Bangladesh J. Mushroom.

2(2): 33-42, 2008 (December)

Effect of Culture Media and Environmental Factors on the Mycelial Growth of Volvariella volvacea
Mahbuba Moonmoon, Md. Nazim Uddin, Abdus Salam Khan and S.M. Ruhul Amin National Mushroom Development and Extension Centre, Savar, Dhaka, Bangladesh

Abstract
Studies on nutritional requirement and environmental factors were carried out on synthetic culture media to evaluate mycelial growth of Volvariella volvacea. Among the different culture media used, the best mycelial growth (0.87 cm/day) and density of mycelium of this mushroom was observed on semi-synthetic, potato dextrose agar (PDA) medium while the least growth (0.57 cm/ day) was recorded on wheat extract yeast agar (WEYA) medium. The optimum temperature that suported the best growth of this fungus was 350 C while the optimum pH was 6.5. In case of light, the best mycelial growth was found in alternation of light (200-280 lux) and dark (0 lux) condition. Among five different carbon sources, sucrose was the best for mycelial growth of V. volvacea. As nitrogen source, peptone was found best for vegetative growth of V. volvacea. No significant variation was observed between the mycelial growth in different C/N ratio (6:1, 12:1, 18:1, 24:1, 36:1, 48:1 and 96:1).

Key words: Volvariella volvacea, culture media, environmental factors, C/N ratio and mycelial growth.

INTRODUCTION Volvariella volvacea is one of the most popular mushrooms in the world, especially in the tropical regions (Zoberi, 1972). In Bangladesh, this mushroom is common during the summer season and commonly known as ‘straw mushroom’. It has been reported to be a very good source of essential vitamins, amino acids, glycogen and phosphorus (Alofe, 1985). Ibekwe et al. (2008), Stamet (1993), Kadiri and Kehinde (1999) reported that mycelial stage of mushroom was significantly influenced by the carbohydrate source, nitrogen source, temperature, pH and light intensity. Usually, potato dextrose agar (PDA), malt extract agar (MEA), yeast extract agar (YEA), cane juice agar (CJA), wheat extract agar (WEA), malt extract yeast agar (MEYA), wheat extract yeast agar (WEYA) and potato dextrose yeast agar (PDYA) media have been used as culture media for mycelial growth of mushroom. Fasola et al. (2007) reported that potato dextrose agar (PDA) is the best culture media for mycelial growth of Volvariella speciosa. Fasola et al. (2007) also reported that the maximum, optimum and minimum mycelial growth of Volvariella speciosa at the pH of 9.0, 6.0 and 3.0, respectively while no growth was observed at pH 2.0 and 10.0. Generally, the growth of this fungus reduced at very strong acidic and alkaline pH. Jonathan and Fasidi (2004) observed very good mycelial growth of Volvariella esculenta at pH 6.0. Moreover, mycelial growth of this fungus is very sensitive to environmental factors. The optimum temperature range for the growth of its mycelium is 30-350 C and mycelium does not grow at all when the temperature is raised to 450 C or falls bellow 150 C (Chadha and Sharma, 2005). Ibekwe et al. (2008) found that the absence of light promotes the yield best, while the alternation of light and

