Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible (IPADS Balcarce), INTA - CONICET, Ruta 226 Km 73.5 (7620), Balcarce, Buenos Aires, Argentina..
Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible (IPADS Balcarce), INTA - CONICET, Ruta 226 Km 73.5 (7620), Balcarce, Buenos Aires, Argentina.; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Ruta 226 Km 73.5 (7620), Balcarce, Buenos Aires, Argentina.; Facultad de Ciencias Agrarias, Universidad Nacional de Mar del Plata (FCA, UNMdP), Ruta 226 Km 73.5 (7620), Balcarce, Buenos Aires, Argentina.
J Microbiol Methods. 2021 May;184:106197. doi: 10.1016/j.mimet.2021.106197. Epub 2021 Mar 10.
This is the first report describing the genetic transformation of Diaporthe caulivora, the soybean stem canker fungus. A simple and 100% efficient protocol of Agrobacterium tumefaciens-mediated transformation used mycelium as starting material and the hygromycin B resistance and green fluorescent protein (GFP) as a selection and reporter agents, respectively. All transgenic isolates were mitotically stable in two independent experiments and polymerase chain reaction with hygromycin B resistance primers confirmed successful T-DNA integration into the fungal genome. Plant-fungus interaction studies, including pathogenicity, latency, and endophytism, as well as further studies of random and targeted mutagenesis will be possible with GFP-expressing isolates of D. caulivora and other species in the Diaporthe / Phomopsis complex.
这是第一篇描述尖孢镰刀菌（大豆茎溃疡病菌）遗传转化的报告。我们建立了一个简单且 100%有效的根癌农杆菌介导的转化方法，以菌丝体为起始材料，潮霉素 B 抗性和绿色荧光蛋白（GFP）分别作为选择和报告标记。在两个独立的实验中，所有的转基因分离株都在有丝分裂中保持稳定，而利用潮霉素 B 抗性引物的聚合酶链反应证实了 T-DNA 成功整合到真菌基因组中。利用 GFP 表达的尖孢镰刀菌和其它尖孢镰刀菌/拟茎点霉复合种群中的种进行植物-真菌互作研究，包括致病性、潜伏性和内生性，以及随机和靶向诱变的进一步研究将成为可能。
