{"paper_id":"2ab1b866-4057-4e95-a027-e8c92b048bca","body_text":"Beyond Static Control: A Dynamic Environmental Regime Informed by Structural Equation Modeling Enhances Yield and Quality of Pleurotus geesteranus | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Beyond Static Control: A Dynamic Environmental Regime Informed by Structural Equation Modeling Enhances Yield and Quality of Pleurotus geesteranus Songtao Lan, Yanting Li, Liping Luo, Enbei Xie, Shichan Pang, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8225452/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Overcoming the trade-off between yield and quality in mushroom cultivation requires a deeper understanding of how environmental factors coordinately regulate morphogenesis. This study introduces a paradigm shift from static environmental control to a phased, dynamic strategy for Pleurotus geesteranus. We proposed that sequentially optimizing conditions for distinct developmental phases—first enhancing stipe elongation under high temperature/CO₂, then initiating pileus expansion under moderated temperature with ventilation and light—would synchronize yield and quality. Validated in a digitally-controlled fruiting cabin (DSMFC), this strategy significantly increased yield (304.2 g/bag), biological efficiency (46.7%), and the premium mushroom ratio (46.0%) over traditional systems. The underlying mechanisms were elucidated via Structural Equation Modeling, which quantified accumulated heat units and CO₂ concentration as the pivotal environmental variables directly influencing stipe morphology and indirectly boosting yield. Economic analysis confirmed commercial viability, highlighting the potential of this targeted environmental programming to advance precision horticulture. Biological sciences/Ecology Earth and environmental sciences/Ecology Earth and environmental sciences/Environmental sciences Biological sciences/Plant sciences Pleurotus geesteranus Dynamic environmental control Two-stage cultivation Yield and quality Structural equation modeling (SEM) Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction The oyster mushroom, Pleurotus geesteranus, is an important commercial edible fungus cultivated worldwide due to its unique flavor, nutritional value, and relatively short growth cycle （ Thongklang,2016;Effiong,et al.，2024;Yu,et al.，2020 ） . Its production plays a significant role in ensuring global food security, increasing farmers' income, and promoting regional economic development ( Song,et al.，2022;Boukary,et al.，2024 ) . However, the full potential of its production is often unrealized due to inadequate environmental control, epitomized by the persistent challenge of simultaneously achieving high yield and superior quality. Current cultivation methods, ranging from forest-based analog systems to simple sheds and traditional mushroom houses, struggle to achieve precise and stable control over key environmental parameters such as temperature, humidity, light, and CO₂ concentration ( Jarial,et al.，2024;Yang,et al.，2024;Hausiku,2022 ) . These traditional approaches often rely on maintaining static environmental conditions, which fail to accommodate the distinct requirements of different developmental stages of mushroom morphogenesis, such as stipe elongation and pileus expansion. Consequently, the commercial quality of fruiting bodies (e.g., shape, cap-to-stem ratio) is often inconsistent, leading to a low proportion of premium-grade mushrooms and severely limiting economic returns ( Ten,et al.，2021;Islam,et al.，2017 ) . This limitation is exacerbated by global climate change, which increases the frequency and intensity of extreme weather events, further highlighting the vulnerability of traditional cultivation models ( Hoegh-Guldberg,et al.，2019;Chen,et al.，2022 ) . The core of this bottleneck lies in the complex, non-linear interactions between environmental factors and mushroom growth and development. Conventional research methods have difficulty systematically deciphering the underlying causal pathways. Therefore, a paradigm shift from \"static control\" to \"dynamic programming\" is urgently needed in edible fungus cultivation. This study explores a novel intelligent cultivation pathway based on developmental biology logic: actively \"programming\" the morphogenesis of P. geesteranusthrough the temporal regulation of environmental conditions. To this end, we developed a Digital Smart Mushroom Fruiting Chamber (DSMFC), integrated with photovoltaic power generation and Internet of Things (IoT) technology, serving as the hardware platform for implementing dynamic control. More critically, we introduced Structural Equation Modeling (SEM) as a powerful analytical tool (Cheng,et al.，2023;De La Croix,et al.，2022). Although SEM has found applications in agricultural environmental research, its potential in deciphering complex systems in edible fungus cultivation remains underexplored (Sunghyoun,et al.,2015;Seung-Mi,et al.,2015;Sakamoto,2018). The strength of SEM lies in its ability to quantify the direct and indirect effect pathways of multiple environmental factors on the final yield, thereby providing a solid mechanistic basis for optimizing control strategies (Lee,et al.,2018;Hair,et al.,2019;Hair,et al.,2016;A,et al.,2020). Using P. geesteranusas a model, this study compares the DSMFC with traditional cultivation environments. By employing principal component analysis, correlation analysis, and SEM (Sa,et al.,2025;Liu,et al.,2023), we aim to:Validate the effectiveness of a novel two-stage dynamic regulation strategy—promoting stipe elongation under high temperature/high CO₂ followed by stimulating pileus development under lower temperature with ventilation and light—in synergistically enhancing yield and quality.Uncover the key pathways through which pivotal environmental factors like accumulated heat units and CO₂ concentration drive morphogenesis and yield formation.Evaluate the economic viability of this intelligent cultivation mode, providing a replicable theoretical framework and technical example for the intelligent upgrading of the edible mushroom industry. 2. Materials and Methods 2.1 Experimental Site and Period The experiment was conducted in March 2025 in Xixiangtang District, Nanning City, Guangxi Zhuang Autonomous Region, China. The specific locations were the Guangxi Vocational University of Agriculture (108°14'40\"E, 22°51'16\"N) for the Traditional Mushroom Cultivation House (TMCH), Glass Greenhouse (GGH), and Passion Fruit Orchard Shade (SPO) treatments, and the Guangxi Minzu University (108°13'58\"E, 22°50'21\"N) for the Digital Smart Mushroom Fruiting Chamber (DSMFC). The experimental area is characterized by a typical tropical monsoon climate at an altitude of 86 meters. During the experimental period (March), the weather was predominantly cloudy with low light intensity, scant rainfall (average approx. 52 mm), and an average air temperature ranging from 15°C (night) to 21°C (day). 2.2 Mushroom Strain and Substrate Preparation The Pleurotus geesteranusstrain used in this study was ‘Jinxiu’, a widely cultivated commercial variety in China. The spawn was purchased from Guangxi Longzhou Beibu Gulf Modern Agriculture Co., Ltd. The substrate formulation was adapted from Ke et al. (2023) and consisted of the following components (by weight): 25.5% cottonseed hulls, 22% corn cobs, 20% wheat bran, 12% sawdust, 8% bagasse, 5% corn flour, and 5% soybean meal, supplemented with 1% light calcium carbonate, 1% gypsum, and 0.5% lime. The corn cobs were pre-wetted for 30 minutes before being thoroughly mixed with other components for 40 minutes (Ke,et al.,2023). The moisture content of the mixture was adjusted to 61-62%, and the initial pH was regulated to 8.0-9.0 (targeting pH 6.0-6.5 post-sterilization). The substrate was packed into high-pressure polypropylene bags (18 cm × 36 cm, 0.04 mm thickness). Each bag was filled to a height of 18 cm, resulting in a net weight of 1.1-1.3 kg. Sterilization was performed using a standard autoclaving cycle, which included a final stage of 121°C at 0.12 MPa for 150 minutes, followed by a 30-minute holding period. After cooling to approximately 90°C, the bag-logs were transferred to a class-10,000 clean room for forced cooling until the core temperature reached ~23°C. Aseptic inoculation was then performed in a class-100 laminar flow cabinet, with each bag receiving 20 mL of liquid spawn before being sealed for mycelial colonization . 2.3 Experimental Design and Fruiting Environment Treatments A completely randomized design was employed. Eighty bag-logs with uniform mycelial growth (≥95% coverage at the incision site) were selected and randomly assigned to four treatment groups (20 biological replicates per group), each corresponding to a distinct fruiting environment:SPO (Shade of Passion Fruit Orchard): Under two-year-old passion fruit vines providing natural shade and a cool microclimate.GGH (Glass Greenhouse): Equipped only with a sunshade net and axial flow fans.TMCH (Traditional Mushroom Cultivation House): Equipped with fans and an Emerson Copeland Scroll air compressor for manual temperature adjustment. DSMFC (Digital Smart Mushroom Fruiting Chamber):A self-developed system utilizing IoT technology for precise multi-factor environmental control, powered by integrated photovoltaic panels, smart sensors, and a closed-loop control module.A multi-functional remote environmental monitor was used to continuously record temperature, relative humidity, CO₂ concentration, and light intensity in each treatment throughout the fruiting period until harvest. 2.4 Two-Stage Dynamic Regulation Strategy in DSMFC Upon completion of the 64-day mycelial colonization period, bag-logs in the DSMFC underwent a precise, two-stage priming management process to induce and support fruiting body development:Primordia Induction Phase: A controlled temperature reduction (2°C/h) was applied to lower the temperature to 12.0 ± 0.5°C for 12 hours to stimulate primordia differentiation. Fruiting Body Development Phase: Following the low-temperature induction, a 1-2 cm segment was aseptically removed from the bag opening. The IoT control system was then activated to maintain the environment at 25.0 ± 1°C and 85-95% relative humidity for the first 20 hours, with CO₂ concentration dynamically controlled by fans and cooling systems until mycelial recovery at the incision site reached ≥95%. Subsequently, to promote pileus development, conditions were adjusted to approximately 20°C, supplemented with blue LED light, and CO₂ concentration was maintained around 2000 ppm. 2.5 Sample Collection and Measurements 2.5.1 Yield, Premium Mushroom Ratio, and Agronomic Traits For each treatment, yield per bag was measured daily from March 11th to 13th using an electronic balance. Harvested mushrooms were graded into Grade-1, Grade-2, and Grade-3 according to the Jiangxi Provincial Standard (DB36/T 824-2023). The premium mushroom ratio was calculated as: Premium Mushroom Ratio (%) = (Weight of Grade-1 mushrooms + Weight of Grade-2 mushrooms) / Total mushroom weight × 100. Grade-2 mushrooms were selected for agronomic trait measurements: individual mushroom weight, pileus weight, and stipe weight were recorded to calculate the pileus-to-stipe ratio. The maximum pileus diameter and minimum pileus width were measured to determine the length-to-width ratio. The diameter of the stipe at its midpoint was measured using a vernier caliper (Yan,et al.,2024;Yu,et al.,2023;Yu,et al.，2021;). 2.5.2 Nutritional Quality Analysis Approximately 200 g of fresh mushroom samples (a mix of pileus and stipe) from each treatment were freeze-dried in liquid nitrogen, ground into a fine powder, and analyzed in triplicate. The contents of crude polysaccharides, crude fat, crude protein, crude fiber, moisture, total saponins, and total phenols were determined using established standard methods as referenced (He,et al.,2012;Jeong,et al.,2025). 2.5.3 Texture Profile Analysis (TPA) Four mushrooms per treatment were randomly selected. The central part of the pileus (approx. 4 cm²) and the mid-section of the stipe (approx. 2 cm length) were prepared as test samples. Textural properties, including hardness, chewiness, cohesiveness, springiness, adhesiveness, and gumminess, were measured using a TMS-PILOT texture analyzer with a 50-mm diameter cylindrical probe. The test conditions were: pre-test speed: 60 mm/min, test speed: 60 mm/min, post-test speed: 60 mm/min, compression strain: 60%, and trigger force: 0.15 N. 2.6 Statistical Analysis Data compilation was performed using Microsoft Office LTSC Professional Plus 2021. Graph plotting and correlation analysis (significance level set at p < 0.05) were conducted using OriginPro 2024. One-way analysis of variance (ANOVA), Principal Component Analysis (PCA), and correlation heatmaps were performed using IBM SPSS Statistics 27. The Partial Least Squares Structural Equation Modeling (PLS-SEM) analysis was conducted using SmartPLS 4(Chen,et al.,2024;Hair,et al.,2021). 3. Results 3.1 Characterization of Environmental Conditions Across Cultivation Systems The environmental parameters across the four cultivation systems exhibited distinct dynamic patterns, which fundamentally shaped the subsequent growth and development of P. geesteranus. 3.1.1 Temperature and Relative Humidity Dynamics The temperature and relative humidity profiles for each treatment throughout the fruiting period (March 9-13) are shown in Fig. 1. The DSMFC treatment demonstrated a well-defined temperature trajectory, initially rising to a peak of 30.3°C at 17:00 on March 10th, then decreasing to a minimum of 14.4°C by 08:00 on March 12th, before stabilizing around 20°C (Fig. 1a). This pattern reflected the intentional two-stage regulatory strategy. While SPO and GGH showed similar diurnal fluctuation trends, SPO maintained the lowest overall temperature. In contrast, GGH experienced the highest daytime temperatures but cooled rapidly at night, approaching SPO levels around 05:00. TMCH provided the most stable thermal environment, consistently maintaining around 20°C. The average temperature was highest in TMCH and lowest in SPO, with both being significantly different from other treatments (Fig. 1c). Relative humidity across all treatments displayed a characteristic diurnal pattern, decreasing during the day and increasing at night (Fig. 1b). Occasional sharp fluctuations in TMCH and DSMFC were attributable to the activation of ventilation fans. The average relative humidity was comparable (approximately 93%) in SPO, TMCH, and DSMFC, but significantly lower (87.7%) in GGH (Fig. 1d). 