After autoclaving. Therefore. But the better mycelial growth of Volvariella diplasia on starch. 5. a lot of information exist on the cultivation of oyster mushroom but. volvacea : Eight different culture media such as potato dextrose agar (PDA). Dhaka during July to October. 8. glucose and fructose (Garcha and Kalra. Volvariella volvacea utilized starch and glucose better than other carbon sources (Ofosu-Asiedu et al. The media was poured into petri plates at 20 ml/plates after adjusted pH 6. very little is known about Volvariella volvacea. the petri plates were inoculated and transferred into incubation room for mycelial growth at 210 to 25o C temperaure. The pH ranges were 4. volvacea : The effect of temperature on growth of this fungus was investigated on PDA medium.0.5% (100 ±5µl) was aseptically added to the medium after it had cooled to prevent bacterial contamination. .0. 6. autoclaved and inoculated with a 0.plates.0. In case of Bangladesh. Savar.. the medium dispensed into Petri. and 9. Study of the different growth media on mycelial growth of V. darkness gave better yield and continuous light gave little growth and yield.5. yeast extract agar (YEA). The medium was adjusted at pH 6. this study is aimed to find out useful information about the culture media.0.5.5.5. Study of the temperature on mycelial growth of V. Although this mushroom has been cultivated from the very beginning of mushroom cultivation in Bangladesh.5. malt extract agar (MEA). Chloramphenicol BP 0. 8. malt extract yeast agar (MEYA). 2008. volvacea : The selected medium.5. Volvariella requires less nitrogen than carbon (Chadha and Sharma. allowed to solidify.0. Study of the different pH on mycelial growth of V.34 Moonmoon et al. cane juice agar (CJA). Threfore. The media were autoclaved for 20 minutes at 121o C temperature and 1kg/cm2 pressure and cooled at normal temperature. nutritional requirements and environmental factors that enhance the optimum growth and yield of Volvariella volvacea. wheat extract yeast agar (WEYA) and potato dextrose yeast agar (PDYA) medium were used to investigate the mycelial growth of Volvariella volvacea (Table 1). 7.6 cm (diameter) disc of vigorously growing . wheat extract agar (WEA). 1979). 6. 1984). Then same size of inocula was transferred from culture grown on PDA to placed in the centre of petri plates of each media.The basal component of each media mixed with agar and the mixture was boiled until the agar dissolved. commonly cultivated in Bangladesh. MATERIALS AND METHODS This experiment was conducted in the tissue culture laboratory of National Mushroom Development and Extension Center (NAMDEC). 2005). The inocula used in different experiments were performed with 4 replications. Then the inoculated petri dishes were transferred into incubation room for mycelial growth at 210 to 25o C temperaure. PDA was adjusted at different pH level with the addition of 1N HCl or NaOH before autoclave. The inocula of Volvariella volvacea were used from the germplasm center of NAMDEC. but unfortunately.5. no works had been done to evaluate the factors that enhance the growth and cultivation of Volvariella volvacea. Sobhanbag. In case of nitrogen sources. 5. 7.

19-22o C. 48:1 and 96:1 were used to investigate the mycelial growth of Volvariella volvacea.51) and urea (pH 7. volvacea : Seven different ratio of carbon (Glucose) and nitrogen (Yeast) source such as 6:1. alternation of light (120-150 lux) and dark (0 lux). yeast (pH 6. 36:1. 12:1. The agar medium was then dispensed into Petri.5 before autoclaved. maltose (pH 6.02).6 cm (diameter) disc of vigorously growing culture of Volvariella volvacea and incubated at 210 to 25o C. Then petri. Glucose.63). 18:1. 28-31o C.47) were used to investigate the mycelial growth of V. In case of nitrogen source.Effect of Culture Media 35 culture of Volvariella volvacea and incubated at 12-15o C. Study of the carbon and nitrogen source on mycelial growth of V. 250 to 300 lux continuous. peptone (pH 6. volvacea : The selected medium (PDA) was employed to evaluate the suitable light for mycelial growth of V. 24:1. YEA: Yeast extract agar. ammonium nitrate (NH4NO3) (pH 6. yeast mixed with 2% agar and the mixture was boiled until the agar dissolved. Same size of inocula grown on PDA was placed in the centre of each media after autoclaving for 20 minutes at 120o C temperature and 1kg/cm2 pressure and cooling. volvacea : Five different carbon source such as glucose (pH 6. MEYA: Malt extract yeast agar.45). Two percent of each carbon source mixed with 2% agar and the mixture was boiled until the agar dissolved. The medium was adjusted at pH 6. CJA: Cane juice agar Study of the light on mycelial growth of V. volvacea. volvacea.57) were used to investigate the mycelial growth of V. WEA: Wheat extract agar.68). alternation of light (40-60 lux) and dark (0 lux).50) and lactose (pH 6. Table 1. alternation of light (250-300 lux) and dark (0 lux) as well as alternation of light(200. The media was poured into petri plates at 20 ml/plates.70). Study of the C/N ratio of culture medium on mycelial growth of V. volvacea.120 to 150 lux. 40 to 60 lux. sodium nitrate (NaNO3) (pH 6. After cooling the Petri. MEA: Malt extract agar. .280 lux) and dark (0 lux) separately at 210 to 25o C. allowed to solidify.65). fructose (pH 6. sucrose (pH 6. Different culture media and their constituents used in this study Composition (g/l) PDA PDYA MEA MEYA YEA WEA WEYA CJA Agar 20 20 20 20 20 20 20 20 Dextrose 20 20 Potato 250 250 Malt powder 50 50 Yeast powder 20 20 20 20 120 120 Wheat powder Cane juice 200 PDA: Potato dextrose agar. 35 and 40o C separately.plates were transferred in to incubation room for mycelial growth at 210 to 25o C temperature.plates. Then the inoculated plates were incubated at different light condition viz: 0 lux (continuous dark). PDYA: Potato dextrose yeast agar. inoculated with a 0.plates were inoculated with the inocula of Volvariella volvacea. WEYA: wheat extract yeast agar. 24-27o C.

