The determinant step in ergot alkaloid biosynthesis by an endophyte of perennial ryegrass

Jinghong Wang; Caroline Machado; Daniel G. Panaccione; Huei Fung Tsai; Christopher L. Schardl

doi:10.1016/j.fgb.2003.10.002

The determinant step in ergot alkaloid biosynthesis by an endophyte of perennial ryegrass

Jinghong Wang, Caroline Machado, Daniel G. Panaccione, Huei Fung Tsai, Christopher L. Schardl

Plant Pathology

Research output: Contribution to journal › Article › peer-review

93 Scopus citations

Abstract

Many cool-season grasses harbor fungal endophytes in the genus Neotyphodium, which enhance host fitness, but some also produce metabolites - such as ergovaline - believed to cause livestock toxicoses. In Claviceps species the first step in ergot alkaloid biosynthesis is thought to be dimethylallyltryptophan (DMAT) synthase, encoded by dmaW, previously cloned from Claviceps fusiformis. Here we report the cloning and characterization of dmaW from Neotyphodium sp. isolate Lp1, an endophyte of perennial ryegrass (Lolium perenne). The gene was then disrupted, and the mutant failed to produce any detectable ergovaline or simpler ergot and clavine alkaloids. The disruption was complemented with the C. fusiformis gene, which restored ergovaline production. Thus, the biosynthetic role of DMAT synthase was confirmed, and a mutant was generated for future studies of the ecological and agricultural importance of ergot alkaloids in endophytes of grasses.

Original language	English
Pages (from-to)	189-198
Number of pages	10
Journal	Fungal Genetics and Biology
Volume	41
Issue number	2
DOIs	https://doi.org/10.1016/j.fgb.2003.10.002
State	Published - Feb 2004

Bibliographical note

Funding Information:
We thank Alfred D. Byrd and Walter Hollin for technical assistance. Alkaloid analyses were conducted during a visit by D.G.P. to AgResearch Grasslands, Palmerston North, New Zealand, in collaboration with Brian Tapper, Geoff Lane, Elizabeth Davies, and Karl Fraser. Authentic standards of ergot alkaloids were provided by Forrest Smith (Auburn University) and Miroslav Flieger (Czech Academy of Science). This work was supported by Grant 2001-35319-10930 from the United States Department of Agriculture National Research Initiative. This is publication No. 03-12-113 of the Kentucky Agricultural Experiment Station, published with approval of the director.

ASJC Scopus subject areas

Microbiology
Genetics

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title = "The determinant step in ergot alkaloid biosynthesis by an endophyte of perennial ryegrass",

abstract = "Many cool-season grasses harbor fungal endophytes in the genus Neotyphodium, which enhance host fitness, but some also produce metabolites - such as ergovaline - believed to cause livestock toxicoses. In Claviceps species the first step in ergot alkaloid biosynthesis is thought to be dimethylallyltryptophan (DMAT) synthase, encoded by dmaW, previously cloned from Claviceps fusiformis. Here we report the cloning and characterization of dmaW from Neotyphodium sp. isolate Lp1, an endophyte of perennial ryegrass (Lolium perenne). The gene was then disrupted, and the mutant failed to produce any detectable ergovaline or simpler ergot and clavine alkaloids. The disruption was complemented with the C. fusiformis gene, which restored ergovaline production. Thus, the biosynthetic role of DMAT synthase was confirmed, and a mutant was generated for future studies of the ecological and agricultural importance of ergot alkaloids in endophytes of grasses.",

author = "Jinghong Wang and Caroline Machado and Panaccione, {Daniel G.} and Tsai, {Huei Fung} and Schardl, {Christopher L.}",

note = "Funding Information: We thank Alfred D. Byrd and Walter Hollin for technical assistance. Alkaloid analyses were conducted during a visit by D.G.P. to AgResearch Grasslands, Palmerston North, New Zealand, in collaboration with Brian Tapper, Geoff Lane, Elizabeth Davies, and Karl Fraser. Authentic standards of ergot alkaloids were provided by Forrest Smith (Auburn University) and Miroslav Flieger (Czech Academy of Science). This work was supported by Grant 2001-35319-10930 from the United States Department of Agriculture National Research Initiative. This is publication No. 03-12-113 of the Kentucky Agricultural Experiment Station, published with approval of the director.",

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AU - Machado, Caroline

AU - Panaccione, Daniel G.

AU - Tsai, Huei Fung

AU - Schardl, Christopher L.

N1 - Funding Information: We thank Alfred D. Byrd and Walter Hollin for technical assistance. Alkaloid analyses were conducted during a visit by D.G.P. to AgResearch Grasslands, Palmerston North, New Zealand, in collaboration with Brian Tapper, Geoff Lane, Elizabeth Davies, and Karl Fraser. Authentic standards of ergot alkaloids were provided by Forrest Smith (Auburn University) and Miroslav Flieger (Czech Academy of Science). This work was supported by Grant 2001-35319-10930 from the United States Department of Agriculture National Research Initiative. This is publication No. 03-12-113 of the Kentucky Agricultural Experiment Station, published with approval of the director.

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N2 - Many cool-season grasses harbor fungal endophytes in the genus Neotyphodium, which enhance host fitness, but some also produce metabolites - such as ergovaline - believed to cause livestock toxicoses. In Claviceps species the first step in ergot alkaloid biosynthesis is thought to be dimethylallyltryptophan (DMAT) synthase, encoded by dmaW, previously cloned from Claviceps fusiformis. Here we report the cloning and characterization of dmaW from Neotyphodium sp. isolate Lp1, an endophyte of perennial ryegrass (Lolium perenne). The gene was then disrupted, and the mutant failed to produce any detectable ergovaline or simpler ergot and clavine alkaloids. The disruption was complemented with the C. fusiformis gene, which restored ergovaline production. Thus, the biosynthetic role of DMAT synthase was confirmed, and a mutant was generated for future studies of the ecological and agricultural importance of ergot alkaloids in endophytes of grasses.

AB - Many cool-season grasses harbor fungal endophytes in the genus Neotyphodium, which enhance host fitness, but some also produce metabolites - such as ergovaline - believed to cause livestock toxicoses. In Claviceps species the first step in ergot alkaloid biosynthesis is thought to be dimethylallyltryptophan (DMAT) synthase, encoded by dmaW, previously cloned from Claviceps fusiformis. Here we report the cloning and characterization of dmaW from Neotyphodium sp. isolate Lp1, an endophyte of perennial ryegrass (Lolium perenne). The gene was then disrupted, and the mutant failed to produce any detectable ergovaline or simpler ergot and clavine alkaloids. The disruption was complemented with the C. fusiformis gene, which restored ergovaline production. Thus, the biosynthetic role of DMAT synthase was confirmed, and a mutant was generated for future studies of the ecological and agricultural importance of ergot alkaloids in endophytes of grasses.

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The determinant step in ergot alkaloid biosynthesis by an endophyte of perennial ryegrass

Abstract

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