cultivation of king tuber mushroom 3/(852786 using … edition/mycoson publication_6.pdf ·...

$: CULTIVATION OF KING TUBER MUSHROOM 3/(852786 USING … edition/MYCOSON PUBLICATION_6.pdf · 1LJHULDQ -RXUQDO RI 0\FRORJ\ 9RO 65 INTRODUCTION Mushroom, according to Chang and Miles$
Nigerian Journal of Mycology Vol. 9 (2017) 64

CULTIVATION OF KING TUBER MUSHROOM (PLEUROTUSTUBERREGIUM) USING CASSAVA (MANIHOT ESCULENTA) PEELS,LOAMY SOIL AND THE COMBINATION OF CASSAVA PEELS AND

LOAMY SOIL AS SUBSTRATES.

OKIGBO, R. N. ; *ANUAGASI, C. L. AND EZENWELU, C. L.Department of Botany, Faculty of Biosciences,Nnamdi Azikiwe University, Awka, Nigeria.

*Corresponding Author’s Email: chillyblack19@yahoo. com

ABSTRACTCultivation of king tuber mushroom (Pleurotus tuberregium) was carried out usingcassava (Manihot esculenta) peels, loamy soil and the combination of cassava peelsand loamy soil as substrates. This project was carried out to determine the bestsubstrates; cassava (Manihot esculenta) peels, loamy soil and the mixture of 70%cassava peels with 30% loamy soil that will produce higher yield of Pleurotustuberregium. The procedure involved were site preparation, substrate preparation,sclerotia preparation, bagging of substrates, planting of sclerotia, irrigation andharvesting of mushrooms. The experiment was laid out using a completelyrandomized design (CRD) with three treatments and ten replicates. The meanresults obtained from different parameters were subjected to statistical analysisusing Analysis of Variance (ANOVA). Means were separated using Duncan’s NewMultiple Range Tests (DNMRT). Loamy soil yielded mushroom, while cassavapeels and the mixture of cassava peel with loamy soil substrates did not yield anymushroom. Loamy soil produced 17 mushrooms in 5 flushes while cassava peelsubstrate and the mixture of cassava peel with loamy soil substrates did not yieldany mushrooms in many flushes. Loamy soil recorded higher sporophore heightof 7. 37± 0. 02cm while the cassava peels and mixture of cassava peels with loamysoil did not record any height for sporophore. Loamy soil gave higher pileusdiameter of 5. 58 ± 0. 05cm while the cassava peels and mixture of cassava peelswith loamy soil did not give pileus diameter. Higher stipe height of 4. 99 ± 0. 02cmwas recorded by loamy soil while the cassava peels and mixture of cassava peelswith loamy soil did not have any record for stipe height. Loamy soil gave higherstipe girth of 2. 64 ± 0. 06cm while the cassava peels and mixture of cassava peelswith loamy soil did not give any stipe girth. Loamy soil gave fresh weight of 6. 27

NigerJ.mycol Vol.9, 64-75


INTRODUCTIONMushroom, according to Chang and

Miles (1991), is a macrofungus with adistinctive fruit body which can be eitherepigeous or hypogenous and largeenough to be seen with the naked eyeand can be picked with hand. Agina andJoshua (2004) defined mushroom asfleshy saprophytic fungi with noticeablefruiting body which may be eitherepigeous or hypogenous and are largeenough to be picked or harvested byhand. According to Kalac (2009), theword mushroom refers only to the fruitbody. Mushrooms exhibit variousshapes and sizes ranging from sessileforms to large globule forms with typicalcap (pileus), stalk (stipe), and root-likestructures (Myers et al., 2000; Okigboand Nwatu, 2015). Unlike green plants,

mushrooms are heterotrophs, andwithout chlorophyll, they cannotgenerate nutrients by photosynthesis, butinstead take nutrients from othersources. Most mushroom species areeither under the Basidiomycota orAscomycota. The two phyla are underthe kingdom fungi (Cho, 2004).Mushrooms according to Okhuoya et al.,(2010), act as primary decomposers ofwood especially deciduous trees andbeech trees.

