Agorbacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum
Wang JY and Li HY
Program and Abstracts, 11th International Citrus Congress (ISC Congress), 26-20 October 2008, Wuhan, China. 333 pages.
2008
บทคัดย่อ
Penicillium digitatum (Pers.:Fr) Sacc. is the causal agent of citrus green mold, which is the most widespread and destructive disease of postharvest citrus. Despite the economic importance of this fungus, little is know about the genetic basis of pathogenicity, which is necessary to develop a disease control strategy. Understanding the molecular mechanism of pathogenesis of P. digitatumis hampered by the lack of a readily accessible genetic tool to efficiently generate and recover mutant genes. Thus, the development of and efficient system to generate tagged insertional mutants in P. digitatumwill greatly aid the efforts to characterize gene(s) involved in pathogenicity. Agrobacterium tumefaciens-mediated transformation (ATMT) is widely used to create insertional mutants in a wide variety of plants. Recently, increasing numbers of fungal species have been transformed with A. tumefaciens. We report a method of A. tumefaciens-mediated genetic transformation of P. digitatum using hygromycin phosphotransferase gene (hph) as a selected marker. A transformation efficiency up to 60 transformants per 106 conidia of P. digitatum was achieved. Southern blot analysis of 6 randomly selected transformants verified the integration of the hph gene into the fungal genome. Therefore showing that the hph gene was randomly integrated as single copy into the fungal genome of P. digitatum. The 6 transformants tested were also shown to be mitotically stable. Thus Agrobacterium tumefaciens-mediated transformation of P. digitatum is a practical genetic tool for conduction insertional mutagenesis in P. digitatum to search for a gene(s) responsible for its pathogenicity and other characters.