1984 and using MSTAT-C computer program. volvacea : Potato dextrose agar (PDA) media. cooled at normal temperature. Means were computed following DMRT using the same computer program. volvacea (Table 2). Method of measuring mycelial growth on Petri plate . The following formula was used to calculate the mycelial growth. sucrose + peptone) were used as culture media to investigate the mycelial growth of V.36 Moonmoon et al. poured into petri plates. Data on mycelium growth rate (cm/day). sucrose sgar. peptone and their combination on mycelial growth of V. Average mycelial growth on each petri. The basal component of each media were mixed with agar. autoclaved. Comparative study of potato dextrose agar (PDA) media. inoculated by V. 1. volvacea and incubated as mentioned before. sucrose + peptone + PDA. The mixtures were boiled to dissolved the agar.plate = [{(OA1+OB1+OC1+OD1)/ 4}/ days + {(A1A2+B1B2+C1C2+D1D2)/ 4}/ days] / 2 Fig. PDA + peptone. days to complete mycelium running (days) were recorded and analyzed following Gomez and Gomez. sucrose. Table 2. peptone agar and their combination (PDA + sucrose. Culture media and their constituents used in this study Culture media Potato dextrose agar (PDA) Sucrose agar Peptone agar Sucrose agar + peptone Potato dextrose agar (PDA) + sucrose Potato dextrose agar (PDA) + peptone Potato dextrose agar (PDA) + sucrose + peptone Potato 250 250 250 250 Composition (g/l) Dextrose Sucrose Peptone 20 20 20 20 20 20 20 20 20 20 20 20 Agar 20 20 20 20 20 20 20 Analysis of data: The experiment was laid out following by completely randomized design (CRD) with 4 replications.

(2007) reported that pH 6. In case of duration of complete mycelium running.50 a 5. commune when PDA was supplemented with 0.5 which was statically similar to pH 6. Fasidi.50 cm/day) was recorded at pH 4. (2007) who reported potato dextrose agar (PDA) as the best culture media for mycelial growth of V.5 to 9.75 cd 7.57 f Cane juice agar (CJA) 0. The lowest mycelial growth (0.50 days) was found in WEYA medium.0 days) was recorded at pH 6. Chandra and Purkayastha (1977) and Jonathan and Fasidi (2004) obtained very good growths of Agaricus campestris and V.76 cd CV (%) 4.5.5 as the best pH level for mycelial growth of V.75 cd 9. Study of the different pH: To determine the suitable pH on PDA media with the pH range of 4.83 ab Yeast extract agar (YEA) 0. volvacea. C and D are the symbol of the density of mycelium and A > B > C > D.73 de Wheat extract agar (WEA) 0. atroumbonata. In case of duration of complete mycelium running.Effect of Culture Media 37 RESULT AND DISCUSSION Study of the different growth media: The effect of mycelial growth and duration of complete mycelium running on eight different culture media is shown in Table 3. The best mycelial growth (0.0 and pH 7.0 bc 5.57 cm/day) was found in WEYA.83 cm/day).5) which was statistically similar to pH 6.5.0 Density of mycelium A B B C D B C C In a column. 1993. Oso.68 e Malt extract yeast agar (MEYA) 0. Study of the different growth media on mycelial growth of V.0. volvacea.0 were used for mycelial growth of V. subnudus and S. The augment of growth by PDA is not a surprise.0 and pH 7. Here A. speciosa.13 Culture Media Days to compelete mycelium run (day) 5. Jonathan and Fasidi (2001a. volvacea Mycelial growth (cm/day) Potato dextrose agar (PDA) 0.80 bc Wheat extract yeast agar (WEYA) 0.87 cm/day) was observed in slightly acidic medium (pH 6. speciosa. the minimum time (5.0. the minimum time (5. 1985.5% yeast extract. Ibekwe et al (2008) reported that the optimum mycelia growth of Pleurotus ostreatus was recorded at pH 6. Likewise. 1996. . esculenta at pH 6. because this medium has been widely reported to support mycelial growth of mushroom (Alofe. This result is not similar with the findings of Chang & Miles (1987) who reported 7.81 b Malt extract agar (MEA) 0.0 d 5.75 ab 5. means followed by a common letter are not significantly different at 5% level by DMRT.87 cm/day) was found in PDA medium which was statistically similar to that on MEYA (0. 1977).50 cd 6. Fasola et al.0 and the maximum time (9. This result is similar with the findings of Fasola et al. B. 2001b) also obtained better mycelial growth of P. The density of mycelium on PDA was higher than other culture media. Huang.5. Significantly the highest mycelial growth (0.25 days) was found at pH 4.0 days) was recorded in PDA medium and the maximum time (7. Table 3.0 is best for mycelial growth of V. The lowest mycelial growth (.57 cd 6. L.87 a Potato dextrose yeast agar (PDYA) 0.

