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Article

Utilization of Giant Mimosa Stalk to Produce Effective Stick Spawn for Reducing Inoculum Costs in Economic Mushroom Farming Systems

by
Orlavanh Xayyavong
1,2,3,
Worawoot Aiduang
2,4,
Kritsana Jatuwong
2,4,* and
Saisamorn Lumyong
2,5,6,*
1
Doctor of Philosophy Program in Applied Microbiology (International Program) in Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand
2
Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand
3
Department of Biology, Faculty of Science, Champasack University, Pakse 16010, Laos
4
Office of Research Administration, Chiang Mai University, Chiang Mai 50200, Thailand
5
Center of Excellence in Microbial Diversity and Sustainable Utilization, Chiang Mai University, Chiang Mai 50200, Thailand
6
Academy of Science, The Royal Society of Thailand, Bangkok 10300, Thailand
*
Authors to whom correspondence should be addressed.
Agriculture 2025, 15(15), 1584; https://doi.org/10.3390/agriculture15151584
Submission received: 25 May 2025 / Revised: 30 June 2025 / Accepted: 21 July 2025 / Published: 23 July 2025
(This article belongs to the Special Issue Edible and Medicinal Mushrooms: Molecular Biology, Cultivation, Active Compounds, Preservation and Processing)
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Versions Notes

Abstract

The high cost of mushroom spawn remains a critical constraint to economically viable mushroom cultivation, particularly for small-scale farmers. This study investigated four spawn types, including stick (giant mimosa stalks, GMS), sawdust, sorghum, and liquid culture as inoculum sources for 10 edible mushroom species. The results indicated that GMS stick spawn provides excellent conditions for the mycelial growth of seven species, outperforming other spawn types in terms of colonization rate and pinhead formation. Mushrooms grown on GMS substrate demonstrated rapid development, with full colonization occurring within 11 to 26 days and pinhead initiation between 18 and 47 days, depending on the species. Among the mushroom species tested, Schizophyllum commune exhibited the fastest growth, reaching full colonization in 11 days and forming pinheads after 18 days of inoculation. In comparison, Auricularia polytricha showed the slowest development. Economically, GMS spawn was the most cost-effective at 0.074 USD per unit, significantly lower than sawdust (0.24 USD), sorghum (0.29 USD), and potato dextrose broth (PDB; 2.80 USD). The conversion from PDB with GMS could reduce industrial inoculum costs from 35,000 USD to 600 USD annually. These findings demonstrate the potential of GMS as an effective, low-cost, and sustainable spawn option that can enhance mycelial growth and support eco-friendly farming practices.

1. Introduction

The cultivation of edible and medicinal mushrooms is obtaining widespread attention worldwide due to their remarkable nutritional value, economic potential, and environmental sustainability [1,2,3,4]. Despite growing interest in mushroom cultivation, the industry continues to face several critical challenges. These include rising costs of substrate raw materials, occasional shortages of high-quality spawn, production delays that increase the risk of contamination, and growing difficulties in postharvest handling and preservation [5,6]. The most significant obstacles for both small-scale farmers and industrial producers are the high cost and the limited efficiency of key inputs, particularly inoculum or mushroom spawn, which is essential for successful cultivation [7,8]. Obtaining access to high-quality, cost-effective inoculum remains a key challenge in improving overall production efficiency.
Typically, several types of mycelium inoculum, such as sticks, liquid, solid or grain, and sawdust spawn, each offer unique benefits and limitations [9]. Among the various spawn types, stick spawn has proven to be a practical and cost-effective choice for indoor, outdoor, semi-commercial, and industrial cultivation [10,11,12,13]. It offers advantages such as ease of handling, extended shelf life, and adaptability to diverse growing systems [14,15]. Yet, the resources typically utilized for manufacturing stick spawn, like wooden sticks made from hardwood or softwood trees, are frequently more expensive, less sustainable, or simply available in specific regions [11,16]. Previous research has explored alternative biomass sources, such as rubber wood [14], corn stalks [17,18,19], banana leaf sheaths, and sugarcane residues [20], to address these constraints. Still, the search for more sustainable, low-cost options remains vital, particularly in connection with specific locations.
Building on this work, the present study explores the use of giant mimosa (Mimosa pigra L.), a fast-growing and invasive plant with a sturdy stalk and wide distribution, as a sustainable and low-cost material for stick spawn production [21,22]. Alongside giant mimosa, other biomass materials reported in prior research were collected for comparative analysis. Each material was subjected to comprehensive physicochemical analysis, including assessments of chemical composition, density, and water absorption, to determine its suitability for supporting mycelial colonization.
The study then examined these materials under both laboratory and field conditions to determine their effectiveness in promoting mushroom mycelium growth. Multiple mushroom species were inoculated using standardized stick spawn preparation and inoculation methods to demonstrate consistency and reliability. Key performance indicators included mycelial growth rate, time to full colonization, and subsequent mushroom yield. An economic analysis was also conducted to compare the cost-effectiveness of giant mimosa-based spawn against conventional types, demonstrating potential cost savings for mushroom farmers.
Finally, this study aims to offer practical solutions that enhance the sustainability and affordability of mushroom cultivation. By transforming an invasive plant into a valuable resource, it not only addresses ecological concerns but also contributes to accessible, eco-friendly agricultural practices that benefit both ecosystems and farming communities.