According to Oei (2003), the historyof mushroom have existed since thelower cretaceous period (approximately130 million years ago) and long beforehuman beings evolved on the planet. Oei(2003) reported that the history ofmushrooms is dated back to the ancientEgyptians of 4600 years ago, which

± 0. 18g while the cassava peels and mixture of cassava peels with loamy soil didnot yield any fresh weight. Higher dry weight of 1. 40 ± 0. 01g was recorded byloamy soil while the cassava peel and mixture of cassava peel with loamy soil didnot record any dry weight. There was a significant difference (p<0. 05) in the freshweight of Pleurotus tuberregium between substrates. There was also a significantdifference (p≤0. 05) in the dry weight of Pleurotus tuberregium between harvests.Cassava peels and the mixture of cassava peels with loamy soil did not support thegrowth of Pleurotus tuberregium and cannot serve as substrate for cultivation.Loamy soil is therefore considered a very good substrate for sporophore productionof Pleurotus tuberregium.

Keywords: Pleurotus tuberregium, Cassava peels, Loamy soil, Substrates,Sclerotia, Fruit body, Sporulation.

Okigbo, Anuagasi & Ezenwelu


interested Pharaoh of Egypt leading todue decree that mushrooms are deliciousand special food meant for royal peopleand therefore shouldn’t be consumed bythe ordinary man or even be touched bythem. Thus, giving the royal family theentire available supply. In some parts ofEurasia, especially in Russia and Nordiccountries, mushrooms are an importantpart of the diet (Aaronson, 2000;Okhuoya et al., 2010).

Pleurotus tuberregium (Fr. ) Sing isa tuberous basidiomycetes, one of theedible species in the genus Pleurotus(Okhuoya and Okogbo, 1990, 1991;Isikhuemhen and Okhuoya, 1996),family, Pleurotaceae and order,Agaricales (Kong, 2004). Ediblemushroom is a mushroom that canpotentially be safely eaten, includingthousands of types of mushrooms thatare regularly harvested. Okigbo andNwatu (2015) reported that ediblemushrooms are recommended by theFood and Agricultural Organization(FAO) as food, contributing to proteinnutrition of developing countriesdependent largely on cereals. Nigeria byvirtue of its vantage tropical location isone of the world’s potential hotspots forvarious forms of biological resourcesincluding mushrooms (Akpaja et al.,2003; Okigbo and Nwatu 2015).

Kalac (2009), opined that mushroomsare also a source of some mineral,

including iron, selenium, potassium andphosphorus. Mushrooms are also easilypreserved, and historically have providedadditional nutrition over winter.According to Boa (2004), mostmushrooms that are sold in thesupermarkets have been commerciallygrown on farm wastes in mushroomsfarms. The composition of mushroomfruit bodies is very rich withcarbohydrates and its concentration isranging between 20 up to or more than70% of the dry content and is usually inthe range of 60 and 140kg/kg. In mosttypes of mushrooms, carbohydrate andproteins are the two main components.Mushrooms unlike plant that storespolysaccharide in the form of starch, storetheirs in the form of glycogen and usuallycontribute to about 5-10% of dry matter(Kalac, 2009). Mushrooms also containhigh level of insoluble fibre which alsoincreases their nutritional value and theirproteins comprise about 30% weight.Protein distribution is usually changeableduring the fungal development (Boa,2004). In addition, the proportion ofessential amino acids makes it of highernutritional value compared to plantproteins (Nedelcheva et al., 2007).

Apart from the nutritional value andthe medicinal uses, many workers (Atlasand Bartha, 1992; Isikhuehmen et al.,2003 and Adenipekun, 2008) havereported the use of Pleurotus species in

Cultivation of King Tuber Mushroom with Cassava (Manihot Esculenta) Peels, Loamy Soil …


bioremediation and mycoremediation.Also, some mushrooms are useful indyeing wool and other natural fibres aswell as in the development of effectivebiological remediation and filtrationtechnologies (Nedelcheva et al., 2007).

Mushroom cultivation can be said tobe the practice of obtaining fruit bodiesartificially by repeating their growingstages (Cho, 2004). One of the strongesttechnical points recently advancingmushroom production in Nigeria besidesimproving food options is theconversion of ordinarily valueless ortoxic wastes of diverse origin to value-added products via perm culture system.Nigeria by virtue of her population sizegenerates several tons of agricultural,industrial, municipal and domesticwastes that overwhelm the nation’swaste disposal system. These arepotentially degradable by mushrooms(Okhuoya and Okogbo, 1991; Okhuoyaet al., 2010; Okigbo and Nwatu, 2015).In view of mushrooms’ popularity (useas food condiment and in medicine), lowcost of artificial substrates (farm wastes)and fast growth rate of some ediblemushrooms such as the king tuberspecies, (Pleurotus tuberregium), itbecame necessary to study the simplestand cheapest substrate that will give thehighest weight yield and proteincontents for its production. This studywas designed to investigate the effect of

using different substrates, especiallywaste farm products of fermentedcassava peels, loamy soil, and themixture of fermented cassava peels andloamy soil substrates in the cultivationand yield of Pleurotus tuberrregium(King tuber mushroom), sporophoresand sclerotia and hence recommend thebest substrate to farmers in developingcountries.