50 b 9.00 d 8. which was statistically similar to 190-220 C.00 c 8.050 cm/day) was recorded in 120-150 C.84 a 0. volvacea.87 a 0.60 df 0.75 days) was found in alternation of light (40-60 lux) and dark (0 lux) condition. In case of duration of complete mycelium running. The highest mycelial growth (1. Study of the different pH of culture media on mycelial growth of V. volvacea Temperature ( 0c) 12-15 19-22 24-27 28-31 35 40 CV (%) Mycelial growth (cm/day) 0. Table 4.5 8.28 Days to compelete mycelium run (day) 9. Study of the temperature: Table 5 shows the effect of temperature on mycelial growth of V.0 days) was recorded at 350 C.75 b 8.76 In a column.51 g 0.5 5.24 Days to compelete mycelium run (day) > 25 a 10.0 7.67 cd 0.443 d 8. the minimum time (4.75 bc 8.0 days) was recorded in alternation of light (200-2850lux) and dark (0 lux) condition and the maximum time (7. means followed by a common letter are not significantly different at 5% level by DMRT.0 CV (%) Mycelial growth (cm/day) 0. The lowest mycelial growth (0.0 c 4. the duration of complete mycelium running was more than 25 days.82 ab 0.0 8.5 6.438 a 0.00 b 6. The best mycelial growth (1. In case of duration of complete mycelium running.0 d 3.050 e 0.438 cm/days) was obtained at 350 C which was significantly higher on compare to all the treatment. Table 5.5 9. Incase of 120-150 C temperature.38 d 0.53 fg 0.63 de 0.00 c 5.0 6. Study of the light: The effect of different light conditions on the mycelial growth is shown in Table 6.55 cm/day) was as found in alternation of light (120-180 lux) and dark (0 lux) condition.0 5. Liao (1990) reported that 12 hours of alternating light gave better than continuous light and darkness for mycelial growth of Coriolus versicolor.085 b 1.25 de 7.00 e 5.91 In a column.5 7.25 a 8.58 eg 0.00 b 7. means followed by a common letter are not significantly different at 5% level by DMRT.070 cm/day) was found in alternation of light (200-280 lux) and dark (0 lux) condition and the lowest mycelial growth (0. This result is similar with that of Cang and Chu (1969) who reported 350 C as the optimum temperature for mycelial growth of V. .735 c 1.25 de 5. Study of the temperature on mycelial growth of V.00 d 6. the minimum time (3. volvacea.0 d 10.38 Moonmoon et al. volvacea pH level 4.