3.1.2 CO₂ Concentration and Light Intensity Profiles The CO₂ concentration and light intensity profiles are critical differentiators among the systems (Fig. 2). The DSMFC treatment was characterized by the highest and most dynamically managed CO₂ environment. The concentration peaked at 9998.55 ppm at 11:00 on March 11th, followed by a decline due to activated ventilation and cooling systems, eventually stabilizing around 2000 ppm (Fig. 2a, b). In contrast, the CO₂ concentrations in TMCH, SPO, and GGH were not significantly different from each other, although TMCH showed greater variability (Fig. 2e).Light intensity was predominantly governed by the natural solar cycle in SPO, resulting in the highest average intensity among the treatments (Fig. 2c, f). TMCH employed continuous artificial lighting at an intensity of approximately 33 lux (Fig. 2d). The DSMFC was maintained in near-darkness (0.002 lux) except for a brief period of blue LED illumination to promote pileus development in the later stage. Consequently, the light intensity in TMCH was significantly higher than in both DSMFC and GGH (Fig. 2f). 3.2 Effects of Cultivation Environments on Yield, Quality Grade, and Morphological Traits The cultivation environment exerted a profound and systematic impact on the yield structure, commercial quality, and morphology of P. geesteranus. The Digital Smart Mushroom Fruiting Cabin (DSMFC) demonstrated superior performance across most evaluated metrics. DSMFC achieved the highest total yield (304.2 g/bag) and biological efficiency (46.7%), which were significantly greater than all other treatments (Fig. 3a, c). This high yield was attributed to a significantly greater number of fruiting bodies per bag (82.2) compared to other environments (Fig. 3b). Crucially, the DSMFC strategy effectively translated this high yield into superior commercial value. It produced the highest proportion of premium-grade mushrooms (Grade-1 + Grade-2) at 46.0%, driven predominantly by a substantial yield of Grade-1 mushrooms (102.0 g/bag) that was nearly absent in traditional systems (Fig. 3e, f, m). In contrast, the traditional facilities (GGH and TMCH) presented a \"high-yield, low-quality\" paradox. While their total yields were comparable to each other and higher than SPO, they severely lacked Grade-1 mushrooms, resulting in low premium ratios (Fig. 3m). The SPO treatment consistently underperformed, yielding the lowest in both total output (132.8 g/bag) and biological efficiency (19.9%). Morphologically, DSMFC-grown mushrooms exhibited the most uniform appearance with the lowest incidence of malformations (Fig. 3d). While no significant differences were detected in pileus dimensions (length, width, weight) across treatments (Fig. 3n,o,p), stipe morphology was distinctly affected. DSMFC promoted the development of significantly longer (5.4 cm) and thinner (7.1 mm diameter) stipes compared to others (Fig. 3q, r). This specific stipe architecture (long and thin), coupled with a lower pileus-to-stipe weight ratio, is a key characteristic associated with higher commercial grade in P. geesteranus. Although individual mushroom weight was slightly higher in GGH, the difference was not statistically significant (Fig. 3v). 3.3 Comprehensive Analysis of Quality, Multivariate Relationships, and Economic Benefits A comprehensive evaluation of the Pleurotus geesteranusfruiting bodies under different cultivation environments revealed systematic differences in nutritional quality, eating texture, the drivers of key traits, and ultimately, economic returns. 3.3.1 Nutritional and Textural Quality Analysis of nutritional components showed that the most significant differences were observed in the moisture content and crude fiber content of the fruiting bodies (Table 1). The moisture content was highest in the DSMFC treatment (86.89 ± 0.34%), significantly exceeding other treatments and indicating better freshness. Conversely, the crude fiber content was significantly higher in the SPO treatment (43.91 ± 1.04%), potentially related to the natural stress environment it experienced. Other nutritional components (e.g., crude protein, crude fat) showed no significant differences among treatments. Table 1. Effects of different treatments on the nutritional quality of Pleurotus geesteranus. Treatment Moisture content (%) Crude protein (%) Crude fat (%) Crude fiber (%) Crude polysaccharide (g/kg) Total saponins (g/kg) Total phenolics (g/kg) SPO 84.05±1.78b 40.56±2.43a 1.13±0.26a 43.91±1.80a 43.22±5.58a 7.69±0.48a 11.86±0.47a GGH 84.61±1.11b 38.8±1.32a 1.02±0.08a 35.85±3.47b 41.35±3.36a 7.92±0.49a 12.14±0.81a TMCH 85.54±0.58ab 40.15±2.25a 1.28±0.55a 36.22±3.31b 39.96±2.61a 7.22±0.22a 12.42±0.41a DSMFC 86.89±0.76a 39.34±2.21a 1.06±0.29a 35.89±3.62b 42.47±6.45a 7.74±0.08a 11.55±1.3a Different lowercase letters within a column indicate significant differences between treatments at the 5% level. 3.3.2 Multivariate Relationships and Driver Analysis Textural profile analysis (TPA) revealed significant treatment-specific variations in the texture of Pleurotus geesteranus (Fig. 4a-f). Stipes generally exhibited greater hardness and chewiness than pilei. The GGH treatment yielded the highest values for stipe hardness and chewiness, while DSMFC resulted in the highest pileus chewiness and gumminess. Stipe springiness was highest in GGH but lowest in DSMFC, which conversely showed the highest stipe adhesiveness. Cohesiveness and pileus springiness remained consistent across treatments. Principal Component Analysis (PCA) indicated that the primary composite factor affecting yield (PC1, 78.9% variance contribution) was strongly positively correlated with premium mushroom weight, premium mushroom count, and total mushroom count (Fig. 4g). PCA of environmental factors showed that accumulated heat units (AHU) and average temperature were the main positive driving variables (Fig. 4i). Correlation heatmaps further confirmed that temperature-related variables and average CO₂ concentration were significantly positively correlated with premium mushroom count and weight (Fig. 4k). Most importantly, Partial Least Squares Structural Equation Modeling (PLS-SEM) quantified the effect pathways of key environmental factors (Fig. 4l). The model revealed that accumulated heat units (AHU) and CO₂ concentration were the core factors driving stipe morphogenesis. They had highly significant positive direct effects on stipe length and diameter, which in turn indirectly promoted the increase in individual mushroom weight. The model explained a large portion of the variance in individual mushroom weight (R² = 0.932), clearly outlining the key regulatory pathway: \"Temperature & CO₂ → Stipe Morphology → Yield\". 3.3.3 Economic Benefits Economic benefit analysis ultimately validated the commercial value of the intelligent cultivation mode (Table 2). Despite higher equipment costs, the DSMFC, through precise environmental control, greatly optimized the yield structure: the Grade-1 mushroom yield reached 102.04 kg per 1000 bags, accounting for 62.1% of the total yield, far surpassing traditional modes. This significant \"premium mushroom price premium effect\" resulted in a net profit for the DSMFC (443.02 USD/1000 bags) that was substantially higher than those of GGH, TMCH, and SPO—by 96.0%, 129.7%, and 203.3%, respectively—fully demonstrating its great potential for market promotion. Table 2. Economic benefit analysis of different treatments (based on 1000 bags). Treatment Grade-1 yield (kg) Grade-2 yield (kg) Grade-3 yield (kg) Grade-1 revenue (USD) Grade-2 revenue (USD) Grade-3 revenue (USD) Total cost (USD) Net profit (USD) SPO 0 42.34 90.48 0.00 71.28 76.17 1.40 146.05 GGH 2.74 64.5 138.72 7.69 108.59 116.77 7.02 226.04 TMCH 4.134 48.54 143.23 11.60 81.72 120.57 21.05 192.85 DSMFC 102.04 37.32 164.84 286.32 62.83 138.76 44.90 443.02 Note: Yield calculations were based on 1,000 mushroom bags. According to the exchange rate, 1 Chinese Yuan (CNY) = 0.1403 US Dollars (USD). The prices for Grade-1, Grade-2, and Grade-3 mushrooms were set at 2.81 USD, 1.68 USD, and 0.84 USD, respectively. The primary production costs included electricity and water expenses. 4. Discussion 4.1 Characteristics of fruiting environments across treatments Environmental conditions are critical factors influencing mushroom growth, significantly affecting morphology and yield（Raudaskoski and Viitanen,1982,Sohi and Upadhyay,1989）. Different Pleurotus species require distinct growth environments, necessitating climate-adapted varieties and facility management strategies based on regional conditions(Yan,et al.,2024). Previous research has identified optimal artificial environmental parameters for Pleurotus morphological development: CO₂ concentration of 0.3% (with deformity rates increasing significantly above 0.5%), temperature of 13-16°C, humidity >80%, and ventilation rate of 0.2-0.5 feet/minute (exposed conditions). This parameter combination effectively suppresses abnormal fruiting body development(Jang,et al.,2003).This study observed that SPO provided the highest light intensity following natural diurnal patterns, with the lowest average temperature but relatively high average humidity (93%). However, it exhibited substantial diurnal fluctuations in both temperature and humidity, showing parallel but overall lower patterns compared to GGH. GGH relied on shading nets and fan ventilation for environmental control. Due to the greenhouse effect of glass enclosure, daytime temperatures exceeded those in TMCH, while radiative cooling at night caused rapid temperature loss, resulting in the largest day-night temperature variation. Concurrently, GGH maintained the lowest humidity levels (average 87.7%), making it difficult to sustain the stable high-humidity environment required by P. geesteranus.TMCH utilized compressors and fans for temperature and humidity regulation, providing the most stable environment with temperatures consistently maintained around 20°C and minimal fluctuations. However, forced ventilation caused \"jump\" oscillations in humidity, though the average humidity remained at 93%. This treatment employed constant artificial lighting at 33 lx.Due to high-density cultivation, DSMFC exhibited gradually increasing temperature and CO₂ concentrations, reaching a maximum temperature of 30.3°C, higher than the temperature settings reported by Lee et al(Lee,et al.,2018). Jhune et al found that low-temperature Pleurotus strains perform optimally at 10°C, while medium-high temperature strains show improved performance with increasing temperature(Jhune,et al.,2011). At 10°C, pileus thickness increased but the harvest period extended, and stipes became longer and thinner with rising temperatures, with some varieties exhibiting temperature stress-induced malformations. In DSMFC, although brief high temperatures occurred, the automatic cooling system activated promptly, preventing significant impacts on fruiting body development.Paul et al.(1983) suggested that high CO₂ concentrations cause stipe elongation and suppressed pileus development. Kinugawa et al.(1994) found that CO₂ concentrations exceeding 6,000 ppm for 7 days led to trumpet-shaped deformities in Pleurotus pileus and reduced yield. In this study, CO₂ concentrations exceeding 9,000 ppm were maintained for 27 hours before the ventilation and cooling systems gradually reduced both CO₂ levels and temperature. Despite these high concentrations, no malformations occurred because the increase was gradual during fruiting body development. However, some delayed-development bags showed shortened stipes and altar-shaped mushrooms, consistent with findings by Jang et al.(2003) and Lin et al.(2022).When stipe length reached approximately 5 cm, blue LED lighting was activated while maintaining temperature around 20°C and CO₂ concentration around 2,000 ppm to promote pileus development. Yang et al identified 25°C and 90% humidity as the optimal combination for summer cultivation of P. geesteranus, with misting humidification significantly reducing malformations and enhancing pileus coloration(Yang,et al.,2023). Building on this, DSMFC achieved initial gradual increases in temperature and CO₂ to promote stipe elongation, followed by cooling, ventilation, and light supplementation, ultimately reducing the fruiting cycle duration and energy consumption. 4.2 Effects of different fruiting environments on yield and agronomic traits of Pleurotus geesteranus The fruiting environment significantly influenced the yield, quality, and agronomic traits of P. geesteranus (Song,et al.,2025). DSMFC demonstrated exceptional performance across most indicators, with its highest yield (304.2 g), biological efficiency (46.7%), and premium mushroom ratio (46.0%) fully validating the effectiveness of its environmental regulation strategy.GGH exhibited high yield but low quality, accompanied by yellowing disease and size heterogeneity issues. These problems were closely associated with its environmental characteristics: high daytime temperatures, low humidity (average 87.7%), and substantial fluctuations in temperature and humidity, leading to significantly reduced market value.TMCH showed similar yield to GGH but also manifested a \"high yield with low value\" phenomenon: near absence of Grade I mushrooms, the lowest premium mushroom ratio (approximately 20%), thick stipes, and deformed pileus, all compromising marketability.The SPO treatment, mimicking ecological cultivation, demonstrated no advantages in either yield or quality. It produced the lowest yield (132.8 g), lowest biological efficiency (19.9%), and the highest number of malformed mushrooms. Fruiting bodies from SPO exhibited the shortest stipes and the highest pileus-to-stipe ratio. Although total yield was low, these mushrooms may possess unique flavor qualities worthy of further investigation.Nakazawa et al, Ohm et al and Terashima et al suggested that many mushrooms exhibit phototropism, and pileus differentiation in basidiomycetes is influenced by light conditions(Nakazawa,et al.,2008;Ohm,et al.,2013;Terashima,et al.,2005). The higher incidence of malformed mushrooms in TMCH and SPO may be attributed to either continuous 24-hour lighting (33 lx) in TMCH or uneven light distribution under tree shade in SPO. Additionally, relatively frequent ventilation or natural environmental fluctuations could contribute to abnormal fruiting body development. 