Culture Substrate Research Articles

Efficacy of slow sand filtration enriched with Trichoderma atroviride in the control of Rhizoctonia solani in soilless culture

Soilless cultivation is increasingly common, but the nutrient-rich drainage from substrate cultivation is often discarded. However, drainage can be safely reused if previously disinfected. Slow sand filtration (SSF) is a low-cost, ecological, and effective method for water disinfection, primarily through biological control. Enhancing SSF with antagonistic microorganisms is not well-studied. Additionally, SSF has not been tested to control Rhizoctonia solani, a phytopathogen that can be spread by irrigation water. Therefore, the objective of his work was to test the efficacy of a slow sand filter improved through the inoculation of the antagonistic fungus Trichoderma atroviride, evaluating its suppression capacity against Rhizoctonia solani spread by the irrigation water in a closed substrate cultivation of cucumber (Cucumis sativus). Five experiments were conducted, testing the presence and absence of a sand filter, T. atroviride, and R. solani in each trial. Median disease severity was expressed on a scale of 1–5. The improved SSF increased disease control percentage by 49% compared to SSF alone and by 86% compared to no disease control method. In some experiments, SSF with T. atroviride totally controlled R. solani. The results confirm that biologically enhanced SSF with T. atroviride can effectively disinfect drainage in closed soilless cultivation systems infected with R. solani.

Rapid Reduction of Phytotoxicity in Green Waste for Use as Peat Substitute: Optimization of Ammonium Incubation Process.

The rapid growth of the horticultural industry has increased demand for soilless cultivation substrates. Peat, valued for its physical and chemical properties, is widely used in soilless cultivation. However, peat is non-renewable, and over-extraction poses serious ecological risks. Therefore, sustainable alternatives are urgently needed. Ammonium incubation, a novel method to reduce phytotoxicity, offers the potential for green waste, a significant organic solid waste resource, to substitute peat. This study optimized the ammonium incubation process to reduce green waste phytotoxicity. It systematically examined different nitrogen salts (type and amount) and environmental conditions (temperature, aeration, duration) affecting detoxification efficiency. Results show a significant reduction in phytotoxicity with ammonium bicarbonate, carbonate, and sulfate, especially carbonate, at 1.5%. Optimal conditions were 30 °C for 5 days with regular aeration. Under these conditions, ammonium salt-treated green waste significantly reduced total phenolic content and stabilized germination index (GI) at a non-phytotoxic level (127%). Using treated green waste as a partial peat substitute in lettuce cultivation showed promising results. This low-cost, low-energy method effectively converts green waste into sustainable peat alternatives, promoting eco-friendly horticulture and environmental conservation.

Valuation of the growth response of oyster (Pleurotus ostreatus) mushroom on partially composted sesame stalk with different blends of wheat straw

Aside from being a significant source of highly nutritious and medicinally valuable mushrooms, mushroom cultivation is an important means of environmental remediation. Oyster mushroom (Pleurotus ostreatus) is a versatile mushroom that can be grown in various environmental conditions and non-composted lignocellulosic wastes. The present study was conducted to assess the growth response of oyster mushroom on sesame stalk. Spawn was prepared from a mycelium culture grown on potato dextrose agar. The spawn was inoculated onto sesame stalk and wheat straw substrates and incubated at room temperature in the dark for 21 days. A Completely Randomized Design (CRD) was used with five treatments, including the control (100% wheat straw), which were triplicated. Data was analyzed using Minitab version 19. Significant (P < 0.05) differences in days taken for mycelial colonization, primordial appearance, maturity and yield was observed between the substrate formulations. 100% SS (100% sesame stalk) was fastest (19.00 days) in mycelial colonization period but statistically parity with 75% SS + 25% WS (75% sesame stalk + 25% wheat straw). 75% SS + 25% WS (75% sesame stalk + 25% wheat straw) was faster (3.33 days) for primordial appearance while 100% SS(100% sesame stalk) was faster (6.67 days) in maturity. The highest yield (1850 g) and biological efficiency (185%) was recorded by 100% SS (100% sesame stalk). Pearson’s correlation showed that cap diameter and stipe length directly correlate yield and biological efficiency and faster substrate colonization leads to quicker maturation. The current study suggests that sesame stalk can be an alternative substrate for cultivation of oyster mushrooms.

Sphagnum moss and peat comparative study: Metal release, binding properties and antioxidant activity.

