The brix% of ASI 52822 resulted in the highest solid solubility. ASI 53399, ASI 50167, and ASI 52822 displayed high carbohydrate contents, whereas GBGA 01, GBGA 02, and ASI 52823 exhibited the half those values. The highest protein content was measured in ASI 52822. The antioxidant activity in the case of the hydrothermal method was lower than that for ascorbic acid, although it revealed more than 65% antioxidant activity. Furthermore, ASI 53399 and ASI 52822 exhibited antioxidant activities of 83.8% and 81.3%, respectively. To confirm the anticancer activity of the Ganoderma applanatum extract, we measured the original extract and also prepared a 10-time extract. The 10-time extract yielded strong anticancer activity, although the original extract did not exhibit any.

CRISPR/Cas9 ribonucleoprotein (RNP)-mediated gene editing has recently been applied to Ganoderma lucidum as a promising tool for functional genomics and strain improvement. However, the multinuclear nature of this basidiomycete can result in genetic mosaicism, raising concerns about the long-term stability of edited strains. In this study, we report the occurrence of revertant phenotypes in CRISPR/Cas9-edited transformants of G. lucidum. Although the edited colonies initially exhibited the expected phenotype, repeated subculturing led to the reappearance of wild-type phenotypes. PCR and sequencing analyses revealed the coexistence of edited and non-edited nuclei, and the progressive loss of edited genotypes over successive generations. These findings demonstrate that multinuclearity is a key factor contributing to the instability of CRISPR/Cas9-based edits in G. lucidum. This brief report provides the first direct documentation of revertant occurrence in edited G. lucidum strains and underscores the need for rigorous selection strategies and novel approaches to secure stable homokaryotic transformants in mushroom genetic engineering.

Despite their historical use, studies on the genetic functions of mushrooms and varietal improvement via biomolecular techniques are limited compared to other organisms. Recent advancements in CRISPR/Cas9 have enabled precise genetic modifications in mushrooms, with RNP-based systems offering high editing efficiency without foreign gene insertion. In this study, we optimized gene-editing conditions for Ganoderma lucidum (Yongji 2) by utilizing RNP/nanoparticle complexes to enhance efficiency. The optimal conditions included a 0.2 M sorbitol buffer (pH 7.0) and a protoplast-to-complex ratio of 10:1. Among eight gRNAs designed for the catA gene, three were identified with high activity, and PEG-mediated transformation resulted in successful gene edits, primarily involving 1 bp deletions. The editing efficiency reached 7–8%, demonstrating that nanoparticle-supported RNP systems are effective for marker-free gene editing in mushrooms. These findings highlight a promising approach for advancing genetic research and varietal improvement in G. lucidum and other mushroom species.

Ganoderma lucidum, a fungus of significant scientific interest that encompasses a diverse array of bioactive compounds, has been extensively investigated for its potential health-promoting and disease-preventing properties. Ganodermanontriol, a triterpenoid, is the principal active component contributing to its biological activities. This study aimed to explore the anti-inflammatory properties of ganodermanontriol and to evaluate its potential as a functional ingredient. The expression of inflammatory mediators, including tumor necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS), and nitric oxide (NO) production was significantly inhibited in ganodermanontriol (1.25–5 μg/mL)-treated compared to that of untreated lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Additionally, the phosphorylation of mitogen-activated protein kinases (MAPK), specifically p38 and c-Jun N-terminal kinase (JNK), was markedly inhibited in ganodermanontriol (3–5 μg/mL)-treated cells. These findings indicate that ganodermanontriol possesses significant potential as both a prophylactic and therapeutic agent for immune-related disorders, owing to its ability to modulate inflammatory responses.

Processing of medicinal plants and mushrooms plays a crucial role in oriental medicine. This study investigated the antioxidant potency and chemical composition of antler-shaped Ganoderma lucidumprocessed under different heating temperatures (120 to 180 °C) and times (10 to 20 minutes). Processed samples were analyzed for their antioxidant activities, including total phenol content, DPPH, ABTS radical scavenging capacity, NO assay, ROS scavenging capacity, and chemical composition changes. Antioxidant potency was highest with processing at 180 °C for 10 min and at 210 °C for 10 min. This enhanced activity is likely attributed to changes in compound functional groups due to heat processing, which in turn influences antioxidant activity. In contrast to antioxidant potency, chemical components, including ganoderic acid D, ganodermanonitriol, and ergosterol, showed decreasing trends with increasing temperature and time.