2. Materials and Methods

2.1. Mushroom Mycelium Source and Preparation

Different pure culture strains of 10 edible mushroom species, including Auricularia polytricha (TMCC-NO8), Ganoderma lucidum (TMCC-NO1), Lentinus polychrous (TMCC-NO1), Lentinus sajor-caju (TMCC-NO1), Lentinus squarrosulus (TMCC-NO3), Pleurotus cornucopiae (CMU-NK1318), Pleurotus pulmonarius (CMU-NK1215), Pleurotus ostreatus (CMU-NK1375), Schizophyllum commune (TMCC-NO2), and Volvariella volvacea (TMCC-NO9), were obtained from the Research Center of Microbial Diversity and Sustainable Utilization, Chiang Mai University, Chiang Mai, Thailand (CMU-strains) and the Department of Agriculture, Bangkok, Thailand (TMCC-strains). All cultures were grown on potato dextrose agar (PDA; Conda, Madrid, Spain) and incubated at 30 °C for 7 days under dark conditions [23].

2.2. Material Source and Preparation

In this experiment, four different types of biomass, including giant mimosa (M. pigra L.) stalk (GMS), rubber tree [Hevea brasiliensis (Willd.) Muell. Arg.] wood (RTW), maize (Zea mays L.) stalks residues (MSR), and banana (Musa acuminata Colla) leaf sheath (BLS) (Figure 1) were used as a material for stick spawn inoculum preparation. Each different type of material was separately collected from local sources in Chiang Mai Province, Thailand, as follows: BLS and MSR were obtained from agricultural farms, GMS from the Ping Riverbank, and RTW from a commercial sawmill. Subsequently, GMS, RTW, MSR, and BLS were manually chopped into 100 mm in length. All materials were dried in a hot air oven at 60 °C until a stable weight was reached.

2.3. Determination of Physicochemical Properties of Raw Biomass Materials Used for Stick Spawn Preparation

2.3.1. Chemical Properties

The physicochemical properties of the raw biomass materials were analyzed using standardized procedures. Each type of material was dried and ground before being analyzed. Electrical conductivity (EC) and pH were determined using a 1:10 (w/v) substrate to distilled water ratio, measured with an Ecoscan COND 6+ Conductivity Meter (EUTECH Instruments, Melaka, Malaysia) and a Sartorius PB-10 pH meter (Sartorius, Göttingen, Germany), respectively. The cellulose, hemicellulose, and lignin contents were determined using detergent fiber analysis according to the methods of the Association of Official Analytical Chemists (AOAC) [24]. Organic matter (OM) and organic carbon (OC) were also determined following AOAC methods, with OM percentage calculated from ash content obtained after burning at 550 °C. Total nitrogen (N) content was measured using the Kjeldahl method as previously described by Cortés-Herrera et al. [25]. Raw material samples were analyzed at the Central Laboratory and the Animal Nutrition Laboratory, Department of Animal and Aquatic Sciences, which are part of the Faculty of Agriculture, Chiang Mai University, Thailand.

2.3.2. Density

The bulk density of each raw biomass material was determined using a standard calculation. Specimens were cut into rectangular shapes measuring approximately 5 cm in length, 1 cm in width, and 0.2 cm in thickness. The samples were first oven-dried to a constant weight, then carefully measured [26]. The density was calculated by dividing the dry weight (kg) by the corresponding volume (m3). This method was confirmed to be stable and consistent across all materials tested.

2.3.3. Determination of Water Absorption of Raw Biomass Material

After determining the density of each raw biomass material, the prepared samples were submerged in water to assess the water absorption capacity, a crucial step in achieving the optimal moisture content for supporting mycelium growth. The determination was modified and conducted following the primary standard protocols, with measurements taken at various time intervals: 0.02 h, 0.03 h, 0.08 h, 0.12 h, 0.33 h, 0.66 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 16 h, and 24 h [27]. Water absorption was calculated using the formula: Water absorption (%) = [(Final mass − Initial mass)/Initial mass] × 100.

2.4. Evaluation of the Suitable Material for Mushroom Mycelial Growth on Stick Spawn

The experiment was designed to evaluate mushroom mycelial growth on various material types. Initially, each type of material (GMS, RTW, MSR, and BLS) was soaked in PDB according to water absorption data described in Section 2.3.3, to achieve a moisture content of approximately 60–65%. The materials were then placed in glass test tubes and autoclaved at 121 °C for 30 min. After cooling, each tube was inoculated with a mycelium plug from each mushroom species and incubated at 30 °C in the dark. Mycelium colonization was determined within a seven-day period, with RTW sticks serving as a control. The mycelial growth rate was calculated by dividing the diameter of the mycelial colony (mm) on the colonized material by the incubation time (days) [28]. Mycelial density was qualitatively categorized as thick (+++), thin (++), and very thin (+), according to Schoder et al. [29].

