Transformation ofGaeumannomyces graminis to benomyl resistance

Henson, Joan M.; Blake, N. K.; Pilgeram, Alice L.

doi:10.1007/bf00569334

Cited by 34 publications

(14 citation statements)

References 13 publications

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“…All published reports of transformation of G. graminis have used PEG‐CaCl 2 ‐mediated protoplast transformation. Benomyl‐ and phleomycin‐resistant transformants of Ggg and Ggt have been obtained using this method, but the transformation efficiency is very low (1–5 transformants/µg DNA per 10 7 protoplasts) (Bowyer et al ., 1995; Henson et al ., 1988; Pilgeram and Henson, 1990), and varies greatly from experiment to experiment (P. Bowyer, personal communication). The stability of the integrated DNA varies.…”

Section: Genetics and Transformation Of G Graminismentioning

confidence: 99%

Gaeumannomyces graminis, the take‐all fungus and its relatives

Freeman

Ward

2004

Molecular Plant Pathology

100

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SUMMARY Take‐all, caused by the fungus Gaeumannomyces graminis var. tritici, is the most important root disease of wheat worldwide. Many years of intensive research, reflected by the large volume of literature on take‐all, has led to a considerable degree of understanding of many aspects of the disease. However, effective and economic control of the disease remains difficult. The application of molecular techniques to study G. graminis and related fungi has resulted in some significant advances, particularly in the development of improved methods for identification and in elucidating the role of the enzyme avenacinase as a pathogenicity determinant in the closely related oat take‐all fungus (G. graminis var. avenae). Some progress in identifying other factors that may be involved in determining host range and pathogenicity has been made, despite the difficulties of performing genetic analyses and the lack of a reliable transformation system.

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Section: Genetics and Transformation Of G Graminismentioning

confidence: 99%

Gaeumannomyces graminis, the take‐all fungus and its relatives

Freeman

Ward

2004

Molecular Plant Pathology

100

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“…Protoplasts were generated in peas medium described by Robinson and Sharon (1999). Protoplasts were prepared as described by Henson et al. (1988) using 2500 ml of cultures shaken at 20–22°C.…”

Section: Methodsmentioning

confidence: 99%

“…(1988) using 2500 ml of cultures shaken at 20–22°C. Ggg was transformed as previously described by Henson et al. (1988), with the following modifications.…”

Section: Methodsmentioning

confidence: 99%

Transformation of Gaeumannomyces graminis with β-Glucuronidase Reporter and Hygromycin Resistance Genes

Park

Radwan

Martin

et al. 2010

Journal of Phytopathology

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Take-all disease is caused by Gaeumannomyces graminis, (Sacc.) Arx & D. Olivier, a soil-borne fungus, which colonizes the root and crown tissue of many members of the Poaceae plant family. This fungus is able to grow along the surface of roots as darkly pigmented runner hyphae, which has the ability to penetrate the root. Here, we describe a genetic transformation of G. graminis var. graminis by using polyethylene glycol (PEG)-based protoplast transformation. Fungus cells were transformed with a plasmid, pHPG, containing the gusA reporter gene that codes for b-glucuronidase (GUS) and the hph gene for hygromycin resistance as the selectable marker. A de novo transformant selection assay was developed to identify the putative transformants that were expressing the hph gene. In addition, the transformed cells maintained the ability to infect the plant tissues. The GUS-expressing fungus can be used to study fungal infection processes including fungal penetration, colonization and the role(s) of melanin during pathogenesis. Thus, this study is the first report of G. graminis var. graminis transformed with a visibly detectable reporter gene that provides a useful tool to a better understanding of host-Gaeumannomyces interactions.

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“…An internal portion of the gene (lacking both the 5' and 3' ends) was cloned into a vector carrying a selectable marker (in this case the hygromycin B phosphotransferase (hph) gene, which confers drug resistance). The vector was introduced into Gga by standard DNA transformation procedures 38 • A number of hygromycin-resistant transformants were isolated in which integration of plasmid DNA by homologous recombination had occurred, giving rise to two incomplete copies of the avenacinase gene with intervening vector DNA sequences. These transformants could be distinguished from ectopic integration events by Southern blot analysis 25 • As expected, disrupted mutants were more sensitive to avenacin A-1 than the wild-type fungus and had lost the ability to deglucosylate this saponin.…”

Section: 'mentioning

confidence: 99%