Int.J.Curr.Microbiol.App.Sci (2017) 6(10): 1172-1177 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 10 (2017) pp. 1172-1177 Journal homepage: http://www.ijcmas.com Original Research Article https://doi.org/10.20546/ijcmas.2017.610.141 Tyrophagus putrescentiae as Causative Agent of Wet Bubble Disease in Agaricus bisporus Itisha*, Rachna Gulati, Manoj, Anita and Surjeet Singh Department of Zoology, Chaudhary Charan Singh Haryana Agriculture University, Hisar – 125004, Haryana, India *Corresponding author ABSTRACT A study was conducted to assess the role of Tyrophagus putrescentiae in dispersal K e yw or ds of Mycogone perniciosa, a causative agent of wet bubble disease in Agaricus Agaricus bisporus, bisporus. T. putrescentiae started feeding as soon as they were released in M. perniciosa culture plates. A continuous significant increase in mite number was Mycogone perniciosa , recorded at each observation period with peak (150.50 mites) after 20 days which Tyrophagus was statistically comparable with 145.50 mites at 18 days. After 15 days, 10 T. putrescentiae , Wet bubble disease. putrescentiae pairs were released from M. perniciosa infected plates to A. bisporus culture plates under aseptic conditions. After 6 days, whitish mouldy growth was Article Info started appearing on A. bisporus culture which soon spread to whole culture plate Accepted: within 18 days. Mites were feeding and multiplying at this stage. The mite number 10 September 2017 significantly increased with increase in observation period on A. bisporus culture nd Available Online: plates till 22 day. At this stage peak in the population was recorded (177.25 10 October 2017 mites). Introduction Mushroom growing is one of the fastest with mushrooms (Tripathi, 2005), out of growing and most technologically which Tarsonemus myceliophagus Hussey, sophisticated horticultural industries in the Tyrophagus lantneri Osb, Caloglyphus world. Three species of edible mushroom, mycophagus (Megnin) (Kumar et al., 2004), white button mushroom (Agaricus bisporus), Tyrophagus putrescentiae Schrank, paddy straw mushroom (Volvariella spp.) and Uroobovella sp. (Rana, 2008) are considered oyster mushroom (Pleurotus sp.) are economically important pests. commercially grown in India. Button mushrooms are subject to a range of diseases Tyrophagus putrescentiae, commonly called and pests which have the capacity to cause as mould mite, copra mite or cheese mite, serious crop losses. Sciarid fly, phorid fly, belong to the family Acaridae of the order spring tails and mites are important arthropod Astigmata. Their small size makes early pests of cultivated mushroom in India. Fifty infestation by mites difficult to detect and four mite species have been found associated enables them to enter packaging and exploit 1172 Int.J.Curr.Microbiol.App.Sci (2017) 6(10): 1172-1177 food residues in very small cracks and A. bisporus was reared on Potato Dextrose crevices. The life cycle consists of the egg, Agar (PDA) medium. Petriplates were larva, protonymph, tritonymph, and adult inoculated with a bit of Mycogone perniciosa stages. Development from the egg to adult under Laminar flow. After five days of takes about 1 to 3 weeks, and could take 118 inoculation of M. perniciosa, 10 pairs of T. days depending on environmental conditions putrescentiae were released in plates under and the food type on which the mite is reared aseptic and penta-replicate conditions. (Kheradmand et al., 2007). Experiments were conducted for 28 days at It is reported as an important vector of 27±1ºC, 95-99 percent relative humidity in dispersing weed fungi throughout mushroom the BOD incubator. Observations on mite cultivation facilities (Okabe et al., 2001; feeding and multiplication were recorded after Czajkowska, 2002). These include Fusariurm each alternative day. After 15 days, 10 pairs sp., Alternaria sp., Geotrichum sp., Mucor sp. of T. putrescentiae were picked from M. and Trichophyton sp. Red pepper mites are perniciosa infected plates and released in A. also considered as vectors of Trichoderma bisporus culture ABL-2 plates. Observations and Hypocrea nigricans. The mites swarming on the T. putrescentiae population build up on the casing and mushrooms bodies spread and development of whitish mouldy growth the spores of Trichoderma spp. (Hussey et al., of mycelium on strain of A. bisporus was 1969). Erban et al., (2016) studied the recorded to observe the transmission of presence of Wolbachia, Cardinium, disease. Counting of mites was done under Bartonella, Blattabacterium and Solitalea in the stereozoom microscope under 10X the eggs of T. putrescentiae which indicated magnification. mother to offspring (vertical) transmission. In white button mushroom, the