AGROBACTERIUM TUMEFACIENS-MEDIATED TRANSFORMATION OF TRICHODERMA HARZIANUM

Article Sidebar

PDF
Published: Nov 1, 2007
Keywords:
Agrobacterium tumefaciens genetic transformation Trichoderma harzianum hygromycin resistance

Main Article Content

Qian Yang*
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin, China
Li-ming Yang
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin, China
Pi-Gang Liu
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin, China
Sen Li
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin, China
Jinzhu Song
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin, China

Abstract

The development of a transformation system is a significant technical hurdle in the study of many filamentous fungi. Agrobacterium tumefaciens has been shown to transform yeasts, fungi and even human cells. By using this transformation system, filamentous fungus Trichoderma harzianum was successfully transformed with an efficiency of 60-110 transformants per 107 spores. PCR and Southern analysis showed that the T-DNA was integrated into the genome. The transformants contained a single copy that was stable through mitotic cell division. The tranformation system mediated by Agrobacterium tumefaciens may prove to be a powerful tool for filamentous fungi transformation and also for functional genomic study due to its high transformation frequency, simplicity of T-DNA integration, and genetic stability of transformants. These findings should facilitate future study of T. harzianum and stimulate wider use of this valuable transformation method in fungal research.


Keywords: Agrobacterium tumefaciens, genetic transformation, Trichoderma harzianum, hygromycin resistance


Corresponding author: E-mail: [email protected]

Article Details

Issue
Vol. 7 No. 2-1 (2007)
Section
Original Research Articles

Copyright Transfer Statement

 The copyright of this article is transferred to Current Applied Science and Technology journal with effect if and when the article is accepted for publication. The copyright transfer covers the exclusive right to reproduce and distribute the article, including reprints, translations, photographic reproductions, electronic form (offline, online) or any other reproductions of similar nature.

 The author warrants that this contribution is original and that he/she has full power to make this grant. The author signs for and accepts responsibility for releasing this material on behalf of any and all co-authors.

Here is the link for download: Copyright transfer form.pdf

 

 

Author Biography

Qian Yang*, Department of Life Science and Engineering, Harbin Institute of Technology, Harbin, China

Department of Life Science and Engineering,
Harbin Institute of Technology, Harbin, 150001 P R China

References

[1] Hansen, G. and Chilton, M.D. 1999 Lessons in Gene Transfer to Plants by a Gifted Microbe. In Plant Biotechnology: new products and applications (J. Hammond, P. McGarvey & V. Yusibov, eds), 21-57. Springer-Verlag, Berlin.
[2] Hynes, M. J. 1996 Genetic Transformation of Filamentous Fungi. Journal of Genetics, 75, 297-311.
[3] Leung, H., Lehtinen, U., Karjalainen, R., Skinner, D., Tooley, P., Leong, S. and Ellingboe, A. 1990 Transformation of the Rice Blast Fungus Magnaporthe grisea to Hygromycin-B Resistance, Current Genetics, 17, 409-411.
[4] Durand, N., Reymond, P. and Fevre, M. 1991 Transformation of Penicillium Roqueforti to Phleomycin-B-Resistance and to Hygromycin-B-Resistance. Current Genetics, 19, 149-153.
[5] Marmeisse, R., Gay, G., Debaud, J. C. & Casselton, L. A. 1992 Genetic transformation of the symbiotic basidiomycete fungus Hebeloma cylindrosporum. Current Genetics, 22, 41-45.
[6] Liu, P. and Yang, Q. 2005 Identification of Genes with a Biocontrol Function in Trichoderma harzianum Mycelium Using the Expressed Sequence Tag Approach, Research in Microbiology, 156, 416-423.
[7] Bundock, P., Dendulkras, A., Bdijersbergen, A.G. and Hooykaas, P.J. 1995 Transkingdom T-DNA Transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae, EMBO J., 14, 3206-3214.
[8] de Groot, M. J. A., Bundock, P., Hooykaas, P., J. and Bdijersbergen, A. G. 1998 Agrobacterium tumefaciens-mediated Transformation of Filamentous Fungi, Nat. Biotechnol, 16, 839-842.
[9] Mullins, E. D., Chen, X., Romaine, P., Raina, R., Geiser, D. M. and Kang, S. 2001 Agrobacterium-mediated Transformation of Fusarium oxysporum: An Efficient tool for Insertional Mutagenesis and Gene Transfer, Phytopathology, 91, 173-180.
[10] Hanif, M., Pardo, A. G., Gorfer, M and Raudaskoski, M. 2002 T-DNA Transfer and Integration in the Ectomycorrhizal Fungus Suilus Bovines using Hygromycin B as a Selectable Marker, Curr Genet., 41, 183-188.
[11] Combier, J. P., Melayah, D., Raffier, C., Gay, G. and Marmeisse, R. 2003 Agrobacterium Tumefaciens-mediated Transformation as a Tool for Insertional Mutagenesis in the Symbiotic Ectomycorrhizal Fungus Hebeloma cylindrosporum, FEMS Microbiol. Lett., 220, 141-148.
[12] Xing-xi, G. and Qian, Y. 2005 Transformation of Gene Cry I A(b) Mediated by Agrobacterium tumefaciens in Chaetomium globosum. Journal of Agro-Environment Science, 24 (1), 22-25.
[13] Xing-xi, G. and Qian, Y. 2005 Agrobacterium Tumerfaciens-medaited Transformation of Chaetomium globosum and its T-DNA Insertional Mutagenesis. Acta Microbiologica Sinica, 45 (1), 129-131.
[14] Xing-xi, G. and Qian, Y., Guan-ling, S. and Hong-xing, J. 2005 T-DNA Insertional Mutagenesis in Filamentous Fungi and its Application Prospect. Journal of Harbin Institute of Technology, 37 (9), 1177-1179.