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.

This study reports the development of a new pure white Flammulina velutipes variety, ‘Seoldan’. The breeding process involved a primary cross between a brown strain (KMCC02267) collected by the Rural Development Administration and a white cultivar ‘Baekwoon’ (KMCC05472). From this cross, a white line ‘Fv-23-553’ was selected and subsequently crossed with a superior pure white resource (KMCC05474), resulting in the novel cultivar Seoldan. Mycelial growth tests showed that both varieties grew best at 25°C; however, Seoldan exhibited faster mycelial growth than the control variety under four temperature conditions except at 20°C. In media tests, Seoldan also demonstrated superior growth on three media types, with the exception of YM medium. In sawdust bottle cultivation, The time from scratching to the first fruiting was 2 days shorter in Seoldan than in the control variety, contributing to an overall reduction in cultivation time. The most distinct morphological trait of Seoldan is its pure white cap color. The average yield was 218.8 ± 10.6 g per bottle, comparable to that of the control. Morphologically, Seoldan produced smaller pileus and thinner stipes than the control. Furthermore, somatic incompatibility tests confirmed that Seoldan is genetically distinct from the control variety. These results indicate that Seoldan is a promising pure white variety with stable productivity, improved cultivation efficiency, and clear genetic differentiation from existing varieties.

Tricholoma matsutake, one of the most famous edible mushroom in Asia, has been cultivated in red pine forest. Because of its difficulty in artificial cultivation, T. matsutake cultivation has relied on foraging in the forest. Under certain environmental conditions, T. matsutake form the Shiro, the condensed mycelium, and develop into fruiting bodies. Among the certain environmental conditions, fungal communities play a major role in the mushroom development. Therefore, fungal community investigations for Bonghwa and Yangyang have been conducted on soil with fairy rings in the past, soil with existing fairy rings, and soil with presumptive fairy rings developing in the future. From the six soil samples, total 163 genera of fungi were detected and species diversity and species abundance of each sample were also analyzed. In result, the species diversity and species abundance of the fairy rings in Yangyang were lower than those in Bonghwa. In comparison with the fairy rings in Yangyang and Bonghwa, the dominance of the genus Tricholoma was higher and that of the genus Motilella was lower in Yangyang. Through the continuous study, establishment of optimal environmental conditions to promote the T. matsutake cultivation is highly expected.

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.

In order to develop a stable production and delicious unique cultivar of beech mushrooms, we generated hybrids using wild resources collected domestically, and we developed excellent strains by identifying the mycelial cultivability of the hybrid strains. The developed strains were cultivated according to the type of spawn and incubation time, and strains with excellent yield and shape were first selected, and second strains with less bitterness and excellent taste were selected through quantitative descriptive analysis. This was verified in farms, and ‘HM6-6’, which had a good reputation in the field, was directly developed cultivars as ‘Maruking’. In addition, the yield was high overall in the 90-day culture of the growth substrate inoculated with solid spawn, and the ‘Maruking’ cultivar, was also best in the 90-day culture. The yield was high overall in the 75-day culture of the liquid spawn, but ‘Maruking’ showed excellent yield when cultured for 70 days.

본 연구는 게임이라는 매체에서 음악이 단순한 배경 요소를 넘어 내러티브와 감정적 몰입을 심화하는 핵심적 요소로 작용할 수 있음을 규명하는 데 있다. 이를 위해 액션 어드벤처 게임 데스 스트랜딩에 개념적 혼성망(CIN) 이론을 적용해 라이선스 음악이 게임 내러티브와 결합하 여 메시지를 전달하는 방식을 분석하였다. 히데오 코지마 게임 감독은 특정 상황에 밴드 로우 로어의 세 곡을 의도적으로 배치하여 주인공의 여정과 주제적 요소를 상징적으로 나타냈다. CIN 이론에 따라 줄거리와 음악 입력 공간의 상호작용을 분석한 결과, 이러한 음악들이 게임 의 주제와 서사적 요소가 긴밀히 결합하여 의미를 형성함을 확인하였다. 본 연구는 음악과 게 임 간의 상호작용을 탐구하는 새로운 관점을 제시하며 CIN 이론을 활용한 게임 음악 분석 분 야에서의 연구 방향과 그 잠재적 효과를 시사한다.

Oyster mushroom is one of the most widely cultivated and consumed mushrooms in Korea, and mechanization and automation of cultivation systems have enabled mass production. Many cultivars have been developed to replace the old ones such as ‘Suhan‘ and ‘Chunchuneutari 2 ho,‘ which have been cultivated for over 20 years. Among these, ‘Soltari‘ was developed in 2015. Although it has excellent quality, its cultivation is challenging and the productivity is somewhat lower. To address these issues, the Mushroom Division at the National Institute of Horticultural and Herbal Science selected the genetic resource KMCC05165 and attempted hybridization between monokaryons from KMCC05165 and ‘Soltari(KMCC04940)’. Through repeated cultivation tests and evaluation of fruiting body characteristics, the superior strain ‘Po-2019-smj22’ was selected and finally named ‘Otari‘. The optimal mycelial growth temperature of ‘Otari’ was between 25 and 30°C and optimal fruiting body growth temperature was between 13 and 18°C. Mycelial growth on PDA medium was best at 25°C, and at the same temperature, mycelial growth was similar across four media: PDA, MEA, MCM, and YM. In 1,100 mL bottle cultivation, the yield was approximately 174 g, which is about 5% higher than the control cultivar ‘Soltari‘, and the number of valid individuals was also higher at about 25. The diameter and height of the pileus were 29.8 mm and 17.6 mm, respectively, slightly smaller than ‘Soltari‘, and the stipe was thin and long with a thickness of 12.2 mm. Additionally, the pileus’ lightness index (L index) was 30.7, indicating a darker brown color compared to 'Soltari.' With excellent mycelial growth, ease of cultivation, and high yield, the new cultivar ‘Otari‘ is expected to be widely adopted by domestic oyster mushroom farms.

