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Article

Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity

by
Naruemon Wannasawang
1,
Thatsanee Luangharn
1,
Anan Thawthong
1,2,
Rawiwan Charoensup
3,4,
Wuttichai Jaidee
4,
Wirongrong Tongdeesoontorn
5,6,
Kevin D. Hyde
1 and
Naritsada Thongklang
1,2,*
1
Center of Excellence in Fungal Research, Mae Fah Luang University, Chiang Rai 57100, Thailand
2
School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand
3
School of Integrative Medicine, Mae Fah Luang University, Chiang Rai 57100, Thailand
4
Medicinal Plant Innovation Center of Mae Fah Luang University, Chiang Rai 57100, Thailand
5
School of Agro-Industry, Mae Fah Luang University, Chiang Rai 57100, Thailand
6
Research Group of Innovative Food Packaging and Biomaterials, Mae Fah Luang University, Chiang Rai 57100, Thailand
*
Author to whom correspondence should be addressed.
Life 2023, 13(9), 1887; https://doi.org/10.3390/life13091887
Submission received: 19 July 2023 / Revised: 29 August 2023 / Accepted: 29 August 2023 / Published: 9 September 2023
(This article belongs to the Section Plant Science)
Browse Figures
Versions Notes

Abstract

:
Ganoderma (Ganodermataceae) has a worldwide distribution and has been widely used in traditional medicines. In this study, we report wild strains of Ganoderma that include two G. sichuanense and one G. orbiforme from northern Thailand. Optimal conditions for mycelium growth were ensured. The most favourable medium was potato sucrose agar for G. sichuanense and oatmeal agar for G. orbiforme and at 25 °C and 30 °C and pH 4–8. All types of cereal grains can be used to promote the growth of the mycelia of Ganoderma species. Fruiting tests were performed. All strains of Ganoderma produce fruiting bodies successfully in bag culture at 28 ± 1 °C with 75–85% relative humidity. Only G. orbiforme produced fruiting bodies in field cultivation at the laboratory scale. In the first flush yields, the G. sichuanense strain MFLUCC 22-0064 gave better production (the B.E was 152.35 ± 6.98 g). This study is the first to document the bag and field cultivation of wild Thai G. orbiforme. Ganoderma species are revealed to contain high amounts of fiber (47.90–52.45% d.b.), protein (12.80–14.67% d.b.), fat (4.90–5.70% d.b.), and carbohydrates (3.16–4.02% d.b.). Additionally, G. sichuanense and G. orbiforme were preliminarily screened for biological activity for inhibition of alpha–glucosidase enzyme activity. The IC50 values of G. orbiforme (MFLUCC 22-0066) was 105.97 ± 1.36 µg/mL and G. sichuanense (MFLUCC 22-0064) was 126.94 ± 0.87 µg/mL. Both strains had better inhibition than acarbose (168.18 ± 0.89 µM). These results on wild strains of Ganoderma will be useful for further studies on the applications of Ganoderma. Later the species can be introduced to domestic markets for cultivation and medicinal use.

1. Introduction

Ganoderma (Ganodermataceae, Basidiomycota) was established by Karsten [1] with Ganoderma lucidum (Curtis) P.Karst. as the type species. Justo et al. [2] treated Ganodermataceae as a synonym of Polyporaceae, while Cui et al. [3] reported that Ganoderma is quite different from Polyporaceae. Ganoderma is distinctively characterized by laccate and nonlaccate basidiocarps, double-walled basidiospores, an apical germinal pore, thin and colourless external wall (exosporium), with a brown to dark brown internal wall (endosporium) [4,5,6]. There are 493 records of Ganoderma species in the Index Fungorum [7]. The morphological characteristics and molecular analysis of Thai wild Ganoderma have been reported [8,9,10]. Members of Ganoderma are cosmopolitan species [11], which are distributed worldwide in subtropical to tropical and temperate regions [12]. Some Ganoderma are regarded as tree pathogens [9,13,14,15] causing white rot diseases on rotting stumps, roots, and living trunks [16]. Ganoderma is economically important, due to the fact that members of the genus are regarded as valuable medicinal mushrooms [17,18]. Several Ganoderma species are known to be prolific sources of highly natural bioactive compounds such as polysaccharides, proteins, steroids, and triterpenoids [19,20,21]. These natural bioactive compounds are used to treat and remedy many pathological diseases [22,23]. In addition, Ganoderma products come in the form of coffee, powder, tea, dietary supplements, spore product, drinks, syrup, toothpaste, soap, lotion, and have been commercialized as effective food and drug supplements for health benefits [24,25,26].
Ganoderma sichuanense is recognized as a medicinal mushroom. This fungus was initially documented in the tropical region of Thailand by Thawthong et al. [10]. This delineation was described in both taxonomical classification and molecular substantiation. Ganoderma sichuanense, a reservoir of bioactive compounds, can address human ailments. The comprehensive exploration of this fungus, such as optimal conditions, fruiting trials, nutritional constituents, and assessing alpha-glucosidase inhibitory effects, still needs to be included in the scientific report, as does Ganoderma orbiforme, a member of the laccate Ganoderma species. Nevertheless, there is a lack of research concerning its medical utility and cultivation in Thailand.
In Thailand, there are limited studies on the cultivation of Ganoderma species. In an attempt to further expand our knowledge of Thai Ganoderma species in this study, we describe Ganoderma sichuanense and Ganoderma orbiforme using morphology and molecular data. Optimal conditions for mycelia, spawning production, proximate and alpha glucosidase inhibitory activity of these fungi are also reported. These two species are likely to be used for cultivation and medicinal purposes as a new native mushroom source in Thailand.

2. Materials and Methods

2.1. Mushroom Collections

Two Ganoderma sichuanense and one G. orbiforme were collected from Chiang Mai Province, Thailand by Anan Thawthong. The fresh basidiomata of Ganoderma were isolated by tissue culture technique on potato dextrose agar (PDA) medium, and incubated at 25 °C for 2 weeks for later use. The mycelium cultures were deposited at the Mae Fah Luang University Culture Collection (MFLUCC) as MFLUCC 22-0066 (G. orbiforme), MFLUCC 22-0064 and MFLUCC 22-0065 (G. sichuanense), while dried samples were deposited at Mae Fah Luang University Fungarium (MFLU) as MFLU22-0111 (G. orbiforme), MFLU22-0109 and MFLU22-0110 (G. sichuanense).

2.2. DNA Extraction, Polymerase Chain Reaction (PCR) and Sequencing

Dried internal tissues from the fruiting bodies were used to extract DNA using the High Pure PCR Template Preparation Kit (Roche), following the manufacturer’s instructions. Total reaction mixtures (25 μL) contained 9.5 μL ddH2O, 12.5 μL of PCR master mix, 1 μL of DNA template, and 1 μL of each primer (10 μM). The primers used in PCR amplification were: ITS4/ITS5 for internal transcribed spacer gene region (ITS); LROR/LR5 for partial large subunit rDNA gene region (LSU) [27,28] and bRPB2-6F/bRPB2-7.1R for partial RNA polymerase II second largest subunit gene (RPB2) [29]. PCR amplification conditions were 3 min at 94 °C, followed by 35 cycles of 95 °C for 30 s, 55 °C for 1 min, 72 °C for 1 min, followed by a final extension at 72 °C for 10 min for ITS and LSU, and 3 min at 94 °C followed by 35 cycles of 94 °C for 1 min, 59.1 °C for 2 min, and 72 °C for 1 min, followed by a final extension at 72 °C for 10 min for RPB2. The PCR products were sequenced by SolGent Co., Ltd. Daejeon, South Korea.

2.3. Phylogenetic Analyses

Sequence accession numbers in the analysis are provided in Table 1. The sequences were subjected to standard BLASTn searches in GenBank to determine the primary identity of the fungal isolates. Tomophagus colossus TC–02 [30] was selected as outgroup taxa. All generated sequences were aligned with the combined datasets of ITS, LSU, and RPB2 and were aligned using the MAFFT v.7.110 online program (http://mafft.cbrc.jp/alignment/server (accessed on 8 February 2023)) [31] and manually adjusted via BioEdit 7.2.3 [32]. Gaps were treated as missing data. Phylogenetic analyses were performed by using PAUP v.4.0b10 [33] for maximum parsimony (MP) and MrBayes v. 3.2.2 [34] for Bayesian analyses. Maximum likelihood analyses (ML) were estimated by using the software on the CIPRES Gateway platform [35] and performed using RAxML-HPC2 on XSEDE (v. 8.2.8) [36], then carried out using the raxmlGUI version v. 1.3.1 [37]. MrModeltest v. 2.3 was used to determine the best-fitting substitution model for each single gene partition and the concatenated dataset for Bayesian analyses [38]. Bayesian inference posterior probabilities (PP) with a GTR+I+G model was used for each partition. Phylogenetic trees were sampled every 100th generation (resulting in 10,000 total trees) in 1,000,000 generations from the running of six simultaneous Markov chains. The first 2000 trees, which contained the burn-in phase of the analysis, were discarded. The remaining 8000 trees were used to calculate the PP in the majority-rule consensus tree. ML and MP bootstrap values, equal to or greater than 70% and Bayesian Posterior Probabilities (BP) equal to or greater than 0.95 are presented above each node (Figure 1). The trees were figured in the FigTree v. 1.4.0 program [39], edited using Microsoft Office PowerPoint 2010 and exported to Adobe Illustrator CS v. 3 (Adobe Systems, San Jose, CA, USA). Sequences derived in this study are deposited in GenBank (http://www.ncbi.nlm.nih.gov, accessed on 8 February 2023).