4.3 Effects of Different Fruiting Environments on Quality and Textural Properties of Pleurotus geesteranus The fruiting environment not only altered the yield and appearance of P. geesteranus but also significantly influenced its nutritional quality and textural properties, which directly determine its commercial value, processing characteristics, and consumer palatability. Regarding nutritional quality, the most notable differences were observed in the moisture content and crude fiber content of the fruiting bodies. Jin et al reported that the moisture content of Pleurotus fruiting bodies varies with flush number, cultivar, and pileus size(Jin,et al.,2021).The DSMFC treatment yielded fruiting bodies with the highest moisture content (86.89%). This elevated moisture content enhanced their fresh and tender appearance but may negatively impact shelf life and drying efficiency. Conversely, fruiting bodies from the SPO treatment contained significantly higher crude fiber (43.91%) compared to other groups. This may be attributed to the stress conditions of stronger light exposure and lower temperatures, which potentially stimulate cellulose synthesis in the cell walls to enhance structural support.No significant differences were detected in crude protein, crude fat, crude polysaccharides, or active components (saponins and polyphenols) among treatments. This indicates that the synthesis of these nutritional compounds is primarily regulated by the cultivation substrate and genetic factors, combined with the short growth cycle of P. geesteranus, which limits substantial differential accumulation of nutrients.Texture profile analysis (TPA) objectively measures the physical properties of mushrooms by simulating oral mastication processes, thereby transforming subjective \"mouthfeel\" into quantifiable and reproducible data. This approach is crucial for scientific research, quality control, and product development(Zivanovic,et al.,2000;Kotwaliwale,et al.,2007;Kortei,et al.,2015).Zou et al found that hardness and chewiness of Pleurotus were significantly positively correlated and jointly determined palatability(Zou,et al.,2024). Stipe cohesiveness showed negative correlations with hardness and adhesiveness. Appropriate enhancement of cohesiveness can optimize texture, while excessively low hardness may lead to structural looseness. As an indicator of cellular binding force, cohesiveness serves as a key parameter for evaluating texture.This study revealed that the GGH treatment produced fruiting bodies with the highest stipe hardness, chewiness, and springiness. This firm and chewy texture likely resulted from the high-temperature and low-humidity stress conditions, leading to more compact cellular structures.The DSMFC treatment exhibited a unique textural profile: the highest chewiness and gumminess in the pileus, but the lowest springiness and gumminess in the stipe. This suggests independent development of pileus and stipe tissues, with the pileus being full and resilient while the stipe tended to be looser and more tender, potentially creating a superior overall mouthfeel experience.The TMCH treatment showed intermediate values for most textural parameters, consistent with its stable temperature-humidity environment but constant weak lighting conditions. 4.4 Relationships among Variables The results from partial least squares structural equation modeling (PLS-SEM), principal component analysis (PCA), and correlation analysis collectively revealed a clear network of pathways governing yield and quality formation in Pleurotus geesteranus, elucidating the complex mechanisms through which key environmental factors directly and indirectly influence final yield（Sa,et al.,2025;Zhao,et al.,2024;Wu,,et al.,2022）.The core pathways of the model identified accumulated heat units (AHU) and mean CO₂ concentration as the two most critical environmental drivers of yield and premium mushroom number. AHU exerted a highly significant positive direct effect on stipe elongation (path coefficient: 0.096), and moderately elongated stipes further supported heavier individual mushroom weight (0.093), forming a crucial pathway for premium yield accumulation.More notably, CO₂ demonstrated strong direct positive effects on stipe morphology (diameter: 0.383; length: 0.260). This indicates that moderately elevated CO₂ levels suppress lateral pileus expansion while redirecting assimilates to the stipe, forming the commercially desirable \"elongated stipe-compact pileus\" morphology. This mechanism enhances the proportion of Grade I mushrooms without compromising individual mushroom weight. These findings were mutually validated by correlation analysis, which showed extremely significant positive correlations between mean CO₂ concentration and both premium mushroom number and Grade I mushroom count.The model also revealed differential responses of various traits to environmental factors. For example, the low coefficient of determination for pileus length (R² = 0.200) suggests that its development is more influenced by microenvironmental fluctuations or genetic factors not included in the model, explaining why no significant differences in pileus size were observed in agronomic traits.Although mean light intensity (PPFD) and relative humidity (RH) did not reach significance in direct pathways, their positive trends and correlations with crude fiber content and pileus width (Figure 10) imply their roles as important regulatory factors. They may function synergistically with other factors (e.g., CO₂) through coupled regulation—for instance, light may suppress excessive elongation while promoting pileus development. 5. Conclusions This study develops and validates a novel two-stage dynamic environmental regulation strategy for Pleutotus geesteranuscultivation, implemented via a Digital Smart Mushroom Fruiting Chamber (DSMFC). The strategy, which sequentially applies high-temperature/high-CO₂ conditions to promote stipe elongation followed by cooler, ventilated, and lit conditions to optimize pileus development, successfully breaks the common trade-off between yield and quality. By employing Structural Equation Modeling, we quantitatively identified accumulated heat units (AHU) and CO₂ concentration as the pivotal environmental drivers of stipe morphogenesis and yield. This strategy significantly enhanced total yield (304.2 g/bag), biological efficiency (46.7%), and the premium mushroom ratio (46.0%), resulting in the highest net profit (443.02 USD/1000 bags). The research underscores that a paradigm shift from static control to dynamic environmental programming is key to advancing precision agriculture in edible mushroom cultivation, providing a replicable framework for simultaneously achieving productivity and economic gains. Declarations Author Contributions: Conceptualization, S.L. and Y.L. (Yanting Li); Methodology, S.L. and L.L.; Project administration, S.L. and L.L.; Writing – original draft, S.L.; Data curation, Y.L. (Yanting Li) and E.X.; Formal analysis, E.X.; Investigation, E.X., Y.Z., Q.Z. and W.C.; Resources, L.L.; Validation, S.P. and Y.Z.; Visualization, S.P.; Writing – review & editing, Y.L. (Yun Li); Funding acquisition, Y.L. (Yun Li). All authors have read and agreed to the published version of the manuscript. Funding: This research was supported by the Guangxi Major Special Project (No. Guike AA24263038), the Introduction of Talents Scientific Research Start-up Project of Guangxi Minzu University (No. 2024KJQD218), and the 2025 University-level Scientific Research Project of Guangxi Vocational University of Agriculture (No. XKJ2540). Data Availability Statement: The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation. The data are not publicly available due to copyright restrictions. Conflicts of Interest: The authors declare no conflicts of interest. References Thongklang, N., 2016. Testing agricultural wastes for the production of Pleurotus ostreatus. Mycosphere 7, 766-772. http://doi.org/10.5943/mycosphere/7/6/6 Effiong, M.E., Umeokwochi, C.P., Afolabi, I.S., Chinedu, S.N., 2024. Comparative antioxidant activity and phytochemical content of five extracts of Pleurotus ostreatus (oyster mushroom). Sci Rep 14, 3794. http://doi.org/10.1038/s41598-024-54201-x Yu, Q., Guo, M., Zhang, B., Wu, H., Zhang, Y., Zhang, L., 2020. Analysis of Nutritional Composition in 23 Kinds of Edible Fungi. Journal of Food Quality. https://doi.org/10.1155/2020/8821315. Song, J.-L., Wang, W.-k., Lu, N., Yan, J., Yuan, W.-d., Wu, Y.-L., 2022. Genetic diversity analysis of Pleurotus pulmonarius(Fr.) Quél. based on agronomic characters and ISSR markers. Plant Genome14, 20128. Boukary, A.A., Olou, A.B., Piepenbring, M., Yorou, N.S., Mushroom cultivation in tropical Africa: successes, challenges, and opportunities. Journal of Agriculture and Food Research. https://doi.org/10.1016/j.jafr.2024.101264 Jarial, R., Jarial, K., Bhatia, J., 2024. Comprehensive review on oyster mushroom species (Agaricomycetes): Morphology, nutrition, cultivation and future aspects. Heliyon 10. http://doi.org/10.1016/j.heliyon.2024.e26539 Yang, Z., Qu, J., Qiao, L., Jiang, M., Zou, X., Cao, W., 2024. Tea and Pleurotus ostreatusintercrop** modulates structure of soil and root microbial communities. Sci. Rep.14, 11295. Hausiku, M.K., 2022. Mushroom Cultivation in Arid Namibia: Cultivation Status, Contribution to Human Health and Future Prospects. https://doi.org/10.1007/978-981-16-6771-8_21 Ten, S.T., Krishnen, G., Khulidin, K.A., Mohamad Tahir, M.A., Hashim, M.H., Khairudin, S., 2021. Automated Controlled Environment Mushroom House. Advances in Agricultural and Food Research Journal. http://doi.org/10.36877/aafrj.a0000230 Islam, M.T., Zakaria, Z., Hamidin, N., Azlan, B.M.I., Mohd, Shi Fern, C., Abdullah, M.A.B., Abd Rahim, S.Z., Muhammad Suandi, M.E., Mat Saad, M.N., Ghazali, M.F., 2017. The Management of Humidifying Treatment for Low Contamination Risks during Indoor Cultivation of Grey Oyster Mushroom (Pleurotus pulmonarius). Matec Web of Conferences 97, 01080. http://doi.org/10.1051/matecconf/20179701080 Hoegh-Guldberg, O., Jacob, D., Taylor, M., Guillén Bolaños, T., Bindi, M., Brown, S., . . . Zhou, G. (2019). The human imperative of stabilizing global climate change at 1.5 °C. Science, 365(6459), Article eaaw6974. Chen, L., Qian, L., Zhang, X., Li, J., Zhang, Z., Chen, X., 2022. Research progress on indoor environment of mushroom factory. International Journal of Agricultural & Biological Engineering 15. http://doi.org/10.25165/j.ijabe.20221501.6872 Cheng, G., Chen, J., Lan, L. Y., et al. Grey correlational and genetic analyses of Pleurotus pulmonariushybrids. Molecular Plant Breeding, 2023, 21(23): 7811-7818. De La Croix, N.J., Didacienne, M., Louis, S., 2022. Fuzzy logic-based shiitake mushroom farm control for harvest enhancement, 2022 10th International Symposium on Digital Forensics and Security (ISDFS). IEEE, pp. 1-6.https://doi.org/10.1109/ ISDFS55398.2022.9800832. Sunghyoun, L., Byeongkee, Y., Hyuckjoo, K., Namkyu, Y., Jongcheon, J., 2015. Technology for Improving the Uniformity of the Environment in the Oyster Mushroom Cultivation House by using Multi-layered Shelves. Protected horticulture and Plant Factory 24, 128-133. http://doi.org/10.12791/ksbec.2015.24.2.128 Seung-Mi, Moon, Sook-Youn, Kwon, Jae-Hyun, Lim, 2015. Improvement of Energy Efficiency of Plants Factory by Arranging Air Circulation Fan and Air Flow Control Based on CFD. Journal of Internet Computing and Services 16, 57-65. http://doi.org/10.25165/j.ijabe.20221501.6872 Sakamoto, Yuichi, 2018. Influences of environmental factors on fruiting body induction, development and maturation in mushroom-forming fungi. Fungal Biology Reviews, S1749461317300568. http://doi.org/10.1016/j.fbr.2018.02.003 Lee, C.J., Lee, S.H., Lee, E.J., Park, H.S., Kong, W., 2018. Analysis of growth environment for precision cultivation management of the oyster mushroom 'Suhan'. Journal of Mushroom 16, 155-161. http://doi.org/10.14480/JM.2018.16.3.155 Hair, J. F., Hult G T, M., Ringle C M, M., Sarstedt, M., Castillo Apraiz, J., Cepeda Carrion, ´ G. A., et al., Manual de Partial least squares structural equation modeling (PLS-SEM) (2nd ed.). Los Angeles [Etc: Omnia Publisher SL Cop. https://doi. org/10.3926/oss.37 Hair, J.F., Hult, G.T.M., Ringle, C.M., Sarstedt, M., 2016. A Primer on Partial Least Squares Structural Equation Modeling (PLS-SEM), 2nd edition. A, Z.J., A, L.P., B, M.W.A., Systematic relationship between soil properties and organic carbon mineralization based on structural equation modeling analysis - ScienceDirect. Journal of Cleaner Production 277. https://doi.org/10.1016/j.jclepro.2020.123338. Sa, Q., Zheng, J., Zhang, K., Wang, Y., 2025. Effects and assessment of the combined application of biogas slurry and chemical fertilizers on greenhouse tomato growth, yield, and soil quality. Scientia Horticulturae 344. https://doi.org/10.1016/j.scienta.2025.114113. Liu, S.Y., Vega, A.R., Dizy, M., 2023. Assessing ultrapremium red wine quality using PLS-SEM. LWT 177, 114560-. https://doi.org/10.1016/j.lwt.2023.114560 Ke, B. R., Lan, Q. X., Lu, Z. H., et al. (2023). Genetic differences and agronomic traits comparison of main cultivated varieties of Pleurotus pulmonarius. Scientia Horticulturae, 11(4), 241–247. Yan, M., Zhai, D., Li, Q., Zhang, M., Jiang, N., Liu, J., Song, C., Shang, X., Chen, H., Yu, H., 2024. Comparative Analysis of Main Agronomic Traits of Different Pleurotus giganteus Germplasm Resources. Life (2075-1729) 14. https://doi.org/10.3390/life14020238 Yu, H.L.,Zhang, M.Y., Li, Q.Z.,Zhang, L.J.,Shang, X.D., Tan, Q., 2023. A new variety of Pleurotus giganteus ’Shen Xun No.1’. Acta Hortic.Sin. 50, 453–454. Yu, H.L.,Zhai, D.D.,Shen, X.F.,Zhang, M.Y.,Wang, Y.X., Shang, X.D.,Zhang, D., Li, Q.Z., 2021. Quantitative character variation andprobability classification of fruiting bodies of Pleurotus giganteus germplasm resources. Acta Edulis Fungi . 28, 42–47. He, J.Z., Ru, Q.M., Dong, D.D., Sun, P.L., 2012. Chemical Characteristics and Antioxidant Properties of Crude Water Soluble Polysaccharides from Four Common Edible Mushrooms. Molecules 17. https://doi.org/10.3390/molecules17044373 Jeong, H., Cho, J., Han, J., Yoon, Y.S., Kim, H.G., Kim, J., Chung, H., 2025. Large-area coverage–transmission near-infrared measurement of dried laver to determine crude protein content. Food Control 168. https://doi.org/10.1016/j.foodcont.2024.110934 Chen, X., Zhang, H., Yu, S., Zhou, C., Teng, A., Lei, L., Ba, Y., Li, F., 2024. Optimizing irrigation and nitrogen application strategies to improve sunflower yield and resource use efficiency in a cold and arid oasis region of Northwest China. Frontiers in Plant Science 15, 16. https://doi.org/10.3389/fpls.2024.1429548. Hair, J. F., Hult, G., Ringle, C. M., & Sarstedt, M., A primer on partial least squares structural equation modeling (PLS-SEM) (3rd ed.). Sage. Raudaskoski, M., Viitanen, H., 1982. Effect of aeration and light on fruit body induction in Schizophyllum commune. Transactions of the British Mycological Society 78, 89-96.https://doi.org/10.1016/S0007-1536(82)80080-6 Sohi, H., Upadhyay, R., 1989. Effect of temperature on mycelial growth of Pleurotus species and their yield performance on selected substrates. Mushroom Science 12(2): 49-56. Yan, M., Zhai, D., Li, Q., Zhang, M., Jiang, N., Liu, J., Song, C., Shang, X., Chen, H., Yu, H., 2024. Comparative Analysis of Main Agronomic Traits of Different Pleurotus giganteus Germplasm Resources. Life (2075-1729) 14. https://doi.org/10.3390/life14020238 Jang, K.Y., Jhune, C.S., Park, J.S., Cho, S.M., Weon, H.Y., Cheong, J.C., Choi, S.G., Sung, J.M., 2003. Characterization of Fruitbody Morphology on Various Environmental Conditions in Pleurotus ostreatus. Mycobiology 31.https://doi.org/4489/MYCO.2003.31.3.145. Lee, C.J., Lee, S.H., Lee, E.J., Park, H.S., Kong, W., 2018. Analysis of growth environment for precision cultivation management of the oyster mushroom 'Suhan'. Journal of Mushroom 16, 155-161. https://doi.org/10.14480/JM.2018.16.3.155 Jhune, C.-S., Yun, H.-S., Jang, K.-Y., Kong, W.-S., Lee, K.-H., Lee, C.-J., Yoo, Y.