The specific objectives of theresearch include; to investigate thecultivation of Pleurotus tuberregiumusing sclerotia popularly known by thesouth easterners of Nigeria as “Osu” incolumn bags in a tropical environment,to investigate and compare the effect ofthe different substrates (loamy soil,cassava peels and 70% fermentedcassava peels mixed with 30% loamysoil) used in the cultivation of P.tuberregiums.

MATERIALS AND METHODSDescription of Study location

This study was conducted at theBotany Department Laboratorypremises, Nnamdi Azikiwe UniversityAwka, Anambra State which liesapproximately between latitudes 6o09’and 6o14’ North and longitude 7o09’ and7o13’ East with a total land area of 8183hectares. Anambra state climate iscomparatively congenial andparticularly calm with a steady



temperature. The mean monthlytemperature in the hottest periods ofFebruary to April is about 330C andaverage annual rainfall of 1828mm. Therain is almost entirely seasonal, most ofit falling between April and October(Okigbo and Nwatu, 2015). TheLaboratory premise was completelycleaned in order to avoid contaminationof the experiment.

Source of MaterialsThe sclerotia of Pleurotus tuberregium

used in this study was obtained in lumpfrom Eke-Awka market in Awka southlocal government Area of AnambraState, Nigeria. Fresh cassava peels werecollected using a sack bag from farmersin “Okukwa” a village in Amansea, inAwka North Local Government Area,Anambra State, while the loamy soil usedwas collected from the premises of HolyFamily Youth Village Hostels, Amanseain Awka North Local Government Area,Anambra State.

Preparation of substratesCassava peels were sorted to remove

sticks and dirt. They were properly driedin an open space under the sun for aboutfour days, and later ground into finepowder and stored in empty clean plasticbucket. Water was poured and allowedto cover the cassava peel substrate. Thebucket was covered and allowed to

ferment for about thirteen days. Atintervals (twenty four hours) within thesoaking period, water was changed toprevent pungent smell from thefermenting substrate. At the end of thefermentation, excessive water wassqueezed out of the substrate. Thesubstrates used were fermented cassavapeels (c), loamy soil (S) and mixture ofcassava peels with loamy soil (CS) inthe ratio of 70%: 30%. Then the loamysoil gathered was sorted out for sticksand dirt and allowed to dry for sometime in open air.

Sclerotia PreparationThe sclerotia “Osu” was also soaked

in a different plastic buckets for thirteendays to make it softer in readiness forfructification according to Oghenekaro etal.(2008) method. Water was changed atintervals before thirteen days elapsed toprevent occurrence of pungent smell fromthe soaked Sclerotia. The softenedsclerotia lump was sliced using a sterileknife into sets of about 20g each using anelectric weighing balance.

Bagging of SubstratesTwo hundred (200) grams of each

substrate was placed in the (19. 7cmlength x 19. 7cm width) polythene bagsperforated under and by the sides foradequate aeration, water drainage andmushroom emergence (Okigbo and



Nwatu, 2015). Each of the substrateswas replicated ten times.

Planting of Sclerotia This was done according to the

method of Oghenekaro et al. (2010). Thesliced Sclerotia were seeded into thebags containing the substrates bycreating a hole of 4cm in the substratesurface and watered enough to createhumid environment required forfructification. The substrate of which theSclerotia were planted on was used tocover up the Sclerotia. The experimentwas watered after planting to createmoist environment that wouldencourage germination of the Sclerotia.The experiment was kept under a closeobservation for sporulation of theSclerotia.

IrrigationThe experiment was watered twice

daily (8am and 6pm) with clean tapwater to ensure that the environment waskept humid until it has taken enoughwater to induce fructification.Harvesting of mushroom

The sporophores were harvestedwhen they were five days old.Harvesting was done by picking in themorning hours. This was done manuallyby holding them between index fingerand thumb on the surface of the substrateand with the other hand holding the base

of the stipe, the sporophore is twisted inclockwise direction and pulled out fromthe substrate. The mushroom was pickedin such a way that the sclerotium is notremoved from the soil and afterharvesting, a soft brush was used toremove stains on the stipe.

