Reduction in solanapyrone phytotoxin production by Ascochyta rabiei transformed with Agrobacterium tumefaciens

• Published in: FEMS microbiology letters Vol. 255; no. 2; pp. 255 - 261
• Main Authors: Mogensen, Estelle Gewiss, Challen, Michael P., Strange, Richard N.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.02.2006; Blackwell Science Ltd; Blackwell; Oxford University Press
• Subjects: Agrobacterium tumefaciens; Agrobacterium tumefaciens - genetics; Ascochyta rabiei; Ascomycota - genetics; Ascomycota - metabolism; Ascomycota - pathogenicity; Biological and medical sciences; Blight; Blotting, Southern; Chickpeas; Cicer - microbiology; Cicer arietinum L; Cinnamates - pharmacology; Deoxyribonucleic acid; DNA; DNA, Bacterial - genetics; Drug Resistance, Bacterial - genetics; Fundamental and applied biological sciences. Psychology; genetic transformation; Growth, nutrition, metabolism, transports, enzymes. Molecular biology; High performance liquid chromatography; Hph gene; Hygromycin; Hygromycin B - analogs & derivatives; Hygromycin B - pharmacology; Laboratory equipment; Liquid chromatography; Microbiology; Mutation; Mycology; Naphthalenes - metabolism; pathogen; Phytotoxins; Plant Diseases - microbiology; polyketide; Polymerase Chain Reaction; Pycnidiospores; Pyrones - metabolism; T-DNA; Toxins; Transformation, Genetic; T‐DNA insertion; pathogen; T-DNA insertion; genetic transformation; L; polyketide; Cicerarietinum L; Genetic transformation; Agrobacterium tumefaciens; Insertion; Fungi; Polyketide; Pathogenic; Bacteria; Fungi Imperfecti; Rhizobiaceae; Ascochyta rabiei; Thallophyta
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.2005.00083.x

abstract Agrobacterium tumefaciens was used to transform Ascochyta rabiei, the causal agent of chickpea blight. Employing a T-DNA containing a hygromycin resistance gene (hph), 908 transformants were obtained from germinated pycnidiospores on a selective medium containing hygromycin. Transformants were confirmed using PCR and Southern analyses and of four of these that were tested, two had integrated multicopies of the hph gene, one had two copies and one had a single insertion. Transformants were tested for the production of solanapyrone A toxin using a microtitre plate assay. Loss of toxin production by transformants was confirmed by reversed phase high-performance liquid chromatography. Sixteen transformants out of 668 tested produced significantly less solanapyrone A than the wild-type strain.