Efficient Agrobacterium tumefaciens-mediated transformation system of Diaporthe caulivora

被引:2
作者
Montoya, Marina R. A. [1 ]
Massa, Gabriela A. [1 ,2 ,3 ]
Colabelli, Mabel N. [3 ]
Ridao, Azucena del Carmen [3 ]
机构
[1] INTA CONICET, Inst Innovac Prod Agr & Desarrollo Sostenible IPA, Ruta 226 Km 73-5, RA-7620 Balcarce, Buenos Aires, Argentina
[2] Consejo Nacl Invest Cient & Tecn, Ruta 226 Km 73-5, RA-7620 Balcarce, Buenos Aires, Argentina
[3] Univ Nacl Mar del Plata, UNMdP, Fac Ciencias Agr, Ruta 226 Km 73-5, RA-7620 Balcarce, Buenos Aires, Argentina
关键词
GFP; Hygromycin; Latent infection; Soybean stem canker; GREEN FLUORESCENT PROTEIN; SOYBEAN STEM CANKER; T-DNA TRANSFER; INSERTIONAL MUTAGENESIS; VERTICILLIUM-DAHLIAE; FUNGUS; GENE; TOOL; PATHOGEN; GFP;
D O I
10.1016/j.mimet.2021.106197
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页数:11
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