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. 2021 Dec 6;1(8):100124. doi: 10.1016/j.crmeth.2021.100124

Figure 4.

Figure 4

Phenotypic effects of crgA- and leuA-targeted disruption

(A) Aerial mycelium and mycelial color of R. microsporus WT strain, an R. microsporus crgA mutant, M. lusitanicus WT strain (CBS 277.49), and an M. lusitanicus crgA mutant.

(B) Growth of R. microsporus wild-type strain (WT) and two crgA mutants (72 h, YPG media).

(C) Analysis of melanin production measured by its absorbance at 405 nm (p value of 0.0008).

(D) leuA mutants (1–8 from gRNA1+Template1 and 9–13 from gRNA2+Template2) after 48 h of growth on YNB media supplemented with leucine (right) and without leucine (left). Mutant growth is compared with the R. microsporus wild-type strain (WT) and the uridine auxotrophic strain UM1. UM6 (1) and UM7 (9) mutants are highlighted in red.

(E) Survival assays of mutant strains generated from R. microsporus. The survival assays were made in groups of eight immunocompetent Swiss mice, which were injected with 1 × 106 spores of the mutants pyrF− (UM1, green), crgA:pyrF (UM3, blue), and leuA::pyrF (UM6, red). In addition, the control WT strain was also injected (black). The survival rate of the mutants crgA:pyrF and leuA::pyrF and the WT strain were compared with the avirulent mutant strain pyrF− by a Mantel-Cox test (p value < 0.0001).