2.5. Evaluation of Mycelial Growth on Different Mushroom Spawn Types

2.5.1. Mushroom Spawn Type

The mushroom spawn was prepared using four different types, including GMS stick, sawdust, sorghum, and liquid spawn (Figure 2). Each spawn type used different base materials, as follows: The stick spawn was prepared by selecting a suitable material for mushroom mycelial growth in a previous experiment (Section 2.4). The substrate spawn was prepared using sawdust as the base material. The liquid spawn was cultivated in PDB following the method of Abdullah et al. [30], and the grain spawn was prepared from sorghum grains. Among these, grain spawn was chosen as the control for comparative analysis due to its widespread use and established reliability in mushroom cultivation, as previously described according to Zhang et al. [18] and Raman et al. [31].

2.5.2. Spawn Preparation and Inoculation

The stick spawn was prepared by immersing selected sticks overnight in PDB. The sticks were then arranged in parallel in a plastic bag, as modified from Liu et al. [11]. For substrate spawn, sawdust was used as a substrate. The moisture content of sawdust was adjusted to 60–65%. Subsequently, 150 g of the moist sawdust substrate was placed into glass bottles. For grain spawn preparation, sorghum grain was used as a substrate. It was thoroughly washed and boiled for 20 min, after that excess water was drained. Then, 150 g of boiled sorghum was placed into each glass bottle according to the method of Girmay et al. [32] and Atila [33] with some modifications. For liquid spawn preparation, 80 mL of PDB was added to 250 mL Erlenmeyer flasks [19].
All spawn substrates were sterilized by autoclaving at 121 °C for 30 min. After cooling to room temperature, each substrate was inoculated with 5 plugs of 5 mm mycelial disc taken from a seven-day-old pure culture of each mushroom species and placed directly on the surface of the substrates. The inoculated substrates were then incubated at 30 °C in the dark for 14–18 days or until complete mycelial colonization.

2.5.3. Substrate Preparation and Inoculation

The cultivation substrate was prepared using sawdust as the main material, which was supplemented with 0.02 kg of calcium sulfate (CaSO4), 0.01 kg of calcium carbonate (CaCO3), 0.003 kg of pumice, 0.002 kg of sodium sulfate (Na2SO4), 0.05 kg of rice bran, and 1 kg of sawdust. All ingredients were mixed, and the moisture content of the substrate mixture was carefully adjusted to 60–65% [34,35]. The 800 g of prepared substrate was then placed into polypropylene bags. A plastic tube was centrally inserted into each bag to serve as an inoculum hole. The filled bags were autoclaved at 121 °C for 60 min. Following sterilization, the substrate was inoculated with each mushroom spawn type from different mushroom species. Each bag was filled with 5 g of grain and substrate spawn, 5 mL of liquid spawn, and 1 piece of stick spawn into the provided center hole. All inoculated bags were then properly sealed and incubated in darkness at 28–30 °C for 14 days or until complete substrate colonization occurred [36,37,38]. The mycelium colonization process was observed by recording the number of days necessary for the substrate to become fully colonized, starting from the first day of inoculation. Additionally, the number of days from inoculation to the appearance of pinhead formation was recorded [39]. The experiment was performed in 10 replications.

2.6. Economic Comparison of Inoculum Types for Mushroom Cultivation to Maximize Profitability

This section delineates the methods employed to assess the economic viability of four inoculum types for mushroom cultivation: GMS sticks, sorghum, sawdust, and liquid spawn. The research sought to determine which inoculum optimized profitability while facilitating efficient mycelial development. A cost-benefit analysis was performed, juxtaposing production costs per unit (bag or bottle) against revenue produced per batch of mushrooms. The unit costs were as follows: GMS sticks at 0.074 USD per bag, sorghum at 0.29 USD per bottle, sawdust at 0.24 USD per bag, and liquid PDB at 2.80 USD per bottle. Revenue and profit were determined depending on the quantity of inoculation bags and the prices of mushrooms. Profit margins were calculated as the disparity between revenue and production expenses.

2.7. Statistical Analysis

The data from all the experiments were analyzed by one-way analysis of variance (ANOVA) using the SPSS software version 17.0 (SPSS Inc.; Chicago, IL, USA for Windows). Duncan’s multiple range test was used to determine significant differences between treatments with p-value of less than 0.05.

