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Applied and Environmental Microbiology, April 2007, p. 2571-2579, Vol. 73, No. 8
0099-2240/07/$08.00+0     doi:10.1128/AEM.00257-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

A Complex Ergovaline Gene Cluster in Epichloë Endophytes of Grasses{triangledown} ,{dagger}

Damien J. Fleetwood,1,2 Barry Scott,2 Geoffrey A. Lane,1 Aiko Tanaka,2 and Richard D. Johnson1*

AgResearch, Grasslands Research Centre, Palmerston North, New Zealand,1 Centre for Functional Genomics, Institute of Molecular BioSciences, Massey University, Palmerston North, New Zealand2

Received 31 January 2007/ Accepted 8 February 2007

Clavicipitaceous fungal endophytes of the genera Epichloë and Neotyphodium form symbioses with grasses of the subfamily Pooideae, in which they can synthesize an array of bioprotective alkaloids. Some strains produce the ergopeptine alkaloid ergovaline, which is implicated in livestock toxicoses caused by ingestion of endophyte-infected grasses. Cloning and analysis of a nonribosomal peptide synthetase (NRPS) gene from Neotyphodium lolii revealed a putative gene cluster for ergovaline biosynthesis containing a single-module NRPS gene, lpsB, and other genes orthologous to genes in the ergopeptine gene cluster of Claviceps purpurea and the clavine cluster of Aspergillus fumigatus. Despite conservation of gene sequence, gene order is substantially different between the N. lolii, C. purpurea, and A. fumigatus ergot alkaloid gene clusters. Southern analysis indicated that the N. lolii cluster was linked with previously identified ergovaline biosynthetic genes dmaW and lpsA. The ergovaline genes are closely associated with transposon relics, including retrotransposons and autonomous and nonautonomous DNA transposons. All genes in the cluster were highly expressed in planta, but expression was very low or undetectable in mycelia from axenic culture. This work provides a genetic foundation for elucidating biochemical steps in the ergovaline pathway, the ecological role of individual ergot alkaloid compounds, and the regulation of their synthesis in planta.

* Corresponding author. Mailing address: AgResearch, Grasslands Research Centre, Private Bag 11008, Palmerston North, New Zealand. Phone: 64 6 351 8090. Fax: 64 6 351 8032. E-mail: richard.johnson{at}agresearch.co.nz

{triangledown} Published ahead of print on 16 February 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, April 2007, p. 2571-2579, Vol. 73, No. 8
0099-2240/07/$08.00+0     doi:10.1128/AEM.00257-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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