2.4. Taxonomy Study

Morphological characteristics were described following the methodology described by Lodge et al. [57]. Macromorphological characteristics were examined using the Leica M125C (Leica, Wetzlar, Germany) digital microscope camera. Colors were recorded following the instructions of Kornerup and Wanscher [58]. Micromorphological characteristics were observed using a compound Nikon Eclipse Ni-E (Nikon, Tokyo, Japan) microscope. Microscopic features and measurements were made from glass slide preparations, staining tissues with 5% potassium hydroxide (KOH) and Melzer’s reagent. The features of the basidiospore, the hyphal system, the color, the sizes, shapes, and photographs were recorded and measured using the Tarosoft Image Framework programme v. 0.9.7. The size of the basidiospore was measured with and without the myxosporium using at least 50 basidiospores from each basidiomata [59]. The basidiospore quotient was followed [Q = L/W] with dimensions are given as (a—) b—c—d (—e), where Q, the quotient of basidiospore length to width (L/W) of a basidiospore inside view, and Qm, the mean of Q values ± SD, were calculated considering the mean value of the lengths and widths of basidiospores [60].

2.5. Optimal Agar Medium for Mycelium Growth

Five different medium types, namely potato dextrose agar (PDA), potato sucrose agar (PSA), corn meal agar (CMA), oatmeal agar (OMA), and malt extract agar (MEA) were tested for the growth of mycelium of two strains of G. sichuanense and one strain of G. orbiforme. The media plates (9 cm) were centrally inoculated with mycelium plugs of approximately 0.5 mm diam and incubated at 25 °C for 8 days in the dark. The medium was melted and washed away with hot water, leaving the fungal mycelia. The growth of the mycelium was obtained by determining the dry weight in triplicate.

2.6. Optimal Temperature and pH for Mycelium Growth

The best agar medium was selected to test the optimal temperature and incubated at 20, 25, 30 and 40 °C. The medium was melted, the agar was washed with hot water and the mycelium was collected. The mycelium growth was obtained by determining the dry weight on day 8 in triplicate.
The suitable pH was determined in potato dextrose broth (PDB) that was adjusted to pH 2, 4, 6, 7 and 8 with 1N HCl or 1N NaOH prior to autoclaving. Appropriate 100 mL of PDB was inoculated with the mycelium, and incubated at 25 °C on a rotary shaker at 120 rpm for 14 days. Mycelial growth was evaluated via the determination of dry weight on day 14 in triplicate.

2.7. Effect of Spawn Production

Five cereal grains, Coix lacryma-jobi (millet), Hordeum vulgare (barley), Oryza sativa (rice berry), Triticum aestivum (wheat) and Zea mays (maize), were used for the testing of Ganoderma species. Grains were washed and soaked overnight, then water was drained off, and grains were boiled for 15 min. Fifty-gram samples of cereal grains in test tubes (25 × 200 mm) were autoclaved at 121 °C for 15 min and left at room temperature to cool. The three mycelial plugs (approximately 0.5 cm diam) were inoculated into test tubes and incubated at 25 °C. Linear mycelium length was measured for 18 days. The experiment was determined in five replicates.

2.8. Fruiting Test of Thai Wild Ganoderma in Bags and Field Cultivation

For bag cultivation, rubber sawdust was used as the main substrate and mixed with supplements following the instructions of Thongklang et al. [61]. Fifty grams of spawn was inoculated in bags and incubated at 25 ± 1 °C in the dark for 110 days. The same temperature and 75–85% relative humidity were used for the fruiting stage. The experiment was determined in 15 replicates. Field cultivation was conducted on the farmland of Mae Fah Luang University. The plot size was 3 m × 1 m for five replicates for each strain. The inoculated sawdust bags were inoculated and covered by soil casing (3 cm thick). During the experiment, relative humidity was maintained at 70% by watering 2 times per day for 170 days.

2.9. Statistical Analysis

Mycelial growth rate in media, temperature, pH, and spawning production of mushroom strains was determined and data were statistically analyzed in terms of variance of means using Duncan’s test with significance for p < 0.05.
The fruiting bodies of wild Thai Ganoderma from both bag and field cultivations were manually harvested, counted, and weighed daily. The total weight of fresh mushrooms per spent substrate was used to calculate yield data [62,63]. In addition, biological efficiency (B.E.) was conducted using the formula: weight of harvest/weight of dry substrate) × 100% [61,64,65].

2.10. Proximate Analysis

Total protein, fat, fiber, and carbohydrates were calculated from the oven-dried powder using standard protocols. The total protein was calculated using the Kjeldahl method. Copper sulfate: potassium sulfate (1:10) was used for the Kjeldahl digestion catalyst. Mushroom samples (0.2–0.5 g) were placed in digestion tubes. Five grams of catalyst was added to the appropriate volume (12–15 mL) of sulfuric acid to each tube with the sample. Then, the sample was digested using VELP Scientifica S30100210 Model DKL 20 Automatic Kjeldahl Digestion (VELP Scientifica, Usmate Velate, Italy) at 420 °C for 45 min. Distillation and titration via the fully automatic Kjeldahl Analyzer–distillation unit with integrated colorimetric titrator were executed and then the percentages of N and protein were calculated automatically. Determination of the fat content was evaluated via the FOSS–Fat Analyzer–Soxtec™ 8000 (FOSS, Hillerød, Denmark). The samples (2–3 g) were put in the thimble and then placed in a Soxhlet extractor. Then, approximately 70 mL of petroleum ether was added using a dispenser. After the program was finished, the cup was heated in the oven at 105 ± 2 °C for 2 h, cooled in the desiccators, and weighed. The percentage of fat was calculated using the following equation:
% Fat = [(wt of cup + fat) − wt of cup]/wt of wet sample × 100
Determination of crude fiber was carried out using Fibertherm (Gerhardt GmbH & Co. KG, Königswinter, Germany). The fiber bag was dried at 105 ± 2 °C for 1 h and then cooled to room temperature in desiccators and checked weight (M1). Then, one gram of sample was weighed on fiber bags (M2). Fiber bags with glass spacers were put into the carousel (M1 = fiber bag without sample). Samples were defatted at a fat content of >10% with acetone or petroleum ether. Then, the fibertherm process was initiated. Subsequently, dry fiber bags were placed in the crucible at 105 ± 2 °C until the mass was constant (at least 4 h) and were cooled in desiccators and weighed (M3). The amount of ash from the sample was determined by drying in the crucible at 525 ± 25 °C for 4 h and cooled down in the desiccator for 30 more minutes. The crude fiber content (%) was calculated from the weight of the ash residue (M4), weight of the sample, and fiberbag via the following equation:
% Crude fiber = (M3 − M1 − M4)/M2 × 100
where
M1 = Weight of fiber bags without sample
M2 = Weight of dry sample
M3 = Weight of crucible and fiber bags after drying at 105 ± 2 °C for at least 4 h and cooling in desiccators
M4 = Weight of crucible and fiber bags after ashed at 525 ± 25 °C for 4 h and cool off in desiccators
Furthermore, the carbohydrate content was estimated from the formula: 100 − (ash + crude fat + crude fiber + protein) The experiment was determined in three replicates.

2.11. The α-Glucosidase Inhibitory Assay

The α-glucosidase inhibitory assay procedure followed a previously described method with modifications. Sample solutions at 200 μg/mL were dissolved with 10% dimethyl sulfoxide (DMSO) in phosphate buffer (pH 6.8), and then 50 μL of each sample was pipetted and mixed with 100 μL α-glucosidase enzyme (0.35 U/mL) in an Eppendorf tube. After preincubation at 37 °C for 10 min, 100 μL of 1.5 mM p-NPG was added, and the samples were further incubated at 37 °C for 20 min. Next, 1000 μL of Na2CO3 (1 M) was added to terminate the reaction. Acarbose was used as a positive control. The absorbance was measured at 405 nm with a microplate reader (PerkinElmer, Inc., Waltham, MA, USA).

3. Results

3.1. Phylogenetic Analyses

Phylogenetic analyzes included 58 taxa, and the tree was inferred from the combination of ITS, LSU, and RPB2 sequences, which comprises 2553 characters with gaps. The maximum parsimonious dataset consisted of 2103 constant, 313 parsimony-informative, and 137 parsimony-uninformative characters. Tree topologies of the ML and MP were similar to the Bayesian analysis. The final ML optimization likelihood value of −14,158.684850. Estimated base frequencies were as follows: A = 0.225017, C = 0.242278, G = 0.264931, T = 0.267773; substitution rates AC = 1.209773, AG = 3.527855, AT = 1.594973, CG = 1.560722, CT = 5.400917, GT = 1.000000.