3. Results and Discussion

3.1. Physicochemical Properties of Raw Biomass Materials Used for Stick Spawn Preparation

Lignocellulosic biomass, which is abundantly available in nature, is primarily composed of cellulose, hemicellulose, and lignin. The composition of these components differs depending on the type and source of raw materials used [40]. The physicochemical properties of raw biomass materials used in this study for stick spawn preparation are presented in Table 1. The results indicate that these properties can vary significantly depending on the type of biomass, thereby leading to influences on the suitability for fungal growth and the development of high-quality spawn. Among the evaluated lignocellulosic materials, GMS demonstrated the highest cellulose (45.10%) and hemicellulose (26.73%) contents. In contrast, RTW displayed the highest lignin content (28.40%), whereas BLS had the lowest levels of cellulose, hemicellulose, and lignin. The findings of this study are consistent with previous reports indicating that lignocellulosic materials typically comprise 35–55% cellulose, 15–40% hemicellulose, and 7–30% lignin [41,42,43]. Although the lignin content of BLS was low, this finding aligns with previous reports indicating that lignin concentration is lower than that typically found in both hardwoods and softwoods [44]. Furthermore, OM, OC, and total N are critical components in mushroom cultivation. Our results showed that the levels of organic OM, OC, and N ranged from 88.31 to 91.49%, 50.55 to 53.06%, and 0.29 to 1.01%, respectively. Among the samples, MSR had the highest levels of OM, with an average of 91.49% and OC, with an average of 54.06%, respectively. While the total N content of the BLS was 1.01% (Table 1). A substrate high in OM and OC demonstrates an abundant amount of carbon sources required for microbial activity. During the colonization phase, mushroom mycelium uses OM and OC to produce new microbial cells, which helps release essential nutrients that support mushroom development and growth [45].
The EC values exhibited a wide range, varying from 4.24 to 80.30 ds m−1, indicating substantial variability in salinity levels across the materials. In the case of pH, it revealed a range from acidic to slightly alkaline, between 5.53 and 8.69. Maize stalk residues exhibited the highest average pH at 8.69, followed by BLS (6.90), GMS (5.97), and RTW, which had the lowest pH at 5.53. The pH of the substrate is a critical factor influencing mushroom cultivation. However, each species of mushroom has a specific optimal pH range that supports its growth, development, and overall yield. Moreover, this optimal pH range can vary not only between species but also across different stages of the mushroom life cycle [46]. Overall, a detailed analysis of the chemical properties of raw biomass materials not only provides information on the substrate’s structural and nutritional properties, but it also serves in the selection or modification of raw materials to improve fungal colonization efficiency and metabolic activity.

3.2. Water Absorption

The water absorption test conducted on various raw materials, including GMS, RTW, MSR, and BLS, provides essential insights into their suitability for use in mushroom spawn, particularly for optimizing the moisture content range (between 60 and 70%) for efficient mycelium growth [47]. Figure 3 shows that all tested materials had a stable and consistent increase in water absorption over time. Notably, among the materials tested, BLS demonstrated the most significant and consistent high rate of water absorption throughout the testing period, eventually reaching a peak absorption of 729.81% at 24 h. This suggests that BLS has a greater capacity for water retention than the other materials. Furthermore, MSR followed closely, showing a maximum absorption of 623.57% at 24 h. According to the findings, although the highest absorption capacities of BLS and MSR may support moisture retention, they could potentially reduce spawn growth. High moisture content in the substrate can affect mycelial respiration by limiting aeration, inhibiting perspiration, thereby rendering it impossible for the fruiting body to form, as well as resulting in the growth of undesirable organisms such as bacteria and nematodes [46].
Giant mimosa stalk and RTW achieved a water absorption level of 170.54% and 96.01%, respectively, after 24 h. Previous study indicates significant differences in the water absorption capacities of the materials over time, which is a crucial factor in preparing an optimal moisture level ranging from 60 to 70% for efficient mycelium cultivation, depending on the mushroom species [46]. Our findings revealed that GMS could absorb over 63.73% of water within 0.66 h, while RTW exhibited a slower absorption rate, reaching only 62.12% after 6 h.
The differences in water absorption capacity across these materials can be attributed to key factors such as porosity, surface area, fiber structure and composition, density, and surface chemistry [48,49,50]. Overall, the findings suggest that although all tested materials exhibit moisture retention capabilities, GMS and RTW provide the most balanced performance, particularly suitable for producing cost-effective and efficient mushroom stick spawn. In contrast, MSR and BLS, despite strong absorption potential, require careful moisture management to prevent excessive water accumulation that could hinder mycelium development. These results demonstrate the critical role of selecting appropriate raw materials to optimize spawn quality and production efficiency.

3.3. Evaluation of Suitable Material for Mushroom Mycelial Growth on Stick Spawn

The experiment determined the mycelial growth of 10 edible mushroom species cultivated on 4 distinct stick spawn materials. Among these, the results revealed that the substrate used had a substantial impact on mycelial growth among the mushroom species evaluated, showing the mycelial growth rate (mm/day) and mycelial density after a seven-day incubation period. Significant differences in the mean mycelial growth rate were observed among the indigenous stick spawn materials used. As the results present in Figure 4, it was found that all mushroom species could be grown on the tested material. However, the stick spawn prepared using GMS and RTW consistently produced the most extensive mycelial growth and thickest, particularly for L. squarrosulus, L. sajor-caju, and S. commune. Furthermore, S. commune and V. volvacea exhibited the highest growth rate, with an average radial growth of approximately 15 mm/day on GMS and MSR, respectively. However, differences in mycelial density were observed, with S. commune displaying a thin (++), and V. volvacea showing a very thin (+) mycelial density. Following this, L. squarrosulus obtained an average growth rate of approximately 13 mm/day on GMS, with thick mycelial density. While L. sajor-caju exhibited a notable growth rate of approximately 14 mm/day on RTW, as well as a thin (++) mycelial density. Moreover, when comparing the effects of different spawn material types on the mycelial growth of P. ostreatus, P. pulmonarius, and A. polytricha, it was found that using GMS as stick spawn resulted in a thick and strong mycelial growth, indicating its effectiveness in promoting high mycelial density. Conversely, the stick spawn prepared using BLS showed the slowest growth performance, with significantly thinner and less developed mycelial growth. The average growth rate was approximately 3 to 5 mm/day, which was observed across most of the species studied.
For the development of mushroom spawn, a variety of lignocellulosic wastes, such as banana leaf-midribs [51], corn stalk [11], and broadleaf tree wood, have been investigated as effective support materials for enhancing mycelial biomass production. Our results suggest that different materials used as substrates for stick spawn preparation can significantly impact the mycelial growth of edible mushroom species. The significantly different results observed in this study align with findings from previous studies, indicating the essential impact of substrate composition, physical and chemical properties of materials, nutrient availability, and moisture content on the colonization efficiency and metabolic activity of mushroom mycelia [52,53]. As a result, among 10 mushroom strains, only 7 strains, namely P. ostreatus, P. pulmonarius, A. polytricha, G. lucidum, L. polychrous, L. squarrosulus, and S. commune, demonstrated significantly higher mycelial growth rates on GMS compared to the others. Based on these results, the seven high-performing strains and GMS were selected for further investigation to compare their responses to different inoculum types.