Recently, active research in Korea and worldwide has begun to focus on gene function and cultivar development using gene editing tools. This research, in addition to studies on edible mushroom, aims to enhance the physical and biochemical characteristics of mushrooms for applications in materials and substance production. For these studies, efficient isolation of protoplasts from the target mushroom is critical. However, several commercial cell wall-lysing enzyme cocktails, including Novozyme234, Glucanex, and Lysing enzymes, have recently been discontinued. In this study, we aimed to identify alternative enzyme systems to replace the discontinued cell wall-lysing enzymes for stable isolation of protoplasts from Ganoderma lucidum. To select an optimal osmotic buffer, enzyme function in 0.6 and 1.2 M Sorbitol, Sucrose, Mannitol, and KCl was assessed. The effect of reaction time was also evaluated. Protoplast isolation efficiency of each alternative enzyme was tested using lysing enzymes from Trichoderma harzianum, Chimax-N, and Yatalase, either individually or in combination. This matrix of studies identified enzymes and optimal conditions that could replace the discontinued lysing enzymes.

This study investigated the microscopic characteristics and genetic relationships of Ganoderma applanatum fruiting bodies. Basidiospores were brown, ellipsoid, and had one or two large vacuoles and a double wall. The surface of basidiospores was smooth or wrinkled and most had numerous small and shallow holes. The length and width of basidiospores of Ganoderma applanatum isolates GBGA-01, GBGA-02, ASI 50167, ASI 52821, ASI 52822, ASI 52823, and ASI 53399 were on average 7.6×4.8 mm, 7.9×4.6 mm, 7.7×4.9 mm, 8.2×5.3 mm, 7.7×5.0 mm, 8.0×4.9 mm, and 7.9×4.9 mm, respectively. In contrast, the basidiospores of Ganoderma lucidum isolate ASI 7125 were 7.7×5.2 mm. Using the universal ITS1/ITS4 primer set, the ITS region of the isolates were amplified and sequenced. The ITS sequences were very closely related to G. applantum isolate GBGA-01, GBGA-02, ASI 50167, ASI 52821, ASI 52822, ASI 52823 and ASI 53399, but were not the same species. Whereas, G. lucidum isolate ASI 7125 belongs to different group.

Owing to its diverse range of bioactive compounds, Ganoderma lucidumhas garnered significant research attention for health promotion and disease prevention. Ganodermanondiol, which has a triterpenoid structure, is one of the major active compounds of G. lucidum. In the present study, the anti-inflammatory effects of ganodermanondiol were investigated to evaluate its usefulness as a functional ingredient. Ganodermanondiol (0.5–2 g/mL) significantly inhibited the production of nitric oxide (NO), the expression of the cytokines tumor necrosis factor (TNF)??and interleukin 6 (IL-6), and the expression of cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) in lipopolysaccharide-induced RAW 264.7 (murine macrophage) cells. Ganodermanondiol (0.5–2 g/mL) also inhibited the phosphorylation of mitogen-activated protein kinase (MAPK) signal molecules, including p38 and c-Jun N-terminal protein kinase (JNK) in RAW 264.7 cells. Ganodermanondiol significantly inhibited the essential factors involved in the inflammatory responses of RAW 264.7 cells and would, therefore, serve as a potential prophylactic and therapeutic agent for immune-related diseases.

Ganoderma applanatum is one of the most popular medicinal mushrooms due to the various biologically active components it produces. In order to study for the possibility of artificially cultivating G. applanatum, we investigated the growth status of G. applanatum mycelium using oak sawdust bottle(850 ml). It took approximately 17~27 days for G. applanatum cultivation and 15~16 days for G. lucidum cultivation with sawdust until the mycelial growth was complete. The weight of dried fruiting bodies showed that G. applanatum GBGA-01 weighed 16.0±7.4 g, and G. lucidum ASI 7125 weighed 7.9±2.7g. The thickness of the pileus was measured as 10.6±2.6 mm for G. applanatum GBGA-01, 16.3±8.7 mm for G. applanatum ASI 52823 and 8.6±3.5 mm for G. lucidum ASI 7125. The color of Ganoderma spp. showed that L degree was 55.5±1.1 for the contex of G. applanatum GBGA-01, 47.7±2.2 for the contex of G. applanatum ASI 52823 and 39.6±2.9 for the contex of G. lucidum ASI 7125.

