Instability of pTEL vectors in B. cinerea under non-selective culture conditions.

Conidia of B. cinerea strains containing vectors pFB2N, pTEL-BcCas9GFP-NLS-Stux2 and pFB2N (stable HygR strain Bcgfp1 (25) and stable FenR strain niaD-fferg27 as controls) were transferred from solid media containing ME+Hyg and ME+Fen, respectively, to the edge of a non-selective ME plate and allowed to grow and sporulate for 7 d. From this plate, conidia were harvested from a ca. 1 cm2 area above the site of inoculation (site A) and from a similar area at 4 cm distance from the inoculation site. The conidia were transferred at low density to either ME+Hyg or ME+Fen plates. After 20 h incubation, the germination rate from 100–500 spores was determined. n.a.: not analysed. (XLSX)