Salicylic Acid Treatment Alleviates the Heat Stress Response by Reducing the Intracellular ROS Level and Increasing the Cytosolic Trehalose Content in Pleurotus ostreatus.

Pleurotus ostreatus is usually cultivated in horticultural facilities that lack environmental control systems and often suffer heat stress (HS). Salicylic acid (SA) is recognized as a plant defense-related hormone. Here, SA treatment (200 µM) induced fungal resistance to HS of P. ostreatus, with decreased malondialdehyde (MDA) content and HSP expression. Further analysis showed that SA treatment in P. ostreatus increased the cytosolic trehalose content and reduced the intracellular reactive oxygen species (ROS) level. Moreover, H2O2 could restore the MDA content and HSP expression of P. ostreatus treated with SA under HS. In addition, trehalose (25 mM) or CaCl2 (5 mM) treatment induced fungal resistance to HS, and CaCl2 treatment increased the cytosolic trehalose content of P. ostreatus under HS. However, inhibiting Ca2+ levels using Ca2+ inhibitors or mutants reversed the trehalose content induced by SA in P. ostreatus under HS. In addition, inhibiting trehalose biosynthesis using Tps-silenced strains reversed the MDA content and HSP expression of P. ostreatus treated with SA under HS. Taken together, these results indicate that SA treatment alleviates the HS response of P. ostreatus by reducing the intracellular ROS level and increasing the cytosolic trehalose content. IMPORTANCE Heat stress (HS) is a crucial environmental challenge for edible fungi. Salicylic acid (SA), a plant defense-related hormone, plays key roles in plant responses to biotic and abiotic stresses. In this study, we found that SA treatment increased the cytosolic trehalose content and induced fungal resistance to HS in P. ostreatus. Further analysis showed that SA can alleviate the HS of P. ostreatus by reducing the intracellular ROS level and increasing the cytosolic trehalose content. Our results help to understand the mechanism underlying the responses of P. ostreatus to HS. In addition, this research provides new insights for the cultivation of P. ostreatus.

Functional Roles of LaeA-like Genes in Fungal Growth, Cellulase Activity, and Secondary Metabolism in Pleurotus ostreatus.

The global regulator LaeA plays crucial roles in morphological development and secondary metabolite biosynthesis in filamentous fungi. However, the functions of LaeA in basidiomycetes are less reported. The basidiomycete Pleurotus ostreatus is a well-known fungus used both in medicine and as food that produces polysaccharides and cellulolytic enzymes. In this study, we characterized three LaeA homologs (PoLaeA1, PoLaeA2, and PoLaeA3) in P. ostreatus. PoLaeA1 showed different expression patterns than PoLaeA2 and PoLaeA3 during different developmental stages. Silencing PoLaeA1 decreased the intracellular polysaccharide (IPS) content by approximately 28-30% and reduced intracellular ROS levels compared with those of the WT strain. However, silencing PoLaeA2 and PoLaeA3 decreased cellulase activity by 31-34% and 35-40%, respectively, and reduced the cytosolic Ca2+ content, compared with those of the WT strain. Further analysis showed that PoLaeA1 regulated IPS biosynthesis through intracellular ROS levels, whereas PoLaeA2 and PoLaeA3 regulated cellulase activity through intracellular Ca2+ signaling. Our results provide new insights into the regulation of polysaccharide biosynthesis and cellulase production in filamentous fungi.

The non-canonical functions of telomerase reverse transcriptase gene GlTert on regulating fungal growth, oxidative stress, and ganoderic acid biosynthesis in Ganoderma lucidum.

The telomerase reverse transcriptase (TERT) is the core catalytic subunit of telomerase. Its canonical function is synthesizing telomeric repeats to maintain telomere length and chromosomal stability. Accumulating evidence suggests that TERT has other important fundamental functions in addition to its catalytic telomere repeat synthesis activity. However, the non-canonical roles of TERT independent of its enzymatic activity are not clear in filamentous fungi. In the present study, we characterized the GlTert gene in Ganoderma lucidum. The non-canonical roles of GlTert were explored using GlTert-silenced strains (Terti8 and Terti25) obtained by RNA interference. Silencing GlTert delayed the fungal growth, decreased the length between hyphal branches, and induced fungal resistance to oxidative stress in G. ludicum. Further examination revealed that the intracellular ROS (reactive oxygen species) levels were increased while the enzyme activities of the antioxidant systems (superoxide dismutase, catalase, glutathione peroxidase, and ascorbate peroxidase) were decreased in GlTert-silenced strains. In addition, silencing GlTert decreased the ganoderic acid (GA) biosynthesis of G. lucidum. Taken together, our results indicate that GlTert plays a fundamental function on fungal growth, oxidative stress, and GA biosynthesis in G. lucidum, providing new insights for the canonical functions of TERT in filamentous fungi. KEY POINTS: • GlTert affected fungal growth and hyphal branching of G. lucidum. • Silencing GlTert increased the intracellular ROS levels of G. lucidum. • GlTert regulated GA biosynthesis of G. lucidum.