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PEG-mediated and Agrobacterium-mediated transformation in the mycopathogen Verticillium fungicola

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Amey, Richard C., Athey-Pollard, Anna, Burns, C. (Claire), Mills, P. R. (Peter R.), Bailey, Andy M. and Foster, Gary D. . (2002) PEG-mediated and Agrobacterium-mediated transformation in the mycopathogen Verticillium fungicola. Mycological Research, Vol.10 (No.1). pp. 4-11. ISSN 0953-7562

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Official URL: http://dx.doi.org/10.1017/S0953756201005251

Abstract

Verticillium fungicola, a severe mycopathogen of the cultivated mushroom Agaricus bisporus, was successfully transformed using both PEG-mediated and Agrobacterium-mediated techniques. PEG-mediated co-transformation was successful with hygromycin B resistance (hph), uidA (β-glucuronidase GUS), and green fluorescent protein (GFP) genes. Agrobacterium-mediated transformation was successful with the hph gene. Transformation frequencies of up to 102 transformants per μg DNA and 4068 transformants per 105 conidia were obtained for PEG-mediated and Agrobacterium-mediated transformation respectively. Expression of integrated genes in co-transformants was stable after 18 months of successive sub-culturing on non-selective medium, and following storage at -80 °C in glycerol. Molecular analysis of PEG-mediated transformants showed integration of the transforming genes into the target genome. Molecular analysis of Agrobacterium-mediated transformants showed integration of transforming DNA as single copies within the target genome. Co-transformants exhibited symptoms of disease in inoculation experiments and were at least as virulent as the wild-type fungus. GFP and GUS expression were observed in-vivo with the GFP-tagged strain showing great potential as a tool in epidemiological and host-pathogen interaction studies. The development of transformation systems for V. fungicola will allow in-depth molecular studies of the interaction of this organism with A. bisporus.

Item Type:

Journal Article

Subjects:

S Agriculture > SB Plant culture

Divisions:

Faculty of Science > Life Sciences (2010- ) > Warwick HRI (2004-2010)

Library of Congress Subject Headings (LCSH):

Verticillium, Fungi imperfecti, Cultivated mushroom -- Diseases and pests, Agrobacterium, Fungi -- Genetics

Journal or Publication Title:

Mycological Research

Publisher:

Cambridge University Press

ISSN:

0953-7562

Official Date:

January 2002

Dates:

Date	Event
January 2002	Published

Volume:

Vol.10

Number:

No.1

Page Range:

pp. 4-11

Identification Number:

10.1017/S0953756201005251

Status:

Peer Reviewed

Access rights to Published version:

Open Access

Funder:

Royal Society (Great Britain), Great Britain. Ministry of Agriculture, Fisheries and Food (MAFF)

References:

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