3.2. Taxonomy

Ganoderma sichuanense J.D. Zhao and X.Q. Zhang [66] (Figure 2).
Basidiome annual, stipitate to short stipitate, laccate to strongly laccate, and woody. Pileus 5–7 × 3–10 cm, up to 1.8 cm thick at the base, suborbicular to orbicular, convex, and flabelliform shape. The surface of the pileus is reddish orange (7B8) when fresh, reddish brown (9E7) when dried, and an irregularly ruptured crust overlying the context. Context up to 1.4 cm thick, dried, brown (6E4), reddish-brown (9E7), and dark brown (7F8) at upper layer, corky when dried. Margin soft when fresh, wavy, white when become, orange (6B8) to reddish-yellow (4B7) to concolous with the pileus. Hymenophore up to 18 mm long, indistinctly stratose. Pores 4–6 per mm, round, angular, up to 119–169 × 123–191 µm ( x ¯ = 146 × 158 μm, n = 50). Pore surface yellow when fresh, turning yellowish-white (3A2) to yellowish–grey (3B2), dull-yellow (3B3-4) when dried, and greyish-brown (7D3) when touched. Stipe 1–3 cm, lateral, some sub-cylindrical to cylindrical, thick, short, laccate, reddish-brown (9E7), concolorous with the pileus. Tubes up to 0.3–5.9 mm in length, brown (7E8). Basidiospores ellipsoid to broadly ellipsoid, some globose and ovoid, truncate at the apex with double wall (ganodermoid), brown (6E5), with a dark brown (7F8) eusporium bearing thick echinulae, (7.9–) 9.0–10.4 (–11.4) × (5.6–) 6.1–7.0 (–7.5) μm ( x ¯ = 9.7 × 6.5 μm, n = 50) with Q = 1.2–1.6 μm, L = 9.7, W = 6.5 (including myxosporium), (5.0–) 5.8–7.7 (–8.6) × (3.2–) 4.0–5.1 (–5.6) μm ( x ¯ = 6.7 × 4.5 μm, n = 50) with Q = 1.1–1.7 μm, L = 6.7, W = 4.5 (excluding outer myxosporium). Pileipellis a hymeniderm, brownish orange (6C8), clavate-like cells, with dextrinoid. Hyphal system trimitic; generative hyphae 1.9–2.7–3.5 μm, n = 20 in width, thin-walled and hyaline; skeletal hyphae 2.3–3.5–4.9 μm, n = 20 in width, thick–walled, branched, brownish orange (6C8); binding hyphae 1.7–3.7–5.6 μm, n = 20 width, thick-walled, branched, light orange (5A5) to light brown (5D5) in Melzer’s reagent.
Specimen examined: THAILAND, Chiang Mai Province, 13 July 2017, MFLUCC 22-0064, MFLU22-0109, original K17-55 and MFLUCC 22-0065, MFLU 22-0110, original K17-69.
Ganoderma orbiforme (Fr.) Ryvarden [5] (Figure 3).
Basidiome annual to perennial, distinctly contracted base, weakly laccate, sessile, and woody when dried. Pileus 4–6 × 5–9 cm, up to 2.6 cm thick at the base, applanate to plano-convex shape with several thick layers. Pileus surface weakly laccate, smooth when young, reddish-brown (9F6-7) at concentrically zones, reddish brown (8F6) to dark brown (8F5) when dried, crust overlies the pileus, brown (7E7-8) when fresh, reddish brown (8E7–8) when dried, concentrically sulcate zones with turberculate bumps and rivulose depressions, differentiated zone at the point of attachment. Margin 1.5–3 mm, undulate and irregularities, reddish brown (8E7), soft, round, white (8A1) to orange-white (5A2) when young, brown (7D8) when become mature, and reddish brown (8E8) when dried. Hymenophore up to 2.8 mm in length with orange-grey (6B2). Pores 4–6 per mm, up to 102.6–132.8 × 79.4–131.7 µm ( x ¯ = 116 × 102 μm, n = 50), with subcircular to circular. Pore surface white (8A1) when fresh, light brown (6D5) when touched, and turning orange-grey (5B2) when dried. Context up to 2 cm thick, soft, brown (6E8) to dark brown (6F8) at the lower layer near the tube layers, and composed of coarse loose fibrils. Tubes up to 0.1–5.6 mm in length with brown (7E5) to reddish brown (8E4) when mature. Basidiospores ellipsoid to oblong ellipsoid, some elongate with double wall (ganodermoid) at maturity, almost colorless, yellowish-brown (5D8), (8.4–) 9.8–10.9 (–11.3) × (4.4–) 5.4–6.3 (–6.6) μm ( x ¯ = 10.3 × 5.8 μm, n = 50), with Q =1.4–2.3 μm, L = 10.3, W = 5.8 (including myxosporium), (6.1–) 7.2–8.5 (–9.2) × (3.3–) 3.7–4.6 (–5.1) μm ( x ¯ = 7.9 × 4.1 μm, n = 50) with Q = 1.4–2.4 μm, L = 7.9, W = 4.1 (excluding outer myxosporium), yellowish brown (5D8), overlaid by a hyaline myxosporium. Pileipellis a hymeniderm, composed of apically acanthus like branched cells with some dextrinoid, with brown (6E8) to dark brown (7F8). Hyphal system trimitic; generative hyphae 2.0–3.3–4.5 μm, n = 20 in width, thin–walled or occasionally, hyaline, occasionally with irregular cuticle cells; skeletal hyphae 3.2–5.4–6.8 μm, n = 20 in width, light brown (5D8, 6D5), thick–walled; binding hyphae 3.5–4.8–6.9 μm, n = 20 in width, light brown (5D8), some brown (6D8), thick–walled, branched, some intertwined skeletal hyphae.
Specimen examined: THAILAND, Chiang Mai Province, 14 July 2017, MFLUCC 22-0066, MFLU22-0111, original K17-76.

3.3. Optimal Conditions for The Growth of The Mycelium of Wild Ganoderma from Thailand

Ganoderma mycelium, G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) and G. orbiforme (MFLUCC 22-0066) grow well in all mediums, however, the most favorable medium was potato sucrose agar (PSA) for G. sichuanense (both strains). The average dried weight of mushroom strains MFLUCC 22-0064 and MFLUCC 22-0065 were 0.1241 ± 0.0029 g and 0.1414 ± 0.0217 g, respectively, while oatmeal agar (OMA) was suitable for growing G. orbiforme (MFLUCC 22-0066). The average dried weight was 0.1496 ± 0.0118 g on day 8 after inoculation.
The optimal temperatures of these three Ganoderma strains are between 25 and 30 °C. G. sichuanense (MFLUCC 22-0064) grew well at 30 °C and followed by 25 °C, and the average dry weight was 0.1295 ± 0.0611 g and 0.0861 ± 0.0192 g, respectively. G. sichuanense (MFLUCC 22-0065) grew well at 30 °C, and the average dry weight was 0.1721 ± 0.0235 g. In addition, G. orbiforme (MFLUCC 22-0066) also prefers both temperatures (25 and 30 °C), and the average dry weight was 0.0707 ± 0.0246 g at 25 °C and 0.0512 ± 0.0006 g at 30 °C, respectively.
All pH values (4–8) were suitable for promoting mycelium growth of G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065), while pH levels 4–6 were suitable for growing G. orbiforme (MFLUCC 22-0066) Table 2.

3.4. Effect of Spawn Production

The data for mycelium growth on different spawn media were investigated and is shown in Table 2. We found that all cereal grain types can be used to promote the mycelial growth of the Ganoderma species studied here. All types of grains can be used to grow G. sichuanense MFLUCC 22-0065, while G. sichuanense (MFLUCC 22-0064) and G. orbiforme (MFLUCC 22-0066) can grow in C. lacryma-jobi, H. vulgare, O. sativa, and T. aestivum. However, the mushroom grew very slowly in the cereal grain of Z. mays.

3.5. Fruiting Test of Thai Wild Ganoderma in Bags and Field Cultivation

The fruiting bodies of G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) and G. orbiforme (MFLUCC 22-0066) were manually harvested and measured daily. The mushrooms were produced at 28 ± 1 °C in 75–85% humidity. The cultivation of a wild strain of G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) was carried out with fifteen replicates. We found that three strains of Ganoderma can produce fruiting bodies on a laboratory scale (Figure 4). The mycelium of G. sichuanense (MFLUCC 22-0064) fully covered the rubber sawdust bags on day 31. The first primordia appeared on 44–61 days. In G. sichuanense (MFLUCC 22-0065) it took 22 days to fully cover the rubber sawdust bag, and the first primordia appeared on day 35–62 days. The average yield of MFLUCC 22-0064 and MFLUCC 22-0065 were 29.43 ± 4.72 g and 33.92 ± 6.98 g in the first flush production, respectively. A strain, G. orbiforme (MFLUCC 22-0066) was successfully cultivated in rubber sawdust bags. The mycelium was fully colonized with the bags on day 22. The first primordia appeared on day 41–58 days. The average yield was 16.21 ± 3.51 g. The yield data and the biological efficiency of three Ganoderma strains are given in Table 3.
Meanwhile, the mycelium of these three strains was inoculated into a sawdust-based medium. After that, the media were fully covered by mycelia, and the bags were placed into the field and covered by soil. After 57–112 days, the first primordia of G. orbiforme (MFLUCC 22-0066) occurred (Figure 5). However, yield production of the strain was low. Unfortunately, G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) fruiting bodies were not produced in field cultivation.

3.6. Nutrition Values of Thai Wild Ganoderma

In this study, the total protein content of Ganoderma species varies between 12 and 15 g/100 g of sample. The highest protein content for G. orbiforme (MFLUCC 22-0066) was 14.67 ± 0.25% of the sample and the lowest was G. sichuanense (MFLUCC 22-0064) 12.80 ± 0.15%. We found that the amount of carbohydrates contained within Ganoderma species is 3–5% of. The highest composition of these three Ganoderma species is fiber. The crude fiber was found to be between 47 and 53% of the fruiting bodies. The heightened fiber was from G. orbiforme (MFLUCC 22-0066) (52.45 ± 0.18%), while two strains of G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) were 48.61 ± 0.67% and 47.90 ± 0.39%, respectively. In addition, the amount of fat was between 4 and 6% in the fruiting bodies of Ganoderma species. The fat content in G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) was 5.70 ± 0.56% and 5.11 ± 0.11%, respectively, while in G. orbiforme (MFLUCC 22-0066) it was 4.90 ± 0.56% (Table 4).

3.7. The α-Glucosidase Inhibitory Activity

The mixed fresh mushroom Ganoderma sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) and G. orbiforme (MFLUCC 22-0066) were extracted with ethanol and water under sonication for 1 h at 60 °C. The ethanol extracts were obtained by evaporation under reduced pressure, producing a yield of 2.29, 1.86, and 1.61%, respectively; water extracts of 0.55, 2.70, and 0.31% were obtained by freeze-drying. The mycelium mushrooms MFLUCC 22-0064, MFLUCC 22-0065, and MFLUCC 22-0066 were extracted with acetone and water under maceration for 24 h. Acetone extracts were obtained by evaporation under reduced pressure, to produce a yield of 2.15, 0.15, and 1.25%, respectively. The water extracts were then freeze-dried into powder. The IC50 value of Thai Ganoderma extract α-glucosidase inhibitory activity is shown in Table 5. All ethanolic extracts from fruiting bodies of three Ganoderma strains demonstrated that the IC50 ranged from 105.97 ± 1.36 to 171.68 ± 2.78. In addition, only the water extract of G. orbiforme (MFLUCC 22-0066) showed an inhibitory effect, and the IC50 value was 124.40 ± 3.18.