-B., 2011. The effect of temperature on morphological of fruiting body and cultivated characteristics of oyster mushroom. Journal of Mushroom 9, 155-160. Paul, S. and Chilton, J. S., 1983. Chapter environmental factors: sustaining the mushroom crop in the mushroom cultivator. pp. 149-157. Kinugawa, K., Suzuki, A., Takamatsu, Y., Kato, M., Tanaka, K., 1994. Effects of concentrated carbon dioxide on the fruiting of several cultivated basidiomycetes (II). Mycoscience 35, 345-352. https://doi.org/10.1007/BF02268504 Jang, K.Y., Jhune, C.S., Park, J.S., Cho, S.M., Weon, H.Y., Cheong, J.C., Choi, S.G., Sung, J.M., 2003. Characterization of Fruitbody Morphology on Various Environmental Conditions in Pleurotus ostreatus. Mycobiology 31. https://doi.org/10.4489/MYCO.2003.31.3.145 Lin, R., Zhang, L., Yang, X., Li, Q., Zhang, C., Guo, L., Yu, H., Yu, H., 2022. Responses of the Mushroom Pleurotus ostreatus under Different CO(2) Concentration by Comparative Proteomic Analyses. J Fungi (Basel) 8. http://doi.org/10.3390/jof8070652 Yang Y., Shen Y., Cao Y., Xu G., Cai W., Effects and Analysis of Temperature and Humidity on the Appearance Quality of Pleurotus pulmonarius During Summer Fruiting. Edible and Medicinal Mushrooms 31, 191-195. Song J., Chen Q., Zhang C., Zong T., Yuan W., Yu B., The effect of cultivation period on the agronomic traits and yield of Pleurotus eryngii. Journal of Northwest A&F University (Natural Science Edition) 53,137-144+156.http://doi.org/10.13207/j.cnki.jnwafu.2025.06.014 Nakazawa, T., Miyazaki, Y., Kaneko, S., Shishido, K., 2008. Stimulative effects of light and a temperature downshift on transcriptional expressions of developmentally regulated genes in the initial stages of fruiting-body formation of the basidiomycetous mushroom Lentinula edodes. FEMS Microbiology Letters. https://doi.org/10.1111/j.1574-6968.2008.01364.x. Ohm, R.A., Aerts, D., Wosten, H.A., Lugones, L.G., 2013. The blue light receptor complex WC-1/2 of Schizophyllum commune is involved in mushroom formation and protection against phototoxicity. Environ Microbiol 15, 943-955. http://doi.org/10.1111/j.1462-2920.2012.02878.x Terashima, K., Yuki, K., Muraguchi, H., Akiyama, M., Kamada, T., 2005. The dst1 gene involved in mushroom photomorphogenesis of Coprinus cinereus encodes a putative photoreceptor for blue light. Genetics 171, 101-108. http://doi.org/10.1534/genetics.104.040048 Jin R., Tan X., Ma W., Dong Y., Li F., 2021. Quality Evaluation of Pleurotus ostreatus Fruiting Bodies. China Vegetables 34, 74-79.http://doi.org/10.16861/j.cnki.zggc.2021.0236 Zivanovic, S., Busher, R.W., Kim, K.S., 2000. Textural Changes in Mushrooms (Agaricus bisporus) Associated with Tissue Ultrastructure and Composition. Journal of Food Science 65, 1404-1408. http://doi.org/10.1111/j.1365-2621.2000.tb10621.x Kotwaliwale, N., Bakane, P., Verma, A., 2007. Changes in textural and optical properties of oyster mushroom during hot air drying. Journal of Food Engineering 78, 1207-1211. http://doi.org/10.1016/j.jfoodeng.2005.12.033 Kortei, N.K., Odamtten, G.T., Obodai, M., Appiah, V., Abbey, L., Oduro-Yeboah, C., Akonor, P.T., 2015. Infuence of gamma radiation on some textural properties of fresh and dried oyster mushrooms (Pleurotus ostreatus). https://csirspace.foodresearchgh.org/handle/1/348 Zou M., Xu P., Lu X., Zhu K., Zhen J., Jin R., 2024. Analysis of the characteristics of Pleurotus ostreatus texture based on multivariate analysis method. Northern Horticulture, 119-126. http://doi.org/doi:10.11937/bfyy.20232781 Sa, Q., Zheng, J., Zhang, K., Wang, Y., 2025. Effects and assessment of the combined application of biogas slurry and chemical fertilizers on greenhouse tomato growth, yield, and soil quality. Scientia Horticulturae 344. http://doi.org/10.1016/j.scienta.2025.114113 Zhao, X., Mak-Mensah, E., Zhao, W., Wang, Q., Zhou, X., Zhang, D., Zhu, J., Qi, W., Liu, Q., Li, X., 2024. Optimized ridge-furrow technology with biochar amendment for alfalfa yield enhancement and soil erosion reduction based on a structural equation model on sloping land. Agricultural Water Management, 298. http://doi.org/10.1016/j.agwat.2024.108866 Wu, W., Han, J., Gu, Y., Li, T., Xu, X., Jiang, Y., Li, Y., Sun, J., Pan, G., Cheng, K., 2022. Impact of biochar amendment on soil hydrological properties and crop water use efficiency: A global meta‐analysis and structural equation model. GCB Bioenergy 14, 657-668. https://doi.org/10.1111/gcbb.12933. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 06 May, 2026 Reviewers agreed at journal 28 Apr, 2026 Reviews received at journal 27 Apr, 2026 Reviewers agreed at journal 26 Apr, 2026 Reviewers agreed at journal 23 Apr, 2026 Reviewers invited by journal 21 Apr, 2026 Editor assigned by journal 14 Apr, 2026 Editor invited by journal 10 Dec, 2025 Submission checks completed at journal 08 Dec, 2025 First submitted to journal 08 Dec, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-8225452\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Article\",\"associatedPublications\":[],\"authors\":[{\"id\":631297516,\"identity\":\"cb1a3a46-0daa-4241-a9ef-9703408a2618\",\"order_by\":0,\"name\":\"Songtao Lan\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guangxi Vocational University of Agricultural\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Songtao\",\"middleName\":\"\",\"lastName\":\"Lan\",\"suffix\":\"\"},{\"id\":631297517,\"identity\":\"12b27433-6b91-411c-8095-09858c74c928\",\"order_by\":1,\"name\":\"Yanting Li\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guangxi Minzu University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Yanting\",\"middleName\":\"\",\"lastName\":\"Li\",\"suffix\":\"\"},{\"id\":631297518,\"identity\":\"6d728fd0-8150-4b2c-9062-e4c38fa11bfb\",\"order_by\":2,\"name\":\"Liping Luo\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guangxi Minzu University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Liping\",\"middleName\":\"\",\"lastName\":\"Luo\",\"suffix\":\"\"},{\"id\":631297519,\"identity\":\"8906332f-c0e3-48d0-94cd-0b3bf7865858\",\"order_by\":3,\"name\":\"Enbei Xie\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guangxi Vocational University of Agricultural\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Enbei\",\"middleName\":\"\",\"lastName\":\"Xie\",\"suffix\":\"\"},{\"id\":631297520,\"identity\":\"7681c38e-0903-44e8-985c-0e3dd69422bb\",\"order_by\":4,\"name\":\"Shichan Pang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guangxi Vocational University of Agricultural\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Shichan\",\"middleName\":\"\",\"lastName\":\"Pang\",\"suffix\":\"\"},{\"id\":631297521,\"identity\":\"f05e0460-bd17-44db-8529-f4e3116a8dc3\",\"order_by\":5,\"name\":\"Yongchen Zhou\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Baise Tea Development Center\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Yongchen\",\"middleName\":\"\",\"lastName\":\"Zhou\",\"suffix\":\"\"},{\"id\":631297522,\"identity\":\"1e362ce3-a5e1-4b39-b8e9-c66a116b9bfd\",\"order_by\":6,\"name\":\"Qingqun Zeng\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Baise Tea Development Center\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Qingqun\",\"middleName\":\"\",\"lastName\":\"Zeng\",\"suffix\":\"\"},{\"id\":631297523,\"identity\":\"bb1c9ce1-f1b3-4b56-baec-3a1dedb5573f\",\"order_by\":7,\"name\":\"Weimei Chen\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Guangxi Vocational University of Agricultural\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Weimei\",\"middleName\":\"\",\"lastName\":\"Chen\",\"suffix\":\"\"},{\"id\":631297524,\"identity\":\"b2759893-4926-498e-a817-2acf683962ab\",\"order_by\":8,\"name\":\"Yun Li\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAyElEQVRIiWNgGAWjYBACPmYQWQHjshGhhQ2s5QxJWkAEYxtJWth5zKQL59Umru0/Y8DwoewwA//sBkIOA2qZue24sdmBMwaMM84dZpC4c4AILbzbjsmZHewxYOZtO8xgIJFAjJY5x3jMDvMYMP8lXktDjZzZMaAWRuK0sBVb8xw7YGx2hq3gYM+5dB6JGwS08PMf3nibp6Yucdv5wxsf/CizluOfQUALAwOHAZA4DGYeAGIeQuqBgP0BkKgjQuEoGAWjYBSMWAAAgso4YDvaKxcAAAAASUVORK5CYII=\",\"orcid\":\"\",\"institution\":\"Guangxi Minzu University\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Yun\",\"middleName\":\"\",\"lastName\":\"Li\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2025-11-28 01:08:09\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-8225452/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-8225452/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":108088321,\"identity\":\"a18f637a-f2f3-4284-9eb0-c1c9fda0fa98\",\"added_by\":\"auto\",\"created_at\":\"2026-04-29 08:56:32\",\"extension\":\"jpg\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":7242729,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDynamics of temperature and relative humidity in different treatments. (a) Time-course curve of temperature. (b) Time-course curve of relative humidity. (c) Comparison of average temperature among treatments. (d) Comparison of average relative humidity among treatments. Different lowercase letters indicate significant differences (p \\u0026lt; 0.05).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Fig.1.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8225452/v1/07e9f3dc99cf73007cb50b81.jpg\"},{\"id\":108181837,\"identity\":\"5cd1cb88-6be6-4d2c-b1a5-0a002cc4568d\",\"added_by\":\"auto\",\"created_at\":\"2026-04-30 08:58:57\",\"extension\":\"jpg\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":6416122,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDynamics of CO₂ concentration and light intensity in different treatments. (a) Time-course curve of CO₂ concentration. (b) Detailed view of CO₂ concentration dynamics in TMCH, SPO, and GGH. (c) Time-course curve of light intensity in SPO. (d) Time-course curve of light intensity in TMCH. (e) Comparison of average CO₂ concentration among treatments. (f) Comparison of average light intensity among treatments. Different lowercase letters indicate significant differences (p \\u0026lt; 0.05).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Fig.2.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8225452/v1/0602007d545723ac76446531.jpg\"},{\"id\":108181747,\"identity\":\"53c5b3df-0c57-4ab6-b4e7-57f693c1d6d8\",\"added_by\":\"auto\",\"created_at\":\"2026-04-30 08:58:53\",\"extension\":\"jpg\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":4167346,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eThe cultivation environment systematically influences the yield, commercial quality, and morphological traits of Pleurotus geesteranus.​\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Fig.3.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8225452/v1/111670976109f9b353b0b8eb.jpg\"},{\"id\":108088323,\"identity\":\"709b4380-8477-4cde-a99c-3ffe240a0bd1\",\"added_by\":\"auto\",\"created_at\":\"2026-04-29 08:56:32\",\"extension\":\"jpg\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":6865246,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eMultivariate analysis reveals the influence of fruiting environments on the textural properties, quality traits, and their key driving factors in Pleurotus geesteranus.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Fig.4.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8225452/v1/ef7ad73145dfbc3563fbee82.jpg\"},{\"id\":108183490,\"identity\":\"bb70f726-c0bc-4c78-a03a-ee719362aeee\",\"added_by\":\"auto\",\"created_at\":\"2026-04-30 09:01:39\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":24948833,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8225452/v1/ba61bf09-1f5b-45d7-a7df-da22cb161409.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Beyond Static Control: A Dynamic Environmental Regime Informed by Structural Equation Modeling Enhances Yield and Quality of Pleurotus geesteranus\",\"fulltext\":[{\"header\":\"1. Introduction\",\"content\":\"\\u003cp\\u003eThe oyster mushroom, Pleurotus geesteranus, is an important commercial edible fungus cultivated worldwide due to its unique flavor, nutritional value, and relatively short growth cycle\\u0026nbsp;\\u003cem\\u003e（\\u003c/em\\u003eThongklang,2016;Effiong,et al.，2024;Yu,et al.，2020\\u003cem\\u003e）\\u003c/em\\u003e. Its production plays a significant role in ensuring global food security, increasing farmers' income, and promoting regional economic development\\u0026nbsp;\\u003cem\\u003e(\\u003c/em\\u003eSong,et al.，2022;Boukary,et al.，2024\\u003cem\\u003e)\\u003c/em\\u003e. However, the full potential of its production is often unrealized due to inadequate environmental control, epitomized by the persistent challenge of simultaneously achieving high yield and superior quality.\\u003c/p\\u003e\\n\\u003cp\\u003eCurrent cultivation methods, ranging from forest-based analog systems to simple sheds and traditional mushroom houses, struggle to achieve precise and stable control over key environmental parameters such as temperature, humidity, light, and CO₂ concentration\\u0026nbsp;\\u003cem\\u003e(\\u003c/em\\u003eJarial,et al.，2024;Yang,et al.，2024;Hausiku,2022\\u003cem\\u003e)\\u003c/em\\u003e. These traditional approaches often rely on maintaining static environmental conditions, which fail to accommodate the distinct requirements of different developmental stages of mushroom morphogenesis, such as stipe elongation and pileus expansion. Consequently, the commercial quality of fruiting bodies (e.g., shape, cap-to-stem ratio) is often inconsistent, leading to a low proportion of premium-grade mushrooms and severely limiting economic returns\\u0026nbsp;\\u003cem\\u003e(\\u003c/em\\u003eTen,et al.，2021;Islam,et al.，2017\\u003cem\\u003e)\\u003c/em\\u003e. This limitation is exacerbated by global climate change, which increases the frequency and intensity of extreme weather events, further highlighting the vulnerability of traditional cultivation models\\u003cem\\u003e(\\u003c/em\\u003eHoegh-Guldberg,et al.，2019;Chen,et al.，2022\\u003cem\\u003e)\\u003c/em\\u003e.\\u003c/p\\u003e\\n\\u003cp\\u003eThe core of this bottleneck lies in the complex, non-linear interactions between environmental factors and mushroom growth and development. Conventional research methods have difficulty systematically deciphering the underlying causal pathways. Therefore, a paradigm shift from \\\"static control\\\" to \\\"dynamic programming\\\" is urgently needed in edible fungus cultivation. This study explores a novel intelligent cultivation pathway based on developmental biology logic: actively \\\"programming\\\" the morphogenesis of P. geesteranusthrough the temporal regulation of environmental conditions.\\u003c/p\\u003e\\n\\u003cp\\u003eTo this end, we developed a Digital Smart Mushroom Fruiting Chamber (DSMFC), integrated with photovoltaic power generation and Internet of Things (IoT) technology, serving as the hardware platform for implementing dynamic control. More critically, we introduced Structural Equation Modeling (SEM) as a powerful analytical tool\\u0026nbsp;(Cheng,et al.，2023;De La Croix,et al.，2022). Although SEM has found applications in agricultural environmental research, its potential in deciphering complex systems in edible fungus cultivation remains underexplored\\u0026nbsp;(Sunghyoun,et al.,2015;Seung-Mi,et al.,2015;Sakamoto,2018). The strength of SEM lies in its ability to quantify the direct and indirect effect pathways of multiple environmental factors on the final yield, thereby providing a solid mechanistic basis for optimizing control strategies\\u0026nbsp;(Lee,et al.,2018;Hair,et al.,2019;Hair,et al.,2016;A,et al.,2020).