~ ) Pergamon 0305-1978194)EOOO59-X Biochemical Systematics and Ecology, Vol. 22, No. 8, pp. 803-806, 1994 Copyright © 1994 Elsevier Science Ltd Printed in Great Britain. All rights reserved 0305-1978/94 $7.00 + 0.00 Chemotaxonomic Discrimination Among the Fungal Genera Tolypocladium, Beauveria and Paecilomyces ZBYNIr:K KADLEC,* PETR ~IMEK,'f ANNA HEYDOVA, t ALEXANDR JEGOROV,*$ VLADIMIR MA'['HA,* ZDENI~K LANDA§ and JACOB EYAL II *Galena Co., Research Unit, Brani~ovsk& 31, 370 05 (~esk~ Bud~jovice, Czech Republic 1Institute of Entomology, Laboratory of Analytical Chemistry, Academy of Sciences of the Czech Republic Breni,~ovsk~i 31, 370 05 (~esk0 Bud~jovice, Czech Republic; §Faculty of Agriculture, 370 05 (~?,esk~ Bud~jovice, Czech Republic; IIW. R. Grace & Co.-Conn., 7379 Route 32, Columbia, MD 21044, U.S.A. Key Word Index--Beauveria; Paecilomyces; Tolypocladium; fungi imperfecti; beauverolides; I]-hydroxy acids; chemotaxonomy. Abstract--Fungi of the genera Beauveria and Paecilomyces were found to produce cyclotetradepsipeptides, beauverolides. Production of beauverolides was not detected at the genus Tolypocladium, Analysis of beauverolides therefore provides a very simple chemotaxonomic test which seems to be suitable for fas discrimination between the genera Beauveria vs Tolypocladium and complementing morphological examination. A GC-MS study of I]-hydroxy acid distribution in the beauverolide hydrolyzates revealed that all strains produce 3'-methyl-I]-hydroxy acids only. Their occurrence thus cannot be used as a taxonomic marker of different species within the genera Beauveria and Paecilomyces. Introduction There is ongoing discussion on the taxonomy of important entomopathogenic fungi of the genera Tolypocladium and Beauveria, classified by some authors as one genus (von Arx, 1986). Although morphological variations between the genera are in some cases difficult to recognize by classical methods, the fungi are also characterized by the production of unique secondary metabolites, which could be employed for their discrimination. To exemplify this feature, we recently demonstrated that genera of Tolypocladium and Beauveria differ in the production of intracellular siderophores (Jegorov et al., 1993). Furthermore, the fungi, and also bacteria, produce cyclic peptides synthesized by multifunctional peptide synthetases (von DShren, 1993). Due to the complexity and magnitude of these multienzymatic systems it is assumed, that one strain could produce only a particular class of these metabolites. Consequently, the cyclic peptides and/or the related enzymes might be used as taxonomic markers of some genera. Cyclosporines, produced by some species of Tolypocladium are a novel group of cyclic undecapeptides with profound immunosuppressive properties (Wenger, 1986). From the taxonomical point, however, the cyclosporines are produced by a number of non-relatedfungi and,.moreover, their production depends strongly on cultivation conditions (Jegorov et al., 1990). Beauverolides represent another group of cyclic peptides isolated from Beauveria bassiana (Elsworth and Grove, 1977, 1980; Grove, 1980) and Isaria felina (Vining and Taber, 1962; Deffieux et al., 1981). Although some species of Isaria are now classified under the genus Paecilomyces, I. felina was chosen as a lectotype of Isaria in order to provide a generic name for synnematous Beauveriaqike fungi (yon Arx, 1970; de Hoog, 1972). These cyclotetradepsipeptides SAuthor to whom correspondence should be addressed. (Received4 May 1994) 803