Co esponding au ho : Phillip N. B. Da Sil a
Copy igh © 2025 Au ho (s) e ain he copy igh o his a icle. This a icle is published unde he e ms o he C ea i e Commons A ibu ion License 4.0.
P oduc ion o whi e oys e mush oom spawn using di e en g ain-based subs a es
Meshach Jaikishun 1 and Phillip N. B. Da Sil a 2, *
1 Facul y o Na u al Sciences, Uni e si y o Guyana, Be bice Campus, Tain, Co en yne, Guyana.
2 Ins i u e o Ma ine and Ri e ine Ecologies and Economies, Uni e si y o Guyana, Be bice Campus, John’s Science Cen e,
Co en yne, Be bice, Guyana.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1531-1538
Publica ion his o y: Recei ed on 08 June 2025; e ised on 12 July 2025; accep ed on 15 July 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.27.1.2679
Abs ac
Spawn p oduc ion is an impo an s ep in mush oom cul i a ion. This s udy in es iga es he use o i e di e en
s e ilized g ain-based subs a es ( ice, ba ley, whea , maize and bi d seed) o p oducing Pleu o us os ea us spawn.
The aim is o de e mine which g ain-based subs a e is mos a o able o spawn p oduc ion. Fa ou abili y decision
was made a e compa ing expe imen al esul s o mean diame e size o he mycelium sp ead, ime aken o he
subs a e o be comple ely colonized and he di e ence be ween esh and d y weigh s o he spawn. Al hough all o he
es ed subs a es suppo ed mycelium g ow h and spawn p oduc ion, he subs a e made wi h co n g ains pe o med
signi ican ly be e in e ms o he h ee iden i ied pa ame e s. The la ge g ain size o co n and p obably i s highe
nu i ional con en , was mos likely esponsible o s imula ing g ea e mycelium g ow h, which was e lec ed in he
g ea e diame e o he colony, less ime aken o colonise he subs a e and g ea e d y weigh o he spawn.
Keywo ds: Mush oom; Spawn; G ains; Subs a e; Mycelium g ow h
1. In oduc ion
Unde na u al condi ions mush ooms, sap ophy ic decompose s, g ow on dead o ganic ma e [2, 12] and diges he
lignocellulosic con en o hei g ow h medium be o e abso bing he diges ed subs ances [2, 12, 23]. Mush ooms a e
success ully g own unde con olled and semi-con olled condi ions [2, 12] and he choice o subs a e di ec ly
in luences he g ow h, yield, and quali y o he mush ooms p oduced [2, 9]. Among he a ious subs a es used o
g owing mush ooms, g ains a e commonly used o spawning [4, 7, 23, 32] mainly because o hei nu i ional
composi ion and ease o accessibili y.
When cul i a ing edible mush ooms, h ee key ac o s a e needed; eliable spawn, good subs a e and a conduci e
en i onmen [2, 19, 29]. While much esea ch has al eady been epo ed on sui able subs a es o spawn p oduc ion
and he cul i a ion o edible mush ooms [13, 19,28], choosing a subs a e is o en based on he a ailabili y o a pa icula
subs a e [5]. Among he desi ed cha ac e is ics o a good subs a e o he cul i a ion o edible mush ooms is ha he
subs a e mus be s e ile and ich in essen ial nu ien s [2, 16, 30].
Spawn p epa a ion is one o he impo an s eps when cul i a ing mush ooms [20] and inocula ion o mycelium in o an
aga medium, ollowed by p opaga ion in a g ain-based subs a e a e impo an ollow-on s eps [3]. P oducing
mush oom spawn equi es ha a s e ile, g ain-based medium be p epa ed and inocula ed wi h mush oom mycelium
[22] and he inocula ed g ain-based medium is hen used o in oduce he ungus o a la ge subs a e o mush oom
cul i a ion [25, 26]. I is impo an ha a en ion is placed on p oducing spawn o high quali y ha in no con amina ed
because he quali y o he spawn can ul ima ely a ec he yield and quali y o he mush oom [1, 11, 33].
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1531-1538
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Hoa & Wang (2015) [10] sugges ed ha he main ac o s in luencing he p oduc ion o spawn, include cul u e media,
empe a u e, ca bon and ni ogen sou ces, g ain sou ces, and lignocellulosic subs a e sou ces. In his s udy, spawn
p oduc ion was e alua ed on i e s e ile g ain-based subs a es ( ice, ba ley, whea , maize and bi d seed) wi h he main
objec i e being o de e mine which one will p oduce be e yields o spawn o Pleu o us os ea us in a con olled
en i onmen .
