Cultural and Nutritional Requirements for the Growth of Medicinal Mushroom Schizophyllum commune Fr.

Author: M. Roja, G. Thiribhuvanamala*, K. Angappan, P. Geetha and D. Amirtham

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Abstract

Schizophyllum commune Fr. is a medicinal mushroom renowned for its potential pharmacological and nutraceutical properties. To enhance the yield and explore its potential values it is necessary to investigate the cultural and nutritional conditions for the growth in detail. The investigation on the cultural characters of all isolates in potato dextrose agar medium showed the maximum colony diameter in isolate 1 (90.00mm) followed by isolate 8 (89.00mm) and isolate 2 (86.66mm). The maximum mycelial mat dry weight was recorded in the isolate 5 (2.33g/100mL) followed by isolate 2 (2.20g/100mL) and isolate 1 (2.00g/100mL). Of the different growth media tested, mushroom complete media (MCM) showed the maximum mycelial growth in all the isolates. Among the different carbon sources tested, sorbitol and mannitol supported the maximum mycelial growth in most of the selected isolates such as isolate 1 and isolate 4 (90.00mm) compared to other carbon sources. Similarly, among the different nitrogen sources tested peptone recorded maximum mycelial growth of (90.00mm) in isolate 2 when compared to all other nitrogen sources. The pH requirement for S. commune isolate 1 for its maximum mycelial growth (89.60mm) recorded in pH 5 and pH 6. And optimum temperature of 25 to 30˚C was quite suitable for the mycelial growth of S. commune where isolate 4 and isolate 8 recorded maximum mycelial growth (90.00mm).

Keywords

Schizophyllum commune, Different media, Carbon source, Nitrogen source, pH, Temperature, Colony diameter

Conclusion

Schizophyllum commune is an edible medicinal mushroom that is furnished with lot of medicinal values related to human health including the antioxidant, antidiabetic, anticancerous potential compounds. Development of cultivation technology is indeed important to tap its ample potential values. Hence, the results emanated from the in vitro investigations on the nutritional requirements like different culture media, carbon nitrogen sources and cultural requirements like different pH and temperature studied will be useful for selection of substrates to take up cultivation and tap more yield.

