGAL DISEASES; fungal diseases of plants; Fungal plant pathogens; FUSARIUM OXYSPORUM F. SP. GLADIOLI; genbank/af158099; genbank/af158100; GENETICA MOLECULAR; GENETIQUE MOLECULAIRE; GLADIOLUS; host range; IMMUNODIAGNOSIS; IMMUNODIAGNOSTIC; infection; INMUNODIAGNOSTICO; latent infections; MALADIE DES PLANTES; MALADIE FONGIQUE; MOLECULAR GENETICS; molecular sequence data; multiplex PCR; NUCLEOTIDE SEQUENCES; pathotypes; PCR; PHYSIOLOGICAL RACES; Phytopathology. Animal pests. Plant and forest protection; PLANT DISEASES; PLANT PATHOGENIC FUNGI; polymerase chain reaction; RACE PHYSIOLOGIQUE; races; random amplified polymorphic DNA technique; RAPDs; RAZAS FISIOLOGICAS; Variation, races, biotypes, parasitic specialization, genetics; YELLOWS; Iridaceae; Monocotyledones; Plant pathogen; Genetic marker; Gladiolus; Species specificity; Random amplified polymorphic DNA marker; Fungi; Polymerase chain reaction; Angiospermae; Flower crop; Spermatophyta; Specific identification; Fungi Imperfecti; Detection; Thallophyta
• ISSN: 0032-0862; 1365-3059
• DOI: 10.1046/j.1365-3059.2000.00421.x

Fusarium oxysporum f.sp. gladioli (FOG) race 1 infects both large‐ and small‐flowered Gladiolus cultivars. Race 2 isolates infect only small‐flowered cultivars but can be present as epiphytes on large‐flowered plants. When 160 arbitrary 10‐mer oligonucleotide primers were tested on FOG by PCR to find RAPD markers specific for race 1, the RAPD primer G12 amplified two discriminating DNA fragments, AB (609 bp) and EF (1196 bp), in race 1 isolates only. Both fragments were cloned and sequenced. Two pairs of race 1‐specific primers for multiplex PCR were designed. Tests of 112 F. oxysporum isolates by PCR showed that, in almost all cases, race 1 isolates of vegetative compatibility group 0340 could be distinguished with these primers. Seven putative race 1 isolates did not react in multiplex PCR; hybridization studies with labelled AB and EF DNA fragments showed that these isolates belong to separate groups. A bioassay was developed to detect corms that were latently infected with FOG race 1. Gladiolus corms were homogenized and incubated for 5 days at 28°C in a semiselective medium to induce growth of Fusarium. Cultivated mycelium was isolated and subjected to the developed multiplex PCR after standard DNA isolation or disruption by microwave treatment.