Peat is the main constituent of cultivation substrates and a precious non-renewable fossil material. Peatlands provide important ecosystem services and allow the absorption and storage of carbon. Protecting peatlands helps tackle climate change and contributes to biodiversity conservation. Due to its importance, it is necessary to implement strategies to reduce the use of peat, such as replacing it with biomass-based alternative growing media constituents, such as Sphagnum moss. In this study, we compared the metal release and binding properties at two different pH, antioxidant activity, and total phenolic content of peat and Sphagnum moss from the Tierra del Fuego (TdF) region of southern Patagonia. Levels of the elements were determined by inductively coupled plasma mass spectrometry (ICP-MS), while the types and amounts of functional groups were characterized and compared using Fourier transform infrared (FTIR) spectroscopy. The total phenol level and antioxidant capacity were assessed using the Folin-Ciocalteu method and 2,2-diphenyl-1-picrylhydrazyl test. There are generally higher concentrations of leachable elements in peat than in Sphagnum moss at pH = 2, except Cs, Rb, Ti, and Zr. In contrast, at pH = 5, levels of all leached elements are highest in Sphagnum moss. Sphagnum moss shows a higher metal adsorption capacity than peat, except for Be, Mn, Tl, and Zn. Finally, the results showed that both matrices contained similar total phenolic contents: 0.018 ± 0.011 mg gallic acid equivalent (GAE) per gram dry sample for peat and 0.020 ± 0.007 mg GAE g-1 for Sphagnum moss. Instead, Sphagnum moss extracts showed a significantly higher antioxidant activity [0.026 ± 0.028 mmol Trolox equivalents (TE) g-1] than that estimated in peat (0.009 ± 0.005 mmol TE g-1). Humic acids, carboxylic acids, and phenolic and lignin groups were identified as the functional groups that mainly determined the antioxidant activity of the Sphagnum moss compared to peat. The present study resulted in an advancement of knowledge of these materials for more thoughtful future use and possible replacements.

Collagen as Bioink for Bioprinting: A Comprehensive Review.

Biomaterials made using collagen are successfully used as a three-dimensional (3D) substrate for cell culture and considered to be promising scaffolds for creating artificial tissues. An important task that arises for engineering such materials is the simulation of physical and morphological properties of tissues, which must be restored or replaced. Modern additive technologies, including 3D bioprinting, can be applied to successfully solve this task. This review provides the latest evidence on advances of 3D bioprinting with collagen in the field of tissue engineering. It contains modern approaches for printing pure collagen bioinks consisting only of collagen and cells, as well as the obtained results from the use of pure collagen bioinks in different fields of tissue engineering.

Suitability of coconut bran and biochar as a composite substrate for lettuce cultivation in aquaponic systems

Growth substrates are essential for aquaponic systems and play an important role in vegetable growth and water quality. In this study, we explored an innovative combination of coconut bran and coconut shell biochar (CSB) as a composite growth substrate for lettuce cultivation in aquaponic systems. The study included the control (100 % coconut bran as the growth substrate) and treatment groups (T1–T5; containing 10 %, 20 %, 30 %, 40 %, and 50 % CSB as the growth substrate, respectively). The substrate properties; lettuce growth performance; and soil enzyme activity, nitrogen content, and abundance of microbial communities in the substrate were analyzed to determine the optimal substrate. Our findings indicated that CSB incorporation significantly altered the properties of the substrate, resulting in increased dry and bulk densities, pH, and water-holding capacity, and decreased electrical conductivity, water-absorption capacity, and porosity. Furthermore, the fresh weight of lettuce was notably increased in the treatment groups. The activities of fluorescein diacetate hydrolase, urease, nitrate reductase, and hydroxylamine reductase initially increased and further decreased, reaching the maximum in the T3 group. Conversely, the activity of nitrite reductase and contents of available nitrogen, nitrate-nitrogen, and ammonium-nitrogen in the substrates initially decreased and further increased, with the minimum values observed in the T3 group. The microbial sequencing results indicated that CSB incorporation significantly increased the microbial diversity and relative abundance of microorganisms associated with nitrogen transformation. Moreover, 30 % CSB incorporation exhibited the greatest effect on lettuce growth, with a 34.5 % and 31.6 % increase in fresh weight compared to the control during the growth and harvest periods, respectively. This study indicated the enormous potential of biochar in the research and development of green technologies for substrate amendment in aquaponic systems.