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.

Background: Osteoarthritis is a common condition with an increasing prevalence and is a common cause of disability. Osteoarthritic pain decreases the quality of life, and simple gait training is used to alleviate it. Knee osteoarthritis limits joint motion in the sagittal and lateral directions. Although many recent studies have activated orthotic research to increase knee joint stabilization, no study has used patellar tendon straps to treat knee osteoarthritis. Objects: This study aimed to determine the effects of patellar tendon straps on kinematic, mechanical, and electromyographic activation in patients with knee osteoarthritis. Methods: Patients with knee osteoarthritis were selected. After creating the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), leg length difference, Q-angle, and thumb side flexion angle of the foot were measured. Kinematic, kinetic, and muscle activation data during walking before and after wearing the orthosis were viewed. Results: After wearing the patellar tendon straps, hip adduction from the terminal stance phase, knee flexion from the terminal swing phase, and ankle plantar flexion angle increased during the pre-swing and initial swing phases. The cadence of spatiotemporal parameters and velocity increased, and step time, stride time, and foot force duration decreased. Conclusion: Based on the results of this study, the increase in plantar flexion after strap wearing is inferred by an increase due to neurological mechanisms, and adduction at the hip joint is inferred by an increase in adduction due to increased velocity. The increase in cadence and velocity and the decrease in gait speed and foot pressure duration may be due to joint stabilization. It can be inferred that joint stabilization is increased by wearing knee straps. Thus, wearing a patellar tendon strap during gait in patients with knee osteoarthritis influences kinematic changes in the sagittal plane of the joint.

The present study aimed at selecting a cost-effective substrate for king oyster mushroom based on the growth characteristics of fruiting body for use as a basic resource to ensure stable production on farms. Compositional analysis of substrates manufactured with different materials in each process revealed that the total carbon content was 42.24–48.22% higher and the total nitrogen content was 1.7–2.29% higher in the processed lot than in the control (i.e., substrate used by the farmhouse; 40.86% and 1.39%, respectively). Meanwhile, the carbon-to-nitrogen ratio was the highest in the control (27.9% vs. 19.12–27.88% in the processed lot). When cultured for 28 days, the mycelium growth was 11.5 and 11.3 mm in substrates 1 and 6, respectively, indicating the fastest growth; meanwhile, the values were 10.1–10.3 mm in the control and substrate 11, showing a similar tendency. Mycelial density did not differ significantly among the processed lots. Yield per bottle was higher in substrates 8 (205.95 g/bottle), 7 (178.51 g/bottle), and 11 (170.63 g/bottle) than in the control (152.2 g/bottle). Fruiting body quality was comparable to controls in all processed lots. Overall, economic effects, such as substrate material prices, should be analyzed, and stability evaluations, such as residual pesticide and harmful microorganisms, should be undertaken along with further detailed examination to ensure safe and stable production on farms.

본 연구는 조사료 원료인 마늘 껍질의 사료적 가치와 메탄 발생량을 조사하기 위해 in vitro 발효 실험을 수행하였다. Garlic husk를 제외하고 국내에서 널리 사용되는 4가지 조사료 원료(Oat hay, Annual ryegrass, Timothy, Tall fescue)를 실험의 처리구로 구성하였다. In vitro 48 시간 발효 후 건물 소화율, 섬유소 소화율, 암모니아태 질소, 휘발성 지방산, pH, 메탄 발생량을 평가하였다. 가스 발생량은 3, 6, 9, 12, 24, 36 및 48 시간에 각각 측정되었다. 최종 가스 발생량에서 Oat hay가 유의적으로 가장 높았다(p<0.01). Garlic husk는 처리구 중 가장 높은 메탄 발생량을 나타내었다(p<0.01). Garlic husk의 건물 소화율은 Oat hay보다 유의적으로 낮았지만 Annual ryegrass, Timothy, Tall fescue보다 높았다(p<0.01). 섬유소 소화율은 Annual ryegrass에서 가장 낮았다(p<0.01). 총 휘발성 지방산 생성량은 Oat hay에서 가장 높았고(p<0.01), Garlic husk, Timothy, Tall fescue는 유사하였다. 본 연구의 in vitro 발효 성상과 메탄 발생량을 고려할 때, Garlic husk는 국내 주요 조사료를 대체할 영양적 가치를 포함하고 있으나 메탄 저감 효과는 미미할 것으로 추정된다.