UFPR, Department of Genetics, Curitiba, PR, Brazil.
J Microbiol Methods. 2010 Feb;80(2):143-7. doi: 10.1016/j.mimet.2009.11.014. Epub 2009 Dec 6.
Guignardia citricarpa, the causal agent of Citrus Black Spot, was successfully transformed via Agrobacterium tumefaciens with cassettes for gfp and bar expression. Transformation is essential to understand the role of genes during interaction between plants and its pathogens. Using a binary plasmid vector based in the pPZP201BK, both germinated conidia and physically fragmented hyphae of G. citricarpa were transformed. Eight independent transformants of G. citricarpa resistant to ammonium glifosinate displayed GFP fluorescence. The majority (93.75%) of the G. citricarpa transformants was mitotically stable and contained a single T-DNA copy ectopically integrated to the chromosome. This is the first report of G. citricarpa transformation and will allow future work on virulence determinants of the fungus and possibly its control.
桔黑腐病菌（Guignardia citricarpa）是引起柑橘黑斑病的病原菌，本研究通过根癌农杆菌介导转化成功获得 GFP 和 bar 表达载体。基因转化是理解植物与其病原菌互作过程中基因功能的重要手段。本研究利用 pPZP201BK 为基础的二元质粒载体，转化萌发的分生孢子和物理切割的菌丝片段，获得了对氨磷汀抗性的 8 个桔黑腐病菌独立转化株，其中大多数（93.75%）转化株具有稳定的有丝分裂能力，且 T-DNA 拷贝以异位的形式整合到染色体上。这是桔黑腐病菌转化的首例报道，为研究该真菌的毒力决定因子及其防治方法奠定了基础。