4. Discussion

Ganoderma has a long history of use as a traditional medicine in Asian countries. However, Ganoderma has been considered a very difficult genus to classify. Ganoderma is currently in a state of taxonomic chaos, since they were initially classified on the basis of their morphological characteristics. Based on this taxonomy and phylogenetic analysis of three Ganoderma strains in this study, two strains are considered to be G. sichuanense and another one is G. orbiforme. Two strains of G. sichuanense are clustered together with other G. sichuanense collections, while G. orbiforme is closer to the collections of G. mastoporum Lloyd [67], recognized G. mastoporum (=G. orbiforme) as a distinct species with lateral or dorsally lateral stipes, which stipe development varies with different growing environments [68]. Wang et al. [50] concluded that the morphological and molecular data of G. mastoporum are conspecific with G. orbiforme, and the latter is the earliest valid name for use. Ganoderma cupreum, G. fornicatum, G. mastoporum, G. orbiforme, G. subtornatum, and the Chinese species described; G. densizonatum and G. limushanense are morphologically conspecific with G. orbiforme. In this study, we report on Thai G. orbiforme with its molecular and morphological evidence for the first time. Morphological characteristics of G. orbiforme are similar to the collections that described by Ryvarden [6] and Hapuarachchi et al. [51].
It is estimated that more than 650 mushroom species are edible; however, only nine species of Ganoderma species are cultivated and include G. applanatum, G. australe, G. curtisii, G. lucidum, G. oregonense, G. resinaceum, G. sinense, G. tenus, G. tropicum, G. tsugae [9,69]. As G. lucidum is not common in nature, the number of wild mushrooms is not sufficient for commercial exploitation. Its cultivation on solid substrates, stationary liquid medium or submerged cultivation has become essential to meet the increasing demands for this mushroom. The annual sale of products derived from G. lucidum is estimated to be more than USD 2.5 billion in Asian countries, including China, Japan, and South Korea. Mushroom growing is also becoming increasingly popular in Thailand. Ganoderma lucidum has a market price of USD 33.0–50.0 per 1 kg as a commercial mushroom in Thailand. In the cultivation trials of this study, the optimal medium for G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) was PSA. Oatmeal agar was suitable to grow G. orbiforme (MFLUCC 22-0066). Luangharn et al. [8] reported that Ganoderma australe was found in Thailand. The optimal conditions of mycelia growth were found in the PDA medium. Furthermore, ref. [9] also reported that PDA, MEA, and YPD are suitable for growing G. tropicum. Thus, we can consume Ganoderma mushrooms, which can be cultivated on several substrates, depending on the species. Various cultivated mushrooms have different optimal temperatures; for example, Volvariella volvacea at 35 °C, Pleurotus eryngii at 25 °C Phlebopus portentosus at 30 °C [70,71] while the temperature for optimal growth of Ganoderma species is between 25 and 30 °C. The three strains G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065) and G. orbiforme (MFLUCC 22-0066) grow well between 25 and 30 °C, the same as G. australe [8], G.lucidum [72], and G. tropicum at 25–28 °C [9].
Mushroom spawn substrates can be cereal grain, sawdust, wood chips, or rope [73,74]. The most common materials are cereal grains. Sorghum was used as the medium for all types of mushrooms [69,75]. Rye was used to grow Agaricus sp. [63,69]. In the case of Ganoderma mushrooms, durum wheat, sorghum, and wheat grain could be used as an alternative for spawn production [8,76,77,78]. In this study, we found that all types of cereal grains (barley, maize, millet, rice berry, wheat) can be used as spawn production. Similarly, Luangharn et al. [8] reported that barley, corn cobs, pearl mille, and sorghum could be used to promote the mycelium growth in spawn production. In the current trend, agricultural wastes were alternative ways to used. It can be utilized as substrates for spawning to decrease the price of mushrooms. Rashad et al. [79] grew the mycelia of Ganoderma in different types of agriculture waste, e.g., broad bean stalks, cotton stalk, maize straw, rice straw, sugarcane bagasse, and wheat straw mixed with wheat bran or corn gluten, and found that cotton stalk sugarcane bagasse rice straw and wheat straw showed the best supplementation for spawn production.
In cultivation trials, three strains of wild Thai Ganoderma, G. sichuanense (MFLUCC 22-0064 and MFLUCC 22-0065), and G. orbiforme (MFLUCC 22-0066) can be grown in rubber sawdust bags and artificially produce the fruiting bodies. Our findings are similar to those of Luangharn et al. [8], who reported that wild Thai Ganoderma australe produced fruiting bodies in rubber sawdust bags at 30 °C, with 60–75% humidity. However, our strains have production at 75–85% humidity. Rubber sawdust is commonly used and is the preferred medium on the commercial scale in Thailand [69]. However, the availability of raw materials of any region is a key factor when choosing agricultural waste for growing the mushrooms. Luangharn et al. [77] noted that Alnus cremastogyne sawdust that is planted in China can be used to grow three species of Ganoderma: Ganoderma leucocontextum, G. resinaceum, and G. gibbosum. Ozcariz-Fermoselle et al. [78] reported that pecan nut waste (Carya illinoinensis) can be used for G. lucidum cultivation in Spain. An important piece of further study is to find local agricultural waste substrates to grow mushrooms that can produce a better yield than sawdust alone in Thailand. As the use of local agricultural waste substrates will be shown as an environmentally friendly alternative, it might increase the benefits of local agricultural waste recycling.
Mushrooms have been used by mankind as a nutritional food. To date, in terms of nutrition, studies from mushrooms are increasing in interest. The demand for functional foods with high nutritional and medicinal value is a hot issue. Mushrooms are a good source of nutritional value. They contain high levels of protein, high fiber, carbohydrates, minerals, vitamins, and have a low fat content [50,80]. Ganoderma is treasured as a functional food. They are found to be a good source of nutrition component. In this study, nutrition of content of wild Thai Ganoderma, G. sichuanense and G. orbiforme were studied. These mushrooms revealed high amounts of fiber (47.90–52.45%), followed by the total protein (12.80–14.67%), fat (4.90–5.70%) and carbohydrates (3.16–4.02%). While Singh et al. [81] report a proximate analysis of four species of Ganoderma (G. applanatum, G. brownie, G. lucidum, and G. philippii) and found that those strains contained the highest carbohydrates (75.5–81.4%), proteins (9.29–13.3%), fiber (4.92–8.07%), and a low fat content (1.62–2.87%).
The α-glucosidase inhibitor has been used as a treatment for type 2 diabetes (T2D) [82]. Ganoderma species have an important economic value due to their medicinal properties and pathogenicity [17,83]. The genus has been used as a medicinal mushroom for a very long time. There are many reports showing α-glucosidase inhibitory activities from Ganoderma species such as Ganoderma hainanense, G. leucocontextum, G. lucidum, and G. resinaceum [82,84,85,86,87]. In this study, the detection of α-glucosidase inhibitory activity in Thai Ganoderma was found. The fruiting bodies of Ganoderma sp. that were extracted by EtOH showed α-glucosidase inhibitory activity. Ganoderma orbiforme (MFLUCC 22-0066) and G. sichuanense (MFLUCC 22-0064) had stronger inhibition than the positive control (acarbose). The IC50 values were 105.97 ± 1.36, and 126.94 ± 0.87 µg/mL, respectively. In addition, the extraction of Ganoderma orbiforme (MFLUCC 22-0066) by water was also successful. The IC50 values were 124.40 ± 3.18 µg/mL, while the acarbose was 168.18 ± 0.89 µM. These preliminary results suggest that the strains of Thai Ganoderma sp. in this study could be further researched and developed.

5. Conclusions

In this study, the most favorable medium was potato sucrose agar (PSA) for G. sichuanense and oatmeal agar (OMA) for G. orbiforme. The suitable temperature and pH of the three strains were 25 °C and 30 °C and pH was 4–8. The most favorable spawn were rice berry, wheat, and barley, respectively. The nutritional value of three strains of wild Thai Ganoderma revealed high amounts of fiber and was followed by total protein. Furthermore, G. orbiforme (MFLUCC 22-0066) and G. sichuanense (MFLUCC 22-0064) showed better inhibition than acarbose. The result of this study is useful as basic information for preventing and treating diabetes. Therefore, these local Ganoderma strains could be industrially cultivated and introduced to the Thai market. However, more research is needed to find local agricultural waste to increase production yields and more research is needed to study glucose uptake in cell lines and clinical trials.

Author Contributions

Conceptualization: N.T. Formal analysis: N.W., T.L. and N.T. Resources: A.T. Writing—original draft preparation: N.W., T.L. and N.T. Writing—review and editing: N.T., R.C., W.J., W.T. and K.D.H. All authors have read and agreed to the published version of the manuscript.