In vitro protein digestibility and mineral accessibility of edible filamentous Fungi cultivated in oat flour

Edible filamentous fungi, a source of mycoprotein, are one of the sustainable alternative protein. This study compares protein digestibility (DH%) and amino acid and mineral accessibility in Rhizopus oligosporus cultivated in oat flour (OatRO) or glucose media (GluRO) by using the INFOGEST in vitro digestion protocol. Fungal total amino acids was higher in GluRO (39.0 ± 1.1 % dw) than OatRO (21.8 ± 1.3 % dw) which was also the case for calcium and magnesium content. After completed gastrointestinal digestion, there were no significant differences between GluRO and OatRO regarding DH% (27.21 ± 10.4 % and 29.4 ± 0.5 %), however, GluRO provided significantly higher amino acid accessibility compared to OatRO (64.3 ± 1.6 % and 55.1 ± 3.1 %). Mineral accessibility of GluRO was for Ca: 37.9 ± 1.8 %, Zn: 9.3 ± 0.4 %, Fe: 38.2 ± 1.9 %, Mg: 66.5 ± 1.4 % and Cu: 24.7 ± 1.3 % and for OatRO; Ca: −40.2 ± 2.4 %, Zn: −4.13 ± 0.15 %, Fe:14.6 ± 1.6 %, Mg: 74.5 ± 3.1 %, and Cu: 55.95 ± 0.8 %. Despite the low phytic acid content, OatRO thus showed antinutrient properties with respect to calcium, and zinc, suggesting that oat-derived fungi had antinutrients other than phytic acid. This study hereby revealed that the cultivation substrate affect amino acid and mineral accessibility of filamentous fungi and calls for deeper evaluations of antinutrients in oat-derived fungi.

‘Unions et germanies’. Armed Mobilisation, Plebeian Politicisation and Historical Memory in the Kingdom of Valencia (fourteenth-seventeenth centuries)

Although the ‘Unions et germanies’ were forbidden, popular movements arose in the Kingdom of Valencia that expressed themselves mainly through judicial means, but sometimes also resorted to arms. Analysis of the actors’ capacity for agency is therefore complex. In particular, the Germania (1519-1522) revalued the participation of the urban majorities in public affairs, based on the armed and legal mobilisation of the city’s artisans. Other grievances throughout the Kingdom of Valencia strengthened the movement. The conflict was radicalised, giving rise to armed actions against privilege that redefined its trajectory. The classist writings of the chroniclers repudiated this collective violence. The adoption of rational choices to achieve objectives reveals the political subjectivity created by the war, which marked the memory of the commoners. A widespread resistance kept alive their banners, which became the cultural substratum of later struggles. The essay rescues the forms assumed by plebeian political identities in the sixteenth-century conflict, inscribing them in the cycle of collective action that began with the Union (1347-1348) and ended with the Second Germania (1693), also comparing the potentialities of these dissimilar experiences.

Response of Yam Yield and Soil Microbial Communities to Soil Fumigation and Substrate Cultivation

Response of Yam Yield and Soil Microbial Communities to Soil Fumigation and Substrate Cultivation

Above- and below-ground morpho-physiological traits indicate that biochar is a potential peat substitute for grapevine cuttings nursery production

The growing demand for grapevine planting materials, due to growing global viticulture, is promoting research studies to improve vineyard sustainability. In greenhouse nurseries, peat is the most common growing medium component used although is an expensive and non-renewable material. Indeed, the reduction of peat exploitation is receiving great attention, and currently, several materials are being investigated as peat substitutes for composing the cultivation substrates. Biochar, a carbon-rich, recalcitrant charred organic co-product of the pyrolysis or gasification process, has emerged as a potentially promising replacement for soilless substrates in nursery plant material propagation. Although several studies carried out at greenhouse nurseries have shown that biochar, can improve plant growth, only a few studies have focused on the production of grapevine plant material. To fulfil this knowledge gap and push forward the sustainability of the nursery sector, we evaluated above and below-ground morpho-physiological traits of one-year-old potted grapevine cuttings growing with 30% volume of four different biochar types (i.e., from pyrolysis and gasification) mixed with commercial peat. The present study shows that biochar can be used in growing media mixes without adverse effects on roots, improves soil water retention and leaf water potential, and improves the effects on soil microbiology.