The ques ions ha guided his esea ch a e:
• How long will i ake o he mush oom spawn o comple ely colonize each subs a e?
• Does he diame e o size o colonies o whi e oys e spawn di e in each subs a e?
• Will he esh and d y weigh di e a he end o he ma u a ion o he spawn un?
2. Me hodology
2.1. S udy loca ion
This esea ch was conduc ed a he Uni e si y o Guyana, John’s Science Campus, Be bice and all labo a o y wo k we e
assessed wi hin he con ines o he labo a o y unde asep ic condi ions ha we e sui able o mush oom inocula ion
and g ow h.
2.2. Subs a e p e e ence and p epa a ion
In his s udy, ice, ba ley, whea , co n and bi d seed we e used as he p ima y g ain subs a es o Pleu o us os ea us
and he p epa a ion o he spawn was done acco ding o a me hod desc ibed by Lali hakuma i (2006) [17], and wi h
modi ica ion o he numbe o hou s he g ains we e soaked (24h s). All o he p ocedu es o mo he cul u e
p epa a ion on po a o dex ose aga (PDA), and subs a e p epa a ion we e ollowed acco ding o Lali hakuma i &
Sub amanian (2004) [18] o he cul i a ion o oys e mush oom in Guyana. The same species o whi e oys e
mush oom (Pleu o us os ea us) was used as he con ol a iable and he manipula i e a iables we e he di e en
g ain-based subs a es made om whea , ba ley, maize, ice and bi d seed.
2.3. Pa ame e s o in e es
The pa ame e s analysed we e diame e (mm) o colony ex ensions in ou (4) s ages a day 4, 8,12 & day 16
espec i ely, ime aken o comple e coloniza ion o spawn un in each subs a e and esh and d y weigh (a e
comple e ma u a ion o spawn un a e 16 days.
2.4. S a is ical analyses
Resul s we e analysed using he SPSS e sion 26 and he da a we e subjec ed o one-way ANOVA and desc ip i e
s a is ics analysis. In e p e a ion o he esul s was based on compa ison o he mean diame e size o he di e en
subs a es, compa ison o he ime aken o he subs a e o be comple ely colonized and compa ison o he di e ence
be ween esh and d y weigh s o he spawn.
3. Resul and Discussion
While spawn p oduc ion was success ul wi h all i e g ain-based subs a es, he co n g ain in used subs a e p oduced
he bes esul s o all h ee pa ame e s ha we e in es iga ed and bi d seed in used subs a e was he leas pe o ming
subs a e in e ms o ime aken o ully colonize he subs a e, he diame e o colony and he d y weigh (Table 1).
Table 1 Summa y o da a collec ed on he in es iga ed pa ame e s
T ea men s
Numbe o
ials
Time aken o colonise
subs a e (days)
Weigh (g)
Diame e o colony (mm)
F esh
D y
Day 4
Day 8
Day 12
Day 14
Co n
T 1
15
200
125
12
49
116
186
T 2
15
200
123
19
74
136
200
T 3
15
200
124
19
68
121
190
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A e age
15.00
200.00
124.00
16.67
63.67
124.33
192.00
Ba ley
T 1
16
200
112
15
44
100
154
T 2
15
200
111
17
54
109
172
T 3
16
200
114
15
41
97
161
A e age
15.67
200.00
112.33
15.67
46.33
102.00
162.33
Rice
T 1
17
200
110
17
48
85
159
T 2
16
200
108
16
39
99
162
T 3
16
200
107
17
49
113
154
A e age
16.33
200.00
108.33
16.67
45.33
99.00
158.33
Whea
T 1
16
200
119
15
49
109
164
T 2
15
200
120
16
50
115
177
T 3
15
200
118
15
46
109
160
A e age
15.33
200.00
119.00
15.33
48.33
111.00
167.00
Bi d Seed
T 1
17
200
105
14
43
80
149
T 2
17
200
110
19
50
103
152
T 3
16
200
102
15
45
99
160
A e age
16.50
200.00
103.50
14.50
44.00
89.50
154.50
3.1. Mean size in diame e o mycelial g ow h on di e en es subs a es
The e was signi ican a ia ion in he diame e o mycelium g ow h, depending on he subs a e used and he ime
allowed o g ow h. The g ow h a e o he spawn un in his expe imen was de e mined by measu ing in millime es
(mm), he diame e o he ex ension o mycelium g ow h o he colony on he subs a e. Based on he esul s, he
subs a e made using co n g ains had he bes colonisa ion by mycelium ollowed by he subs a e made om g ains o
whea , ba ley, ice and bi d seed in ha o de (Figu e 1).