\\u003c/p\\u003e\\n\\u003cp\\u003eUsing P. geesteranusas a model, this study compares the DSMFC with traditional cultivation environments. By employing principal component analysis, correlation analysis, and SEM (Sa,et al.,2025;Liu,et al.,2023), we aim to:Validate the effectiveness of a novel two-stage dynamic regulation strategy—promoting stipe elongation under high temperature/high CO₂ followed by stimulating pileus development under lower temperature with ventilation and light—in synergistically enhancing yield and quality.Uncover the key pathways through which pivotal environmental factors like accumulated heat units and CO₂ concentration drive morphogenesis and yield formation.Evaluate the economic viability of this intelligent cultivation mode, providing a replicable theoretical framework and technical example for the intelligent upgrading of the edible mushroom industry.\\u003c/p\\u003e\"},{\"header\":\"2. Materials and Methods\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003e2.1 Experimental Site and Period\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe experiment was conducted in March 2025 in Xixiangtang District, Nanning City, Guangxi Zhuang Autonomous Region, China. The specific locations were the Guangxi Vocational University of Agriculture (108°14'40\\\"E, 22°51'16\\\"N) for the Traditional Mushroom Cultivation House (TMCH), Glass Greenhouse (GGH), and Passion Fruit Orchard Shade (SPO) treatments, and the Guangxi Minzu University (108°13'58\\\"E, 22°50'21\\\"N) for the Digital Smart Mushroom Fruiting Chamber (DSMFC). The experimental area is characterized by a typical tropical monsoon climate at an altitude of 86 meters. During the experimental period (March), the weather was predominantly cloudy with low light intensity, scant rainfall (average approx. 52 mm), and an average air temperature ranging from 15°C (night) to 21°C (day).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.2 Mushroom Strain and Substrate Preparation\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe Pleurotus geesteranusstrain used in this study was ‘Jinxiu’, a widely cultivated commercial variety in China. The spawn was purchased from Guangxi Longzhou Beibu Gulf Modern Agriculture Co., Ltd.\\u0026nbsp;The substrate formulation was adapted from Ke et al. (2023) and consisted of the following components (by weight): 25.5% cottonseed hulls, 22% corn cobs, 20% wheat bran, 12% sawdust, 8% bagasse, 5% corn flour, and 5% soybean meal, supplemented with 1% light calcium carbonate, 1% gypsum, and 0.5% lime. The corn cobs were pre-wetted for 30 minutes before being thoroughly mixed with other components for 40 minutes (Ke,et al.,2023). The moisture content of the mixture was adjusted to 61-62%, and the initial pH was regulated to 8.0-9.0 (targeting pH 6.0-6.5 post-sterilization).\\u003c/p\\u003e\\n\\u003cp\\u003eThe substrate was packed into high-pressure polypropylene bags (18 cm × 36 cm, 0.04 mm thickness). Each bag was filled to a height of 18 cm, resulting in a net weight of 1.1-1.3 kg. Sterilization was performed using a standard autoclaving cycle, which included a final stage of 121°C at 0.12 MPa for 150 minutes, followed by a 30-minute holding period.\\u003c/p\\u003e\\n\\u003cp\\u003eAfter cooling to approximately 90°C, the bag-logs were transferred to a class-10,000 clean room for forced cooling until the core temperature reached ~23°C. Aseptic inoculation was then performed in a class-100 laminar flow cabinet, with each bag receiving 20 mL of liquid spawn before being sealed for mycelial colonization .\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.3 Experimental Design and Fruiting Environment Treatments\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eA completely randomized design was employed. Eighty bag-logs with uniform mycelial growth (≥95% coverage at the incision site) were selected and randomly assigned to four treatment groups (20 biological replicates per group), each corresponding to a distinct fruiting environment:SPO (Shade of Passion Fruit Orchard): Under two-year-old passion fruit vines providing natural shade and a cool microclimate.GGH (Glass Greenhouse): Equipped only with a sunshade net and axial flow fans.TMCH (Traditional Mushroom Cultivation House): Equipped with fans and an Emerson Copeland Scroll air compressor for manual temperature adjustment. DSMFC (Digital Smart Mushroom Fruiting Chamber):A self-developed system utilizing IoT technology for precise multi-factor environmental control, powered by integrated photovoltaic panels, smart sensors, and a closed-loop control module.A multi-functional remote environmental monitor was used to continuously record temperature, relative humidity, CO₂ concentration, and light intensity in each treatment throughout the fruiting period until harvest.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.4 Two-Stage Dynamic Regulation Strategy in DSMFC\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eUpon completion of the 64-day mycelial colonization period, bag-logs in the DSMFC underwent a precise, two-stage priming management process to induce and support fruiting body development:Primordia Induction Phase: A controlled temperature reduction (2°C/h) was applied to lower the temperature to 12.0 ± 0.5°C for 12 hours to stimulate primordia differentiation.\\u003c/p\\u003e\\n\\u003cp\\u003eFruiting Body Development Phase: Following the low-temperature induction, a 1-2 cm segment was aseptically removed from the bag opening. The IoT control system was then activated to maintain the environment at 25.0 ± 1°C and 85-95% relative humidity for the first 20 hours, with CO₂ concentration dynamically controlled by fans and cooling systems until mycelial recovery at the incision site reached ≥95%. Subsequently, to promote pileus development, conditions were adjusted to approximately 20°C, supplemented with blue LED light, and CO₂ concentration was maintained around 2000 ppm.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.5 Sample Collection and Measurements\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.5.1 Yield, Premium Mushroom Ratio, and Agronomic Traits\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eFor each treatment, yield per bag was measured daily from March 11th to 13th using an electronic balance. Harvested mushrooms were graded into Grade-1, Grade-2, and Grade-3 according to the Jiangxi Provincial Standard (DB36/T 824-2023). The premium mushroom ratio was calculated as: Premium Mushroom Ratio (%) = (Weight of Grade-1 mushrooms + Weight of Grade-2 mushrooms) / Total mushroom weight × 100.\\u003c/p\\u003e\\n\\u003cp\\u003eGrade-2 mushrooms were selected for agronomic trait measurements: individual mushroom weight, pileus weight, and stipe weight were recorded to calculate the pileus-to-stipe ratio. The maximum pileus diameter and minimum pileus width were measured to determine the length-to-width ratio. The diameter of the stipe at its midpoint was measured using a vernier caliper (Yan,et al.,2024;Yu,et al.,2023;Yu,et al.，2021;).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.5.2 Nutritional Quality Analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eApproximately 200 g of fresh mushroom samples (a mix of pileus and stipe) from each treatment were freeze-dried in liquid nitrogen, ground into a fine powder, and analyzed in triplicate. The contents of crude polysaccharides, crude fat, crude protein, crude fiber, moisture, total saponins, and total phenols were determined using established standard methods as referenced (He,et\\u0026nbsp;al.,2012;Jeong,et\\u0026nbsp;al.,2025).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.5.3 Texture Profile Analysis (TPA)\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eFour mushrooms per treatment were randomly selected. The central part of the pileus (approx. 4 cm²) and the mid-section of the stipe (approx. 2 cm length) were prepared as test samples. Textural properties, including hardness, chewiness, cohesiveness, springiness, adhesiveness, and gumminess, were measured using a TMS-PILOT texture analyzer with a 50-mm diameter cylindrical probe. The test conditions were: pre-test speed: 60 mm/min, test speed: 60 mm/min, post-test speed: 60 mm/min, compression strain: 60%, and trigger force: 0.15 N.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.6 Statistical Analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eData compilation was performed using Microsoft Office LTSC Professional Plus 2021. Graph plotting and correlation analysis (significance level set at p \\u0026lt; 0.05) were conducted using OriginPro 2024. One-way analysis of variance (ANOVA), Principal Component Analysis (PCA), and correlation heatmaps were performed using IBM SPSS Statistics 27. The Partial Least Squares Structural Equation Modeling (PLS-SEM) analysis was conducted using SmartPLS 4(Chen,et al.,2024;Hair,et al.,2021).\\u003c/p\\u003e\"},{\"header\":\"3. Results\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003e3.1 Characterization of Environmental Conditions Across Cultivation Systems\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe environmental parameters across the four cultivation systems exhibited distinct dynamic patterns, which fundamentally shaped the subsequent growth and development of P. geesteranus.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e3.1.1 Temperature and Relative Humidity Dynamics\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe temperature and relative humidity profiles for each treatment throughout the fruiting period (March 9-13) are shown in Fig. 1. The DSMFC treatment demonstrated a well-defined temperature trajectory, initially rising to a peak of 30.3\\u0026deg;C at 17:00 on March 10th, then decreasing to a minimum of 14.4\\u0026deg;C by 08:00 on March 12th, before stabilizing around 20\\u0026deg;C (Fig. 1a). This pattern reflected the intentional two-stage regulatory strategy. While SPO and GGH showed similar diurnal fluctuation trends, SPO maintained the lowest overall temperature. In contrast, GGH experienced the highest daytime temperatures but cooled rapidly at night, approaching SPO levels around 05:00. TMCH provided the most stable thermal environment, consistently maintaining around 20\\u0026deg;C. The average temperature was highest in TMCH and lowest in SPO, with both being significantly different from other treatments (Fig. 1c).\\u003c/p\\u003e\\n\\u003cp\\u003eRelative humidity across all treatments displayed a characteristic diurnal pattern, decreasing during the day and increasing at night (Fig. 1b). Occasional sharp fluctuations in TMCH and DSMFC were attributable to the activation of ventilation fans. The average relative humidity was comparable (approximately 93%) in SPO, TMCH, and DSMFC, but significantly lower (87.7%) in GGH (Fig. 1d).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e3.1.2 CO₂ Concentration and Light Intensity Profiles\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe CO₂ concentration and light intensity profiles are critical differentiators among the systems (Fig. 2). The DSMFC treatment was characterized by the highest and most dynamically managed CO₂ environment. The concentration peaked at 9998.55 ppm at 11:00 on March 11th, followed by a decline due to activated ventilation and cooling systems, eventually stabilizing around 2000 ppm (Fig. 2a, b). In contrast, the CO₂ concentrations in TMCH, SPO, and GGH were not significantly different from each other, although TMCH showed greater variability (Fig. 2e).Light intensity was predominantly governed by the natural solar cycle in SPO, resulting in the highest average intensity among the treatments (Fig. 2c, f). TMCH employed continuous artificial lighting at an intensity of approximately 33 lux (Fig. 2d). The DSMFC was maintained in near-darkness (0.002 lux) except for a brief period of blue LED illumination to promote pileus development in the later stage. Consequently, the light intensity in TMCH was significantly higher than in both DSMFC and GGH (Fig. 2f).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e3.2 Effects of Cultivation Environments on Yield, Quality Grade, and Morphological Traits\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe cultivation environment exerted a profound and systematic impact on the yield structure, commercial quality, and morphology of P. geesteranus. The Digital Smart Mushroom Fruiting Cabin (DSMFC) demonstrated superior performance across most evaluated metrics.\\u003c/p\\u003e\\n\\u003cp\\u003eDSMFC achieved the highest total yield (304.2 g/bag) and biological efficiency (46.7%), which were significantly greater than all other treatments (Fig. 3a, c). This high yield was attributed to a significantly greater number of fruiting bodies per bag (82.2) compared to other environments (Fig. 3b). Crucially, the DSMFC strategy effectively translated this high yield into superior commercial value. It produced the highest proportion of premium-grade mushrooms (Grade-1 + Grade-2) at 46.0%, driven predominantly by a substantial yield of Grade-1 mushrooms (102.0 g/bag) that was nearly absent in traditional systems (Fig. 3e, f, m).\\u003c/p\\u003e\\n\\u003cp\\u003eIn contrast, the traditional facilities (GGH and TMCH) presented a \\u0026quot;high-yield, low-quality\\u0026quot; paradox. While their total yields were comparable to each other and higher than SPO, they severely lacked Grade-1 mushrooms, resulting in low premium ratios (Fig. 3m). The SPO treatment consistently underperformed, yielding the lowest in both total output (132.8 g/bag) and biological efficiency (19.9%).\\u003c/p\\u003e\\n\\u003cp\\u003eMorphologically, DSMFC-grown mushrooms exhibited the most uniform appearance with the lowest incidence of malformations (Fig. 3d). While no significant differences were detected in pileus dimensions (length, width, weight) across treatments (Fig. 3n,o,p), stipe morphology was distinctly affected. DSMFC promoted the development of significantly longer (5.4 cm) and thinner (7.1 mm diameter) stipes compared to others (Fig. 3q, r). This specific stipe architecture (long and thin), coupled with a lower pileus-to-stipe weight ratio, is a key characteristic associated with higher commercial grade in P. geesteranus. Although individual mushroom weight was slightly higher in GGH, the difference was not statistically significant (Fig. 3v).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e3.3 Comprehensive Analysis of Quality, Multivariate Relationships, and Economic Benefits\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eA comprehensive evaluation of the Pleurotus geesteranusfruiting bodies under different cultivation environments revealed systematic differences in nutritional quality, eating texture, the drivers of key traits, and ultimately, economic returns.