Soybean molasses as a new and low-cost substrate for gluconic acid production by Aspergillus niger

Soybean molasses (SM), a sugar-rich by-product obtained from manufacturing soy protein concentrate, is an interesting source of fermentable sugars for biotechnological processes. For the first time, SM was used as a new and low-cost substrate in Aspergillus niger cultures to produce gluconic acid (GA), a carboxylic acid obtained by the oxidation of glucose and with a wide range of applications. The effect of SM composition and sugar content on GA batch production was studied in a stirred-tank bioreactor using two SM samples with different total sugar concentrations of 46 % and 52 % (w/w) and different sugar profiles. GA productivity improved as the total sugar concentration increased, whereas yield was unaffected. Agitation rate increase positively affected yield and productivity in SM medium with the highest sugar concentration. Under these conditions, the highest GA titer (79 g·L−1) was obtained. Oxalic acid and fructose syrup were other added-value compounds found from the submerged fermentation of SM. This work proved the potential of A. niger in the bioconversion of SM with different compositions into GA, providing a novel strategy for low-cost and eco-friendly GA production from SM.

Effects of the rice-mushroom rotation pattern on soil properties and microbial community succession in paddy fields.

Currently, straw biodegradation and soil improvement in rice-mushroom rotation systems have attracted much attention. However, there is still a lack of studies on the effects of rice-mushroom rotation on yield, soil properties and microbial succession. In this study, no treatment (CK), green manure return (GM) and rice straw return (RS) were used as controls to fully evaluate the effect of Stropharia rugosoannulata cultivation substrate return (SRS) on soil properties and microorganisms. The results indicated that rice yield, soil nutrient (organic matter, organic carbon, total nitrogen, available nitrogen and available potassium) and soil enzyme (urease, saccharase, lignin peroxidase and laccase) activities had positive responses to the rice-mushroom rotation. At the interannual level, microbial diversity varied significantly among treatments, with the rice-mushroom rotation significantly increasing the relative alpha diversity index of soil bacteria and enriching beneficial microbial communities such as Rhizobium, Bacillus and Trichoderma for rice growth. Soil nutrients and enzymatic activities were significantly correlated with microbial communities during rice-mushroom rotation. The fungal-bacterial co-occurrence networks were modular, and Latescibacterota, Chloroflexi, Gemmatimonadota and Patescibacteria were closely related to the accumulation of nutrients in the soil. The structural equation model (SEM) showed that fungal diversity responded more to changes in soil nutrients than did bacterial diversity. Overall, the rice-mushroom rotation model improved soil nutrients and rice yields, enriched beneficial microorganisms and maintained microbial diversity. This study provides new insights into the use of S. rugosoannulata cultivation substrates in the sustainable development of agroecosystems.

Hydroponic cultivation enhances biomass and medicinal safety of Houttuynia cordata Thunb: An integrated physiological and multi-omics analysis

Houttuynia cordata Thunb. (H. cordata) is a significant medicinal plant in China, known for its strong effects in healthcare and disease treatment. However, wild-harvested H. cordata often accumulates heavy metal contaminants, and currently, there is no environmentally safe method for its cultivation. This study aims to explore the influence of cultivation substrates on the growth, development, and medicinal components of H. cordata, and to elucidate the potential mechanisms underlying the increased biomass of H. cordata under hydroponic conditions using multi-omics techniques. Methodologically, we employed soilless cultivation to ensure medicinal safety and used multi-omics analysis to reveal the differential metabolites and genes associated with various cultivation methods. Our findings indicate that soilless cultivation effectively reduces H. cordata’s accumulation of benzene while maintaining medicinal component levels comparable to traditional soil cultivation. Hydroponically grown H. cordata exhibits significantly higher biomass compared to other methods. The integration of multi-omics analysis shows that differential metabolites and genes in hydroponically grown H. cordata are primarily enriched in nitrogen metabolism and photosynthetic pathways. Additionally, we found that increased nitric oxide content significantly enhances H. cordata ’s photosynthesis and gene expression, promoting leaf and root growth. In conclusion, soilless cultivation reduces benzene metabolite levels in H. cordata while preserving its medicinal properties, and hydroponics facilitates its growth and development. This study lays a theoretical foundation for the industrial production of environmentally friendly, uncontaminated medicinal H. cordata.