References

INTRODUCTION Schizophyllum commune Fr. is a medicinal mushroom that grows naturally on decaying woods, thus the name white rot fungus. This mushroom is also known as Split gill mushroom, and the name Schizophyllum commune is derived from the Greek terms Schiza, which means "split," and commune, which means "common" (Mahajan 2022). The fruiting body of S. commune is tiny flabellilform (fan shaped) white stipeless cap with hairs. It is consumed as food and medicine in number of nations, including Korea, Malaysia, China, Thailand, Vietnam, and North East India due to its high medicinal properties. S. commune mushroom possess a storey of potential compounds that have antimicrobial, anticancerous, antidiabetic activities against many human diseases (Chandrawanshi et al., 2017). Extracellular melanin by S. commune showed antibacterial, antifungal activity and anti cell proliferation activity against human epidermoid larynx carcinoma cell lines (Arun et al., 2015). The polysaccharide schizophyllan from S.commune is known for its high medicinal value. Chandrawanshi et al. (2019) reported that different solvent extracts of S. commune possess antidiabetic activity. The bioactive compounds from the mushroom S. commune is also reported to have antimicrobial property against plant pathogens. Dutta et al. (2019) reported that the active compound schizostatin from S. commune is responsible for the antifungal activity against the plant pathogens of pepper. Considering its importance, the cultivation technique is required for the large scale production and for its other industrial use. Thiribhuvanamala et al. (2020) reported that S. commune was a good source of lignin degrading fungus that has high lignocellulolytic machinery and scope in industrial applications. Thus the cultivation could be done on substrates that are rich in lignin content. Good substrates for the growth of the mushroom include paddy straw, wheat straw, saw dust etc., (Singh et al., 2021; Dasanayaka and Wijeyaratne 2017). All the fungi require good cultural and nutritional conditions for their growth. Limited studies were carried out in the field of cultural and nutritional requirements of S. commune. S. commune have little gastronomic appeal due to its rough texture and small fruiting body size. In order to increase the yield and tap its bioactive compounds from S. commune which could be in pharmaceutical and nutraceutical wide applications, the cultural and nutritional requirements is investigated in detail. MATERIALS AND METHODS The cultures of S. commune - Isolate 1, Isolate 2, Isolate 3, Isolate 4, Isolate 5, Isolate 6 and Isolate 7 obtained from Directorate of Mushroom Research, Solan; Isolate 8 and Isolate 9 from Department of Plant Pathology, TNAU, Coimbatore and Isolate 10 from Indira Gandhi Krishi Vishwavidyalaya, Raipur were used in this study. A. Colony diameter measurement Under in vitro conditions, the cultural characteristics of ten different S.commune mushroom isolates were investigated. In a Petri plate, 15mL of sterilized Potato Dextrose Agar media was poured and allowed to solidify. A 9 mm mycelial disc was taken from a 7-day old culture of different isolates and placed in the centre of a Petri plate aseptically. The plates were incubated at room temperature for further observations. On 3rd, 5th and 7th days after inoculation, growth characters such as colony diameter, colony colour and morphology were recorded. B. Estimation of Biomass production. To estimate the biomass production (fresh and dry weight) of different S. commune isolates, a 9 mm mycelial disc of S. commune mushroom isolates from a 7-day old culture was inoculated in Potato dextrose broth and incubated at room temperature. After 10 days of inoculation, the mycelial mat was separated from the broth using Whatman No. 1 filter paper and oven dried at 50 to 55°C to determine the amount of biomass produced. Fresh and dry weights of mycelial mats were recorded for each isolate. C. Effect of different media for the growth of S. commune Six different culture media (Czapek Dox (CPZ), Malt extract agar (MEA), Mushroom complete media (MCM), Oat meal agar (OMA), Sabouraud dextrose agar (SDA) and Yeast-malt extract (YME) were evaluated for the growth of S. commune isolates as described by Imtiaj et al. (2008). These media was prepared with their respective composition and sterilized in an autoclave. After sterilization the media poured into the sterilized petriplate and 9 mm diameter disc of culture was taken from a 7 days old culture grown on PDA medium and placed in the centre of each plate of six different culture media. After 7 days of incubation period at room temperature, mycelial growth was recorded. D. Effect of different carbon and nitrogen sources for the growth of S. commune Based on the results of the cultural growth characters of S. commune in PDA media, the five S. commune isolates (Isolate 1, Isolate 2, Isolate 4, Isolate 5 and Isolate 8) were chosen for further tests. The effect of various carbon and nitrogen sources on the growth of selected S. commune isolates was tested. Different carbon source viz., fructose, lactose, sorbitol, mannitol and dextrin was added to the basal medium at the rate of 4% and different nitrogen sources such as sodium nitrtate, calcium nitrate, potassium nitrate, glycine and petone was added to PDA medium at the rate of 1%. The medium containing different carbon and nitrogen sources was sterilized in an autoclave at 121°C for 15 minutes. A 9mm mycelial disc of 7 days old culture of selected isolates was placed separately in the centre of the Petri plate. The plates were incubated at room temperature for colony diameter measurement and morphology (Adejoye et al., 2007). E. Effect of different Temperature and pH for the growth of S. commune Based on the results of the cultural growth characters of S. commune in PDA media, the five S. commune isolates (Isolate 1, Isolate 2, Isolate 4, Isolate 5 and Isolate 8) were chosen for further tests. The effects of different temperature and pH for the selected S. commune isolates were tested. PDA media was prepared and it was sterilized in an autoclave at 121°C for 15 minutes. A 9mm mycelial disc of 7 days old culture of selected isolates was placed separately in the centre of the Petri plate. The plates were incubated at 20°C, 25°C, 30°C, 35°C and 40°C temperatures to observe the suitable temperature for the mycelial growth. Similarly the PDA media was prepared