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The effect of culture media and pH on the mycelial growth of Trametes versicolor was determined (Table 1). Commercial medium, malt extract agar (MEA), and potato dextrose agar (PDA) were determined to be optimal for the luxuriant mycelial growth of T. versicolor with an average mycelial growth rate of 10.27 mm d−1 and 8.64 mm d−1, respectively. In addition, coconut water gulaman (CWG), an indigenous culture medium also favored the excellent mycelial growth of this mushroom (8.36 mm d−1). These three-culture media (MEA, PDA, CWG) show high mycelial growth with thick mycelial density.
Table 1. Mycelial growth of Trametes versicolor with different intrinsic and extrinsic factors.
Growth rate of
mycelia (mm d−1)Mycelial
densityIntrinsic factors Culture media CWG 8.36 ± 0.94ab +++ PSG 3.80 ± 0.57d ++ CGDG 3.93 ± 0.60d ++ RBDG 5.91 ± 0.73cd +++ PDA 8.64 ± 1.18ab +++ MEA 10.27 ± 0.61a +++ SDA 6.64 ± 0.10bc +++ pH 5.0 7.69 ± 0.21b ++++ 6.0 11.91 ± 4.52a ++++ 7.0 14.26 ± 2.11a ++++ 8.0 14.18 ± 2.19a ++++ Aeration Extrinsic factors Sealed 17.45 ± 0.6a ++++ Unsealed 11.51 ± 0.23b ++++ Illumination Lighted 16.97 ± 1.79a ++++ Dark 13.36 ± 6.76b ++++ Temperature Refrigerated 2.00 ± 0.00b No growth Air-conditioned 17.36 ± 1.12a ++++ Room temperature 18.00 ± 0.00a ++++ Means with similar superscripts are statistically comparable from each other using Tukeys HSD and t-test at 5% level of significance. Coconut water Gulaman (CWG), Potato Sucrose Gulaman (PSG), Corn Grits Decoction Gulaman (CGDG), Rice Bran Decoction Gulaman (RBDG), Potato Dextrose Agar (PDA), Malt Extract Agar (MEA), Saboraud Dextrose Agar (SDA). Refrigerated condition (9 °C), Air-conditioned (25 °C), Room Temperature (32 °C). (+) very thin, (++) thin, (+++) thick, (++++) very thick. After establishing the optimum culture medium, MEA was used to determine the optimum pH for the mycelial growth of T. versicolor. Based on the data obtained, a wide range of pH from 6.0 to 8.0 were determined to be optimal for the mycelial growth of T. versicolor with an average mycelial growth of 11.91, 14.26, and 14.18-mm d−1, respectively. In addition, a very thick mycelial density was also evaluated.
Influence of extrinsic factors
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Aeration, illumination, and temperature conditions for the growth of T. versicolor were also determined in this study. Using the optimal intrinsic factors MEA adjusted to pH 7, the aeration condition was determined. Two sets of aeration conditions unsealed and sealed (with parafilm) conditions are shown in Table 1. With an average mycelial growth rate of 17.45 mm per d−1, it was determined that T. versicolor grew best under sealed conditions. Moreover, two sets of illumination conditions (light and dark) were also evaluated in this study using the optimum medium, pH, and aeration condition (MEA, pH 7, sealed). Based on the data obtained, lighted conditions (16.97 mm d−1) were found to be more suitable for the luxuriant mycelial growth of T. versicolor compared to dark conditions (13.36 mm d−1). However, a very thick mycelial density was evaluated for both sets of aeration and illumination conditions.
Temperature was the last extrinsic factor determined in this study. Three sets of temperatures namely refrigerated (9 °C), air-conditioned (25 °C), and room temperature (32 °C) conditions were evaluated. T. versicolor exhibits a luxuriant mycelial growth in a wide range of temperatures from 32 °C. Both air-conditioned (17.36 mm d−1) and room temperature (18.0 mm d−1) show excellent mycelial growth and very thick mycelial density. Hence, the optimal culture conditions for both intrinsic and extrinsic factors of T. versicolor were established in this study. Mycelial growth of T. versicolor on a different culture condition is shown in Fig. 2.
Figure 2.
Mycelial growth of Trametes versicolor on different culture conditions. (a) Culture media (a1, SDA; a2, MEA; a3, PDA, a4, CWG; a5, RBDG, a6, PSG; a7, CGDG). (b) pH condition (b1, pH 5; b2, pH 6; b3, pH 7; b4, pH 8). (c) Aeration conditions (c1, Sealed; c2, Unsealed). (d) Illumination conditions (d1, Light; d2, Dark). (e) Temperature conditions (e1, Refrigerated; e2, Air conditioned; e3, Room temperature).
Evaluation of the fruiting body production of T. versicolor
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After evaluating the nutritional and physical conditions of T. versicolor mycelial growth, this mushroom was subjected to fructification using the Center for Tropical Mushroom Research and Development (CTMRD) formulation of rice straw and sawdust (Fig. 3). Table 2 shows the parameters of fructification for T. versicolor. Under favorable environmental conditions, incubation days for mycelial colonization of fruiting bags were observed. The duration of incubation, days of primordia initiation, cap diameter, and yield per bag were determined. In addition, biological efficiency was determined.
Figure 3.
Fruiting bodies of T. versicolor grown in 70% rice straw and 30% sawdust substrate formulation.
Table 2. Fructification parameters of Trametes versicolor.