Digital Photoinduced Topographical Microsculpting of Hydrogels

AbstractHydrogels are soft materials widely used as a substrate or matrix for cell culture to mimic the physicochemical, structural, and mechanical properties of the cell's native microenvironment. Recently, a growing body of evidence has shown that substrate topography is a key determinant of cell behavior. These studies, however, mostly rely on the use of hard elastomeric materials, particularly as introducing topography in soft hydrogels is technically challenging, labor intensive, and often does not provide a flexible design space. Here, a new approach is presented that allows robust creation of topographies on hydrogels using a maskless and contactless UV‐sculpting approach. The sculpted topographies are highly programmable: the features can be designed using any digital drawing software, while the dimensions and resolution can be easily tuned via UV exposure dose or photoinitiator concentration. Microscale topographies (2–100 µm in size) can be created on a wide range of hydrogels with diverse chemical characters and mechanical properties (e.g., stiffness). As a proof of concept, this work shows that adherent cells on softer hydrogels align better to topographical cues than on stiffer hydrogels, underlining the importance of studying cell behavior in physiologically more relevant multi‐cue environments.

Long-Term Culture of Human Pluripotent Stem Cells in Xeno-Free Condition Using Functional Polymer Films.

Human pluripotent stem cells (hPSCs), encompassing human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), hold immense potential in regenerative medicine, offering new opportunities for personalized cell therapies. However, their clinical translation is hindered by the inevitable reliance on xenogeneic components in culture environments. This study addresses this challenge by engineering a fully synthetic, xeno-free culture substrate, whose surface composition is tailored systematically for xeno-free culture of hPSCs. A functional polymer surface, pGC2 (poly(glycidyl methacrylate-grafting-guanidine-co-carboxylic acrylate)), offers excellent cell-adhesive properties as well as non-cytotoxicity, enabling robust hESCs and hiPSCs growth while presenting cost-competitiveness and scalability over Matrigel. This investigation includes comprehensive evaluations of pGC2 across diverse experimental conditions, demonstrating its wide adaptability with various pluripotent stem cell lines, culture media, and substrates. Crucially, pGC2 supports long-term hESCs and hiPSCs expansion, up to ten passages without compromising their stemness and pluripotency. Notably, this study is the first to confirm an identical proteomic profile after ten passages of xeno-free cultivation of hiPSCs on a polymeric substrate compared to Matrigel. The innovative substrate bridges the gap between laboratory research and clinical translation, offering a new promising avenue for advancing stem cell-based therapies.

Feasibility study on heterotrophic utilization of galactose by Chlorella sorokiniana and promotion of galactose utilization through mixed carbon sources culture

BackgroundThe development of alternative carbon sources is important for reducing the cost of heterotrophic microalgae cultivation. Among cheap feedstocks, galactose is one of the most abundant sugars and can be easily obtained from many natural biomasses. However, it is generally difficult to be utilized by microalgae. In addition, the mechanism of its low utilization efficiency in heterotrophic cultivation is still unknown.ResultsAmong seven tested carbon sources, only glucose and acetate could be efficiently utilized by C. sorokiniana in heterotrophic cultivation while there were no apparent signs of utilization of other carbohydrates, including galactose, in regular heterotrophic cultivation. However, galactose could be utilized in cultures with high inoculation sizes. This confirmed that C. sorokiniana has a complete pathway for transporting and assimilating galactose under dark conditions, but the rate of galactose utilization is quite low. In addition, the galactose utilization was greatly enhanced in mixotrophic cultures, which indicated that galactose utilization could be enhanced by additional pathways that can enhance cell growth. Based on above results, a mixed carbon source culture strategy was proposed to improve the utilization rate of galactose, and a significant synergistic effect on cell growth was achieved in cultures using a mixture of galactose and acetate.ConclusionsThis study indicated that the galactose metabolism pathway may not be inherently deficient in Chlorophyta. However, its utilization rate was too low to be detected in regular heterotrophic cultivation. Mixed carbon source culture strategy was confirmed effective to improve the utilization rate of galactose. This study contributes to a deeper understanding of the utilization ability of difficultly utilized substrates in the heterotrophic cultivation of microalgae, which is of great significance for reducing the cost of heterotrophic cultivation of microalgae.

Cultivation of earthworms and analysis of associated bacterial communities during earthworms’ growth using two types of agricultural wastes

Earthworm cultivation can effectively promote the resource utilization of agricultural waste. The efficient utilization of agricultural waste by earthworms mainly depends on the microbial communities in the guts. This study used silkworm excrement and cow manure as substrates for earthworm cultivation and investigated the associated bacterial communities during earthworms’ growth. The survival rate of earthworms remained above 89% after 21 days of feeding with the two substrates. Proteobacteria, Actinobacteria, and Firmicutes constituted the predominant bacterial communities in earthworm growth, accounting for over 81% of the relative abundance in both guts and vermicompost. The bacteria richness and diversity in the foregut and midgut of earthworm were lower than those in the hindgut. The prediction function of intestinal bacterial communities of earthworms cultured with two substrates mainly involved biosynthesis, decomposition and energy production.

