Fig. 4: Mutation generations of PtTryp2 and characters of mutants.

a Schematic presentation of PtTryp2 protein. The target site (vertical arrow) for CRISPR/Cas9-based knockout is located within the conserved functional domain (green pentagon), with PAM motif shown in orange font. Red rectangle on the left depicts signal peptide; RPT: internal repeat 1; b Alignment of partial PtTryp2 sequences of the CRISPR/Cas9-generated mutants showing frameshift indels compared to wild type. The frequency by which the sequence was detected within the same colony is indicated in parenthesis. Font color coding: Black, WT sequence; Orange, functional domain containing target for CRISPR/Cas9; Purple, PAM sequence; Blue, Inserted bases; Red dashes, deleted bases. c PtTryp2 expression patterns of knockout and overexpression mutants under different conditions. FC fold change. Data are presented as mean values ± SD (n = 3 biologically independent samples). d PtTryp2 expression of knockout mutants exhibited no response to ambient N and P fluctuation. Data are presented as mean values ± SD (n = 3 biologically independent samples). The comparisons between the averages of the two groups were evaluated using the one-tailed Student’s t test. The p values with significance (p ≤ 0.05) are shown. e Growth curves of different PtTryp2 mutants under different N and P conditions. Nutrient conditions in c–e are indicated by HNHP (Nutrient-replete), LNHP (N-depleted, P-replete), HNLP (N-replete, P-depleted), and LNLP (Nutrient-depleted). Data are presented as mean values ± SD (n = 3 biologically independent samples). Source data are provided as a Source Data file.