ASIAN J. EXP. BIOL. SCI.

VOL 2(2) 2011:328-331

Society of Applied Sciences

ORIGINAL ARTICLE

Influence of Some Growth Regulators on Biomass Production And Sporophore Yield of Milky Mushroom (Calocybe Indica)
B. K. Pani
Department of Plant Pathology, Orissa University of Agriculture and Technology, Bhubaneswar-751 003, Orissa ABSTRACT The effect of growth regulators on milky mushroom (Calocybe indica) was studied both in vitro and in vivo. In the submerged culture, Indole Acetic Acid (IAA), Indole Butryic Acid (IBA), Naphthalene Acetic Acid (NAA), Kinetin, 2,4-D and Maleic Hydrazide (MH) were tested at 10, 25, and 50 ppm. All but MH increased the biomass production. GA was most effective in inducing significantly higher mycelia yield at all doses, maximum dry biomass (851.4 mg) being recorded at 50 ppm. It was followed by Kinetin (675.1mg, 10 ppm). Higher dose (50 ppm) of NAA was toxic while lower concentrations (10, 25 ppm) were significantly stimulating. Unlike GA, the increase in concentration of Kinetin decreased the rate of vegetative multiplication. IAA also sustained appreciable dry weights (390.8- 432.2 mg). In case of IBA, the increase in mycelia harvest over control was statistically insignificant. Field trials revealed that spraying 50 ppm GA on the emerging primordia increased the number as well as size of mushrooms and produced significantly higher yield (82.2 % BE) compared to control (67.8 % BE). Increase in sporophore yields, though not statistically significant than control, was also recorded with 10 ppm Kinetin (76.0 % BE), 25 ppm IAA (73.1 % BE) and 25 ppm NAA (71.6 % BE). KEYWORDS: Growth regulator, Calocybe indica, mushroom, yield

INTRODUCTION Calocybe indica is a tropical edible mushroom of Indian origin [1]. It is popularly known as milky mushroom or white summer mushroom /summer white mushroom. In some places they are called Kuduk and dudhi chhata. In Orissa it is known as dudha chhatu. The sporophores are robust, fleshy and milky white in appearance even after flattening. In rainy season, wild forms of C. indica are sold in Calcutta and their edibility was confirmed [1-2]. Natural occurrence of the mushroom in the plains of Tamilnadu and Rajasthan has also been reported [3-4]. Unlike the button mushroom no special compost is required for its cultivation. As it grows in hot humid climate, the mushroom is highly suitable for cultivation in the plains of India almost throughout the year except in few places. Simple production techniques, sustainable yield, long shelf- life, low capital investment, ability to thrive on a variety of substrates in a wide range of temperature and humidity conditions as well as attractive colour and shape have caught the imagination of the growers as well as consumers. Beginning from its first artificial cultivation in 1976 in a mixture of soil, sand and maize meal (12:6:1) in soil jars [5], many of the several workers [4, 6-10] have attempted to improve upon its yield potential. However, only limited success has been achieved in popularizing this mushroom vis--vis other cultivated mushrooms such as button mushroom (Agaricus spp.), oyster mushroom (Pleurotus spp.) and paddy straw mushroom (Volvariella spp.). Commercial milky mushroom growers remain mostly confined to Tamilnadu particularly in the districts of Erode, Salem, Coimbatore, Trichy, Madurai and other districts [6]. Though the effect of growth regulators aimed at higher yield has been investigated in few other mushroom species [11-15], similar literature pertaining to C. indica is almost lacking. The objective of this study was to improve the biomass production and sporophore yield of C. indica by use of some growth regulators.

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Influence Of Some Growth Regulators On Biomass Production And Sporophore Yield Of Milky Mushroom (Calocybe Indica) ....................B. K. Pani