Study of the C/N ratio of culture medium: The effect of different C/N ratio on the mycelial growth of V. Study of the C/N ratio of culture medium on mycelial growth of V.0 f 6.610 de 0. (2008) where sucrose was found to be the best for mycelial propagation of A.7075 c 0. Table 7. means followed by a common letter are not significantly different at 5% level by DMRT. the minimum time (8. In case of duration of complete mycelium running.0 a 5.0 days) was recorded at 36:1 C/N ratio.095 a 1. volvacea Light 0 Lux 40. the minimum time (4.0 a 3. 2005).50 bc 4.705 cm/day) of V. Study of the carbon source: Among the five different carbon sources used to find out the optimal culture condition.75 a 5.72 Days to compelete mycelium run 6.115 a 1.0 e 6.99 a 20. .6375 d 1. we did not find any significant variation of mycelial growth of V.75 d 7.8525 b 0.5825 de 0.7500 c 0. volvacea is shown in Table 7. In case of duration of complete mycelium running.190 cm/day).75 b 7.79 cm/day) was found in 6:1. It is also similar to that of Shim et al.0 a 4.75 a 5. volvacea was found in sucrose and the most unsuitable source is lactose (0.25 days) was found in lactose.55 e 0. cylindracea and lactose was the most unfavorable carbon source to mycelial growth.0 b 5. volvacea. This result is similar to that of Imtiaj et al.0 a 5.79 a 1.Effect of Culture Media 39 Table 6.85 Days to complete mycelium run 5.055 a 1.0 a 4. Although the highest mycelial growth (1.115 cm/day) was shown at 36:1 C/N ratio and the lowest (0.0 c-d 6. (1997.027 a 1.04 In a column.60 Lux Alternation of light (40-60 lux) and dark (0 lux) 120-180 Lux Alternation of light (120-180 lux) and dark (0 lux) 250-300 Lux Alternation of light (250-300 lux) and dark (0 lux) Alternation of light (200-280 lux) and dark (0 lux) CV (%) Mycelial growth rate (cm/day) 0. maltose and fructose but the maximum time (18.070 a 5.92 In a column. the best mycelial growth (0. Study of the light on mycelial growth of V. means followed by a common letter are not significantly different at 5% level by DMRT. C/N ratio 6:1 12 : 1 18 : 1 24 : 1 36 : 1 48 : 1 96 : 1 CV (%) Mycelial growth rate (cm/day) 0. volvacea in this experiment.045 a 0.0 b 5.0 days) was recorded in sucrose which was statistically similar to glucose.

volvacea.25 a 20.0612 b 5. But the most unsuitable source is urea. volvacea : The comparative effect of potato dextrose agar (PDA) media.417 c 0. peptone and their combination on mycelial growth of V.25 b 18.0 42.97 Days to complete mycelium run (day) 6. means followed by a common letter are not significantly different at 5% level by DMRT. means followed by a common letter are not significantly different at 5% level by DMRT.50 b 6.0 c 24. speciosa.0582 b 0.2-15. auricular. sucrose. Luo (1993) reported that peptone is the preferred nitrogen source for mycelial growth of A. Study of the nitrogen source on mycelial growth of V.705 cm/day) of V.0 33. subnudus. The lowest mycelial growth (0. . This result is similar to that of Kadiri and Fasidi (1994) where peptone was found to be the best nitrogen source for mycelial propagation of L.01 Days to complete mycelium run (day) 8.50 c 5. Table 9. But. sucrose.1475 cm/day) was found in ammonium nitrate (NH4NO3). Nitrogen source Peptone Ammonium nitrate (NH4NO3) Sodium nitrate (NaNO3) Yeast Urea CV (%) Nitrogen (%) 14. Comparative study of potato dextrose agar (PDA) media.537 c .147 d 0.84 In a column. Ofosu-Asiedu et al.75 b 8.0 days) was recorded in peptone which was statistically similar to yeast and the maximum time (24. (2007) reported that PDA is the best culture media for mycelial growth of V.0 * 47. Here * means unknown.75 days) was found in ammonium nitrate (NH4NO3). the best mycelial growth (0. The density of mycelium on PDA was more than sucrose.1 40.0 Mycelial growth (cm/day) 0.705 a 0. volvacea is shown in Table 10. peptone and their combination.00 b 10.535 b .1 Mycelial growth (cm/day) 0. Table 8. (1984) reported that the best mycelial growth of V.1 42. Study of the nitrogen source: Among the five different nitrogen sources used to find out the optimal culture condition.87 cm/day) was found in PDA media.62 In a column. But there was no result found in urea. peptone and their combination on the mycelial growth of V.190 d 8.6125 cm/day) was found in the combination of PDA.705 a 0. Significantly the highest mycelial growth (0. sucrose and peptone.40 Moonmoon et al.0 42.50 b 8. the minimum time (6. volvacea was found in peptone and the lowest mycelial growth (0. Fasola et al.5 35.75 a 7. volvacea was obtained on media containing peptone or potassium nitrate. In case of duration of complete mycelium running. volvacea Carbon source Glucose Maltose Sucrose Fructose Lactose CV (%) Carbon (%) 40. Study of the carbon source on mycelial growth of V.

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