Evaluating the Mode of Antifungal Action of Heat-Stable Antifungal Factor (HSAF) in Neurospora crassa

Xiaodong Liu,Xianzhang Jiang,Haowen Sun,Lina Qin,Jianzhong Huang,Yuhang Luo,Jiawen Du

doi:10.3390/jof8030252

Abstract

Heat-stable antifungal factor (HSAF) isolated from Lysobacter enzymogenes has shown a broad-spectrum of antifungal activities. However, little is known about its mode of action. In this study, we used the model filamentous fungus Neurospora crassa to investigate the antifungal mechanism of HSAF. We first used HSAF to treat the N. crassa strain at different time points. Spore germination, growth phenotype and differential gene expression analysis were conducted by utilizing global transcriptional profiling combined with genetic and physiological analyses. Our data showed that HSAF could significantly inhibit the germination and aerial hyphae growth of N. crassa. RNA-seq analysis showed that a group of genes, associated with cell wall formation and remodeling, were highly activated. Screening of N. crassa gene deletion mutants combined with scanning electron microscopic observation revealed that three fungal cell wall integrity-related genes played an important role in the interaction between N. crassa and L. enzymogens. In addition, Weighted Gene Co-Expression Network Analysis (WGCNA), accompanied by confocal microscopy observation revealed that HSAF could trigger autophagy-mediated degradation and eventually result in cell death in N. crassa. The findings of this work provided new insights into the interactions between the predatory Lysobacter and its fungal prey.

Highlights

Heat-stable antifungal factor (HSAF) is an antimycotic compound isolated from a biological control strain Lysobacter enzymogenes
We observed that the sizes of the colonies of N. crassa were reduced with an increased concentration gradient of HSAF (Figure 1B,C)
As reported for other crop and clinical fungal pathogens [16–18], these data suggested that a certain amount of HSAF could inhibit the germination and polarized growth of N. crassa, enabling it to be a reasonable model to investigate the antifungal mechanism of HSAF

Summary

Introduction

Heat-stable antifungal factor (HSAF) is an antimycotic compound isolated from a biological control strain Lysobacter enzymogenes. It belongs to polycyclic tetramate macrolactams (PTMs) and has shown a broad-spectrum of antifungal activities (Figure 1A) [1]. Lysobacter enzymogenes is a Gram-negative soil bacterium belonging to the genus Lysobacter and the Xanthomonadaceae family This bacterium has twitching motility and can produce a variety of extracellular enzymes, including chitinase, protease, cellulase and β-1,3-glucanase [2–5] and produce antibiotic compounds such as β-lactams and HSAF [6,7]. It has an obvious antagonistic effect on many kinds of microorganisms including fungi, oomycetes, nematodes as well as bacteria [8–11].

Methods

Results

Conclusion