MATERIALSAND METHODS The pure culture of Calocybe indica was raised by tissue culture from a healthy and large- sized sporophore collected from the first flush of the cultivated mushroom crop. The cultures were maintained on potato dextrose agar (PDA) medium with periodic sub-culturing at 21 days intervals. For this study, 5 mm mycelia disc cut out from the margin of an actively growing colony was used as inoculum. Growth regulators such as Indole Acetic Acid (IAA), Indole Butryic Acid (IBA), Gibberellic Acid (GA), Naphthalene Acetic Acid (NAA), Kinetin, 2,4-D and Maleic hydrazide (MH) in concentrations of 10, 25 and 50 ppm were added separately to each of the conical flasks containing PDA medium. These flasks were tightly plugged with non-absorbent cotton, properly labelled and sterilized in autoclave at 10 lb p.s.i. for 30 minutes. The media were aseptically inoculated with the fungal inoculums and incubated at room 0 temperature (25 -32 C) for 21 days. The dry mycelia mats were harvested as per standard procedure [16]. Three replications were maintained for each treatment. Medium devoid of any supplementation served as control. Data were subjected to statistical analysis. After ascertaining their efficacy in the liquid culture, selected growth regulators such as GA, Kinetin, IAA and IBA were tested at doses of 50, 10, 25 and 25 ppm, respectively for their effect on mushroom production. Standard method of milky mushroom cultivation in chopped paddy straw with multi-layer spawning in high density polythene bags (60 cm X 40 cm, 100 gauge) was followed [8]. After complete colonization of substrates, one end of the cylindrical bag was opened and rolled outward to expose the top surface. No casing was done. The chemicals were separately sprayed on the emerging buttons daily once in alternate days for two days. Care was taken to withhold the routine spraying of 0 water on the substrate on the days of application of growth regulators. A temperature of 25- 35 C and humidity of 8090 % were maintained throughout the cropping period. Sufficient light and controlled ventilation were allowed in the cropping room. Mature sporophores were harvested just before flattening from two flushes and fresh weights were immediately recorded. Each treatment was replicated thrice. Biological efficiency (BE) was computed as a ratio between fresh weight of harvested mushrooms and dry weight of substrate taken in each bag and was expressed as a per cent. Data pertaining to yield were statistically analyzed. RESULTS AND DISCUSSION All the growth regulators except MH enhanced the biomass production of the fungus in liquid culture (Table 1). GA was the best sustaining significantly higher mycelia yield at all concentrations, maximum dry weight being recorded at 50 ppm. Similar findings were also reported in other edible fungi [17, 18]. The next effective was Kinetin which also significantly enhanced mycelia yields over control at all specified concentrations. At the lowest dose (10 ppm), maximum dry biomass (675.1 mg) was recovered. Unlike GA, increase in concentration of kinetin decreased the rate of vegetative multiplication. However, inhibition of mushroom growth in response to GA and Kinetin has been observed [19]. IAA also sustained appreciable fungal dry weights (390.8-432.2 mg) which corroborates similar reports [19]. In case of IBA, the increase in mycelia harvest over control was statistically in significant. Higher concentration of NAA (50 ppm) was toxic while lower concentrations (10-25 ppm) were significantly stimulating. It was also recorded that 2, 4-D significantly favoured increased biomass production. The stimulatory effect of this chemical on the growth of Volvariella volvacea [15], Lentinus subnudus and Schizophyllum commune [13, 14] has been reported. MH caused growth retardation at all doses.
Table 1: Effect of growth regulators on the growth of Calocybe indica in vitro
M y c e lia l d r y w e i g h t ( m g ) a G r o w t h R e g u la to r s 10 ppm I n d o le A c e tic A c id ( I A A ) I n d o le B u tr y c A c id ( I B A ) G ib b e r llic A c id ( G A ) N a p th a le n e A c e tic A c id ( N A A ) K in e ti n 2, 4 D M a lic H y d r a z id e C o n tr o l 4 3 0 .2 2 3 7 .4 7 2 5 .0 3 9 7 .3 6 7 5 .1 3 1 1 .8 1 9 7 .4 2 1 7 .5 25 ppm 4 3 2 .2 2 3 9 .0 7 9 4 .6 3 2 8 .7 5 7 5 .8 3 1 5 .2 1 8 0 .7 2 1 7 .5 3 9 0 .8 2 3 5 .6 8 5 1 .4 1 9 9 .6 4 7 6 .1 3 0 2 .8 1 5 5 .3 2 1 7 .5 50 ppm

CD (0.05) a = average of three replications

ASIAN J. EXP. BIOL. SCI. VOL 2(2) 2011

43.57

49.41

26.93

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Influence Of Some Growth Regulators On Biomass Production And Sporophore Yield Of Milky Mushroom (Calocybe Indica) ....................B. K. Pani