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e3.3.1 Nutritional and Textural Quality\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAnalysis of nutritional components showed that the most significant differences were observed in the moisture content and crude fiber content of the fruiting bodies (Table 1). The moisture content was highest in the DSMFC treatment (86.89 \\u0026plusmn; 0.34%), significantly exceeding other treatments and indicating better freshness. Conversely, the crude fiber content was significantly higher in the SPO treatment (43.91 \\u0026plusmn; 1.04%), potentially related to the natural stress environment it experienced. Other nutritional components (e.g., crude protein, crude fat) showed no significant differences among treatments.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 1.\\u0026nbsp;\\u003c/strong\\u003eEffects of different treatments on the nutritional quality of Pleurotus geesteranus.\\u003c/p\\u003e\\n\\u003cdiv align=\\\"center\\\"\\u003e\\n \\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"135%\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003eTreatment\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003eMoisture content (%)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003eCrude protein (%)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003eCrude fat\\u003c/p\\u003e\\n \\u003cp\\u003e(%)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003eCrude fiber\\u003c/p\\u003e\\n \\u003cp\\u003e(%)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 15.625%;\\\"\\u003e\\n \\u003cp\\u003eCrude polysaccharide (g/kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003eTotal saponins (g/kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.5417%;\\\"\\u003e\\n \\u003cp\\u003eTotal phenolics\\u003c/p\\u003e\\n \\u003cp\\u003e(g/kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003eSPO\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e84.05\\u0026plusmn;1.78b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e40.56\\u0026plusmn;2.43a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003e1.13\\u0026plusmn;0.26a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e43.91\\u0026plusmn;1.80a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 15.625%;\\\"\\u003e\\n \\u003cp\\u003e43.22\\u0026plusmn;5.58a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e7.69\\u0026plusmn;0.48a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.5417%;\\\"\\u003e\\n \\u003cp\\u003e11.86\\u0026plusmn;0.47a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003eGGH\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e84.61\\u0026plusmn;1.11b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e38.8\\u0026plusmn;1.32a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003e1.02\\u0026plusmn;0.08a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e35.85\\u0026plusmn;3.47b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 15.625%;\\\"\\u003e\\n \\u003cp\\u003e41.35\\u0026plusmn;3.36a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e7.92\\u0026plusmn;0.49a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.5417%;\\\"\\u003e\\n \\u003cp\\u003e12.14\\u0026plusmn;0.81a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003eTMCH\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e85.54\\u0026plusmn;0.58ab\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e40.15\\u0026plusmn;2.25a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003e1.28\\u0026plusmn;0.55a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e36.22\\u0026plusmn;3.31b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 15.625%;\\\"\\u003e\\n \\u003cp\\u003e39.96\\u0026plusmn;2.61a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e7.22\\u0026plusmn;0.22a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.5417%;\\\"\\u003e\\n \\u003cp\\u003e12.42\\u0026plusmn;0.41a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003eDSMFC\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e86.89\\u0026plusmn;0.76a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e39.34\\u0026plusmn;2.21a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.4167%;\\\"\\u003e\\n \\u003cp\\u003e1.06\\u0026plusmn;0.29a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e35.89\\u0026plusmn;3.62b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 15.625%;\\\"\\u003e\\n \\u003cp\\u003e42.47\\u0026plusmn;6.45a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 12.5%;\\\"\\u003e\\n \\u003cp\\u003e7.74\\u0026plusmn;0.08a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.5417%;\\\"\\u003e\\n \\u003cp\\u003e11.55\\u0026plusmn;1.3a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n \\u003c/table\\u003e\\n\\u003c/div\\u003e\\n\\u003cp\\u003eDifferent lowercase letters within a column indicate significant differences between treatments at the 5% level.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e3.3.2 Multivariate Relationships and Driver Analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eTextural profile analysis (TPA) revealed significant treatment-specific variations in the texture of Pleurotus geesteranus (Fig. 4a-f). Stipes generally exhibited greater hardness and chewiness than pilei. The GGH treatment yielded the highest values for stipe hardness and chewiness, while DSMFC resulted in the highest pileus chewiness and gumminess. Stipe springiness was highest in GGH but lowest in DSMFC, which conversely showed the highest stipe adhesiveness. Cohesiveness and pileus springiness remained consistent across treatments.\\u003c/p\\u003e\\n\\u003cp\\u003ePrincipal Component Analysis (PCA) indicated that the primary composite factor affecting yield (PC1, 78.9% variance contribution) was strongly positively correlated with premium mushroom weight, premium mushroom count, and total mushroom count (Fig. 4g). PCA of environmental factors showed that accumulated heat units (AHU) and average temperature were the main positive driving variables (Fig. 4i). Correlation heatmaps further confirmed that temperature-related variables and average CO₂ concentration were significantly positively correlated with premium mushroom count and weight (Fig. 4k).\\u003c/p\\u003e\\n\\u003cp\\u003eMost importantly, Partial Least Squares Structural Equation Modeling (PLS-SEM) quantified the effect pathways of key environmental factors (Fig. 4l). The model revealed that accumulated heat units (AHU) and CO₂ concentration were the core factors driving stipe morphogenesis. They had highly significant positive direct effects on stipe length and diameter, which in turn indirectly promoted the increase in individual mushroom weight. The model explained a large portion of the variance in individual mushroom weight (R\\u0026sup2; = 0.932), clearly outlining the key regulatory pathway: \\u0026quot;Temperature \\u0026amp; CO₂ \\u0026rarr; Stipe Morphology \\u0026rarr; Yield\\u0026quot;.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e3.3.3 Economic Benefits\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eEconomic benefit analysis ultimately validated the commercial value of the intelligent cultivation mode (Table 2). Despite higher equipment costs, the DSMFC, through precise environmental control, greatly optimized the yield structure: the Grade-1 mushroom yield reached 102.04 kg per 1000 bags, accounting for 62.1% of the total yield, far surpassing traditional modes. This significant \\u0026quot;premium mushroom price premium effect\\u0026quot; resulted in a net profit for the DSMFC (443.02 USD/1000 bags) that was substantially higher than those of GGH, TMCH, and SPO\\u0026mdash;by 96.0%, 129.7%, and 203.3%, respectively\\u0026mdash;fully demonstrating its great potential for market promotion.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 2.\\u0026nbsp;\\u003c/strong\\u003eEconomic benefit analysis of different treatments (based on 1000 bags).\\u003c/p\\u003e\\n\\u003cdiv align=\\\"center\\\"\\u003e\\n \\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"125%\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003eTreatment\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003eGrade-1 yield (kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003eGrade-2 yield (kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003eGrade-3 yield (kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003eGrade-1 revenue (USD)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003eGrade-2 revenue (USD)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003eGrade-3 revenue (USD)\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003eTotal cost (USD)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003eNet profit (USD)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003eSPO\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003e0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e42.34\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e90.48\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e0.00\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e71.28\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e76.17\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e1.40\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e146.05\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003eGGH\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003e2.74\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e64.5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e138.72\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e7.69\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e108.59\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e116.77\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e7.02\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e226.04\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003eTMCH\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003e4.134\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e48.54\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e143.23\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e11.60\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e81.72\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e120.57\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e21.05\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e192.85\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003eDSMFC\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 9.27835%;\\\"\\u003e\\n \\u003cp\\u003e102.04\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e37.32\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e164.84\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e286.32\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e62.83\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 13.4021%;\\\"\\u003e\\n \\u003cp\\u003e138.76\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e44.90\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd nowrap=\\\"\\\" style=\\\"width: 10.3093%;\\\"\\u003e\\n \\u003cp\\u003e443.02\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n \\u003c/table\\u003e\\n\\u003c/div\\u003e\\n\\u003cp\\u003eNote: Yield calculations were based on 1,000 mushroom bags. According to the exchange rate, 1 Chinese Yuan (CNY) = 0.1403 US Dollars (USD). The prices for Grade-1, Grade-2, and Grade-3 mushrooms were set at 2.81 USD, 1.68 USD, and 0.84 USD, respectively. The primary production costs included electricity and water expenses.\\u003c/p\\u003e\"},{\"header\":\"4. Discussion\",\"content\":\"\\u003cp\\u003e4.1 Characteristics of fruiting environments across treatments\\u003c/p\\u003e\\n\\u003cp\\u003eEnvironmental conditions are critical factors influencing mushroom growth, significantly affecting morphology and yield（Raudaskoski and Viitanen,1982,Sohi and Upadhyay,1989）. Different Pleurotus species require distinct growth environments, necessitating climate-adapted varieties and facility management strategies based on regional conditions(Yan,et al.,2024). Previous research has identified optimal artificial environmental parameters for Pleurotus morphological development: CO₂ concentration of 0.3% (with deformity rates increasing significantly above 0.5%), temperature of 13-16\\u0026deg;C, humidity \\u0026gt;80%, and ventilation rate of 0.2-0.5 feet/minute (exposed conditions). This parameter combination effectively suppresses abnormal fruiting body development(Jang,et al.,2003).This study observed that SPO provided the highest light intensity following natural diurnal patterns, with the lowest average temperature but relatively high average humidity (93%). However, it exhibited substantial diurnal fluctuations in both temperature and humidity, showing parallel but overall lower patterns compared to GGH. GGH relied on shading nets and fan ventilation for environmental control. Due to the greenhouse effect of glass enclosure, daytime temperatures exceeded those in TMCH, while radiative cooling at night caused rapid temperature loss, resulting in the largest day-night temperature variation. Concurrently, GGH maintained the lowest humidity levels (average 87.7%), making it difficult to sustain the stable high-humidity environment required by P. geesteranus.TMCH utilized compressors and fans for temperature and humidity regulation, providing the most stable environment with temperatures consistently maintained around 20\\u0026deg;C and minimal fluctuations. However, forced ventilation caused \\u0026quot;jump\\u0026quot; oscillations in humidity, though the average humidity remained at 93%. This treatment employed constant artificial lighting at 33 lx.Due to high-density cultivation, DSMFC exhibited gradually increasing temperature and CO₂ concentrations, reaching a maximum temperature of 30.3\\u0026deg;C, higher than the temperature settings reported by Lee et al(Lee,et al.,2018). Jhune et al found that low-temperature Pleurotus strains perform optimally at 10\\u0026deg;C, while medium-high temperature strains show improved performance with increasing temperature(Jhune,et al.,2011). At 10\\u0026deg;C, pileus thickness increased but the harvest period extended, and stipes became longer and thinner with rising temperatures, with some varieties exhibiting temperature stress-induced malformations. In DSMFC, although brief high temperatures occurred, the automatic cooling system activated promptly, preventing significant impacts on fruiting body development.Paul et al.(1983) suggested that high CO₂ concentrations cause stipe elongation and suppressed pileus development. Kinugawa et al.(1994) found that CO₂ concentrations exceeding 6,000 ppm for 7 days led to trumpet-shaped deformities in Pleurotus pileus and reduced yield. In this study, CO₂ concentrations exceeding 9,000 ppm were maintained for 27 hours before the ventilation and cooling systems gradually reduced both CO₂ levels and temperature. Despite these high concentrations, no malformations occurred because the increase was gradual during fruiting body development. However, some delayed-development bags showed shortened stipes and altar-shaped mushrooms, consistent with findings by Jang et al.(2003) and Lin et al.(2022).When stipe length reached approximately 5 cm, blue LED lighting was activated while maintaining temperature around 20\\u0026deg;C and CO₂ concentration around 2,000 ppm to promote pileus development. Yang et al identified 25\\u0026deg;C and 90% humidity as the optimal combination for summer cultivation of P. geesteranus, with misting humidification significantly reducing malformations and enhancing pileus coloration(Yang,et al.,2023). Building on this, DSMFC achieved initial gradual increases in temperature and CO₂ to promote stipe elongation, followed by cooling, ventilation, and light supplementation, ultimately reducing the fruiting cycle duration and energy consumption.