Data CollectionThe growth of mushroom was

recorded on weekly basis. Data werecollected from the different replicatesand the mean of each set of datacalculated. The experiment wasdesigned to determine the best substratefor the cultivation of P. tuberregium.The yield of fruit bodies were harvestedfrom the different substrates at the endof the experiment and the followingparameters were measured: Height (cm),Stipe girth (cm), Number of fruit bodiesfor each treatment after sprouting,Diameter of pileus (cm), Fresh weightand dry weight. The fruit bodies wereweighed immediately after harvest usingelectronic weighing balance. Afterrecording the weight, they were thendried in an oven at 75oC for 3 to 4 hours,then, the mean weights were alsorecorded. The measurements fromdifferent substrates were summed up andtheir mean was determined.

Experimental Layout/ Data AnalysisAll treatments for the experiment



were laid out using a CompletelyRandomized Design (CRD) and eachtreatment was replicated ten times. Theresults obtained were analyzed usinganalysis of variance (ANOVA) to testfor significance. Means were separatedusing Duncan’s New Multiple RangeTests (DNMRT) and analyses werecarried out using Statistical AnalysisSystem (SAS) software.

RESULTSTable 1 shows the number of

harvested Pleurotus tuberregiumcultivated in loamy soil, cassava peeland loamy soil plus cassava peelsubstrates. Findings indicate that the soilsubstrate gave a total of 17 harvests ofP. tuberregium after five harvest timeswhile the cassava peel treatment and thesand plus cassava peel treatment did notshow any harvest.

Table 2 shows the mean height (cm)and pileus diameter per harvest of P.tuberregium cultivated in loamy soil,cassava peel and loamy soil plus cassavapeel as substrates. It was observed inTable 2, that the soil substrate gave thehighest height and pileus diameter of P.tuberregium in the fifth harvest. Themean height of sporophore recorded was7. 37±0. 02cm and higher pileusdiameter of 5. 58±0. 05cm. The cassavapeel substrate and the soil plus cassavapeel substrate did not record any growth.

Between harvests, the height of P.tuberregium for loamy soil was different(p<0. 05). The pileus diameter was alsosignificantly different between harvests(p<0. 05). Table 3 shows the stipe height(cm) and stipe girth per harvest of P.tuberregium cultivated in loamy soil,cassava peel and loamy soil plus cassavapeel as substrates. The table indicatesthat loamy soil gave the highest stipeheight of 4. 99±0. 02cm and higher stipegirth of 2. 64±0. 06cm. The cassava peelsubstrate and the soil plus cassava peelsubstrate did not record any growth.Between harvests, the stipe height of P.tuberregium for the sand substrate wassignificantly different (p<0. 05). Thestipe girth was also significantlydifferent between harvest time (p<0. 05).

Table 4 shows the fresh and dryweight per harvest of P. tuberregiumcultivated in loamy soil, cassava peeland loamy soil plus cassava peel assubstrates. From the results, loamy soilgave the highest fresh weight of 6.27±0. 18g and higher dry weight of 1.40±0. 01g. Between harvests, the stipeheight of P. tuberregium for the sandsubstrate was significantly different(p<0. 05). The stipe girth was alsosignificantly different between harvesttime (p<0. 05).



Table 1: Number of harvested Pleurotus tuberregium cultivated in the loamysoil, cassava peel and loamy soil (30%) + cassava peel (70%) substrates

Table 2: Height and pileus diameter of Pleurotus tuberregium cultivated in theloamy soil, cassava peel and loamy soil (30%) + cassava peel (70%) substrates

Results are in Means ± Standard deviation*columns followed by the same letter are not significantly different according toDuncan’s New Multiple Range Tests.



Table 3: Stipe height and stipe girth of Pleurotus tuberregium cultivated inthe loamy soil, cassava peel and loamy soil (30%) + cassava peel (70%)

Results are in Means ± Standard deviation*Columns followed by the same letter are not significantly different according to Duncan’s NewMultiple Range Tests.