It was also noted (Table 2) that spraying of GA 50 ppm increased the number as well as size of sporophores and produced significantly higher mushroom yield (82.2 % BE) vis--vis control (67.8% BE). The result assumes significance in view of the fact that primordial mortality is a limiting factor in the successful cultivation of many mushroom species including C. indica. Many of the emerging primordial withers away during the cultivation process while only a few develop into mature harvestable sporophores. More research is required in this regard to further reduce the primordial mortality in mushrooms. The yield enhancing effect of GA on milky mushroom has been reported earlier [20]. The increase in sporophore yield, though statistically insignificant, were recorded with kinetin 10 ppm (76.0 % BE), IAA 25 ppm (73.1% BE) and NAA 25 ppm (71.6 % BE).The beneficial effect of these growth regulators have been demonstrated in other edible mushrooms [11, 12].The effectiveness of 100 ppm Kinetin in increasing the yield of C. indica has also been reported [9].
Table 2: Effect of growth regulators on the production of Calocybe indica
Growth regulators Dose (ppm) Sporophore No. Pileus diameter (cm) Stipe length (cm) Yield (g) Avg. wt. of sporophore (g) BE (%)

Gibberllic Acid Kinetin Indole Acetic Acid Naphthalene Acetic Acid Control

50 10 25 25 -

8 7 6 6 6

15.2 15.1 15.4 14.5 14.1

16.8 16.7 16.3 16.3 16.2

822.6 760.6 731.3 716.0 678.0

102.8 108.6 121.8 119.3 113.0

111.88

82.2 76.0 73.1 71.6 67.8

CD (0.05) Each of the observation was the average of three replications.

ACKNOWLEDGEMENT The invaluable guidance and infrastructural facilities rendered by Professor S.R. Das, former Head, Department of Plant Pathology, College of Agriculture, Orissa University of Agriculture and Technology, Bhubaneswar and Professor H.K. Patra, Post Graduate Department of Botany, Utkal University, Bhubaneswar during the period of investigation are highly acknowledged. REFERENCES
[1]. Purkayastha, R.P. and Chandra,A. (1974).Anew species of edible mushroom from India. Trans. British Mycol. Soc., 62: 415-418 [2]. Purkayastha, R.P. (1976).Anew method of cultivation of Calocybe indica - an edible summer mushroom. Taiwan Mushrooms, 3: 14-18. [3]. Doshi, A., Sidan, N. and Chakravorthy, B.P. (1989). Cultivation of summer mushroom, Calocybe indica Purkayastha and Chandra in Rajasthan. In: Mushroom Science. XIII (Part-II). Proc.12th International Congress on the Science and Cultivation of Edible Fungi. Braunschweig, Germany [4]. Krishnamoorthy,A.S. (1995). Ph.D. thesis, TamilnaduAgricultural University, Coimbatore, India, pp. 222 [5]. Purkayastha, R.P. and Chandra, A. (1976). A new technique for in vitro production of Calocybe indica - An edible mushroom from India. Indian Mushroom Journal, 40: 112-113 [6]. Krishnamoorthy, A.S. (2003). Commercial prospects of milky mushroom, (Calocybe indica) on tropical plains of India. In: Current Vistas in Mushroom Biology and Production (R.C. Upadhyay, S.K. Singh and R.D. Rai, eds.) pp.131-135 [7]. Chakravorthy, D. K., Sarkar, B.B. and Kundu, B.M. (1981). Cultivation of a tropical edible mushroom, Calocybe indica. Indian Agriculturist, 25 (1): 57-60 [8]. Pani, B.K. and Das, S.R. (1998). Seasonal productivity of summer white mushroom (Calocybe indica P. & C.) in Orissa. Science and Culture, 40(7&8): 177-178 [9]. Theradimani, M., Meena, B. and Krishnamoorthy, A.S. (2001). Innovative technique for improvement of sporophore size and yield of milky mushroom (Calocybe indica). Mushroom Research, 10(1); 23-26 [10].Tandan, Gayatri and Sharma, V.P. (2006).Yield performance of Calocybe indica on various substrates and supplements. Mushroom 330 ASIAN J. EXP. BIOL. SCI. VOL 2(2) 2011

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Correspondence to Author : B. K. Pani, Department of Plant Pathology, Orissa University of Agriculture and Technology, Bhubaneswar -751 003, Orissa . E mail: dr.bkp1965@indiatimes. com

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