and adjusted to different pH viz., pH 5 to 9 to test its effect on mycelial growth of S. commune (Kumar et al., 2017). RESULT AND DISCUSSION A. Colony diameter measurement The colony diameter was measured at 3, 5 and 7 days after inoculation. The maximum mycelial diameter at 5 DAI was recorded in isolate 1 (90.00mm) followed by isolate 8 (89.00mm), isolate 2 (88.66mm), isolate 5 (87.33mm) and isolate 4 (86.66mm). The colony morphology of the S. commune was pure white mycelium with varying texture such as radiating, cottony, fluffy, flat mycelium depending upon the isolates. Conclusively all the cultures attained the full plate mycelial coverage at 7DAI. This shows that the full plate mycelial coverage could be attained after 5 to 7 DAI. The data on colony diameter is included in the Table 1. B. Estimation of Biomass Production The maximum mycelial dry weight was recorded from isolate 5 (2.33g/100mL) followed by isolate 2 (2.20g /100mL), isolate1 (2.00g /100mL), isolate 10 (1.96g /100mL) and isolate 4 (1.033g /100mL). The reduced mycelia dry weight was recorded in isolate 8 (0.16 g / 100mL), isolate 9 (0.20 g / 100mL) and isolate 7 (0.30 g / 100mL). The fresh and dry weight recorded for all the isolates is included in the Table 1. C. Effect of different culture media for the growth of S. commune Among the tested culture media, Mushroom complete media supported the growth of S. commune followed by Malt extract agar, Oat meal agar, Sabouraud dextrose agar, Yeast malt extract agar medium. CPZ medium was found to be poor performing medium. The mycelial growth was dense in MCM and Oat meal agar medium whereas sparse mycelial growth was observed in Malt extract agar medium and SDA medium and moderately thin mycelial growth in Yeast extract agar medium and poor growth in CPZ medium. All the tested isolates recorded maximum colony diameter (90.00mm) at 7DAI in MCM. Whereas in MEA the maximum colony diameter was observed in isolate 5 (90.00mm) and isolate 4 (89.66mm). The maximum colony diameter (89.33mm) in OMA was recorded in isolate 1 and isolate 10. The colony diameter in SDA was maximum in isolate 1 (87.33mm) and isolate 10 (85.33mm). YME recorded the maximum colony diameter in isolate 8 (78.33mm) and isolate 4 (76.66mm). Observations showed that there was no growth even after the 15 days of incubation in CPZ medium after a certain growth of the mycelium. Conclusively MCM was found to be the best media for S. commune next to PDA medium based on the mycelial growth. This is similar to the report by Imtiaj et al. (2008) where they found that MCM supported compact mycelial density. Kumar et al. (2017) reported that S. commune strains grow moderately in MEA medium which is similar to this report. Czapek Dox did not support the mycelial growth of Macrolepiota procera (Shim et al., 2005) and Phellinus spp. Hur et al. (2008) which is similar to our study. The colony diameter of different isolates in different media is mentioned in the Table 2. D. Effect of different carbon and nitrogen sources for the growth of S. commune Investigation on different carbon sources revealed that sorbitol, fructose, mannitol were most suitable carbon sources for the growth of the selected S. commune isolates and recorded the maximum colony diameter in most of the isolates which is similar to the report by (Alam et al. (2010). Sorbitol and mannitol recorded the maximum colony diameter in isolate 1 and isolate 4 (90.00mm). Contrarily, lactose showed maximum mycelial growth and dextrin showed the moderate mycelial growth compared to other carbon sources as reported by Imtiaj et al. (2008). Different nitrogen sources like peptone, sodium nitrate and calcium nitrate showed maximum mycelial growth as compared to glycine and potassium nitrate which recorded the moderate mycelial growth. The peptone as nitrogen source recorded the maximum colony diameter in isolate 2 (90.00) and isolate 4 (88.33mm). Similar to our study, Niederpruem et al. (1964) reported peptone as good nitrogen source for the growth of S. commune. Deshaware et al. (2021) indicated that peptone and sodium nitrate was suitable nitrogen sources for the growth of Cantharellus cibarius. In our study, calcium nitrate was also suitable for the growth of mycelium which is similar to the report by Alam et al. (2010); Imtiaj et al. (2008) where they mentioned that calcium nitrate was the most suitable nitrogen sources for the growth of S. commune. The colony growth diameter for carbon and nitrogen sources after 5 days of inoculation is included in the Table 3. E. Effect of different Temperature and pH for the growth of S. commune Based on the investigation on different temperatures, 25˚C and 30˚C was found to be optimum temperatures for the good mycelial growth. The maximum colony diameter at 25˚C was recorded in the isolate 4 (90.00mm) and isolate 8 (90.00mm). The maximum colony diameter at 30˚C was recorded in the isolate 4 (90.00mm) and isolate 8 (90.00mm).These observations clear that 25˚C to 30˚C serves as an optimum temperature for the growth of S. commune. This was analog to the findings by Adejoye et al. (2007); Kumar et al. (2017) as they reported that 25˚C to 30˚C serves as a suitable temperature for the growth of S. commune. Whereas moderate mycelial growth was observed in case of 20˚C. Very slow and poor growth was observed in case of 35˚C and 40˚C. The investigation on the different pH showed that pH 5 and pH6 was most suitable for the growth of S. commune and the maximum colony diameter was observed in the isolate 1 (89.60mm) and isolate 4 (89.60mm) at pH 5. In case of pH 6 the maximum colony diameter was recorded in isolate 1 (89.60mm) and isolate 5 (89.60mm). The rest of the pH also showed good mycelial growth. Emayavarman et al. (2021) reported that the maximum mycelial growth was observed at pH 8 and pH 6 in elm oyster mushroom. The colony growth diameter of different temperature and pH was included in the Table 4. Statistical Analysis: The design of experiments i.e. CRD and statistical analyses were followed as suggested by Gomez and Gomez (1984). Statistical software used for the analysis of data is AGRES (Developed by the Department of Physical science, TNAU, Coimbatore).

How to cite this article

M. Roja, G. Thiribhuvanamala, K. Angappan, P. Geetha and D. Amirtham (2022). Cultural and Nutritional Requirements for the Growth of Medicinal Mushroom Schizophyllum commune Fr.. Biological Forum – An International Journal, 14(2a): 32-37.