Use of REMI and Agrobacterium-mediated transformation to identify pathogenicity mutants of the biocontrol fungus Coniothyrium minitans

CW Rogers; MP Challen; Jonathan Green; JM Whipps

doi:10.1016/j.femsle.2004.10.022

Use of REMI and Agrobacterium-mediated transformation to identify pathogenicity mutants of the biocontrol fungus Coniothyrium minitans

CW Rogers, MP Challen, Jonathan Green, JM Whipps

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Abstract

Restriction enzyme mediated integration (REMI) and Agrobacterium-mediated transformation (ATMT) were used to transform protoplasts or germinated conidia of the mycoparasite Coniothyrium minitans to hygromycin resistance. Using REMI, up to 32 transformants mug DNA(-1) were obtained, while 37.8 transformants 5 x 10(5) germlings(-1) were obtained using ATMT. Single-copy integrations occurred in 8% and 40% of REMI and ATMT transformants, respectively. A novel microtitre plate-based test was developed to expedite screening of 4000 REMI and ATMT C. minitans transformants. Nine pathogenicity mutants that displayed reduced or no pathogenicity on sclerotia of Sclerotinia sclerotiorum were identified.

Original language	English
Pages (from-to)	207-214
Number of pages	8
Journal	FEMS Microbiology Letters
Volume	241
Issue number	2
DOIs	https://doi.org/10.1016/j.femsle.2004.10.022
Publication status	Published - 1 Jan 2004

Keywords

Agrobacterium tumefaciens-mediated transformation
pathogenicity
REMI
Coniothyrium minitans
mycoparasitism

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10.1016/j.femsle.2004.10.022

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title = "Use of REMI and Agrobacterium-mediated transformation to identify pathogenicity mutants of the biocontrol fungus Coniothyrium minitans",

abstract = "Restriction enzyme mediated integration (REMI) and Agrobacterium-mediated transformation (ATMT) were used to transform protoplasts or germinated conidia of the mycoparasite Coniothyrium minitans to hygromycin resistance. Using REMI, up to 32 transformants mug DNA(-1) were obtained, while 37.8 transformants 5 x 10(5) germlings(-1) were obtained using ATMT. Single-copy integrations occurred in 8% and 40% of REMI and ATMT transformants, respectively. A novel microtitre plate-based test was developed to expedite screening of 4000 REMI and ATMT C. minitans transformants. Nine pathogenicity mutants that displayed reduced or no pathogenicity on sclerotia of Sclerotinia sclerotiorum were identified.",

keywords = "Agrobacterium tumefaciens-mediated transformation, pathogenicity, REMI, Coniothyrium minitans, mycoparasitism",

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AU - Challen, MP

AU - Green, Jonathan

AU - Whipps, JM

PY - 2004/1/1

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AB - Restriction enzyme mediated integration (REMI) and Agrobacterium-mediated transformation (ATMT) were used to transform protoplasts or germinated conidia of the mycoparasite Coniothyrium minitans to hygromycin resistance. Using REMI, up to 32 transformants mug DNA(-1) were obtained, while 37.8 transformants 5 x 10(5) germlings(-1) were obtained using ATMT. Single-copy integrations occurred in 8% and 40% of REMI and ATMT transformants, respectively. A novel microtitre plate-based test was developed to expedite screening of 4000 REMI and ATMT C. minitans transformants. Nine pathogenicity mutants that displayed reduced or no pathogenicity on sclerotia of Sclerotinia sclerotiorum were identified.

KW - Agrobacterium tumefaciens-mediated transformation

KW - pathogenicity

KW - REMI

KW - Coniothyrium minitans

KW - mycoparasitism

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Use of REMI and Agrobacterium-mediated transformation to identify pathogenicity mutants of the biocontrol fungus Coniothyrium minitans

Abstract

Keywords

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