Fig. 1

Prediction pathway of coumarin synthesis in Metarhizium anisopliae"

Table 1

Primers used for qRT-PCR"

Fig. 2

The construction of the vector A. Construction of overexpression vector pBARGPE1-p5c; B. pBARGPE1-p5cr carrying the target gene was identified by PCR; C. Overexpression vector single colonies were identified by PCR; D. Construction of donor in CRISPR-HDR-mediated p5cr deletion approach; E. up-kozak-EGEP-down fragment verified by PCR; F. Validation of donor pUC19-up-EGEP-down; G. Construction of CRISPR/Cas9 p5cr deletion vector pF332-sgRNA-p5cr-BenA; H. Critical element of pF332-sgRNA-p5cr-BenA; I. Validation of the linearized fragment of intermediate vector plasmid pF332-sgRNA-p5cr (16 513 bp); J. The pF332-sgRNA-p5cr-BenA vector was digested and verified by PCR"

Table 2

Primers for vector construction and validation"

Fig. 3

Correlation between the production of SW and the expression of genes p5cr, sdh, p450 and pks, respectively in W M. anisopliae A. SW levels in samples from fermentation day 1-7; B. Expression level of p5cr, sdh, p450 and pks genes days 1-7; C. Correlation between SW level and gene p5cr, sdh, p450 and pks expression level, respectively"

Fig. 4

Phylogenetic tree of p5cr gene sequence between Metarhizium and A. oxytropis constructed on a swiss-model using homology modeling"

Fig. 5

Inhibitory effect of ben on M. anisopliae A. The growth characteristics of M. anisopliae at different ben concentrations; B. Dose-dependent inhibition of M. anisopliae"

Fig. 6

Preparation of protoplasts A. The number of enzymatically interpreted protoplasts under different enzyme combinations; B. The number of protoplasts released by the complex enzyme at different enzymatic digestion times; C. Identification of protoplast viability"

Fig. 7

Identification of gene mutations mediated by nonhomologous end joining repair"

Fig. 8

PCR gel electrophoresis of single colonies selected by the overexpressed vector pBARGPE1-p5cr"

Fig. 9

Gene expression detection and determination of SW, PA, Lys level in W, PBARGPE1-p5cr and Δp5cr strains A. Expression levels of sdh gene in W, pBARGPE1-p5cr, and Δp5cr strains; B. Expression levels of p5cr gene in W, pBARGPE1-p5cr, and Δp5cr strains; C. Expression levels of pks gene in W, pBARGPE1-p5cr, and Δp5cr strains; D. Expression levels of p450 gene in W, pBARGPE1-p5cr, and Δp5cr strains; E. Lys levels in W, pBARGPE1-p5cr, and Δp5cr strains; F. PA levels in W, pBARGPE1-p5cr, and Δp5cr strains; G. SW levels in W, pBARGPE1-p5cr, and Δp5cr strains"