Funding

We gratefully acknowledge Mae Fah Luang University for grant entitled “Optimal conditions for domestication and biological activities of selected species of Ganoderma” (grant No. 621C15035) for financial support.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Karsten, P.A. Enumeratio Boletinarum et Polyporarum Fennicarum systematae novo dispositorum. Rev. Mycol. 1881, 3, 16–19. [Google Scholar]
  2. Justo, A.; Miettinen, O.; Floudaas, D.; Ortiz-Santana, B.; Sjökvist, E.; Lindner, D.; Nakasone, K.; Niemela, D.; Nakasone, K.; Niemelö, T.; et al. A revised family-level classification of the Polyporales (Basidiomycota). Fungal Biol. 2017, 121, 798–824. [Google Scholar] [CrossRef]
  3. Cui, B.K.; Li, H.J.; Ji, X.; Zhou, J.L.; Song, J.; Si, J.; Yang, Z.L.; Dai, Y.C. Species diversity, taxonomy and phylogeny of Polyporaceae (Basidiomycota) in China. Fungal Divers. Int. J. Mycol. 2019, 97, 137–392. [Google Scholar] [CrossRef]
  4. Moncalvo, J.M.; Ryvarden, L. A Nomenclatural Study of the Ganodermataceae Donk; Fungiflora: Oslo, Norway, 1997; Volume 11, 114p, Available online: https://www.fungiflora.no/_files/ugd/b2cc6b_c203c189159b4c47862b64d1feb6e934.pdf (accessed on 23 January 2023).
  5. Núñez, M.; Ryvarden, L. East Asian Polypores: Ganodermataceae and Hymenochaetaceae; Fungiflora: Oslo, Norway, 2000; p. 168. Available online: www.mykoweb.com/systematics/literature/East%20Asian%20Polypores%20Vol%202.pdf (accessed on 23 January 2023).
  6. Ryvarden, L. Neotropical Polypores Part 1 Introduction, Ganodermataceae & Hymenochaetaceae; Fungiflora: Oslo, Norway, 2004; Volume 19, 229p, Available online: https://www.fungiflora.no/_files/ugd/b2cc6b_f5908c953a45494189c48f11f272b001.pdf (accessed on 23 January 2023).
  7. Index Fungorum. 2023. Available online: http://www.indexfungorum.org/names/names.asp (accessed on 20 June 2023).
  8. Luangharn, T.; Karunarathna, S.C.; Khan, S.; Xu, J.C.; Mortimer, P.E.; Hyde, K.D. Antibacterial activity, optimal culture conditions and cultivation of the medicinal Ganoderma australe, new to Thailand. Mycosphere 2017, 8, 1108–1123. [Google Scholar] [CrossRef]
  9. Luangharn, T.; Karunarathna, S.C.; Mortimer, P.E.; Hyde, K.D.; Thongklang, N.; Xu, J. A new record of Ganoderma tropicum (Basidiomycota, Polyporales) for Thailand and first assessment of optimum conditions for mycelia production. MycoKeys 2019, 51, 65–83. [Google Scholar] [CrossRef]
  10. Thawthong, A.; Hapuarachchi, K.K.; Wen, T.C.; Raspé, O.; Thongklang, N.; Kang, J.C.; Hyde, K.D. Ganoderma sichuanense (Ganodermataceae, Polyporales) new to Thailand. MycoKeys 2017, 22, 27–43. [Google Scholar] [CrossRef]
  11. Cilerdzic, J.; Kosanic, M.; Stajić, M.; Vukojevic, J.; Ranković, B. Species of Genus Ganoderma (Agaricomycetes) Fermentation Broth: A Novel Antioxidant and Antimicrobial Agent. Int. J. Med. Mushrooms 2016, 18, 397–404. [Google Scholar] [CrossRef]
  12. Cao, Y.; Yuan, H.S. Ganoderma mutabile sp. nov. from southwestern China based on morphological and molecular data. Mycol. Prog. 2013, 12, 121–126. [Google Scholar] [CrossRef]
  13. Pilotti, C.A. Stem rots of oil palm caused by Ganoderma boninense: Pathogen biology and epidemiology. Mycopathologia 2005, 159, 129–137. [Google Scholar] [CrossRef]
  14. Li, M.J.; Yuan, H.S. Type studies on Amauroderma species described by J.D. Zhao et al. and the phylogeny of species in China. Mycotaxon 2015, 130, 79–89. [Google Scholar] [CrossRef]
  15. Tchotet-Tchoumi, J.M.; Coetzee, M.P.A.; Rajchenberg, M.; Wingfield, M.J.; Roux, J. Three Ganoderma species, including Ganoderma dunense sp. nov., associated with dying Acacia cyclops trees in South Africa. Australas. Plant Pathol. 2018, 47, 431–447. [Google Scholar] [CrossRef]
  16. Pilotti, C.A.; Sanderson, F.R.; Aitken, E.A.B.; Armstrong, W. Morphological variation and host range of two Ganoderma species from Papua New Guinea. Mycopathologia 2004, 158, 251–265. [Google Scholar] [CrossRef]
  17. Dai, Y.C.; Yang, Z.L.; Cui, B.K.; Yu, C.J.; Zhou, L.W. Species Diversity and Utilization of Medicinal Mushrooms and Fungi in China (Review). Int. J. Med. Mushrooms 2009, 11, 287–302. [Google Scholar] [CrossRef]
  18. Hapuarachchi, K.K.; Karunarathna, S.C.; McKenzie, E.H.C.; Wu, X.L.; Kakumyan, P.; Hyde, K.D.; Wen, T.C. High phenotypic plasticity of Ganoderma sinense (Ganodermataceae, Polyporales) in China. Asian J. Mycol. 2018, 2, 1–47. Available online: https://asianjournalofmycology.org/pdf/AJOM_2_1_1.pdf (accessed on 8 February 2023). [CrossRef]
  19. Teng, B.S.; Wang, C.D.; Yang, H.J.; Wu, J.S.; Zhang, D.; Zheng, M.; Fan, Z.H.; Pan, D.; Zhou, P. A protein tyrosine phosphatase 1B activity inhibitor from the fruiting bodies of Ganoderma lucidum (Fr.) Karst and its hypoglycemic potency on streptozotocin-induced type 2 diabetic mice. J. Agric. Food Chem. 2011, 59, 6492–6500. [Google Scholar] [CrossRef]
  20. De Silva, D.D.; Rapior, S.; Fons, F.; Bahkali, A.H.; Hyde, K.D. Medicinal mushrooms in supportive cancer therapies: An approach to anti-cancer effects and putative mechanisms of action. Fungal Divers. Int. J. Mycol. 2012, 55, 1–35. [Google Scholar] [CrossRef]
  21. De Silva, D.D.; Rapior, S.; Sudarman, E.; Stadler, M.; Xu, J.C.; Alias, S.A.; Hyde, K.D. Bioactive metabolites from macrofungi: Ethnopharmacology, Biological activities, Chemistry. Fungal Divers. Int. J. Mycol. 2013, 62, 1–40. [Google Scholar] [CrossRef]
  22. Richter, C.; Wittstein, K.; Kirk, P.M.; Stadler, M. An assessment of the taxonomy and chemotaxonomy of Ganoderma. Fungal Divers. Int. J. Mycol. 2015, 71, 1–15. [Google Scholar] [CrossRef]
  23. Hapuarachchi, K.K.; Karunarathna, S.C.; Phengsintham, P.; Kakumyan, P.; Hyde, K.D.; Wen, T.C. Amauroderma (Ganodermataceae, Polyporales)—Bioactive compounds, beneficial properties and two new records from Laos. Asian J. Mycol. 2018, 1, 121–136. [Google Scholar] [CrossRef]
  24. Chang, S.T.; Buswell, J.A. Ganoderma lucidum (Curt.: Fr.) P. Karst. (Aphyllophoromycetideae)-A Mushrooming Medicinal Mushroom. Int. J. Med. Mushrooms 1999, 1, 139–146. [Google Scholar] [CrossRef]
  25. Lai, T.; Gao, Y.; Zhou, S. Global marketing of medicinal ling zhi mushroom Ganoderma lucidum (W. Curt.: Fr.) Lloyd (Aphyllophoromycetideae) products and safety concerns. Int. J. Med. Mushrooms 2004, 6, 189–194. [Google Scholar] [CrossRef]
  26. Singh, S.K.; Doshi, A.; Pancholy, A.; Pathak, R. Biodiversity in wood–decay macro–fungi associated with declining arid zone trees of India as revealed by nuclear rDNA analysis. Eur. J. Plant Pathol. 2013, 136, 373–382. [Google Scholar] [CrossRef]
  27. Vilgalys, R.; Hester, M. Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. J. Bacteriol. 1990, 172, 4238–4246. [Google Scholar] [CrossRef] [PubMed]
  28. White, T.J.; Bruns, T.; Lee, S.; Taylor, J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protoc. A Guide Methods Appl. 1990, 38, 315–322. [Google Scholar] [CrossRef]
  29. Liu, Y.J.; Whelen, S.; Hall, B.D. Phylogenetic relationships among ascomycetes: Evidence from an RNA polymerase II subunit. Mol. Biol. Evol. 1999, 16, 1799–1808. [Google Scholar] [CrossRef] [PubMed]
  30. Zhou, L.W.; Cao, Y.; Wu, S.H.; Vlasák, J.; Li, D.W.; Li, M.J.; Dai, Y.C. Global diversity of the Ganoderma lucidum complex (Ganodermataceae, Polyporales) inferred from morphology and multilocus phylogeny. Phytochemistry 2015, 114, 7–15. [Google Scholar] [CrossRef]
  31. Katoh, K.; Standley, D.M. MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Mol. Biol. Evol. 2013, 30, 772–780. [Google Scholar] [CrossRef]
  32. Hall, T.A. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. Ser. 1999, 41, 95–98. Available online: https://www.academia.edu/download/29520866/1999hall1.pdf (accessed on 8 February 2023).
  33. Swofford, D.L. PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods) 2002, Version 4.0 Beta Version 10. Sinauer Associates. Available online: https://www.researchgate.net/publication/271205405_PAUP_Phylogenetic_Analysis_Using_Parsimony_and_Other_Methods_Version_40b10 (accessed on 8 February 2023).
