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Applied and Environmental Microbiology, September 2002, p. 4574-4582, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4574-4582.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Isolation and Characterization of Differentially Expressed Genes in the Mycelium and Fruit Body of Tuber borchii

Isabelle Lacourt,1 Sébastien Duplessis,2 Simona Abbà,1 Paola Bonfante,1* and Francis Martin2

Dipartimento di Biologia Vegetale, Università di Torino and Sezione di Torino, Istituto di Protezione delle Piante-CNR, 10125 Turin, Italy,1 UMR INRA/UHP "Interactions Arbres/Micro-Organismes," Centre de Recherches de Nancy, 54280 Champenoux, France2

Received 5 April 2002/ Accepted 4 June 2002

The transition from vegetative mycelium to fruit body in truffles requires differentiation processes which lead to edible fruit bodies (ascomata) consisting of different cell and tissue types. The identification of genes differentially expressed during these developmental processes can contribute greatly to a better understanding of truffle morphogenesis. A cDNA library was constructed from vegetative mycelium RNAs of the white truffle Tuber borchii, and 214 cDNAs were sequenced. Up to 58% of the expressed sequence tags corresponded to known genes. The majority of the identified sequences represented housekeeping proteins, i.e., proteins involved in gene or protein expression, cell wall formation, primary and secondary metabolism, and signaling pathways. We screened 171 arrayed cDNAs by using cDNA probes constructed from mRNAs of vegetative mycelium and ascomata to identify fruit body-regulated genes. Comparisons of signals from vegetative mycelium and fruit bodies bearing 15 or 70% mature spores revealed significant differences in the expression levels for up to 33% of the investigated genes. The expression levels for six highly regulated genes were confirmed by RNA blot analyses. The expression of glutamine synthetase, 5-aminolevulinic acid synthetase, isocitrate lyase, thioredoxin, glucan 1,3-ß-glucosidase, and UDP-glucose:sterol glucosyl transferase was highly up-regulated, suggesting that amino acid biosynthesis, the glyoxylate cycle pathway, and cell wall synthesis are strikingly altered during morphogenesis.

* Corresponding author. Mailing address: Dipartimento di Biologia Vegetale, Università di Torino, and Sezione di Torino, Istituto di Protezione delle Piante-CNR, Viale Mattioli 25, 10125 Turin, Italy. Phone: 39 011 670 7446. Fax: 39 011 670 7459. E-mail: p.bonfante{at}csmt.to.cnr.it.

Applied and Environmental Microbiology, September 2002, p. 4574-4582, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4574-4582.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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