Agrobacterium-mediated gene transfer and enhanced green fluorescent protein visualization in the mycorrhizal ascomycete Tuber borchii: a first step towards truffle genetics |
| |
Authors: | Benedetto Grimaldi Michiel A de Raaf Patrizia Filetici Simone Ottonello Paola Ballario |
| |
Institution: | (1) Dipartimento di Genetica e Biologia Molecolare, Universitá di Roma La Sapienza , P. le Aldo Moro 5, Roma, 00185, Italy;(2) Department of Life Sciences, Avans Hogeschool, Breda, The Netherlands;(3) Dipartimento di Biochimica e Biologia Molecolare, Università di Parma, Parco Area delle Scienze 23/A, Parma, 43100, Italy;(4) Istituto di Biologia e Patologia Molecolari, CNR, Università di Roma La Sapienza , P. le Aldo Moro 5, Roma, 00185, Italy |
| |
Abstract: | Mycorrhizal ascomycetes are ecologically and commercially important fungi that have proved impervious to genetic transformation so far. We report here on the successful transient transformation of Tuber borchii, an ectomycorrhizal ascomycete that colonizes a variety of trees and produces highly prized hypogeous fruitbodies known as truffles . A hypervirulent Agrobacterium tumefaciens strain bearing the binary plasmid pBGgHg was used for transformation. The genes for hygromycin resistance and the enhanced green fluorescent protein (EGFP), both under the control of vector-borne promoters, were employed as selection markers. Patches of dark and fluorescent hyphae were observed upon fluorescence microscopic examination of hygromycin-resistant mycelia. The presence of EGFP was confirmed by both confocal microscopy and PCR analysis. The lack in the transformed mycelia of the DNA coding for kanamicin resistance (a trait encoded by a vector-borne gene located outside of the T-DNA region) indicates that Agrobacterium-mediated gene transfer correctly occurred in T. borchii. |
| |
Keywords: | Mycorrhizal fungi Tuber borchii Truffles Agrobacterium tumefaciens Genetic transformation EGFP |
本文献已被 PubMed SpringerLink 等数据库收录! |
|