Chaure P, Gurr S J, Spanu P
Department of Plant Sciences, University of Oxford, South Parks Rd, Oxford, OX1 3RB, UK.
Nat Biotechnol. 2000 Feb;18(2):205-7. doi: 10.1038/72666.
Barley powdery mildew, Erysiphe graminis f.sp. hordei, is an obligate biotrophic pathogen and as such cannot complete its life cycle without a living host. The inability to transform this fungus and manipulate its genome has constrained research towards understanding its life cycle and pathogenicity. Here we describe an in planta transformation system based on delivery of DNA using a gold-particle gun and selection using benomyl or bialaphos. Using this method, we consistently obtained stable transformants with efficiencies comparable to other filamentous fungi. Stable expression of the beta-glucuronidase in E. graminis was demonstrated by co-transforming the uidA gene with the selectable markers.
大麦白粉病，即禾本科布氏白粉菌大麦专化型（Erysiphe graminis f.sp. hordei），是一种专性活体营养型病原菌，因此没有活的寄主就无法完成其生命周期。无法转化这种真菌并操纵其基因组限制了对其生命周期和致病性的研究。在此，我们描述了一种基于使用金粒子枪递送DNA并使用苯菌灵或双丙氨膦进行筛选的植物体内转化系统。使用这种方法，我们始终能够获得稳定的转化体，其效率与其他丝状真菌相当。通过将uidA基因与选择标记共转化，证明了β-葡萄糖醛酸酶在禾本科布氏白粉菌中的稳定表达。
