FIG 1.

Sequence feature, subcellular localization, and transcriptional profile of CFP in B. bassiana. (A) Sequence features and phylogenetic ties of CFP homologs found in insect-pathogenic Cordycipitaceae (Bba, B. bassiana; Bbr, B. brongniartii; Cfu, Cordyceps fumosorosea) and Clavicipitaceae (Man, Metarhizium anisopliae; Mbr, Metarhizium brunneum; Mgu, Metarhizium guizhouense; Mro, Metarhizium robertsii). Each abbreviated fungal name is followed by the NCBI accession code of a CFP homolog, the length of its amino acid sequence, and its sequence identify to that of Bba CFP. Scale, branch length proportional to genetic distance assessed with the maximum likelihood method in MEGA7 at http://www.megasoftware.net/. (B and C) LSCM images (bar, 5 μm) for subcellular localization of GFP-tagged CFP fusion protein in hyphae and conidia (produced at L:D 12 h:12 h [light] or in the dark). Rows 1 to 5 are the hyphae stained with DAPI after being collected from a normal SDBY culture (control), the culture triggered for 12 h in CDB free of carbon and nitrogen sources, the cultures triggered for 90 min with 4 mM H₂O₂ and 0.4 M NaCl in one-fourth SDBY, and the culture in PBS irradiated at the UVB dose of 1 J/cm², respectively. (D) Nuclear versus cytoplasmic green fluorescence intensity (N/C-GFI) ratios of the fusion protein in the hyphae of rows 1 to 5. (E and F) Western blots for signal of the fusion protein detected in the nuclear protein extracts from the 60-h-old SDBY cultures of the WT and transgenic strain (TS) and secreted into the supernatants of the cultures. (G) Relative transcript (RT) levels of cfp in the WT cultures during 7-day incubation on SDAY or 4-day incubation in TPB with respect to a standard at the end of 2-day incubation. Error bars, standard deviations (SD) of the means from 10 hyphae (D) or three cDNA samples (F).