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Applied and Environmental Microbiology, January 2003, p. 616-624, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.616-624.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Genetic Diversity of Isolates of Glomus mosseae from Different Geographic Areas Detected by Vegetative Compatibility Testing and Biochemical and Molecular Analysis

Manuela Giovannetti,1* Cristiana Sbrana,2 Patrizia Strani,1 Monica Agnolucci,1 Valeria Rinaudo,1 and Luciano Avio2

Dipartimento di Chimica e Biotecnologie Agrarie, Università di Pisa,1 Istituto di Biologia e Biotecnologia Agraria, C.N.R., Pisa, Italy2

Received 22 May 2002/ Accepted 6 September 2002

We detected, for the first time, the occurrence of vegetative incompatibility between different isolates of the arbuscular mycorrhizal fungal species Glomus mosseae. Vegetative compatibility tests performed on germlings belonging to the same isolate showed that six geographically different isolates were capable of self-anastomosing, and that the percentage of hyphal contacts leading to fusions ranged from 60 to 85%. Successful anastomoses were characterized by complete fusion of hyphal walls, protoplasm continuity and occurrence of nuclei in the middle of hyphal bridges. No anastomoses could be detected between hyphae belonging to different isolates, which intersected without any reaction in 49 to 68% of contacts. Microscopic examinations detected hyphal incompatibility responses in diverse pairings, consisting of protoplasm retraction from the tips and septum formation in the approaching hyphae, even before physical contact with neighboring hyphae. Interestingly, many hyphal tips showed precontact tropism, suggesting that specific recognition signals may be involved during this stage. The intraspecific genetic diversity of G. mosseae revealed by vegetative compatibility tests was confirmed by total protein profiles and internal transcribed spacer-restriction fragment length polymorphism profiles, which evidenced a higher level of molecular diversity between the two European isolates IMA1 and BEG25 than between IMA1 and the two American isolates. Since arbuscular mycorrhizal fungi lack a tractable genetic system, vegetative compatibility tests may represent an easy assay for the detection of genetically different mycelia and an additional powerful tool for investigating the population structure and genetics of these obligate symbionts.

* Corresponding author. Mailing address: Dipartimento di Chimica e Biotecnologie Agrarie, Via del Borghetto 80, 56124 Pisa, Italy. Phone: 39-050-571561. Fax: 39-050-571562. E-mail: mgiova{at}agr.unipi.it.

Applied and Environmental Microbiology, January 2003, p. 616-624, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.616-624.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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