Volvariella volvacea, the Chinese mushroom, is a high-temperature grass-rot fungus with great production potential, yet its low yield limits industrial development. Exogenous sodium acetate (NaAc) has been shown to increase yield by promoting indole-3-acetic acid (IAA) synthesis during the primordium stage, but the underlying mechanism remains unclear. In this study, the aldehyde dehydrogenase gene VvALDH, highly expressed at the primordium stage, was cloned and functionally characterized. VvALDH encodes a 1509 bp cDNA with a conserved aldehyde dehydrogenase domain. Using Agrobacterium-mediated transformation, overexpression lines showed a 4.76-fold increase in VvALDH expression, accompanied by higher biomass (38%), yield (83%), and IAA content (34%), while RNAi lines showed opposite trends. These results demonstrate that VvALDH promotes IAA biosynthesis, enhances primordium differentiation, and increases yield. Further analysis revealed its involvement in multiple IAA biosynthetic pathways, including indolepyruvate, tryptamine, and tryptophan side-chain oxidase pathways. This work clarifies the molecular basis of NaAc-mediated yield improvement and provides a theoretical foundation for genetic and cultivation strategies in V. volvacea.

Cyclic diguanylate (c-di-GMP) affects bacterial physiological and biochemical functions like biofilm, motility, virulence, and bacterial secretion systems. GGDEF/EAL-domain proteins, participating in c-di-GMP synthesis and degradation, are widely present in Pseudomonas, with various structures and functions. Pseudomonas glycinae MS82 is a rhizosphere bacterium that protects mushroom against the pathogenic fungi. Although 14 genes encoding GGDEF/EAL-domain proteins have been identified in the genome of MS82, c-di-GMP regulation is poorly understood as a facilitator or repressor of physiological phenotypes. Here, PafQ and PafR, encoding the proteins with the tandem GGDEF/EAL domain, were functionally analyzed and found to regulate antifungal activity. Individual deletion mutants of PafQ and PafR were constructed in P. glycinae MS82 through biparental conjugation and homologous recombination. Subsequently, antifungal activity, biofilm formation, motility, and expression of the genes related to antifungal substance synthesis were examined and contrasted with those of wild-type P. glycinae MS82. Most phenotypes of physiological activities were significantly reduced after knocking out PafQ or PafR. In other members of the genus Pseudomonas, homologous genes of PafQ and PafR possess different functions in c-di-GMP regulation. In P. glycinae, the positive regulation of PafQ and PafR on fungistatic substance synthesis, biofilm formation, and motility is crucial in the biocontrol of mushroom diseases.

Pleurotus ostreatus cultivation is often affected by pest infestations, which contaminate the bag by eating nutrients and mycelium. This contamination eventually leads to a decline in the quality and yield of edible mushrooms and affects farmers’ income. Therefore, pesticides are commonly used for pest control. To examine the impact of insecticides on the growth of P. ostreatus, this study quantified the activities of antioxidant enzymes, including catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), and phenylalanine deaminase (PAL), in the mushroom under different insecticide treatments. Additionally, transcriptome sequencing was performed to investigate the underlying regulatory mechanisms. The findings indicated that dinotefuran, diflubenzuron, chlorantraniliprole, and beta-cypermethrin treatments resulted in a significant reduction in catalase and peroxidase activities in P. ostreatus. Conversely, the application of beta-cypermethrin and chlorantraniliprole significantly enhanced PAL and SOD activities in the mycelium. PAL activity was significantly increased in all the mixed substrates, whereas only spray treatments with diflubenzuron resulted in a significant increase in PAL activity. SOD activity in the substrates was reduced by diflubenzuron in the mixed treatment and chlorantraniliprole in the spray treatment. In contrast, all other treatments resulted in a significant increase in SOD activity in the substrates. Transcriptome sequencing revealed that differential genes were predominantly enriched in valine, leucine, and isoleucine degradation, fatty acid degradation, tyrosine metabolism, ascorbate and aldarate metabolism, and histidine metabolism, among others. These biological processes are hypothesized to be involved in the growth regulatory effects of insecticides on the mycelium and ascospores of P. ostreatus. The reliability of the transcriptomic data was also validated through quantitative real-time polymerase chain reaction.