3.4. Comparison of Mycelial Growth on Different Mushroom Spawn Types

Mushroom spawn plays a crucial role in the mushroom cultivation process, serving as the material that provides mycelium into the cultivating substrate. Spawn is essentially a carrier, such as grains, sawdust, liquid, and sticks, that has been colonized with mushroom mycelium. Its primary function is to ensure rapid, uniform colonization of the substrate, promoting healthy growth and high yields [11,17,53]. The comparative investigation of different spawn types revealed significant differences in both mycelial colonization rates and the time required for pinhead formation among different mushroom species. The effects of different spawn types on colonization rate and pinhead formation of mushroom species are summarized in Table 2. Among the investigated spawn types, stick spawn consistently exhibited the most rapid performance. Specifically, stick spawn showed the shortest time for both mycelial colonization rate and the earliest pinhead formation among all investigated mushroom species compared to other spawn types. Full substrate colonization was achieved within a range of 11.40 ± 0.54 to 26.80 ± 0.83 days, while pinhead formation occurred within a range of 18.20 ± 0.44 to 47.00 ± 0.70 days, depending on both the mushroom species and the type of spawn used. Among the tested mushroom species, S. commune showed the fastest colonization, completing substrate colonization in 11.40 ± 0.54 days. Pleurotus ostreatus and P. pulmonarius followed, with colonization periods of 16.00 ± 0.70 and 15.80 ± 0.83 days, respectively. Similarly, L. squarrosulus and L. polychrous showed colonization rates of 14.00 ± 0.70 and 17.20 ± 0.83 days, respectively. In contrast, G. lucidum and A. polytricha showed longer colonization rate times of 21.00 ± 0.70 and 26.80 ± 0.83 days, respectively. In terms of pinhead formation, S. commune also showed the shortest time, averaging 18.20 ± 0.44. Pleurotu pulmonarius and P. ostreatus were taken 23.80 ± 0.57 and 26.70 ± 0.83 days, respectively. The results revealed that S. commune achieved the shortest duration for both complete substrate colonization and pinhead formation, while A. polytricha demonstrated the longest time when inoculated with sawdust, sorghum, and liquid spawn (Table 2).
The findings of this study indicate the potential of stick spawn produced from GMS as the most effective inoculum for mushroom cultivation, significantly reducing colonization durations and accelerating pinhead formation across various mushroom species. These results are consistent with the results reported by Liu et al. [11], who revealed that stalk chip and stick spawn reduced spawn running time, thereby enhancing substrate colonization efficiency. This suggests that this type of inoculum can be a potential, long-term alternative to conventional spawn preparation techniques. Laeid and Sangsila [54] found that mixing sawdust, rice straw, maize, and giant mimosa in a 1:2:1:1 ratio resulted in a biological efficiency (BE) of approximately 68%, which was higher than the control using sawdust alone (~66%). This suggests that the inclusion of GMS enhances mycelial growth and yield in mushroom species.
Moreover, the utilization of GMS-derived stick spawn as an inoculum is an influential technique for increasing both crop production and environmental sustainability. Giant mimosa sticks not only improve mycelial growth and expedite up-pinhead formation, but they also have significant long-term effects that improve the overall productivity and economic viability of mushroom production systems.
Moreover, the utilization of GMS-derived stick spawn presents a dual benefit: it not only enhances mycelial vigor and shortens the developmental timeline to fruiting body formation but also contributes positively to the long-term sustainability of mushroom production systems. The incorporation of agricultural waste materials such as giant mimosa sticks into spawn production adds value to otherwise underutilized biomass, reduces reliance on traditional materials, and promotes circular agricultural practices. Collectively, these advantages underline the role of GMS stick spawn as an innovative and environmentally responsible approach to improving yield efficiency and the economic viability of commercial mushroom cultivation.