Recently, with the increasing global demand for Ganoderma lucidum as a material for functional beverages, varieties with enhanced functionality are needed. As part of this effort, 10 strains were selected from a collection of 160 G. lucidum strains, based on their quantity and fruiting body characteristics. These included four strains collected in Korea, three in Japan, and three of unknown origin. Approximately 44% of the 1,290 hybrid strains were secured. After two rounds of bag cultivation in oak sawdust, three strains (G-20-4, G-20-5, and G-20-7) were ultimately selected for single-timbered cultivation. Among these, G-20-4 demonstrated the earliest harvesting with a large size and a bright cap. Additionally, this strain exhibited the highest levels of total glucan, β-glucan, total triterpenoid, total polyphenol, and total flavonoid content.

High-income mushroom crops require complex farming. Therefore, we conducted a test to identify the optimum temperature for the production of antler-shaped Ganoderma lucidum using the King Oyster mushroom cultivation facility. T-N showed 0.28% of oak sawdust and 2.2% of nutritional source. The pH of oak sawdust was 6.0, indicating weak acidity, and that of rice steel was 6.6, indicating neutrality in nutrition source. Study on the quality characteristics of mushrooms showed that the number of days at 25°C were 5~6 and those at 30°C were 3~5; the representative length at 25°C was 57.5 mm and that at 30°C was 92.2 mm; the biological weight at 25 °C was 43 g, which was greater than that at 30°C.

한약재로 사용되는 더덕과 도라지에 표고의 유용성분을 함유한 소재를 개발하고자 더덕표고균사발효물(FCLM)과 더덕도라지균사발효물(FPLM)을 제조하였다. 두 가지 발효물의 일반성분, 유리당, 유기산, β-glucan, ergothioneine, ergosterol, vitamin D2 함량 및 3T3-L1 지방전구세포의 분화에 미치는 영향을 연구하였다. 일반성분 분석결과, FCLM에서 조섬유와 조지방함량이 FPLM에 비하여 높게 나타났으며, FPLM은 조단백질과 가용성무질소물의 함량 이 FCLM에 비하여 높게 나타났다. 유리당은 FCLM과 FPLM 모두 arabinose, glucose, sucrose 3종이 검출되었으며, 총 유리당 함량은 FPLM에서 높게 나타났다. 유기 산 함량은 발효 전 더덕과 도라지에 비하여 발효 후 FCLM과 FPLM에서 낮게 나타났다. β-Glucan 함량은 FCLM과 FPLM 두 발효물에서 대조구로 사용한 표고보다 높은 함량을 보였다. 항산화 물질로 알려진 ergothioneine의 함량은 FCLM이 FPLM보다 높게 나타났다. Ergosterol 함량은 대조구로 사용한 표고에서 가장 높게 나타났으며, 발효물들은 비슷한 함량을 나타내었다. Vitamin D2는 발효물들에서만 검출되었으며, FPLM (0.58±0.01 mg%)가 FCLM (0.47±0.01 mg%)보다 높은 함량을 보였다. 더덕과 도라지 표고균사발효물인 FCLM과 FPLM는 3T3-L1 지방전구세포에 대하여 발효 전 더덕과 도라지에 비하여 높은 수준의 세포 생존율을 나타내었다. 또한 FCLM과 FPLM은 3T3-L1 지방전구세포 분화를 발효 전 더덕과 도라지에 비하여 높은 수준으로 억제하여, 비만 억제 효과 가 있는 것으로 기대된다.

Anti-melanogenesis and skin anti-wrinkle effects of methanol (ME) and hot water (HE) extracts from the fruiting bodies of Ganoderma applanatum were investigated in this study. The total phenolic contents of the ME and HE of the mushroom were 11.68 and 3.15 μg GAEs/mg, respectively, whereas the total flavonoid contents of the ME and HE were 21.82 and 2.69 μg QEs/mg, respectively. The survival rate of B16-F10 murine melanoma cells treated with 750 μg ME and HE were 83.46% and 85.54%, respectively, thereby suggesting that mushroom extracts were slightly cytotoxic at the tested concentration. The in vitro tyrosinase inhibition by ME (83.15%) and HE (83.44%) was significantly lower than that of kojic acid (99.61%), the positive control, at 2.0 mg/mL. Although the inhibition of cellular melanin synthesis in B16-F10 melanoma cells by 2.0 mg/mL of ME (50.24%) and HE (51.24%) was lower than that of arbutin (64.84%), the inhibition by both ME and HE was higher than 50%. Collagenase inhibition by HE was comparable to 2.0 mg/mL epigallocatechin (EGCG), the positive control; however, elastase inhibition by ME and HE was lower than that of EGCG at the concentration tested. The results showed that the fruiting bodies of G. applanatum had good anti-tyrosinase, good anti-collagenase, and moderate anti-elastase activities, which might be useful for developing novel skin-whitening and anti-wrinkle agents.