Sanghuangporus, a medicinal mushroom, has gained significant attention due to its beneficial properties. Phenolic acids are among the major bioactive compounds in Sanghuangporus, known for their antioxidant and anti-inflammatory activities. To precisely quantify the phenolic acid content, we developed a method utilizing ultra-high-performance liquid chromatography with triple quadrupole (UHPLC-QqQ). This study optimized the UHPLC-QqQ conditions to simultaneously separate and detect eight phenolic acids in Sanghuangporus baumii (Pilát) L.W. Zhou and Y.C. Dai, including chlorogenic acid, p-coumaric acid, caffeic acid, cryptochlorogenic acid, protocatechuic acid, ferulic acid, sinapic acid, and syringic acid. The separation process utilized a ZORBAX Eclipse Plus C18 column using 0.01% formic acid and 2 mmol/L ammonium formate in water as the aqueous phase and methanol containing 0.01% formic acid and 2 mmol/L ammonium formate as the organic phase. Calibration curves were constructed using standard solutions to quantitatively determine the phenolic acid content. The results showed significant variation in phenolic acid content among S. baumii fruiting bodies, with Protocatechuic acid, p-coumaric acid, and caffeic acid being the most abundant. This method is valuable for quantifying phenolic acid compounds under different cultivation conditions. It provides excellent sensitivity, selectivity, and reproducibility for the quantification of phenolic acids in Sanghuangporus, contributing to a better understanding of its chemical composition and potential health benefits. This approach represents a novel technical means for the simultaneous analysis of compound phenolic acids in Sanghuangporus fruiting bodies.

A rapid, precise, and dependable method for quantifying flavonoids in the fruiting bodies of Sanghuangporus was established using ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). Separation was achieved using a ZORBAX Eclipse Plus C18 column (1.8 μm, 3.0 mm × 100 mm) with a 15 min gradient of a mobile phase consisting of 0.01% aqueous formic acid and 2 mm/L ammonium formate (mobile phase A), and 0.01% formic acid and 2 mm/L ammonium formate in methanol (mobile phase B). A mass spectrometry analysis was performed using the multiple reaction monitoring (MRM) mode with an electrospray ion source. This method enabled the simultaneous detection of 10 flavonoids (sakuranetin, quercitrin, myricitrin, kaempferol, luteolin, rutin, hyperoside, kaempferol-3-O-rutinoside, catechin, and catechin gallate) in the fruiting bodies of Sanghuangporus. Additionally, we applied this method to analyze the flavonoid content in fruiting bodies of various Sanghuangporus species. The results revealed substantial variations in flavonoid content, up to a 100-fold difference, among different species, with myricitrin, hyperoside, and rutin identified as the most abundant flavonoids. This protocol serves as a valuable tool for quantifying flavonoid compounds in different Sanghuangporus species or under diverse cultivation conditions, particularly for identifying species with high levels of specific flavonoid compounds.

Camellia shells are the main by-product of camellia seed processing and are usually incinerated or disposed of as agricultural waste. In this study, camellia shells were employed in the Pleurotus pulmonarius cultivation process using five distinct formulae substituting for cottonseed shells. Our results show that as the substitution rate of camellia shells increased from 0% to 35%, the protein content in P. pulmonarius significantly increased from 34.05% to 53.35%. The polysaccharide content reached a peak value of 5.62% at 30% substitution of camellia shells. The DPPH free radical scavenging rate reached its maximum of 82.70% at 20% substitution of camellia shells. Furthermore, increases in the total amino acid contents in P. pulmonarius were positively correlated with the substitution rate of camellia shells. Considering the yield characteristics, the formula of 20% camellia shell substitution tested in this study appears to be optimal for P. pulmonarius cultivation. These findings not only provide a substrate to enhance the nutritional quality of P. pulmonarius but also demonstrate a novel approach for the ecological utilization of camellia shells.

An integrated transcriptomic and metabolomic analysis was conducted to elucidate the molecular mechanisms underlying heat stress response in Pleurotus pulmonarius, a commercially important edible mushroom sensitive to elevated temperatures. Through comparative RNA-seq and UPLC-MS based metabolomics, we identified 4,388 differentially expressed genes (DEGs) and 1,282 differentially accumulated metabolites (DAMs) in mycelia exposed to 42 °C for 6 h compared with control conditions. Integrated pathway analysis revealed that pyrimidine metabolism, tryptophan metabolism, and cysteine/methionine metabolism were coordinately reprogrammed at both transcriptional and metabolic levels, forming a synergistic regulatory network. These pathways collectively provide essential precursors for nucleic acid synthesis, modulate auxin (IAA)-mediated stress signaling, and sustain antioxidant capacity, respectively. Our findings establish a systems-level framework for understanding heat adaptation in P. pulmonarius and highlight key metabolic nodes that may serve as targets for molecular breeding of thermotolerant cultivars.