incidence of wet Statistical analysis bubble disease caused by the fungus, Mycogone perniciosa is a serious problem Critical Difference (CD) was calculated for (Gahukar, 2014) whose carriers are presumed Tyrophagus putrescentiae population on fungi to be pests but no detailed study is available. to see the effect of observation periods. The It produces symptoms like dense white Software ‘OPSTAT’, developed at the growth on gills; swollen stems and caps from Computer Centre, College of Basic Sciences which reddish-brown liquid oozes, chlorine and Humanities, CCS Haryana Agricultural like smell and crinkled, walnut-like bodies on University, Hisar, was used for the analysis. the surface of the casing. T. putrescentiae can spread viruses causing diseases in mushrooms Results and Discussion (Hussey et al., 1969) and are also responsible for the dispersal of weed and pathogenic The data pertaining to T. putrescentiae fungal spores, so the present study has been population build up on M. perniciosa is undertaken to assess the role of Tyrophagus presented in Table 1. T. putrescentiae started putrescentiae in wet bubble disease dispersal. feeding as soon as they were released in culture plates. Eggs were laid by T. Materials and Methods putrescentiae, a continuous significant increase in number at each observation period To assess the role of T. putrescentiae in (CD= 6.62; p= 0.05). It was 28.00, 35.00, transmission of wet bubble disease, culture of 61.00, 71.75, 84.75, 101.75, 115.50, 129.25, pathogenic fungi (Mycogone perniciosa) and 145.50 mites and 150.50 at 2, 4, 6, 8, 10, 12, 1173 Int.J.Curr.Microbiol.App.Sci (2017) 6(10): 1172-1177 14, 16, 18 and 20 day. A peak in population II). The colour of culture changed from pale (150.50 mites) was recorded after 20 days brown to white. The statistical analysis which was statistically comparable with showed significant effect of observation 145.50 mites at 18 days. Thereafter, a gradual period on population buildup of T. but significant decline in population was putrescentiae on A. bisporus culture (CD= recorded with 137.25, 123.75 and 111.00 6.73; p= 0.05) (Table 2). The mite number mites at 22, 24 and 26 day respectively. significantly increased with increase in observation period till 22nd day. At this stage After 15 days, 10 T. putrescentiae pairs were peak in the population was recorded (177.25 released from M. perniciosa infected plates to mites). Afterwards, it gradually declined to A. bisporus culture plates under aseptic the lowest population count of 105.50 mites at conditions. After 6 days, whitish mouldy 32 day. The mite count was statistically non- growth was started appearing on A. bisporus significant at zero and second day, 20th and culture (Plate I). It soon spread to whole 26th days, 18th and 28th days, 16th and 30th culture plate within 18 days. Mites were days. Mite number at 14th and 32nd days was feeding and multiplying at this stage (Plate also found to be at par with each other. Table.1 Population of Tyrophagus putrescentiae on Mycogone perniciosa Observation (days after 10 pairs of mite/ strain of release of mites) Mycogone perniciosa 0 20.00 ± 0.00 2 28.25 ± 1.43 4 35.00 ± 1.78 6 61.00 ± 2.82 8 71.75 ± 4.55 10 84.75 ± 3.22 12 101.75 ± 1.93 14 115.50 ± 2.39 16 129.25 ± 1.93 18 145.50 ± 2.10 20 150.50 ± 1.44 22 137.25 ± 1.93 24 123.75 ± 1.88 26 111.00 ± 2.34 SE(m) 2.31 CD(p = 0.05) 6.62 1174 Int.J.Curr.Microbiol.App.Sci (2017) 6(10): 1172-1177 Table.2 Population of Tyrophagus putrescentiae on Agaricus bisporus Observation (days 10 pairs of mite/ strain of after release of mites) Agaricus bisporus 0 20.00 ±0.00a 2 26.00 ±0.81a 4 37.25 ±1.54 6 50.25 ±2.05 8 62.50 ±1.75 10 74.00 ±2.48 12 89.25 ±1.54 14 102.75 ±1.93e 16 113.50 ±2.95d 18 129.50 ±3.12c 20 150.50 ±1.70b 22 177.25 ±2.21 24 164.50 ±2.10 26 152.25 ±1.79b 28 131.25±3.70c 30 118.75 ±3.35d 32 105.50 ±3.66e SE(m) 2.36 CD(p = 0.05) 6.73 Plate.1 Appearance of wet bubble disease on A. bisporus Plate II: Spreading of wet bubble disease Most of the mites feed on wild fungus In nature, the spores of coprophilous fungi (contaminants in cultivated mushrooms) and survive their passage through the animal help in dispersal of their spores (Price, 1976). digestive system when consumed by Wild fungi act as competitive fungi in herbivorous (Richardson, 2001). Basidiospores cultivated mushroom beds thus affecting the ingested by the mites will not be digested. overall yield (Okabe, 1999). The life cycle of C. Faecal pellets containing a number of fungal cinerea along with the mite T. putrescentiae spores are formed and excreted. The spores in starts with the mite feeding on the mushroom the faecal pellets easily germinate and form tissues including the spores (González, 2008).