Table 4: Fresh and dry weight of Pleurotus tuberregium cultivated in the loamysoil, cassava peel and loamy soil (30%) + cassava peel (70%) substrates

Results are in Means ± Standard deviation*Columns followed by the same letter are not significantly different according to Duncan’s NewMultiple Range Tests.



DISCUSSIONPleurotus tuberregium (Fr. ) Sing.

has been successfully grown forsporophore production on three differentsubstrates (fermented cassava peels,loamy soil and combination offermented cassava peel with loamy soil).This study demonstrated that P.tuberregium showed a preference forsoil substrate in terms of number ofmushrooms harvested, height ofmushroom, pileus diameter, stem height,stem girth, fresh and dry weight overcassava peel and cassava peel plus soilsubstrates. This supports the work ofOkigbo and Nwatu (2015) who reportedthat soil substrate yielded moresporophore than oil palm fruit fibre. Thiscan be due to high water holdingcapacity while still allowing water toflow freely, high nutrient and humuscontent of Loamy soil. Cassava peelsubstrate and the combination of cassavapeels with loamy soil substrates did notsupport the growth of mushroomsthereby disagreeing with the studies ofGbolagade (2006) and Oluranti et al.,(2012) who recommended cassava peelsas good substrates for growing P.tuberregium. According to Oboh (2006)and Nambisan (2011), the inability ofCassava peels to support the growth ofthe mushroom can be due to the highcontent of cyanogenic glycosides andphytates in fresh cassava peels whichmay be toxic to mycelium growth.

Similarly, Nebiyu and Getachew (2011)revealed that extracts from fresh cassavapeels often inhibit the mycelium growthof certain fungi.

However, the critical limits of thegrowth and yield of P. tuberregium onthe loamy soil substrate were higher thanthose reported by Adedokun (2014) onsandy soil substrate. This result can bedue to higher nutrient contents of loamysoil. Additionally, this studydemonstrated that the growth and yieldof P. tuberregium showed a significantdifference between harvest times. Fromthe study, growth and yield of P.tuberregium increased with time ofharvest until the fifth harvest after whichthere was no more growth. Thesefindings are consistent with Oghenekaroet al. (2008) who observed thatmushroom gradually adapt to substratesand then increased in growth withharvest time until the basic nutrientswere exhausted.

Conclusion and RecommendationIt is then concluded that with this

study, fresh cassava peels substrate andthe mixture of fresh cassava peel withsoil substrate are not good substrates forgrowing P. tuberregium ifunsupplemented. Alternatively, loamysoil substrate is good for growing P.tuberregium but maximum yield can beobtained when supplemented withinorganic fertilizers. Further research



should be carried out so as to investigatethe possibility of supplementingfermented cassava substrate and usingother types of substrates for growing P.tuberregium.

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Adenipekun, C.O. (2008). Bioremediationof engine-oil polluted soil byPleurotus tuberregium SingeraNigerian white rot-fungus. AfricanJournal of Biochemistry, 7:55-58.

Agina, S. E. and V. I. Joshua, (2004).Mushroom mother spawnpreparation with different grains andsawdust types. Nigerian Journal ofBotany, 17:128-131.

Akpaja, E. O., Isikhuemhen, O. S. andOkhuoya, J. A. (2003). Ethnom-ycology and uses of edible andmedicinal mushrooms among theIgbo people of Nigeria.International Journal of MedicinalMushrooms, 5:313-319.

Atlas, R. M. and Bartha, R. C. (1992).Hydrocarbon biodegradation andsoil spill bioremediation. In:Marshal, K. (ed). AdvancedMicrobiology and Ecology, 12: 287-338.

Chang, S. T. and Miles, P. G. (1991).Recent trends in world productionof edible mushrooms. Themushroom Journal, 503: 15-30

Chiejina, N. V. and Olufokunbi, J. O.(2010). Effects of differentsubstrates on the yield of Pleurotustuberregium. African Journal ofBiotechnology, 9:1573-1577.

Chitamba, J., Dube, F., Chrota, W. andHandeseni, M. (2012). Evaluation ofSubstrate Productivity and MarketQuality of Oyster Mushroom(Pleurotus ostreatus) Grown onDifferent Substrates. InternationalJournal of Agriculture, 7:100-106.

Isikhuemhen, O., Anoliefo, G. andOghale, O. (2003). Bioremediationof crude oil polluted soil by thewhite rot fungus Pleurotustuberregium (Fr. ) Sing. Environm-ental Science Pollution Research,10:108-112.

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