Disruption of Cerevisin via Agrobacterium tumefaciens‐mediated transformation affects microsclerotia formation and virulence of Verticillium dahliae

Abstract

Agrobacterium tumefaciens-mediated transformation (ATMT) was used to obtain a large number of Verticillium dahliae (Vd991) T-DNA insertion mutants that were randomly integrated. Insertion mutants that produced significantly fewer microsclerotia were chosen for further analysis. Mutant T0065 was identified as having the Cerevisin gene interrupted by T-DNA, and it was named cerevisin. The cerevisin protein showed a high amino acid sequence similarity with the vacuolar protease B. The mutant strain cerevisin displayed decreased production of microsclerotia and conidia, significantly reduced growth rate and reduction in virulence compared to the wild type. Moreover, the composition of secreted proteins differed between the cerevisin mutant and the wild type. Loss of function of Cerevisin decreased secretion of proteins of low molecular weight (14-25 kDa). Upon treatment with the secreted proteins of the mutant, the degree of leaf wilting decreased, indicating that Cerevisin is implicated in production of these proteins, which are putative pathogenicity factors of V. dahliae. The results suggest that Cerevisin is involved in controlling multiple processes of development and metabolism, plays an important role in vegetative growth and microsclerotia formation and affects virulence of V. dahliae.

Identification of pathogenicity-related genes and the role of a subtilisin-related peptidase S8 (PePRT) in authophagy and virulence of Penicillium expansum on apples

“…Consistent with these results, deletion of Proteinase B homologous genes in V. dahliae (Vd991) also resulted in a significant decrease in the production of conidia, significant reduction in growth rate, and a reduction in virulence, relative to the wild-type (He et al 2015). Targeted disruption of the cellular serine proteinase ALP2-encoding gene in Aspergillus fumigatus also resulted in a slightly decreased rate of vegetative growth and a more than 80 % reduction in sporulation in the alp2-negative mutants, which was correlated with an approximate 50 % reduction in the median diameter of conidiophore vesicles (Reichard et al 2000).…”

“…dahliae and Aspergillus fumigatus also exhibited a reduction in the radial growth rate (He et al 2015, Reichard et al 2000. Growth studies also indicated that the colony morphology of the ΔPeprt null mutant was different from that of the WT and the ectopic mutant (Fig.…”

The Subtilisin-Like Protease Bcser2 Affects the Sclerotial Formation, Conidiation and Virulence of Botrytis cinerea

“…Subtilisin-like serine proteases are reported to be involved in the sporulation of M. oryzae and C. parasitica [25,34], and impacted the sclerotial development of V. dahlia [21]. Our results indicated that Bcser1 is essential neither for sporulation nor sclerotial formation in B. cinerea.…”

“…Subtilisin-like serine proteases are also reported to play important roles in the virulence of many pathogenic fungi [20]. Orthologous of Bcser1 and Bcser2 are widely present in pathogenic fungi, and they also have been reported to play important roles in the virulence of pathogenic fungi [12,21,22,34,37]. For example, disruption of Spm1, an orthologous of Bcser2 in the rice blast fungus M. oryzae, resulted in a significant reduction in virulence on seedlings of rice and barley [34].…”

“…For example, disruption of the subtilase Cerevisin affects microsclerotia formation and virulence in Verticillium dahliae. Furthermore, the results showed that Cerevisin is involved in secretion of low molecular weight (14-25 kDa) proteins and controlling multiple processes of development and metabolism [21]. In Pseudogymnoascus destructans, the fungus responsible for white nose syndrome (WNS) in bats, a dominant subtilase PdSP1 was identified and considered to be potentially involved in the WNS host-pathogen interaction [12].…”