Incubation period (d) Days of primordia initiation (d) Cap diameter (mm) Yield per bag (g bag−1) Biological efficiency (%) 19.70 ± 0.48 36.90 ± 14.76 22.11 ± 3.59 37.68 ± 15.70 7.54 ± 3.14 Mycochemicals found in T. versicolor
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Mycochemical screening was carried out to determine the presence of secondary metabolites such as steroids, terpenoids, flavonoids, glycosidase, tannins, saponins, and alkaloids present in T. versicolor. Table 3 presents the mycochemical components present in the aqueous extract of the fruiting body of T. versicolor. Among the seven mycochemicals tested, terpenoids, flavonoids, tannins, saponins, and alkaloids were found present in T. versicolor.
Table 3. Mycochemical analysis of an aqueous extract of the fruiting body of T. versicolor.
Mycochemicals Reaction Findings Alkaloids Turbidity was formed + Flavonoids Lighter brown coloration* + Glycosidase No reaction − Steroids No reaction − Saponins Frothing formation + Tannins Brownish with lighter green coloration + Terpenoids Brown coloration interface + * Initial color of the extract was brown, thus positive yellow coloration results in a lighter brown. Note: (+) present; (−) absent. Cytotoxic effect of T. versicolor
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T. versicolor's cytotoxic activity was evaluated using the brine shrimp lethality assay. After being exposed to various concentrations of T. versicolor ethanolic and methanolic extract for 24 h, the mean percent mortality and LC50 values of A. salina nauplii are shown in Table 4. It can be noticed that at 10,000 ug/ml concentrations of both ethanolic and methanolic extract, the highest mortality rate of 96.67% and 93.3%, respectively was recorded. Moreover, the lowest mortality rate was recorded at 1 µg/ml accounting for 16.67% and 10.0% mortality for the ethanolic and methanolic extracts, respectively. Based on the data obtained, the cytotoxic effect of ethanolic extract is higher than methanolic extract. Analysis of variance shows that there are no significant differences in the mortality rate for 10,000 µg/ml and 1,000 µg/ml for ethanolic extract. However, significant differences were observed at 10,000 µg/ml and 1,000 µg/ml for methanolic extract. Additionally, both ethanolic and methanolic extracts of T. versicolor exhibited high toxicity levels with LC50 values of 70.93 µg/ml and methanolic extract of 74.43 µg/ml (Fig. 4). The data consist of corrected %mortality values that have been probit converted and are compared to the log of treatment concentration. For each concentration, the average percent mortality was calculated from three (3) replicates.
Table 4. Cytotoxic effect and LC50 values of ethanolic and methanolic extract of T. versicolor on brine shrimp nauplii after 24 h of exposure.
Extract Concentration
(µg/ml)Mortality
(%)LC50
(µg/ml)Toxicity level* Ethanol 0 0.00e 70.93 Highly toxic 1 16.67de 10 40.00cd 100 53.33bc 1,000 73.33ab 10,000 96.67a Methanol 0 0.00d 74.43 Highly toxic 1 10.00d 10 36.67c 100 46.67bc 1,000 66.67b 10,000 93.33a Means with similar superscripts are statistically comparable from each other using Tukey's HSD and t-test at 5% level of significance.
* Toxicity level was based on the study of Mendoza et al.[28]. -
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request
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About this article
Cite this article
Fabros JA, Lazo MKM, Magpantay JES, Abon MD, Dulay RMR, et al. 2023. The effect of nutritional and physical factors on the growth of Trametes versicolor (L.) Lloyd and its mycochemical and cytotoxic properties. Studies in Fungi 8:18 doi: 10.48130/SIF-2023-0018
The effect of nutritional and physical factors on the growth of Trametes versicolor (L.) Lloyd and its mycochemical and cytotoxic properties
- Received: 14 August 2023
- Accepted: 04 October 2023
- Published online: 22 November 2023
Abstract: Trametes versicolor (L.) Lloyd is an agaricomycetous fungi characterized by its fan-shaped or shelf-like fruiting bodies with a thin leathery and velvety upper surface that displays concentric bands of brown-black and white color and undersurface cap features with numerous tiny spores. In this study, the culture conditions for the mycelial growth as well as fruiting body production was established. The mycochemical compositions and the cytotoxic activity were also elucidated. Optimization study of the secondary mycelia shows that T. versicolor grew well on malt extract agar (MEA), potato dextrose agar (PDA), and coconut water gulaman (CWG) culture media. In terms of pH of the medium, pH 6.0 to 8.0 supports the best mycelial growth. On the other hand, sealed, lighted conditions incubated at 25 to 32 °C were the requirements for the optimum growth of T. versicolor. After 37 d incubation, the fruiting body production of T. versicolor was determined. T. versicolor produced 37.68 g bag−1 which is equivalent to 7.65% biological efficiency (BE) on a substrate consisting of rice straw and sawdust at a 7:3 ratio by volume. Moreover, qualitative mycochemical analysis of the aqueous extract of T. versicolor revealed the presence of different mycochemicals such as terpenoids, flavonoids, tannins, saponins, and alkaloids. In terms of the cytotoxic effect of T. versicolor, the LC50 values of ethanol and methanol extracts were calculated, showing that they had a high toxicity level of 70.93 and 74.43 µg/ml, respectively against brine shrimp nauplii after 24 h of incubation. Overall, the optimum culture condition for mycelial growth and fruiting body production, mycochemical compounds, and cytotoxic effect of T. versicolor tabulated in this study provide significant data that elucidated the value of this mushroom in the pharmaceutical industries.
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Key words:
- Mycochemical /
- Cytotoxicity /
- Biological efficiency /
- Trametes versicolor /
- Optimization