Lettuce (Lactuca sativa L.) Cultures and the Bioactivity of Their Root Microflora Are Affected by Amended Soil.

This study aimed to highlight the positive effects of various recycled organic substrates on lettuce plants (Lactuca sativa L.) and to promote sustainable waste management practices, contributing to the concept of a circular economy. Over a two-month period, the growth potential and rhizosphere microflora of lettuce plants grown in soil amended with different recycled substrates were investigated. All data were compared, and the effects of the culture substrates were evaluated. All groups containing soil improvers offered a significant increase in the number of leaves per plant and, in two cases, an increase in dry biomass as well as an increase in the concentration of all leaf pigments. Both MDA and H2O2 concentrations were the lowest in two groups containing soil improvers (VG 5% and PLUS 10%). At the end of the culture period, isolation and culture of bacteria from the plant rhizosphere were performed. Different bacterial strains were isolated and tested for the production of antimicrobial agents against six microbial indicators (B. subtilis, E. coli, S. aureus, S. cerevisiae, C. albicans, and P. aeruginosa). The greater percentage of the isolated strains showed an ability to inhibit the growth of the B. subtilis index. Most of the strains with antimicrobial activity were isolated from the soil samples of the plain soil group and the soil amended with the commercial fertilizer. Three of the isolated strains originating from the Ginagro 5% group are multiproducers as they inhibit the growth of three microbial indicators or more.

Loop-mediated isothermal amplification (LAMP) assay coupled with gold nanoparticles for colorimetric detection of Trichoderma spp. in Agaricus bisporus cultivation substrates

One of the significant challenges in organic cultivation of edible mushrooms is the control of invasive Trichoderma species that can hinder the mushroom production and lead to economic losses. Here, we present a novel loop-mediated isothermal amplification (LAMP) assay coupled with gold nanoparticles (AuNPs) for rapid colorimetric detection of Trichoderma spp. The specificity of LAMP primers designed on the tef1 gene was validated in silico and through gel-electrophoresis on Trichoderma harzianum and non-target soil-borne fungal and bacterial strains. LAMP amplification of genomic DNA templates was performed at 65 °C for only 30 min. The results were rapidly visualized in a microplate format within less than 5 min. The assay is based on salt-induced aggregation of AuNPs that is being prevented by the amplicons produced in case of positive LAMP reaction. As the solution color changes from red to violet upon nanoparticle aggregation can be observed with the naked eye, the developed LAMP-AuNPs assay can be easily operated to provide a simple initial screening for the rapid detection of Trichoderma in button mushroom cultivation substrate.

The role of deadwood substrates in promoting moss growth: Decay class and particle size effects

AbstractPlagiomnium acutum has a high value of landscape application and medicinal value, but there is a lack of related research on propagation and cultivation techniques. The deadwood substrate has rich nutrients and superior water retention properties, which will be conducive to promoting the growth of moss. Nevertheless, the underlying mechanisms by which deadwood influences moss growth are not yet fully unclear. In this study, we pulverized deadwood from five decay classes of Pinus massoniana into three distinct particle sizes. Through a pot experiment, we investigated the effects of decay class and physicochemical properties on the growth and physiology of Plagiomnium acutum, aiming to identify the most suitable growth substrate. The results indicated that both the decay class and particle diameter of deadwood significantly affect the substrate's physicochemical characteristics and the growth indexes of P. acutum, with the decay class exerting a more pronounced effect. The water‐holding porosity, water‐holding capacity, total nitrogen, total phosphorus, total potassium and lignin content of the substrate positively affected the growth of P. acutum, while the bulk density, void ratio, total carbon, carbon‐to‐nitrogen ratio, condensed tannin content and cellulose content had negative impacts. A comprehensive evaluation using a fuzzy membership function indicated that deadwood with higher decay classes (IV and V) was more conducive to the growth of P. acutum. Specifically, substrates from decay class IV with particle sizes of 10–20 mm provided the most favorable conditions for P. acutum and were recommended as the optimal cultivation substrate. The results of this study provide theoretical basis and technical support for the propagation and cultivation of P. acutum, and provide a foundation for further development of the industrial, pharmaceutical and environmental biotechnology potential of P. acutum.