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The grass parasitesClaviceps purpurea andClaviceps fusiformis produce ergot alkaloids (EA) in planta and in submerged culture. Whereas EA synthesis (EAS) inC. purpurea proceeds via clavine intermediates to lysergic acid and the complex ergopeptines,C. fusiformis produces only agroclavine and elymoclavine. InC. purpurea the EAS gene (EAS ) cluster includesdmaW (encoding the first pathway step),cloA (elymoclavine oxidation to lysergic acid), and thelpsA/lpsB genes (ergopeptine formation). We analyzed the correspondingC. fusiformis EAS cluster to investigate the evolutionary basis for chemotypic differences between theClaviceps species. Other than three peptide synthetase genes (lpsC and the tandem paralogueslpsA1 andlpsA2 ), homologues of allC. purpurea EAS genes were identified inC. fusiformis , including homologues oflpsB andcloA , which inC. purpurea encode enzymes for steps after clavine synthesis. Rearrangement of the cluster was evident aroundlpsB , which is truncated inC. fusiformis . This and several frameshift mutations render CflpsB a pseudogene (CflpsB Ψ ). No obvious inactivating mutation was identified in CfcloA . AllC. fusiformis EAS genes, including CflpsB Ψ and CfcloA , were expressed in culture. Cross-complementation analyses demonstrated that CfcloA and CflpsB Ψ were expressed inC. purpurea but did not encode functional enzymes. In contrast, CpcloA catalyzed lysergic acid biosynthesis inC. fusiformis , indicating thatC. fusiformis terminates its EAS pathway at elymoclavine because thecloA gene product is inactive. We propose that theC. fusiformis EAS cluster evolved from a more complete cluster by loss of somelps genes and by rearrangements and mutations inactivatinglpsB andcloA .

applied and environmental microbiology

Comparison of Ergot Alkaloid Biosynthesis Gene Clusters in <i>Claviceps</i> Species Indicates Loss of Late Pathway Steps in Evolution of <i>C. fusiformis</i>

Comparison of Ergot Alkaloid Biosynthesis Gene Clusters in Claviceps Species Indicates Loss of Late Pathway Steps in Evolution of C. fusiformis