\\u003c/p\\u003e\\n\\u003cp\\u003e4.2 Effects of different fruiting environments on yield and agronomic traits of Pleurotus geesteranus\\u003c/p\\u003e\\n\\u003cp\\u003eThe fruiting environment significantly influenced the yield, quality, and agronomic traits of P. geesteranus\\u0026nbsp;(Song,et al.,2025). DSMFC demonstrated exceptional performance across most indicators, with its highest yield (304.2 g), biological efficiency (46.7%), and premium mushroom ratio (46.0%) fully validating the effectiveness of its environmental regulation strategy.GGH exhibited high yield but low quality, accompanied by yellowing disease and size heterogeneity issues. These problems were closely associated with its environmental characteristics: high daytime temperatures, low humidity (average 87.7%), and substantial fluctuations in temperature and humidity, leading to significantly reduced market value.TMCH showed similar yield to GGH but also manifested a \\u0026quot;high yield with low value\\u0026quot; phenomenon: near absence of Grade I mushrooms, the lowest premium mushroom ratio (approximately 20%), thick stipes, and deformed pileus, all compromising marketability.The SPO treatment, mimicking ecological cultivation, demonstrated no advantages in either yield or quality. It produced the lowest yield (132.8 g), lowest biological efficiency (19.9%), and the highest number of malformed mushrooms. Fruiting bodies from SPO exhibited the shortest stipes and the highest pileus-to-stipe ratio. Although total yield was low, these mushrooms may possess unique flavor qualities worthy of further investigation.Nakazawa et al, Ohm et al and Terashima et al suggested that many mushrooms exhibit phototropism, and pileus differentiation in basidiomycetes is influenced by light conditions(Nakazawa,et al.,2008;Ohm,et al.,2013;Terashima,et al.,2005). The higher incidence of malformed mushrooms in TMCH and SPO may be attributed to either continuous 24-hour lighting (33 lx) in TMCH or uneven light distribution under tree shade in SPO. Additionally, relatively frequent ventilation or natural environmental fluctuations could contribute to abnormal fruiting body development.\\u003c/p\\u003e\\n\\u003cp\\u003e4.3 Effects of Different Fruiting Environments on Quality and Textural Properties of Pleurotus geesteranus\\u003c/p\\u003e\\n\\u003cp\\u003eThe fruiting environment not only altered the yield and appearance of P. geesteranus but also significantly influenced its nutritional quality and textural properties, which directly determine its commercial value, processing characteristics, and consumer palatability. Regarding nutritional quality, the most notable differences were observed in the moisture content and crude fiber content of the fruiting bodies. Jin et al reported that the moisture content of Pleurotus fruiting bodies varies with flush number, cultivar, and pileus size(Jin,et al.,2021).The DSMFC treatment yielded fruiting bodies with the highest moisture content (86.89%). This elevated moisture content enhanced their fresh and tender appearance but may negatively impact shelf life and drying efficiency. Conversely, fruiting bodies from the SPO treatment contained significantly higher crude fiber (43.91%) compared to other groups. This may be attributed to the stress conditions of stronger light exposure and lower temperatures, which potentially stimulate cellulose synthesis in the cell walls to enhance structural support.No significant differences were detected in crude protein, crude fat, crude polysaccharides, or active components (saponins and polyphenols) among treatments. This indicates that the synthesis of these nutritional compounds is primarily regulated by the cultivation substrate and genetic factors, combined with the short growth cycle of P. geesteranus, which limits substantial differential accumulation of nutrients.Texture profile analysis (TPA) objectively measures the physical properties of mushrooms by simulating oral mastication processes, thereby transforming subjective \\u0026quot;mouthfeel\\u0026quot; into quantifiable and reproducible data. This approach is crucial for scientific research, quality control, and product development(Zivanovic,et al.,2000;Kotwaliwale,et al.,2007;Kortei,et al.,2015).Zou et al found that hardness and chewiness of Pleurotus were significantly positively correlated and jointly determined palatability(Zou,et al.,2024). Stipe cohesiveness showed negative correlations with hardness and adhesiveness. Appropriate enhancement of cohesiveness can optimize texture, while excessively low hardness may lead to structural looseness. As an indicator of cellular binding force, cohesiveness serves as a key parameter for evaluating texture.This study revealed that the GGH treatment produced fruiting bodies with the highest stipe hardness, chewiness, and springiness. This firm and chewy texture likely resulted from the high-temperature and low-humidity stress conditions, leading to more compact cellular structures.The DSMFC treatment exhibited a unique textural profile: the highest chewiness and gumminess in the pileus, but the lowest springiness and gumminess in the stipe. This suggests independent development of pileus and stipe tissues, with the pileus being full and resilient while the stipe tended to be looser and more tender, potentially creating a superior overall mouthfeel experience.The TMCH treatment showed intermediate values for most textural parameters, consistent with its stable temperature-humidity environment but constant weak lighting conditions.\\u003c/p\\u003e\\n\\u003cp\\u003e4.4 Relationships among Variables\\u003c/p\\u003e\\n\\u003cp\\u003eThe results from partial least squares structural equation modeling (PLS-SEM), principal component analysis (PCA), and correlation analysis collectively revealed a clear network of pathways governing yield and quality formation in Pleurotus geesteranus, elucidating the complex mechanisms through which key environmental factors directly and indirectly influence final yield（Sa,et al.,2025;Zhao,et al.,2024;Wu,,et al.,2022）.The core pathways of the model identified accumulated heat units (AHU) and mean CO₂ concentration as the two most critical environmental drivers of yield and premium mushroom number. AHU exerted a highly significant positive direct effect on stipe elongation (path coefficient: 0.096), and moderately elongated stipes further supported heavier individual mushroom weight (0.093), forming a crucial pathway for premium yield accumulation.More notably, CO₂ demonstrated strong direct positive effects on stipe morphology (diameter: 0.383; length: 0.260). This indicates that moderately elevated CO₂ levels suppress lateral pileus expansion while redirecting assimilates to the stipe, forming the commercially desirable \\u0026quot;elongated stipe-compact pileus\\u0026quot; morphology. This mechanism enhances the proportion of Grade I mushrooms without compromising individual mushroom weight. These findings were mutually validated by correlation analysis, which showed extremely significant positive correlations between mean CO₂ concentration and both premium mushroom number and Grade I mushroom count.The model also revealed differential responses of various traits to environmental factors. For example, the low coefficient of determination for pileus length (R\\u0026sup2; = 0.200) suggests that its development is more influenced by microenvironmental fluctuations or genetic factors not included in the model, explaining why no significant differences in pileus size were observed in agronomic traits.Although mean light intensity (PPFD) and relative humidity (RH) did not reach significance in direct pathways, their positive trends and correlations with crude fiber content and pileus width (Figure 10) imply their roles as important regulatory factors. They may function synergistically with other factors (e.g., CO₂) through coupled regulation\\u0026mdash;for instance, light may suppress excessive elongation while promoting pileus development.\\u003c/p\\u003e\"},{\"header\":\"5. Conclusions\",\"content\":\"\\u003cp\\u003eThis study develops and validates a novel two-stage dynamic environmental regulation strategy for Pleutotus geesteranuscultivation, implemented via a Digital Smart Mushroom Fruiting Chamber (DSMFC). The strategy, which sequentially applies high-temperature/high-CO₂ conditions to promote stipe elongation followed by cooler, ventilated, and lit conditions to optimize pileus development, successfully breaks the common trade-off between yield and quality. By employing Structural Equation Modeling, we quantitatively identified accumulated heat units (AHU) and CO₂ concentration as the pivotal environmental drivers of stipe morphogenesis and yield. This strategy significantly enhanced total yield (304.2 g/bag), biological efficiency (46.7%), and the premium mushroom ratio (46.0%), resulting in the highest net profit (443.02 USD/1000 bags). The research underscores that a paradigm shift from static control to dynamic environmental programming is key to advancing precision agriculture in edible mushroom cultivation, providing a replicable framework for simultaneously achieving productivity and economic gains.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cbr\\u003e\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAuthor Contributions:\\u003c/strong\\u003eConceptualization, S.L. and Y.L. (Yanting Li); Methodology, S.L. and L.L.; Project administration, S.L. and L.L.; Writing – original draft, S.L.; Data curation, Y.L. (Yanting Li) and E.X.; Formal analysis, E.X.; Investigation, E.X., Y.Z., Q.Z. and W.C.; Resources, L.L.; Validation, S.P. and Y.Z.; Visualization, S.P.; Writing – review \\u0026amp; editing, Y.L. (Yun Li); Funding acquisition, Y.L. (Yun Li). All authors have read and agreed to the published version of the manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding:\\u0026nbsp;\\u003c/strong\\u003eThis research was supported by the Guangxi Major Special Project (No. Guike AA24263038), the Introduction of Talents Scientific Research Start-up Project of Guangxi Minzu University (No. 2024KJQD218), and the 2025 University-level Scientific Research Project of Guangxi Vocational University of Agriculture (No. XKJ2540).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eData Availability Statement:\\u0026nbsp;\\u003c/strong\\u003eThe raw data supporting the conclusions of this article will be made available by the authors, without undue reservation. The data are not publicly available due to copyright restrictions.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflicts of Interest:\\u003c/strong\\u003e The authors declare no conflicts of interest.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eThongklang, N., 2016. Testing agricultural wastes for the production of Pleurotus ostreatus. Mycosphere 7, 766-772. http://doi.org/10.5943/mycosphere/7/6/6\\u003c/li\\u003e\\n\\u003cli\\u003eEffiong, M.E., Umeokwochi, C.P., Afolabi, I.S., Chinedu, S.N., 2024. Comparative antioxidant activity and phytochemical content of five extracts of Pleurotus ostreatus (oyster mushroom). Sci Rep 14, 3794. http://doi.org/10.1038/s41598-024-54201-x\\u003c/li\\u003e\\n\\u003cli\\u003eYu, Q., Guo, M., Zhang, B., Wu, H., Zhang, Y., Zhang, L., 2020. Analysis of Nutritional Composition in 23 Kinds of Edible Fungi. Journal of Food Quality. https://doi.org/10.1155/2020/8821315.\\u003c/li\\u003e\\n\\u003cli\\u003eSong, J.-L., Wang, W.-k., Lu, N., Yan, J., Yuan, W.-d., Wu, Y.-L., 2022. Genetic diversity analysis of Pleurotus pulmonarius(Fr.) Qu\\u0026eacute;l. based on agronomic characters and ISSR markers.\\u0026nbsp;Plant Genome14, 20128.\\u003c/li\\u003e\\n\\u003cli\\u003eBoukary, A.A., Olou, A.B., Piepenbring, M., Yorou, N.S., Mushroom cultivation in tropical Africa: successes, challenges, and opportunities. Journal of Agriculture and Food Research. https://doi.org/10.1016/j.jafr.2024.101264\\u003c/li\\u003e\\n\\u003cli\\u003eJarial, R., Jarial, K., Bhatia, J., 2024. Comprehensive review on oyster mushroom species (Agaricomycetes): Morphology, nutrition, cultivation and future aspects. Heliyon 10. http://doi.org/10.1016/j.heliyon.2024.e26539\\u003c/li\\u003e\\n\\u003cli\\u003eYang, Z., Qu, J., Qiao, L., Jiang, M., Zou, X., Cao, W., 2024. Tea and Pleurotus ostreatusintercrop** modulates structure of soil and root microbial communities.\\u0026nbsp;Sci. Rep.14, 11295.\\u003c/li\\u003e\\n\\u003cli\\u003eHausiku, M.K., 2022. Mushroom Cultivation in Arid Namibia: Cultivation Status, Contribution to Human Health and Future Prospects. https://doi.org/10.1007/978-981-16-6771-8_21\\u003c/li\\u003e\\n\\u003cli\\u003eTen, S.T., Krishnen, G., Khulidin, K.A., Mohamad Tahir, M.A., Hashim, M.H., Khairudin, S., 2021. Automated Controlled Environment Mushroom House. Advances in Agricultural and Food Research Journal. http://doi.org/10.36877/aafrj.a0000230\\u003c/li\\u003e\\n\\u003cli\\u003eIslam, M.T., Zakaria, Z., Hamidin, N., Azlan, B.M.I., Mohd, Shi Fern, C., Abdullah, M.A.B., Abd Rahim, S.Z., Muhammad Suandi, M.E., Mat Saad, M.N., Ghazali, M.F., 2017. The Management of Humidifying Treatment for Low Contamination Risks during Indoor Cultivation of Grey Oyster Mushroom (Pleurotus pulmonarius). Matec Web of Conferences 97, 01080. http://doi.org/10.1051/matecconf/20179701080\\u003c/li\\u003e\\n\\u003cli\\u003eHoegh-Guldberg, O., Jacob, D., Taylor, M., Guill\\u0026eacute;n Bola\\u0026ntilde;os, T., Bindi, M., Brown, S., . . . Zhou, G. (2019). The human imperative of stabilizing global climate change at 1.5 \\u0026deg;C.\\u0026nbsp;Science,\\u0026nbsp;365(6459), Article eaaw6974.\\u003c/li\\u003e\\n\\u003cli\\u003eChen, L., Qian, L., Zhang, X., Li, J., Zhang, Z., Chen, X., 2022. Research progress on indoor environment of mushroom factory. International Journal of Agricultural \\u0026amp; Biological Engineering 15. http://doi.org/10.25165/j.ijabe.20221501.6872\\u003c/li\\u003e\\n\\u003cli\\u003eCheng, G., Chen, J., Lan, L. Y., et al. Grey correlational and genetic analyses of Pleurotus pulmonariushybrids.\\u0026nbsp;Molecular Plant Breeding, 2023, 21(23): 7811-7818.\\u003c/li\\u003e\\n\\u003cli\\u003eDe La Croix, N.J., Didacienne, M., Louis, S., 2022. Fuzzy logic-based shiitake mushroom farm control for harvest enhancement, 2022 10th International Symposium on Digital Forensics and Security (ISDFS). IEEE, pp. 1-6.https://doi.org/10.1109/ ISDFS55398.2022.9800832.