3.5. Cost-Benefit Analysis and Economic Impact Assessments

The utilization of GMS as a raw material for stick spawn production presents a highly cost-effective alternative to traditional inoculum sources in economic mushroom farming systems. When compared to sawdust, sorghum, and liquid inocula (PDB), GMS stick spawn demonstrates a much lower production cost and higher economic efficiency across all farm sizes (Table 3).
At the production level, the cost per GMS stick spawn unit is only 0.074 USD (2.5 Baht) per bag, whereas traditional inocula cost 0.24 USD (8 Baht) for sawdust, 0.29 USD (10 Baht) for sorghum, and 2.80 USD (95 Baht) for liquid inocula (PDB). When analyzed in terms of inoculation efficiency, GMS stick spawn provides approximately 90 bags per batch, while sawdust, sorghum, and PDB inocula yield 100, 45, and 47 bags per batch, respectively. The lower per-bag cost of GMS stick spawn (0.0009 USD or 0.03 Baht) makes it the most cost-efficient choice for farmers.
For small-scale mushroom farms producing 1000 to 30,000 bags annually, the estimated inoculum cost using GMS sticks ranges from 0.97 to 29.52 USD (33.78–1013.33 Baht), whereas traditional inocula cost significantly more: sawdust (3.08–92.35 USD), sorghum (6.87–206.06 USD), and liquid PDB (58.28–1749.42 USD). The cost advantages of GMS stick spawn become even more pronounced in medium-scale farms (30,001–200,000 bags/year), where the total annual inoculum cost using GMS sticks is 29.26–195.06 USD, compared to sawdust (92.38–615.98 USD), sorghum (206.15–1374.60 USD), and PDB (1750.28–11,688.84 USD). For large-scale operations (200,001–600,000 bags/year), GMS stick spawn continues to outperform other inoculum sources in cost efficiency, with estimated expenses ranging from 194.96 to 584.86 USD, compared to 615.77 to 1847.31 USD (sawdust), 1374.85 to 4122.53 USD (sorghum), and 11,670.92 to 35,014.60 USD (PDB). At the industrial scale (>600,000 bags/year), GMS stick spawn provides the greatest cost-saving potential, as costs for traditional inocula escalate beyond 35,000 USD (1.2 million Baht).
The economic analysis highlights that GMS stick spawn significantly reduces inoculation costs, making it an ideal alternative for mushroom farmers seeking sustainable and cost-efficient solutions. The affordability and high yield potential of GMS stick spawn enable mushroom farms ranging from small-scale to industrial production to optimize their operations while minimizing expenses. This innovative approach not only lowers the financial burden on farmers but also enhances profitability and long-term sustainability in the mushroom farming industry.

4. Conclusions

The study analyzed mycelial growth and mushroom optimization on various agricultural residues, including GMS, RTW, MSR, and BLS. It found that substrate selection significantly impacts edible mushrooms’ growth performance and quality. Giant mimosa stalk invasive species provided the most favorable conditions, yielding the fastest mycelial growth rates and producing healthy mushrooms comparable with RTW in terms of commercial use. Giant mimosa sticks showed strong potential as an alternative substrate, supporting consistent growth and demonstrating viability in regions where traditional substrates may not be readily available. While some mushroom species grow well on MSR, the mycelium tends to be very thin, suggesting that MSR is unsuitable as a standalone inoculum substrate. Nonetheless, when paired with other substrates for mushroom cultivation, maize stalks could still be useful. Banana leaf sheaths consistently underperformed, producing slower growth rates and underdeveloped mushrooms, indicating a lack of necessary nutrients. Giant mimosa sticks outperformed other inocula, making them a highly effective and efficient inoculum for mushroom cultivation. The stick inoculum offers the highest profit margins, making it the most cost-effective alternative for commercial-scale mushroom cultivation. This study focuses on the importance of selecting the appropriate substrate and inoculum to optimize mushroom yield, quality, and economic returns. Future research will explore mushroom yield, nutritional value, and biological activities associated with using GMS as substrates, offering a holistic approach to invasive utilization and environmental sustainability.

Author Contributions

Conceptualization, O.X. and S.L.; methodology, O.X. and W.A.; investigation, O.X., K.J., and S.L.; software, O.X., W.A., K.J., and S.L.; validation, O.X., W.A., K.J., and S.L.; formal analysis, O.X. and W.A.; data curation, O.X., K.J., and S.L.; writing—original draft preparation, O.X., W.A., and K.J.; writing—review and editing, O.X., K.J., and S.L.; supervision, K.J. and S.L.; project administration, O.X., W.A., and K.J.; funding acquisition, S.L. All authors have read and agreed to the published version of the manuscript.

Funding

This work was partially supported by Chiang Mai University, Thailand. Express gratitude to the CMU Proactive Researcher program through grants 815/2567, Chiang Mai, Thailand. The author would like to express sincere thanks to the Asian Development Bank (ADB) for the financial support provided through Grant No. 0500-LAO(SF), which supported the author’s doctoral studies. Additional funding for this project was provided by the National Research Council of Thailand (NRCT) via Grant No. N21A670880, Bangkok, Thailand.

Institutional Review Board Statement

Not applicable.

Data Availability Statement

Data are contained within the article.

Acknowledgments

This research was partially supported by Chiang Mai University, Chiang Mai, Thailand. Special appreciation is also extended to Peter Bennewirtz for his valuable assistance in substrate collection.

