Alternative Identification Test Relying upon Sexual Reproductive Abilities of Candidalusitaniae Strains Isolated from Hospitalized Patients

doi:10.1128/JCM.39.11.3906-3914.2001

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Journal of Clinical Microbiology, November 2001, p. 3906-3914, Vol. 39, No. 11
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.11.3906-3914.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Alternative Identification Test Relying upon Sexual Reproductive Abilities of Candida lusitaniae Strains Isolated from Hospitalized Patients

Fabienne François,¹ Thierry Noël,¹^,^* Régis Pépin,² Annie Brulfert,¹ Christiane Chastin,¹ Anne Favel,³ and Jean Villard¹

Laboratoire des Sciences Végétales, Faculté de Pharmacie, Université René Descartes-Paris 5, Paris 75006,¹ Laboratoire d'Écologie Microbienne, UMR 5557, Université Claude Bernard-Lyon 1, Villeurbanne 69100,² and Laboratoire de Botanique, Cryptogamie, et Biologie Cellulaire, Faculté de Pharmacie, Université d'Aix-Marseille, Marseille 13000,³ France

Received 1 November 2000/Returned for modification 15 December 2000/Accepted 19 August 2001

The in vitro mating ability of Candida lusitaniae (teleomorph Clavispora lusitaniae) clinical isolates has been investigated.Studying the effects of culture conditions, we showed that ammoniumion depletion in the medium is a major trigger of the sexual cycle.Moreover, a solid support is required for mating, suggesting arole for adhesion factors in addition to the mating type generecognition function. Monitoring of mating and meiosis efficiencywith auxotrophic strains showed great variations in ascosporeyields, which appeared to be strain and temperature dependent,with an optimal range of 18 to 28°C. The morphogenetic eventstaking place from mating to ascospore release were studied byscanning and electron microscopy, and the ultrastructure of theconjugation canal, through which intercellular nuclear exchangesoccur, was revealed. Labeling experiments with a lectin-fluorochromesystem revealed that the nuclear transfer was predominantly polarized,thus allowing a distinction between the nucleus donor and thenucleus acceptor strains. The direction of the transfer dependedon the strain combination used, rather than on the genotypes ofthe strains, and did not appear to be controlled by the matingtype genes. Finally, we demonstrated that all of the 76 clinicalisolates used in this study were able to reproduce sexually whenmated with an opposite mating type strain, and we identified a1:1 MATa/MAT $alpha$ ratio in the collection. These results support theidea that there is no anamorph state in C. lusitaniae. Accordingly,the mating type test, which is easy to use and can usually becompleted within 48 h, is a reliable alternative identificationsystem for C. lusitaniae.

^* Corresponding author. Mailing address: Laboratoire des Sciences Végétales, Faculté de Pharmacie, 4 Ave. de l'Observatoire,75270 Paris Cedex 06, France. Phone: 33 1 53 73 96 41. Fax: 331 53 73 96 40. E-mail: noel{at}pharmacie.univ-paris5.fr.

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