\\u003c/li\\u003e\\n\\u003cli\\u003eSunghyoun, L., Byeongkee, Y., Hyuckjoo, K., Namkyu, Y., Jongcheon, J., 2015. Technology for Improving the Uniformity of the Environment in the Oyster Mushroom Cultivation House by using Multi-layered Shelves. Protected horticulture and Plant Factory 24, 128-133. http://doi.org/10.12791/ksbec.2015.24.2.128\\u003c/li\\u003e\\n\\u003cli\\u003eSeung-Mi, Moon, Sook-Youn, Kwon, Jae-Hyun, Lim, 2015. Improvement of Energy Efficiency of Plants Factory by Arranging Air Circulation Fan and Air Flow Control Based on CFD. Journal of Internet Computing and Services 16, 57-65. http://doi.org/10.25165/j.ijabe.20221501.6872\\u003c/li\\u003e\\n\\u003cli\\u003eSakamoto, Yuichi, 2018. Influences of environmental factors on fruiting body induction, development and maturation in mushroom-forming fungi. Fungal Biology Reviews, S1749461317300568. http://doi.org/10.1016/j.fbr.2018.02.003\\u003c/li\\u003e\\n\\u003cli\\u003eLee, C.J., Lee, S.H., Lee, E.J., Park, H.S., Kong, W., 2018. Analysis of growth environment for precision cultivation management of the oyster mushroom 'Suhan'. Journal of Mushroom 16, 155-161. http://doi.org/10.14480/JM.2018.16.3.155\\u003c/li\\u003e\\n\\u003cli\\u003eHair, J. F., Hult G T, M., Ringle C M, M., Sarstedt, M., Castillo Apraiz, J., Cepeda Carrion, \\u0026acute; G. A., et al., Manual de Partial least squares structural equation modeling (PLS-SEM) (2nd ed.). Los Angeles [Etc: Omnia Publisher SL Cop. https://doi. org/10.3926/oss.37\\u003c/li\\u003e\\n\\u003cli\\u003eHair, J.F., Hult, G.T.M., Ringle, C.M., Sarstedt, M., 2016. A Primer on Partial Least Squares Structural Equation Modeling (PLS-SEM), 2nd edition.\\u003c/li\\u003e\\n\\u003cli\\u003eA, Z.J., A, L.P., B, M.W.A., Systematic relationship between soil properties and organic carbon mineralization based on structural equation modeling analysis - ScienceDirect. Journal of Cleaner Production 277. https://doi.org/10.1016/j.jclepro.2020.123338.\\u003c/li\\u003e\\n\\u003cli\\u003eSa, Q., Zheng, J., Zhang, K., Wang, Y., 2025. Effects and assessment of the combined application of biogas slurry and chemical fertilizers on greenhouse tomato growth, yield, and soil quality. Scientia Horticulturae 344. https://doi.org/10.1016/j.scienta.2025.114113.\\u003c/li\\u003e\\n\\u003cli\\u003eLiu, S.Y., Vega, A.R., Dizy, M., 2023. Assessing ultrapremium red wine quality using PLS-SEM. LWT 177, 114560-. https://doi.org/10.1016/j.lwt.2023.114560\\u003c/li\\u003e\\n\\u003cli\\u003eKe, B. R., Lan, Q. X., Lu, Z. H., et al. (2023). Genetic differences and agronomic traits comparison of main cultivated varieties of Pleurotus pulmonarius.\\u0026nbsp;Scientia Horticulturae,\\u0026nbsp;11(4), 241\\u0026ndash;247.\\u003c/li\\u003e\\n\\u003cli\\u003eYan, M., Zhai, D., Li, Q., Zhang, M., Jiang, N., Liu, J., Song, C., Shang, X., Chen, H., Yu, H., 2024. Comparative Analysis of Main Agronomic Traits of Different Pleurotus giganteus Germplasm Resources. Life (2075-1729) 14. https://doi.org/10.3390/life14020238\\u003c/li\\u003e\\n\\u003cli\\u003eYu, H.L.,Zhang, M.Y., Li, Q.Z.,Zhang, L.J.,Shang, X.D., Tan, Q., 2023. A new variety of Pleurotus giganteus \\u0026rsquo;Shen Xun No.1\\u0026rsquo;. Acta Hortic.Sin. 50, 453\\u0026ndash;454.\\u003c/li\\u003e\\n\\u003cli\\u003eYu, H.L.,Zhai, D.D.,Shen, X.F.,Zhang, M.Y.,Wang, Y.X., Shang, X.D.,Zhang, D., Li, Q.Z., 2021. Quantitative character variation andprobability classification of fruiting bodies of Pleurotus giganteus germplasm resources. Acta Edulis Fungi . 28, 42\\u0026ndash;47.\\u003c/li\\u003e\\n\\u003cli\\u003eHe, J.Z., Ru, Q.M., Dong, D.D., Sun, P.L., 2012. Chemical Characteristics and Antioxidant Properties of Crude Water Soluble Polysaccharides from Four Common Edible Mushrooms. Molecules 17. https://doi.org/10.3390/molecules17044373\\u003c/li\\u003e\\n\\u003cli\\u003eJeong, H., Cho, J., Han, J., Yoon, Y.S., Kim, H.G., Kim, J., Chung, H., 2025. Large-area coverage\\u0026ndash;transmission near-infrared measurement of dried laver to determine crude protein content. Food Control 168. https://doi.org/10.1016/j.foodcont.2024.110934\\u003c/li\\u003e\\n\\u003cli\\u003eChen, X., Zhang, H., Yu, S., Zhou, C., Teng, A., Lei, L., Ba, Y., Li, F., 2024. Optimizing irrigation and nitrogen application strategies to improve sunflower yield and resource use efficiency in a cold and arid oasis region of Northwest China. Frontiers in Plant Science 15, 16. https://doi.org/10.3389/fpls.2024.1429548.\\u003c/li\\u003e\\n\\u003cli\\u003eHair, J. F., Hult, G., Ringle, C. M., \\u0026amp; Sarstedt, M., A primer on partial least squares structural equation modeling (PLS-SEM) (3rd ed.). Sage.\\u003c/li\\u003e\\n\\u003cli\\u003eRaudaskoski, M., Viitanen, H., 1982. Effect of aeration and light on fruit body induction in Schizophyllum commune. Transactions of the British Mycological Society 78, 89-96.https://doi.org/10.1016/S0007-1536(82)80080-6\\u003c/li\\u003e\\n\\u003cli\\u003eSohi, H., Upadhyay, R., 1989. Effect of temperature on mycelial growth of Pleurotus species and their yield performance on selected substrates. Mushroom Science 12(2): 49-56.\\u003c/li\\u003e\\n\\u003cli\\u003eYan, M., Zhai, D., Li, Q., Zhang, M., Jiang, N., Liu, J., Song, C., Shang, X., Chen, H., Yu, H., 2024. Comparative Analysis of Main Agronomic Traits of Different Pleurotus giganteus Germplasm Resources. Life (2075-1729) 14. https://doi.org/10.3390/life14020238\\u003c/li\\u003e\\n\\u003cli\\u003eJang, K.Y., Jhune, C.S., Park, J.S., Cho, S.M., Weon, H.Y., Cheong, J.C., Choi, S.G., Sung, J.M., 2003. Characterization of Fruitbody Morphology on Various Environmental Conditions in Pleurotus ostreatus. Mycobiology 31.https://doi.org/4489/MYCO.2003.31.3.145.\\u003c/li\\u003e\\n\\u003cli\\u003eLee, C.J., Lee, S.H., Lee, E.J., Park, H.S., Kong, W., 2018. Analysis of growth environment for precision cultivation management of the oyster mushroom 'Suhan'. Journal of Mushroom 16, 155-161. https://doi.org/10.14480/JM.2018.16.3.155\\u003c/li\\u003e\\n\\u003cli\\u003eJhune, C.-S., Yun, H.-S., Jang, K.-Y., Kong, W.-S., Lee, K.-H., Lee, C.-J., Yoo, Y.-B., 2011. The effect of temperature on morphological of fruiting body and cultivated characteristics of oyster mushroom. Journal of Mushroom 9, 155-160.\\u003c/li\\u003e\\n\\u003cli\\u003ePaul, S. and Chilton, J. S., 1983. Chapter environmental factors: sustaining the mushroom crop in the mushroom cultivator. pp. 149-157.\\u003c/li\\u003e\\n\\u003cli\\u003eKinugawa, K., Suzuki, A., Takamatsu, Y., Kato, M., Tanaka, K., 1994. Effects of concentrated carbon dioxide on the fruiting of several cultivated basidiomycetes (II). Mycoscience 35, 345-352. https://doi.org/10.1007/BF02268504\\u003c/li\\u003e\\n\\u003cli\\u003eJang, K.Y., Jhune, C.S., Park, J.S., Cho, S.M., Weon, H.Y., Cheong, J.C., Choi, S.G., Sung, J.M., 2003. Characterization of Fruitbody Morphology on Various Environmental Conditions in Pleurotus ostreatus. Mycobiology 31. https://doi.org/10.4489/MYCO.2003.31.3.145\\u003c/li\\u003e\\n\\u003cli\\u003eLin, R., Zhang, L., Yang, X., Li, Q., Zhang, C., Guo, L., Yu, H., Yu, H., 2022. Responses of the Mushroom Pleurotus ostreatus under Different CO(2) Concentration by Comparative Proteomic Analyses. J Fungi (Basel) 8. http://doi.org/10.3390/jof8070652\\u003c/li\\u003e\\n\\u003cli\\u003eYang Y., Shen Y., Cao Y., Xu G., Cai W., Effects and Analysis of Temperature and Humidity on the Appearance Quality of Pleurotus pulmonarius During Summer Fruiting. Edible and Medicinal Mushrooms 31, 191-195.\\u003c/li\\u003e\\n\\u003cli\\u003eSong J., Chen Q., Zhang C., Zong T., Yuan W., Yu B., The effect of cultivation period on the agronomic traits and yield of Pleurotus eryngii. Journal of Northwest A\\u0026amp;F University (Natural Science Edition) 53,137-144+156.http://doi.org/10.13207/j.cnki.jnwafu.2025.06.014\\u003c/li\\u003e\\n\\u003cli\\u003eNakazawa, T., Miyazaki, Y., Kaneko, S., Shishido, K., 2008. Stimulative effects of light and a temperature downshift on transcriptional expressions of developmentally regulated genes in the initial stages of fruiting-body formation of the basidiomycetous mushroom Lentinula edodes. FEMS Microbiology Letters. https://doi.org/10.1111/j.1574-6968.2008.01364.x.\\u003c/li\\u003e\\n\\u003cli\\u003eOhm, R.A., Aerts, D., Wosten, H.A., Lugones, L.G., 2013. The blue light receptor complex WC-1/2 of Schizophyllum commune is involved in mushroom formation and protection against phototoxicity. Environ Microbiol 15, 943-955. http://doi.org/10.1111/j.1462-2920.2012.02878.x\\u003c/li\\u003e\\n\\u003cli\\u003eTerashima, K., Yuki, K., Muraguchi, H., Akiyama, M., Kamada, T., 2005. The dst1 gene involved in mushroom photomorphogenesis of Coprinus cinereus encodes a putative photoreceptor for blue light. Genetics 171, 101-108. http://doi.org/10.1534/genetics.104.040048\\u003c/li\\u003e\\n\\u003cli\\u003eJin R., Tan X., Ma W., Dong Y., Li F., 2021. Quality Evaluation of Pleurotus ostreatus Fruiting Bodies. China Vegetables 34, 74-79.http://doi.org/10.16861/j.cnki.zggc.2021.0236\\u003c/li\\u003e\\n\\u003cli\\u003eZivanovic, S., Busher, R.W., Kim, K.S., 2000. Textural Changes in Mushrooms (Agaricus bisporus) Associated with Tissue Ultrastructure and Composition. Journal of Food Science 65, 1404-1408. http://doi.org/10.1111/j.1365-2621.2000.tb10621.x\\u003c/li\\u003e\\n\\u003cli\\u003eKotwaliwale, N., Bakane, P., Verma, A., 2007. Changes in textural and optical properties of oyster mushroom during hot air drying. Journal of Food Engineering 78, 1207-1211. http://doi.org/10.1016/j.jfoodeng.2005.12.033\\u003c/li\\u003e\\n\\u003cli\\u003eKortei, N.K., Odamtten, G.T., Obodai, M., Appiah, V., Abbey, L., Oduro-Yeboah, C., Akonor, P.T., 2015. Infuence of gamma radiation on some textural properties of fresh and dried oyster mushrooms (Pleurotus ostreatus). https://csirspace.foodresearchgh.org/handle/1/348\\u003c/li\\u003e\\n\\u003cli\\u003eZou M., Xu P., Lu X., Zhu K., Zhen J., Jin R., 2024. Analysis of the characteristics of Pleurotus ostreatus texture based on multivariate analysis method. Northern Horticulture, 119-126. http://doi.org/doi:10.11937/bfyy.20232781\\u003c/li\\u003e\\n\\u003cli\\u003eSa, Q., Zheng, J., Zhang, K., Wang, Y., 2025. Effects and assessment of the combined application of biogas slurry and chemical fertilizers on greenhouse tomato growth, yield, and soil quality. Scientia Horticulturae 344. http://doi.org/10.1016/j.scienta.2025.114113\\u003c/li\\u003e\\n\\u003cli\\u003eZhao, X., Mak-Mensah, E., Zhao, W., Wang, Q., Zhou, X., Zhang, D., Zhu, J., Qi, W., Liu, Q., Li, X., 2024. Optimized ridge-furrow technology with biochar amendment for alfalfa yield enhancement and soil erosion reduction based on a structural equation model on sloping land. Agricultural Water Management, 298. http://doi.org/10.1016/j.agwat.2024.108866\\u003c/li\\u003e\\n\\u003cli\\u003eWu, W., Han, J., Gu, Y., Li, T., Xu, X., Jiang, Y., Li, Y., Sun, J., Pan, G., Cheng, K., 2022. Impact of biochar amendment on soil hydrological properties and crop water use efficiency: A global meta‐analysis and structural equation model. GCB Bioenergy 14, 657-668. https://doi.org/10.1111/gcbb.12933.\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"scientific-reports\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"scirep\",\"sideBox\":\"Learn more about [Scientific Reports](http://www.nature.com/srep/)\",\"snPcode\":\"\",\"submissionUrl\":\"\",\"title\":\"Scientific Reports\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Scientific Reports\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true},\"keywords\":\"Pleurotus geesteranus, Dynamic environmental control, Two-stage cultivation, Yield and quality, Structural equation modeling (SEM)\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-8225452/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-8225452/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"Overcoming the trade-off between yield and quality in mushroom cultivation requires a deeper understanding of how environmental factors coordinately regulate morphogenesis. This study introduces a paradigm shift from static environmental control to a phased, dynamic strategy for Pleurotus geesteranus. We proposed that sequentially optimizing conditions for distinct developmental phases—first enhancing stipe elongation under high temperature/CO₂, then initiating pileus expansion under moderated temperature with ventilation and light—would synchronize yield and quality. Validated in a digitally-controlled fruiting cabin (DSMFC), this strategy significantly increased yield (304.2 g/bag), biological efficiency (46.7%), and the premium mushroom ratio (46.0%) over traditional systems. The underlying mechanisms were elucidated via Structural Equation Modeling, which quantified accumulated heat units and CO₂ concentration as the pivotal environmental variables directly influencing stipe morphology and indirectly boosting yield. Economic analysis confirmed commercial viability, highlighting the potential of this targeted environmental programming to advance precision horticulture.\",\"manuscriptTitle\":\"Beyond Static Control: A Dynamic Environmental Regime Informed by Structural Equation Modeling Enhances Yield and Quality of Pleurotus geesteranus\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-04-29 08:56:20\",\"doi\":\"10.21203/rs.3.rs-8225452/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2026-05-06T15:48:43+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"296331366687716972919736571671625313555\",\"date\":\"2026-04-28T06:53:10+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2026-04-27T12:04:55+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"25692432851445229841532434802445519734\",\"date\":\"2026-04-26T04:39:38+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"169357208128008048156054122019293390222\",\"date\":\"2026-04-23T11:18:13+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2026-04-21T04:16:10+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2026-04-14T19:41:56+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvited\",\"content\":\"\",\"date\":\"2025-12-10T10:51:44+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2025-12-08T13:27:36+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Scientific Reports\",\"date\":\"2025-12-08T13:13:37+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"scientific-reports\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"scirep\",\"sideBox\":\"Learn more about [Scientific Reports](http://www.nature.com/srep/)\",\"snPcode\":\"\",\"submissionUrl\":\"\",\"title\":\"Scientific Reports\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Scientific Reports\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"cc4cfe73-8b80-4c8d-a1e5-ed1e750a3aca\",\"owner\":[],\"postedDate\":\"April 29th, 2026\",\"published\":true,\"recentEditorialEvents\":[{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2026-05-06T15:48:43+00:00\",\"index\":116,\"fulltext\":\"\"}],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[{\"id\":67193079,\"name\":\"Biological sciences/Ecology\"},{\"id\":67193080,\"name\":\"Earth and environmental sciences/Ecology\"},{\"id\":67193081,\"name\":\"Earth and environmental sciences/Environmental sciences\"},{\"id\":67193082,\"name\":\"Biological sciences/Plant sciences\"}],\"tags\":[],\"updatedAt\":\"2026-04-29T08:56:20+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-04-29 08:56:20\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-8225452\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-8225452\",\"identity\":\"rs-8225452\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}