Conflicts of Interest

The authors declare no conflicts of interest.

References

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Figure 1. Four different types of biomass used as a material for stick spawn inoculum: (A) GMS, (B) MSR, (C) BLS, and (D) RWT.
Figure 1. Four different types of biomass used as a material for stick spawn inoculum: (A) GMS, (B) MSR, (C) BLS, and (D) RWT.
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Figure 2. Schematic representation of mushroom spawn types and preparation stages. Four spawn types—GMS stick spawn, sawdust spawn, sorghum grain spawn, and liquid spawn—shown stepwise, including (i) raw material, (ii) preparation and containerization, and (iii) fully colonized, ready-to-use spawn.
Figure 2. Schematic representation of mushroom spawn types and preparation stages. Four spawn types—GMS stick spawn, sawdust spawn, sorghum grain spawn, and liquid spawn—shown stepwise, including (i) raw material, (ii) preparation and containerization, and (iii) fully colonized, ready-to-use spawn.
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Figure 3. Water absorption capacity of raw materials at different time intervals. “*” denotes significant differences were determined using Duncan’s multiple range test (p ≤ 0.05).
Figure 3. Water absorption capacity of raw materials at different time intervals. “*” denotes significant differences were determined using Duncan’s multiple range test (p ≤ 0.05).
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Figure 4. Mycelial growth rates (mm/day) and mycelial density assessment (+ = very thin, ++ = thin, +++ = thick) of mushroom species on four stick substrates: giant mimosa stalk (blue), rubber tree wood (gray), maize stalk residues (orange), and banana leaf sheath (green). Bars show meaning with standard deviations. Different letters indicate significant differences among substrates within each mushroom species (Duncan’s test, p ≤ 0.05).
Figure 4. Mycelial growth rates (mm/day) and mycelial density assessment (+ = very thin, ++ = thin, +++ = thick) of mushroom species on four stick substrates: giant mimosa stalk (blue), rubber tree wood (gray), maize stalk residues (orange), and banana leaf sheath (green). Bars show meaning with standard deviations. Different letters indicate significant differences among substrates within each mushroom species (Duncan’s test, p ≤ 0.05).
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Table 1. Physicochemical properties of raw biomass materials used for stick spawn preparation.
Table 1. Physicochemical properties of raw biomass materials used for stick spawn preparation.
PropertiesRaw Material
GMSRTWMSRBLS
Cellulose (%)45.10 ± 0.53 a39.46 ± 0.50 b40.02 ± 0.84 b33.42 ± 0.90 c
Hemicellulose (%)26.73 ± 1.38 a26.61 ± 1.60 a25.90 ± 1.47 a21.44 ± 0.84 b
Lignin (%)22.12 ± 1.41 b28.40 ± 0.95 a9.59 ± 1.75 c4.89 ± 1.54 d
OM (%)89.45 ± 0.71 ab87.15 ± 0.98 b91.49 ± 0.70 a88.31 ± 1.55 b
OC (%)51.88 ± 1.01 a50.55 ± 0.83 a53.06 ± 0.77 a51.22 ± 1.19 a
Total N (%)0.76 ± 0.04 b0.29 ± 0.07 c0.83 ± 0.02 b1.01 ± 0.05 a
C/N ratio68.26 ± 0.84 b174.31 ± 0.69 a63.93 ± 0.59 c50.71 ± 0.36 d
EC (ds m−1)12.42 ± 0.36 c4.24 ± 0.15 d35.23 ± 0.84 b80.30 ± 1.16 a
pH5.97 ± 0.72 bc5.53 ± 0.20 c8.69 ± 0.03 a6.90 ± 0.10 b
Density (kg m3)421 ± 5.34 b606 ± 7.13 a97 ± 10.06 c118 ± 9.91 c
Note: Mean ± SD values in the same row that have different lowercase superscripts are significantly different (p ≤ 0.05).
Table 2. Colonization rate and pinhead formation of mushroom species inoculated with different spawn types.
Table 2. Colonization rate and pinhead formation of mushroom species inoculated with different spawn types.
Mushroom SpeciesSpawn Types
GMS StickSawdustSorghumLiquid (PDB)
FC (days)PH (days)FC (days)PH (days)FC (days)PH (days)FC (days)PH (days)
P. ostreatus16.00 ± 0.70 c26.70 ± 0.83 c21.80 ± 0.83 a33.60 ± 0.89 a20.6 ± 0.54 b30.40 ± 0.54 b20.20 ± 0.83 b33.00 ± 0.70 a
P. pulmonarius15.80 ± 0.83 b23.80 ± 0.57 d21.40 ± 0.54 a33.60 ± 0.28 a22.20 ± 0.83 a29.00 ± 0.57 b21.60 ± 0.54 a26.20 ± 0.57 c