  34. Huelsenbeck, J.P.; Ronquist, F. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 2001, 17, 754–755. [Google Scholar] [CrossRef] [PubMed]
  35. Miller, M.A.; Pfeiffer, W.; Schwartz, T. Creating the CIPRES Science Gateway for inference of large phylogenetic trees. In Proceedings of the 2010 Gateway Computing Environments Workshop (GCE), New Orleans, LA, USA, 14 November 2010; pp. 1–8. [Google Scholar] [CrossRef]
  36. Stamatakis, A. RAxML-VI-HPC: Maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics 2006, 22, 2688–2690. [Google Scholar] [CrossRef]
  37. Silvestro, D.; Michalak, I. raxmlGUI: A graphical front-end for RAxML. Org. Divers. Evol. 2012, 12, 335–337. [Google Scholar] [CrossRef]
  38. Nylander, J.A. MrModeltest 2.0. Program Distributed by the Author; Evolutionary Biology Centre, Uppsala University: Uppsala, Sweden, 2004. [Google Scholar]
  39. Rambaut A Fig Tree Version 1.4.0. 2012. Available online: http://tree.bio.ed.ac.uk/software/figtree/ (accessed on 24 January 2023).
  40. Sun, Y.F.; Xing, J.H.; He, X.L.; Wu, D.M.; Song, C.G.; Liu, S.; Vlasák, J.; Gates, G.; Gibertoni, T.B.; Cui, B.K. Species diversity, systematic revision and molecular phylogeny of Ganodermataceae (Polyporales, Basidiomycota) with an emphasis on Chinese collections. Stud. Mycol. 2022, 101, 287–415. [Google Scholar] [CrossRef]
  41. Xing, H.; Sun, Y.F.; Han, Y.L.; Cui, B.K.; Dai, Y.C. Morphological and molecular identification of two new Ganoderma species on Casuarina equisetifolia from China. MycoKeys 2018, 34, 93–108. Available online: https://mycokeys.pensoft.net/article/22593/download/pdf/ (accessed on 8 February 2023). [CrossRef] [PubMed]
  42. Wu, S.H.; Chern, C.L.; Wei, C.L.; Chen, Y.P.; Akiba, M.; Hattori, T. Ganoderma bambusicola sp. nov. (Polyporales, Basidiomycota) from southern Asia. Phytotxa 2020, 456, 75–85. [Google Scholar] [CrossRef]
  43. Luangharn, T.; Karunarathna, S.C.; Dutta, A.K.; Paloi, S.; Promputtha, I.; Hyde, K.D.; Xu, J.C.; Mortimer, P.E. Ganoderma (Ganodermataceae, Basidiomycota) species from the greater Mekong subregion. J. Fungi 2021, 7, 819. [Google Scholar] [CrossRef] [PubMed]
  44. Coetzee, M.P.A.; Marincowitz, S.; Muthelo, V.G.; Wingfield, M.J. Ganoderma species, including new taxa associated with root rot of the iconic Jacaranda mimosifolia in Pretoria, South Africa. IMA Fungus 2015, 6, 249–256. [Google Scholar] [CrossRef]
  45. He, J.; Luo, Z.L.; Tang, S.M.; Li, Y.J.; Li, S.H.; Su, H. Phylogenetic analyses and morphological characters reveal two new species of Ganoderma from Yunnan province, China. MycoKeys 2021, 84, 141–162. [Google Scholar] [CrossRef] [PubMed]
  46. Tchotet-Tchoumi, J.M.; Coetzee, M.P.A.; Rajchenberg, M.; Roux, J. Taxonomy and species diversity of Ganoderma species in the Garden Route National Park of South Africa inferred from morphology and multilocus phylogenies. Mycologia 2019, 111, 730–747. [Google Scholar] [CrossRef]
  47. Crous, P.W.; Wingfield, M.J.; Richardson, D.M.; Leroux, J.J.; Strasberg, D.; Edwards, J.; Roets, F.; Hubka, V.; Taylor, P.W.; Heykoop, M.; et al. Fungal planet description sheets: 400–468. Persoonia 2016, 36, 316–458. [Google Scholar] [CrossRef] [PubMed]
  48. Song, J.; Xing, J.H.; Decock, C.; He, X.L.; Cui, B.K. Molecular phylogeny and morphology reveal a new species of Amaurderma (Basidiomycota) from China. Phytotaxa 2016, 260, 47–56. [Google Scholar] [CrossRef]
  49. Xing, J.H.; Song, J.; Decock, C.; Cui, B.K. Morphological characters and phylogenetic analysis reveal a new species within the Ganoderma lucidum complex from South Africa. Phytotaxa 2016, 266, 115–124. [Google Scholar] [CrossRef]
  50. Wang, D.M.; Wu, S.H.; Yao, Y.J. Clarification of the concept of Ganoderma orbiforme with high morphological plasticity. PLoS ONE 2014, 9, e98733. [Google Scholar] [CrossRef]
  51. Hapuarachchi, K.K.; Karunarathna, S.C.; Phengsintham, P.; Yang, H.D.; Kakumyan, P.; Hyde, K.D.; Wen, T.C. Ganodermataceae (Polyporales): Diversity in Greater Mekong Subregion countries (China, Laos, Myanmar, Thailand and Vietnam). Mycosphere 2019, 10, 221–309. [Google Scholar] [CrossRef]
  52. Kinge, T.R.; Mih, A.M. Ganoderma ryvardense sp. nov. Associated with Basal Stem Rot (BSR) Dis. Oil Palm Cameroon. Mycosphere 2011, 2, 179–188. Available online: https://www.mycosphere.org/pdfs/MC2_2_No8.pdf (accessed on 15 January 2023).
  53. Liu, H.; Guo, L.J.; Li, S.L.; Fan, L. Ganoderma shanxiense, a new species from northern China based on morphological and molecular evidence. Phytotaxa 2019, 406, 129–136. [Google Scholar] [CrossRef]
  54. Raja, H.A.; Baker, T.; Little, J.G.; Oberlies, N.F. DNA barcoding for identification of consumer-relevant mushrooms: A partial solution for product certification? Food Chem. 2017, 214, 383–392. [Google Scholar] [CrossRef]
  55. Cao, Y.; Wu, S.H.; Dai, Y.C. Species clarification of the prize medicinal Ganoderma mushroom “Lingzhi”. Fungal Divers. 2012, 56, 49–62. [Google Scholar] [CrossRef]
  56. Crous, P.W.; Wingfield, M.J.; Roux, J.J.L.; Richardson, D.M.; Strasberg, D.; Shivas, R.G.; Alvarado, P.; Edwards, J.; Moreno, G.; Sharma, R.; et al. Fungal Planet Description Sheets: 371–399; Naturalis Biodiversity Center: Leiden, The Netherlands, 2015; pp. 371–399. [Google Scholar] [CrossRef]
  57. Lodge, D.J.; Ammirati, F.J.; O’Dell, T.E.; Mueller, G.M. Collecting and Describing Macrofungi. In Biodiversity of Fungi Inventory and Monitoring Methods; Mueller, G.M., Bills, G.F., Foster, M.S., Eds.; Elsevier Academic Press: London, UK, 2004; pp. 128–154. Available online: https://www.fpl.fs.usda.gov/documnts/pdf2004/fpl_2004_lodge001.pdf (accessed on 15 January 2023).
  58. Kornerup, A.; Wanscher, J.H. Methuen Handbook of Colour 1978; Eyre Methuen: Methuen, MA, USA, 1981. [Google Scholar]
  59. Miettinen, O.; Larsson, K.H. Trechispora elongata Species Nova from North Europe. Mycotaxon 2006, 96, 193–198. Available online: https://helda.helsinki.fi/bitstream/handle/10138/42882/Miettinen2006Trechispora_elongata.pdf?sequence=2 (accessed on 16 February 2023).
  60. Tulloss, R.E. Amaniteae: Amanita, Limacella, & Torrendia. By Pierre Neville & Serge Poumarat, etc. Book Review. Mycotaxon 2005, 92, 474–484. [Google Scholar]
  61. Thongklang, N.; Keokanngeun, L.; Taliam, W.; Hyde, K.D. Cultivation of a wild strain of Auricularia cornea from Thailand. Curr. Res. Environ. Appl. Mycol. 2020, 10, 120–130. [Google Scholar] [CrossRef]
  62. Royse, D.J. Effects of fragmentation, supplementation and the addition of phase II compost to 2nd break compost on mushroom (Agaricus bisporus) yield. Bioresour. Technol. 2010, 101, 188–192. [Google Scholar] [CrossRef]
  63. Thongklang, N.; Sysouphanthong, P.; Callac, P.; Hyde, K.D. First cultivation of Agaricus flocculosipes and a novel Thai strain of A. subrufescens. Mycosphere 2014, 5, 814–820. [Google Scholar] [CrossRef]
  64. Abdul Razak, D.L.; Abdullah, N.; Johari, N.M.K.; Vikineswary, S. Comparative study of mycelia growth and sporophore yield of Auricularia polytricha (Mont) Sacc on selected palm oil wastes as fruiting substrate. Appl. Microbiol. Biotechnol. 2013, 97, 3207–3213. [Google Scholar] [CrossRef]
  65. Liang, C.H.; Wu, C.Y.; Lu, P.L.; Kuo, Y.C.; Liang, Z.C. Biological efficiency and nutritional value of the culinary-medicinal mushroom Auricularia cultivated on a sawdust basal substrate 129 supplement with different proportions of grass plants. Saudi J. Biol. Sci. 2019, 26, 263–269. [Google Scholar] [CrossRef] [PubMed]
  66. Zhao, J.D.; Xu, L.W.; Zhang, X.Q. Taxonomic studies on the family Ganodermataceae of China II. Acta Mycol. Sin. 1983, 2, 159–167. (In Chinese) [Google Scholar]
  67. Lloyd, J.U. The cultivation of hydrastis. J. Am. Pharmaceut. Assoc. 1912, 1, 5–12. [Google Scholar] [CrossRef]
  68. Moncalvo, J.M. Systematics of Ganoderma. In Ganoderma Diseases of Perennial Crops; Flood, J., Bridge, P.D., Holderness, M., Egham, M., Eds.; CABI Bioscience: Wallingford, UK, 2000; pp. 23–45. [Google Scholar]