Figu e 1 Mean size in diame e o mycelium g ow h a e 4, 8, 12 and 16 days o di e en subs a es
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1531-1538
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On he di e en g ain in used aga media, he a e age diame e o mycelium g ow h anged om 14.5 mm o 16.67 mm
a e 4 days, 44 mm o 63.67 mm a e 8 days, 89.5 mm o 124.33 mm a e 12 days, and 154.5 mm o 192 mm a e 16
days (Table 1), wi h he highes g ow h obse ed on co n subs a e, and he lowes on bi dseed in used subs a e.
The e we e signi ican di e ences obse ed o he diame e o mycelium g ow h a e day 8 (p = 0.052), 12 days (p =
0.029) and 16 days (p = 0.001), while he e was no signi ican di e ence in diame e o mycelium g ow h when he
di e en subs a es we e compa ed a day 4 (p = 0.927).
These di e ences in he diame e o mycelium g ow h we e likely associa ed wi h he size o he g ain used o p epa e
he g ain-in used subs a e (Elhami & Ansa i (2008) [7], since la ge sized g ains we e epo ed o ha e mo e nu ien s
a ailable o mycelium g ow h [34].
Tinoco e al. (2000) [34] also sugges ed ha subs a es wi h a la ge su ace a ea and po e may suppo a highe
mycelium g ow h a e and Elhami & Ansa i (2008) [7] also no ed ha la ge seeds may con ain mo e nu ien s o
suppo mycelium g ow h. This may ha e accoun ed o he g ea e colony ex ension and he a iance seen in he wid h
o colony ex ension o spawn g own on di e en subs a es [7].
Na h e al. (2011) [21] epo ed ha la ge g ained subs a es ha e la ge ai spaces which may inc ease en ila ion,
hus imp o ing ae obic espi a ion o he mycelium, since espi a ion is di ec ly ela ed o he oxygen concen a ion
o he subs a e. This oo migh ha e accoun ed o he highe g ow h a e in he spawn and as e a e o comple e
coloniza ion obse ed o he co n g ain-in used subs a e ea men when compa ed o he o he subs a es.
3.2. Compa ison o he ime aken o comple e coloniza ion o subs a e
Gene ally, ac o s such as empe a u e, humidi y, pH, subs a e composi ion, p esence o speci ic nu ien s, ligh ,
inoculum size and ai quali y ha e been epo ed o in luence he a e o mycelium g ow h o spawn un, he ime aken
mycelium o colonize a subs a e [24, 34]. The spawn un can he e o e be aken as being indica i e o he heal h and
igo o he spawn and subs a e [7, 10, 34].
G ains p o ide a nu ien - ich subs a e o mycelium g ow h [27]. The esul s (Figu e 2) show ha he co n-in used
subs a e was he i s o be ully colonised in he leas numbe o days. This was ollowed by ba ley, ice and bi d seeds
in ha o de . This may likely be a ibu ed o he ac ha he g ains o co n we e he la ges and acco ding o Tinoco
e al. (2000) [34] la ge g ains should ha e a highe a ailabili y o nu ien s o acili a e mycelium g ow h [27]. Bi d seed-
in used subs a e, being he smalles o he g ains used in his esea ch, ook he longes ime o be ully colonized.
Figu e 2 Time aken o g ain-in used subs a es o be ully colonised
While his cu en esea ch did no in es iga e en i onmen al ac o s ha could ha e a ec ed he a e o mycelial
g ow h, his could be an a ea o u he in es iga ion in Guyana, because di e en local en i onmen al condi ions may
ha e di e en in luences on he ou come o he expe imen .
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1531-1538
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3.3. Compa ison o esh and d y weigh s o spawn
Mycelium g ow h is indica i e o success ul spawn p oduc ion [6, 31] and spawn p oduc ion is he e y impo an i s
s ep o cul i a ing mush ooms [31].
Measu ing he d y weigh o he mycelium is a good indica o o e alua e mycelium g ow h and spawn iabili y when
compa ing he e ec i eness o subs a es and g ow h condi ions du ing spawn p oduc ion. The d y weigh e lec s he
amoun o ungal biomass p oduced and is also a good measu e o how well he subs a e is u ilised by he mycelium [6,
31].