A. polytricha26.80 ± 0.83 c47.00 ± 0.70 d35.80 ± 0.83 a55.40 ± 0.89 b31.40 ± 0.89 b50.40 ± 0.54 c31.80 ± 0.83 b59.60 ± 0.89 a
L. polychrous17.20 ± 0.83 b43.40 ± 1.15 b21.00 ± 1.22 a45.00 ± 1.87 ab19.60 ± 1.14 a43.20 ± 1.48 b21.00 ± 1.22 a45.80 ± 0.83 a
G. lucidum21.00 ± 0.70 c30.00 ± 0.70 b24.20 ± 0.44 a33.60 ± 1.14 ab22.60 ± 1.34 b34.80 ± 0.83 a22.00 ± 0.70 bc35.80 ± 0.83 a
L. squarrosulus14.00 ± 0.70 c42.40 ± 1.51 c15.40 ± 0.54 b45.20 ± 1.48 ab16.40 ± 1.14 ab43.40 ± 2.07 bc17.40 ± 1.14 a46.80 ± 1.30 a
S. commune11.40 ± 0.54 b18.20 ± 0.44 c14.00 ± 1.00 a21.00 ± 0.70 ab14.60 ± 0.54 a20.00 ± 1.22 bc14.20 ± 0.83 a23.00 ± 0.70 a
Note: Mean ± SD values in the same row that have different lowercase superscripts are significantly different (p ≤ 0.05). Full colonization time (FC), Pinhead formation (PH).
Table 3. Estimation of primary cost analysis calculation for GMS stick in comparison with different types of inocula across varying farm sizes. Estimation of the primary cost calculation for GMS sticks in relation to various inoculum types across different farm sizes.
Table 3. Estimation of primary cost analysis calculation for GMS stick in comparison with different types of inocula across varying farm sizes. Estimation of the primary cost calculation for GMS sticks in relation to various inoculum types across different farm sizes.
PerformanceGMS StickTraditional Products Used
SawdustSorghumLiquid (PDB)
Analysis of Producing Income and Cost
Cost (USD)0.074 USD
2.5 Bath		0.24 USD
28 Bath		0.29 USD
10 Bath		2.80 USD
95 Bath
Volume per unit85–95 sticks/bag 500–600 g/bag100–150 g/bottle120–200 mL/bottle
Amount of inoculation (Approx. bags) ≈90≈100≈45≈47
Average inoculum cost per bag (USD)0.0009 USD
0.03 Bath		0.0024 USD
0.08 Bath		0.0071 USD
0.24 Bath		0.060 USD
2.02 Bath
Cost analysis across farm sizes
Small scale
(1000–30,000 bags/year)		≈0.97–29.52 USD
≈33.78–1013.33 Baht		≈3.08–92.35 USD
≈106.67–3200.00 Baht		≈6.87–206.06 USD
≈238.10–7142.86 Baht		≈58.28–1749.42 USD
≈2021.28–60.638.30 Bath
Medium scale
(30,001–200,000 bags/year)		≈29.26–195.06 USD
≈1013.37–6755.55 Baht		≈92.38–615.98 USD
≈3200.11–21,333.33 Baht		≈206.15–1374.60 USD
≈7143.10–47,619.05 Baht		≈1750.28–11,688.84 USD
≈60,640.32–404,255.32 Bath
Large scale
(200,001–600,000 bags/year)		≈194.96–584.86 USD
≈6755.59–20,266.66 Baht		≈615.77–1847.31 USD
≈213,33.44–64,000 Bath		≈1374.85–4122.53 USD
≈47,619.29–142,857.14 Baht		≈11,670.92–35,014.6 USD
≈404,257.34–1,212,765.96 Bath
Industrial scale
(>600,000 bags/year)		>584.86 USD
>20,266.67 Baht		>1847.31 USD
>64,000 Bath		>4122.53 USD
>142,857.14 Baht		>35,014.6 USD
>1,212,765.96 Bath
Note: The farm size data are based on National Science and Technology Development Agency (NSTDA), Thailand [55].
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Xayyavong, O.; Aiduang, W.; Jatuwong, K.; Lumyong, S. Utilization of Giant Mimosa Stalk to Produce Effective Stick Spawn for Reducing Inoculum Costs in Economic Mushroom Farming Systems. Agriculture 2025, 15, 1584. https://doi.org/10.3390/agriculture15151584

AMA Style

Xayyavong O, Aiduang W, Jatuwong K, Lumyong S. Utilization of Giant Mimosa Stalk to Produce Effective Stick Spawn for Reducing Inoculum Costs in Economic Mushroom Farming Systems. Agriculture. 2025; 15(15):1584. https://doi.org/10.3390/agriculture15151584

Chicago/Turabian Style

Xayyavong, Orlavanh, Worawoot Aiduang, Kritsana Jatuwong, and Saisamorn Lumyong. 2025. "Utilization of Giant Mimosa Stalk to Produce Effective Stick Spawn for Reducing Inoculum Costs in Economic Mushroom Farming Systems" Agriculture 15, no. 15: 1584. https://doi.org/10.3390/agriculture15151584

APA Style

Xayyavong, O., Aiduang, W., Jatuwong, K., & Lumyong, S. (2025). Utilization of Giant Mimosa Stalk to Produce Effective Stick Spawn for Reducing Inoculum Costs in Economic Mushroom Farming Systems. Agriculture, 15(15), 1584. https://doi.org/10.3390/agriculture15151584

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