  69. Thawthong, A.; Karunarathna, S.C.; Thongklang, N.; Chukeatirote, E.; Kakumyam, P.; Chamyuang, S.; Rizal, L.M.; Mortimer, P.E.; Xu, J.; Callac, P.; et al. Discovering and Domesticating Wild Tropical Cultivatable Mushrooms. Chiang Mai J. Sci. 2014, 41, 731–764. Available online: https://www.researchgate.net/publication/266022771_Discovering_and_Domesticating_Wild_Tropical_Cultivatable_Mushrooms (accessed on 4 March 2023).
  70. Zervakis, G.; Philippoussis, A.; Ioannidou, S.; Diamantopoulou, P. Mycelium growth kinetics and optimal temperature conditions for the cultivation of edible mushroom species on lignocellulosic substrates. Folia Microbiol. 2001, 46, 231–234. [Google Scholar] [CrossRef]
  71. Thongklang, N.; Lumyong, S.; Bussaban, B.; Hyde, K.D. Potential to cultivate Phlebopus portentosus; optimal conditions for mycelia growth and inoculum production. Maejo Int. J. Sci. Technol. 2010, 5, 413–425. Available online: https://mijst.mju.ac.th/vol5/413-425.pdf (accessed on 8 February 2023).
  72. Kapoor, P.; Sharma, B.M. Studies on Different Growth Parameters of Ganoderma lucidum. Int. J. Sci. Environ. Tech. 2014, 3, 1515–1524. Available online: https://citeseerx.ist.psu.edu/document?repid=rep1&type=pdf&doi=a6c5e137ce012d991bd1f544ed36c042413a7722 (accessed on 4 March 2023).
  73. Mendonça, M.D.; Kasuya, M.C.; Cadorin, A.; Vieira, A.J. Agaricus blazei Cultivation for a Living in Brazil. In Mushroom Growers’ Handbook 2: Shiitake Cultivation; MushWorld-Heineart Inc: Seoul, Republic of Korea, 2005; Available online: https://www.goba.eu/wp-content/uploads/2015/06/Mushroom_Growers_Handbook_2_-_Shiitake_Cultivation.pdf (accessed on 4 March 2023).
  74. Sharma, V.P.; Kumar, S. Spawn production technology. In Mushrooms: Cultivation, Marketing and Consumption; Singh, M., Vijay, B., Kamal, S., Wakchaure, G.C., Eds.; Directorate of Mushroom Research (ICAR): Himachal Pradesh, India, 2011; pp. 31–42. Available online: https://www.academia.edu/download/48415834/spawn.pdf (accessed on 12 March 2023).
  75. Thongklang, N.; Luangharn, T. Testing agricultural wastes for the production of Pleurotus ostreatus. Mycosphere 2016, 7, 766–772. [Google Scholar] [CrossRef]
  76. Alheeti, A.; Muslat, M.M.; Ayyash, L.M.; There, R.M. Isolation, Identification and Organic Production of Mushrooms Ganoderma lucidum (Curt.:Fr) Karst (Reishi). Indian J. Ecol. 2020, 47, 231–235. Available online: https://www.researchgate.net/publication/349699150_Isolation_Identification_and_Organic_Production_of_Mushrooms_Ganoderma_lucidum_CurtFr_Karst_Reishi (accessed on 12 March 2023).
  77. Luangharn, T.; Mortimer, P.E.; Karunarathna, S.C.; Hyde, K.D.; Xu, J. Domestication of Ganoderma leucocontextum, G. resinaceum, and G. gibbosum collected from Yunnan Province, China. Biosci. Biotechnol. Res. Asia 2020, 17, 7–26. [Google Scholar] [CrossRef]
  78. Ozcariz-Fermoselle, M.V.; Fraile-Fabero, R.; Girbés-Juan, T.; Arce-Cervantes, O.; Oria de Rueda-Salgueiro, J.A.; Azul, A.M. Use of lignocellulosic wastes of pecan (Carya illinoinensis) in the cultivation of Ganoderma lucidum. Rev. Iberoam. Micol. 2018, 35, 103–109. [Google Scholar] [CrossRef]
  79. Rashad, F.M.; Kattan, M.H.E.; Fathy, H.M.; El-Fattah, D.A.A.; Tohamy, M.E.; Farahat, A.A. Recycling of agro-wastes for Ganoderma lucidum mushroom production and Ganoderma post mushroom substrate as soil amendment. Waste Manag. 2019, 88, 147–159. [Google Scholar] [CrossRef]
  80. Kalač, P. Chemical composition and nutritional value of European species of wild growing mushrooms: A review. Food Chem. 2009, 113, 9–16. [Google Scholar] [CrossRef]
  81. Singh, R.; Ghumman, N.K.; Shri, R.; Singh, A.P.; Dhingra, G.S. Proximate composition and element contents of selected species of Ganoderma with reference to dietary intakes. Environ Monit Assess 2020, 192, 270. [Google Scholar] [CrossRef]
  82. Fatmawati, S.; Shimizua, K.; Kondo, R. Ganoderol B: A potent α-glucosidase inhibitor isolated from the fruiting body of Ganoderma lucidum. Phytomedicine 2011, 18, 1053–1055. [Google Scholar] [CrossRef]
  83. Dai, Y.C.; Cui, B.K.; Yuan, H.S.; Li, B.D. Pathogenic wood–decaying fungi in China. For. Pathol. 2007, 37, 105–120. [Google Scholar] [CrossRef]
  84. Chen, X.Q.; Zhao, J.; Chen, L.X.; Wang, S.F.; Wang, Y.; Li, S.P. Lanostane triterpenes from the mushroom Ganoderma resinaceum and their inhibitory activities against α-glucosidase. Phytochemistry 2018, 149, 103–115. [Google Scholar] [CrossRef]
  85. Wang, S.M.; Han, J.J.; Ma, K.; Jin, T.; Bao, L.; Pei, Y.F.; Liu, H.W. New α-glucosidase inhibitors with p-terphenyl skeleton from the mushroom Hydnellum concrescens. Fitoterapia 2015, 98, 149–155. [Google Scholar] [CrossRef] [PubMed]
  86. Ma, L.F.; Yan, J.J.; Lang, H.Y.; Jin, L.C.; Qiu, F.J.; Wang, Y.J.; Xi, Z.F.; Shan, W.G.; Zhan, Z.J.; Ying, Y.M. Bioassay-guided isolation of lanostane-type triterpenoids as alpha-glucosidase inhibitors from Ganoderma hainanense. Phytochem. Lett. 2019, 29, 154–159. [Google Scholar] [CrossRef]
  87. Costa, T.M.; Mayer, D.A.; Siebert, D.A.; Micke, G.A.; Alberton, M.D.; Tavares, L.B.B.; de Oliveira, D. Kinetics analysis of the inhibitory effects of alpha-glucosidase and identification of compounds from Ganoderma lipsiense mycelium. Appl. Biochem. Biotechnol. 2020, 191, 996–1009. [Google Scholar] [CrossRef] [PubMed]
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Figure 1. Phylogram of Ganoderma species obtained from maximum likelihood (RAxML) of combined ITS dataset. Bootstrap values (BS) from maximum likelihood (ML, left) and maximum parsimony (MP, middle) greater than 70% and Bayesian posterior probabilities (PP), greater than 0.95, are indicated above the nodes as MLBS/MPBS/PP. The tree is rooted with Tomophagus colossus TC-02. Black arrow indicates the values of MLBS/MPBS/PP of Ganoderma tropicum.
Figure 1. Phylogram of Ganoderma species obtained from maximum likelihood (RAxML) of combined ITS dataset. Bootstrap values (BS) from maximum likelihood (ML, left) and maximum parsimony (MP, middle) greater than 70% and Bayesian posterior probabilities (PP), greater than 0.95, are indicated above the nodes as MLBS/MPBS/PP. The tree is rooted with Tomophagus colossus TC-02. Black arrow indicates the values of MLBS/MPBS/PP of Ganoderma tropicum.
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Figure 2. Morphological characteristics of Ganoderma sichuanense (MFLUCC 22-0064) from the artificially cultivated: (a) mature fruiting body with upper surface; (b) mature fruiting body with lower surface; (c) upper surface under Pileipellis; (d) Pileipellis; (e–h) context hyphae; (i–k) pores; (l) basidiospores with pore hyphal system; (m,n) basidiospores. Scale bars: (a,b) = 3 cm, (c,d,f,h) = 50 μm, (e,j,k) = 100 μm, (g,l,m) = 30 μm, (i) = 200 μm, (n) = 10 μm.
Figure 2. Morphological characteristics of Ganoderma sichuanense (MFLUCC 22-0064) from the artificially cultivated: (a) mature fruiting body with upper surface; (b) mature fruiting body with lower surface; (c) upper surface under Pileipellis; (d) Pileipellis; (e–h) context hyphae; (i–k) pores; (l) basidiospores with pore hyphal system; (m,n) basidiospores. Scale bars: (a,b) = 3 cm, (c,d,f,h) = 50 μm, (e,j,k) = 100 μm, (g,l,m) = 30 μm, (i) = 200 μm, (n) = 10 μm.
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Figure 3. Morphological characteristics of Ganoderma orbiforme (MFLUCC 22-0066) from the artificially cultivated. (a) Mature fruiting body with upper surface, (b) pore characteristics, (c) upper surface under Pileipellis, (d) Pileipellis, (e) sections of pores, (f–h) context hyphae, (i) sections of tubes layers, (j,k) basidiospores. Scale bars: (a) = 3 cm, (b) = 1000 μm, (c) = 500 μm, (d,g) = 50 μm, (e) = 100 μm, (f) = 200 μm, (h) = 30 μm, (i) = 300 μm, (j,k) = 5 μm.
Figure 3. Morphological characteristics of Ganoderma orbiforme (MFLUCC 22-0066) from the artificially cultivated. (a) Mature fruiting body with upper surface, (b) pore characteristics, (c) upper surface under Pileipellis, (d) Pileipellis, (e) sections of pores, (f–h) context hyphae, (i) sections of tubes layers, (j,k) basidiospores. Scale bars: (a) = 3 cm, (b) = 1000 μm, (c) = 500 μm, (d,g) = 50 μm, (e) = 100 μm, (f) = 200 μm, (h) = 30 μm, (i) = 300 μm, (j,k) = 5 μm.