Subs a e composi ion, spawn ype and en i onmen al condi ions in luence he d y weigh o mycelium [8, 14, 15] and
some subs a es end o suppo as e mycelium g ow h and highe biomass p oduc ion, esul ing in di e en
subs a es yielding di e en d y weigh s o mycelium [8].
In his s udy, mycelium g ow h was obse ed on all o he g ain in used subs a es. The e was a signi ican di e ence
(p- alue = 0.00) be ween he esh weigh and he d y weigh o he spawn ob ained om he di e en es subs a es.
The esul s (Figu e 3) shows ha s a ing wi h a simila esh weigh (200 gm) o all subs a es, spawn om co n
in used subs a e had he highes d y weigh (124.00g), ollowed by whea (119.00g), ba ley (112.33g), ice (108.33g)
and he leas d y weigh was measu ed o bi d seed (105.67g). This sugges s ha o he subs a es used, he co n
in used subs a e was he mos success ul in e ms o suppo ing spawn p oduc ion o Pleu o us os ea us.
Figu e 3 F esh weigh and d y weigh o p epa ed spawn
The di e ences may likely ha e been a ibu ed o he ype o subs a e used o each ea men . Tinoco e al., (2001)
[34] sugges ed ha la ge g ains would no mally ha e highe nu i ional con en when compa ed wi h smalle g ains,
and his could p obably con ibu e o g ea e spawn g ow h om subs a es wi h la ge g ains, accoun ing o he
highe esh biomass weigh o spawn om co n g ain in used subs a e.
A e comple e ma u a ion, he spawn was o en d ied o cons an d y weigh . Compa ison o he d y weigh o he
spawn om he di e en es subs a es e ealed ha co n g ain subs a e p oduced spawn wi h he highes d y weigh
ollowed by whea , ba ley, ice and bi d seeds in ha o de . This may be a ibu ed o he ac ha as epo ed by Elhami
e al. (2008) [7] & Tinoco e al. (2001) [34], la ge g ains ha e mo e nu i ional con en han smalle g ains o suppo
g ea e yield o spawn p oduc ion.
4. Conclusion and Recommenda ions
All ou subs a es we e success ul in g owing spawn o P. os ea us, howe e , he co n g ain in used subs a e
appea ed o be he bes medium, ou o hose used in his s udy, o spawn un. A compa ison o esul s o mean size in
diame e o he mycelium g ow h o he di e en subs a es, he ime aken o he subs a e o be comple ely colonized
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 27(01), 1531-1538
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and he di e ence be ween esh and d y weigh s o he spawn p oduced all indica ed ha co n g ain in used subs a e
p oduced he bes esul s.
The sequence o subs a e sui abili y de e mined in his s udy was consis en wi h he p oposed heo e ical assump ion
ha la ge g ains, wi h mo e nu ien a ailabili y, would be be e as a subs a e o spawn p oduc ion, once o he
en i onmen al condi ions we e a o able.
The e ec o di e en ac o s on mycelium g ow h and spawn un should be unde aken. I will be bene icial o explo e
o he locally a ailable ma e ials o use as subs a es o spawn p oduc ion o P. os ea us. Fu u e s udies should also
in es iga e he use o di e en combina ions o g ains and nu ien o i ica ion o boos spawn p oduc ion.
Compliance wi h e hical s anda ds
Acknowledgmen s
Since es g a i ude is exp essed o he Uni e si y o Guyana, o p o iding he esou ces needed o success ully
comple e his s udy. Special and hea el hanks a e ex ended o all con ibu o s o hei con inuous and unwa e ing
suppo owa ds his esea ch.
Disclosu e o con lic o in e es
The au ho s ce i y ha his submission is o iginal wo k and is no unde e iew a any o he publica ion. The au ho s
he eby decla e ha his manusc ip does no ha e any con lic o in e es .
Decla a ion o compe ing in e es
The au ho s decla e ha hey ha e no known compe ing inancial in e es s o pe sonal ela ionships ha could ha e
appea ed o in luence he wo k epo ed in his pape .
Da a a ailabili y
Da a will be made a ailable on eques .
S a emen o in o med consen
The au ho s decla e ha in o med consen was ob ained om all indi idual pa icipan s included in he s udy. All wo k
u ilized in his s udy was ully ci ed and e e enced, so au ho s o p io esea ches a e gi en hei due c eden ials o
hei wo k.
Funding
This esea ch did no ecei e any speci ic g an om unding agencies in he public, comme cial, o no - o -p o i
sec o s.
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