The role of Jacalin-related lectin gene AOL_s00083g511 in the development and pathogenicity of the nematophagous fungus Arthrobotrys oligospora

Fus3 regulates asexual development and trap morphogenesis in the nematode-trapping fungus Arthrobotrys oligospora

“… 67 The 5′- and 3′-flanking regions of the fus3 gene (2,064 and 1,927 bp) and the hygromycin resistance cassette ( hph ) were individually amplified by PCR with paired primers ( Table S1 ). The three DNA fragments were isolated using a PCR product recovery kit (TaKaRa) and then co-transformed into the S. cerevisia e FY834 strain by electroporation 68 , 69 with linearized pRS426 (digested with Eco RI and Xho I) to generate the pRS426- fus3 - hph recombinant plasmid. Complete disruption sequences were amplified using primers fus3 -5f/ fus3 -3r ( Table S1 ) and transformed into A. oligospora protoplasts as previously described.…”

“…Complete disruption sequences were amplified using primers fus3 -5f/ fus3 -3r ( Table S1 ) and transformed into A. oligospora protoplasts as previously described. 66 , 69 Then, positive transformants grown on PADS medium were verified by PCR and Southern blotting. 70 Genomic DNA of the WT and Δ fus3 mutant strains were extracted using a plant genomic DNA Kit (TaKaRa) and digested using Stu I for Southern blotting analysis.…”

Function discovery of a non-ribosomal peptide synthetase-like encoding gene in the nematode-trapping fungus Arthrobotrys oligospora

“…Protoplast suspension (100 μL) was mixed well with 10 μg of the knockout fragment PCR product, placed in an ice bath for 40 min, 700 μL of KTC buffer was added, mixed well, and incubated at 28°C for 1 h. Subsequently, the above suspension was spread on a TB3 plate (200 g/L sucrose, 3 g/L tryptone, 3 g/L yeast extract, and 0.75% agar), which was incubated at a constant temperature of 28°C for 18 h to allow regeneration. The TB3 plate was covered with a layer of TB3 containing 150 μg/mL of thaumycin B medium as the upper plate for transformer screening and incubated at 28°C for approximately 7 days to allow transformants to grow ( Dong et al, 2021 ).…”

“…The mycelium was collected and ground in liquid nitrogen, and the genomic DNA of the transformants was extracted using a fungal genome extraction kit. Positive transformants were screened using the validation primers YZ-F/YZ-R ( Supplementary Table S1 ) and via PCR using genomic DNA as the template ( Dong et al, 2021 ).…”