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Figure 4. First cultivation of Ganoderma species from Thailand at 28 ± 1 °C in 75–85% humidity. Ganoderma sichuanense MFLUCC 22-0064 (a,b), G. sichuanense MFLUCC 22-0065 (c,d) and Ganoderma orbiforme MFLUCC 22-0066 (e,f).
Figure 4. First cultivation of Ganoderma species from Thailand at 28 ± 1 °C in 75–85% humidity. Ganoderma sichuanense MFLUCC 22-0064 (a,b), G. sichuanense MFLUCC 22-0065 (c,d) and Ganoderma orbiforme MFLUCC 22-0066 (e,f).
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Figure 5. First field cultivation of wild Thai Ganoderma orbiforme (MFLUCC 22-0066) (a–d).
Figure 5. First field cultivation of wild Thai Ganoderma orbiforme (MFLUCC 22-0066) (a–d).
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Table 1. Taxa used in this study and their GenBank accession numbers for ITS DNA sequence data.
Table 1. Taxa used in this study and their GenBank accession numbers for ITS DNA sequence data.
Fungal SpeciesVoucherGenBank Accession No.References
ITSLSURPB2
Ganoderma acaciicolaCui 16815MZ354895MZ355005MZ245384[40]
G. acontextumJV 0611 21GKF605667--[41]
G. angustisporumCui 13817MG279170--[41]
G. angustisporumCui 14578MG279171--[41]
G. bambusicolaWu1207-151MN957781-LC517944[42]
G. calidophilumMFLU 19-2174MN398337--[43]
G. casuarinicolaDai 16336MG279173--[41]
G. casuarinicolaHKAS 104639MK817650MK817654MK840868[9]
G. destructansCBS 139793NR 132919--[44]
G. destructansDai 16431MG279177-MG367512[41]
G. dianzhongenseL4230MW750236--[45]
G. dianzhongenseL4231MW750237-MZ467043[45]
G. dunenseCMW42157MG020255--[46]
G. ecuadorienseASL799KU128524KX228350-[47]
G. ecuadoriensePMC126KU128525KU128529-[47]
G. eickeriCMW 49692MH571690--[46]
G. enigmaticumDai 15970KU572486-MG367513[48]
G. esculentumL4935MW750242-MW839004[45]
G. esculentumL4946MW750243--[45]
G. gibbosumMFLU 19-2176MN396311-MN423118[43]
G. hochiminhenseMFLU 19-2224MN398324MN396390-[43]
G. hochiminhenseMFLU 19-2225MN396662MN396391-[43]
G. knysnamenseCMW 47755MH571681--[46]
G. knysnamenseCMW 47756MH571684--[46]
G. leucocontextumDai 15601KU572485-MG367516[49]
G. leucocontextumGDGM 44303KJ027607--[14]
G. lucidumCui 14404MG279181-MG367519[41]
G. lucidumMFLU 19-2162MN396341-MN423138[43]
G. mastoporumTNM F0018835JX840351--[50]
G. mastoporumTNM-F0018783JX840352--[50]
G. multipileumCWN 04670KJ143913-KJ143972[30]
G. multipileumDai 9447KJ143914-KJ143973[30]
G. myanmarenseMFLU19-2167MN396330MN428672-[43]
G. myanmarenseMFLU19-2169-MN398325-[43]
G. orbiformeMFLUCC 22-0066OP303318OP303348OP407740This study
G. orbiformeTH15061260MK345448--[51]
G. orbiformeGACP14081185MK313109--[51]
G. ryvardeniiHKAS 58053HM138671--[52]
G. ryvardeniiHKAS 58054HM138672--[52]
G. shanxienseBJTC FM423MK764268-MK783940[53]
G. shanxienseHSA539MK764269-MK789681[53]
G. sichuanenseMFLUCC 22-0064MW246109OP303349-This study
G. sichuanenseMFLUCC 22-0065MW246111OP303350-This study
G. sichuanenseB1 1406706KT693255--[54]
G. sichuanenseDai3583JQ781868--[55]
G. sichuanenseDai12374JQ781867--[55]
G. subflexipesCui 17257MZ354922MZ355129MZ245396[40]
G. subflexipesCui 16804MZ354973-MZ345747[40]
G. thailandicumHKAS104640MK848681MK849879MK875831[9]
G. thailandicumHKAS104641MK848682MK849880MK875832[9]
G. tongshanenseCui 17168MZ354975MZ355024-[40]
G. tropicumYuan 3490JQ781880--[55]
G. tropicumHKAS 97486MH823539--[43]
G. wiiroenseUMN-20-GHAKT952361--[56]
G. wiiroenseUMN-21-GHAKT952363--[56]
G. williamsianumDai 16809MG279183--[41]
G. yunlingenseCui 16288MZ354915MZ355077-[40]
Tomophagus colossusTC-02KJ143923--[30]
Table 2. Effect of media, temperature, pH, and cereal grains media for growing three strains of Ganoderma from Thailand.
Table 2. Effect of media, temperature, pH, and cereal grains media for growing three strains of Ganoderma from Thailand.
ParameterG. sichuanenseG. orbiforme
MFLUCC 22-0064MFLUCC 22-0065MFLUCC
22-0066
Agar
medium		PDA0.0692 ± 0.0110 b0.0640 ± 0.0099 b0.1186 ± 0.0095 b
PSA0.1241 ± 0.0029 a0.1414 ± 0.0217 a0.0415 ± 0.0124 d
CMA0.0313 ± 0.0051 c0.0567 ± 0.0050 bc0.0156 ± 0.0079 e
OMA0.0363 ± 0.0028 c0.0392 ± 0.0056 c0.1496 ± 0.0118 a
MEA0.0666 ± 0.0030 b0.0676 ± 0.0088 b0.0819 ± 0.0058 c
Temperature (°C)200.0352 ± 0.0033 bc0.0478 ± 0.0073 c0.0224 ± 0.0005 b
250.0861 ± 0.0192 ab0.1282 ± 0.0064 b0.0707 ± 0.0246 a
300.1295 ± 0.0611 a0.1721 ± 0.0235 a0.0512 ± 0.0006 a
400.0028 ± 0.0002 c0.0021 ± 0.0003 d0.0005 ± 0.0002 b
pH20.0421 ± 0.0022 d0.0390 ± 0.0063 c0.1147 ± 0.0451 c
40.1899 ± 0.0172 a0.2063 ± 0.0335 a0.4495 ± 0.0236 a
60.1264 ± 0.0186 c0.1282 ± 0.0218 b0.4333 ± 0.0129 a
70.1433 ± 0.0242 bc0.1807 ± 0.0500 a0.0575 ± 0.0126 c
80.1570 ± 0.0298 b0.2221 ± 0.0219 a0.3185 ± 0.1238 b
Spawn ProductionC. lacryma-jobi9.0000 ± 0.5244 a8.3400 ± 0.5273 a10.2600 ± 0.5367 a
H. vulgare7.5600 ± 1.2681 a6.8000 ± 1.4124 a10.4000 ± 0.2236 a
O. sativa10.5000 ± 0.0000 a8.7200 ± 3.7579 a8.2200 ± 4.0801 a
T. aestivum7.8400 ± 2.0852 a9.2400 ± 1.4690 a8.3000 ± 0.5431 a
Z. mays3.7800 ± 4.3672 b8.2200 ± 3.4745 a4.2000 ± 1.8574 b
Note: Values are the mean ± SD of the effects of the temperature of the media and the pH were indicated from the growth of dried weight (g) while the effect of the spawn production was performed from the length of the linear mycelium (mm) Number followed by different lowercase letters in column are significantly different according to Duncan’s multiple range test at p < 0.05.
Table 3. Comparison of the first flush yields of Thai Ganoderma species in bag cultivation.
Table 3. Comparison of the first flush yields of Thai Ganoderma species in bag cultivation.
ContentG. sichuanenseG. orbiforme
MFLUCC
22-0064		MFLUCC
22-0065		MFLUCC
22-0066
Primordia after inoculation (days)44–6135–6241–58
Average weight (g/bag)29.43 ± 4.7233.92 ± 6.9816.21 ± 3.51
Yield data (g/kg−1) 36.7942.4120.26
Biological efficiency (B.E.)132.19 ± 4.72152.35 ± 6.9872.79 ± 3.51
Table 4. Proximate composition of three wild Ganoderma from Thailand (g/100 g of sample).
Table 4. Proximate composition of three wild Ganoderma from Thailand (g/100 g of sample).
Ganoderma SpeciesProximate Analysis (%)
ProteinFatFiberCarbohydrates
MFLUCC 22-006412.80 ± 0.155.70 ± 0.5648.61 ± 0.674.02 ± 0.11
MFLUCC 22-006513.74 ± 0.485.11 ± 0.1147.90 ± 0.393.72 ± 0.31
MFLUCC 22-006614.67 ± 0.254.90 ± 0.5652.45 ± 0.183.16 ± 0.43
Table 5. Screening α-glucosidase inhibitory activity of Thai Ganoderma extract.
Table 5. Screening α-glucosidase inhibitory activity of Thai Ganoderma extract.
Ganoderma SpeciesFruiting BodiesMycelium
EtOH ExtractWater ExtractAcetone ExtractWater Extract
IC50 (µg/mL)IC50 (µg/mL)
MFLUCC 22-0064126.94 ± 0.87inactiveinactiveinactive
MFLUCC 22-0065171.68 ± 2.78inactiveinactiveinactive
MFLUCC 22-0066105.97 ± 1.36124.40 ± 3.18inactiveinactive
Acarbose (µg/mL)168.18 ± 0.89
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Wannasawang, N.; Luangharn, T.; Thawthong, A.; Charoensup, R.; Jaidee, W.; Tongdeesoontorn, W.; Hyde, K.D.; Thongklang, N. Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity. Life 2023, 13, 1887. https://doi.org/10.3390/life13091887

AMA Style

Wannasawang N, Luangharn T, Thawthong A, Charoensup R, Jaidee W, Tongdeesoontorn W, Hyde KD, Thongklang N. Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity. Life. 2023; 13(9):1887. https://doi.org/10.3390/life13091887

Chicago/Turabian Style

Wannasawang, Naruemon, Thatsanee Luangharn, Anan Thawthong, Rawiwan Charoensup, Wuttichai Jaidee, Wirongrong Tongdeesoontorn, Kevin D. Hyde, and Naritsada Thongklang. 2023. "Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity" Life 13, no. 9: 1887. https://doi.org/10.3390/life13091887

APA Style

Wannasawang, N., Luangharn, T., Thawthong, A., Charoensup, R., Jaidee, W., Tongdeesoontorn, W., Hyde, K. D., & Thongklang, N. (2023). Study of Optimal Conditions to Grow Thai Ganoderma, Fruiting Test, Proximate and Their Alpha Glucosidase Inhibitory Activity. Life, 13(9), 1887. https://doi.org/10.3390/life13091887

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