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MATERIALS AND METHODS
STUDY MATERIAL
Cultures of P7eurotus f7orida,P. ostreatus and P.
sajor-caju were obtained from The Mushroom Centre of the
IGKVV, Raipur, for these studies. At the outset single spore
cultures were developed for all the three species according
to the method described below.
The sporophores were first obtained on paddy straw
and basidiospores were collected. Single spore cultures were
then obtained on malt extract agar mRdium using the method
of Suman (1990) in each case. Pure cultures were subse-
quently transferred to slants which were stored ·at 4°C
until used for the entire work.
MASTER CULTURE PREPARATION
Master cultures were prepared from isolated
pure cultures of all three P7eurotus species. Maize grains
were used as substrate which was procersed as described
below.
Maize grains were first ground to small pieces
and steeped in water for 2 hours. Soaked grains were then
steam cooked and cooled. Gypsum and Calcium carbonate (2%
W/W and 5% W/W respectively) were mixed thoroughly with the
cooked grains on wet weight basis. Prepared substrate
was then sterilised for 1 hour.
Actively growing mycelial discs of three P7eurotus
cultures were inoculated to 10 ml malt extract agar medium in
seperate conical flasks and incubated at 25~Z0c for mycelial
growth. The sterilised grains were then aseptically trans-
fer red over the growing mycelium and again incubated
till the mycelium covered the grains surface. Master cultures
were prepared in this way in sufficient quantity required
fer the proposed research work.
SPAWN PREPARATION:
Coarsely ground maize grains were processed in
si"-1i lar way as described for master cultur~ preparation.
Prepared grains were filled in glucose bottles (200 g
0achJ and ~lerilised for 2 hours for 2 consecutive days.
Inoculation was done with equal quantities of master culture
,,.,,(] lx)ttles were incubated at 25t.~C for spawn growth.
R0qltiro~ number of spawn bottles were prepared from master
c•'~'•'d and used at one time in each experiment.
Spawn Substrate Studies :
Fi .8 cereals (Maize, Wheat, Jowar, Bajra and Paddy)
~nd seven pulses (Lathyrus, Lentil, Gram, Moong, Urd, Soy
bean and Arr>~t·) were taken for the study. Grains were proc
e~sed c2nsidering their natures and inoculated with the same
amo~nt of master cultures. Observations were recorded for
pen:ent2.::J"- colonisati:m of the substrate by the developing
,opavm aftE)i" 5, 10, 15 and 20 days of incubation taking the
date ,_,f it:ocu~ation as a zero day. Three replicates were
CULTIVATION AND HARVESTiriG
Chopped paddy straw was used as raw material for
cult i .;a.+~ ion. The st:·avJ was first steeped in water for 18
hour·s. Excess wa~Gr was then decanted and the soa~ed straw
was given a hot water treatment for one hour. Such
pasteurised straw was then spread on raised wire mesh for
draining the excess water and cooling. Spawning was done, at
a rate of 5% of dry straw in three layers. Polythene bags of
size 30x45 em were used to prepare cultivation beds. One kg
straw was filled in the bed volume. The cultivation was also
done in wooden trays (30x45x7 em) in some of the experiments
which were covered with polythene sheet for moisture censer-
vat ion.
Polythene bags and wooden trays filled with kg
spawned compost were kept in a thatched hut at prevailing
room temperature and relative humidity (Table) for spawn run
during different periods of cultivation over the ·year.
Polythene covers were removed after completion of spawn run
and beds were kept on rack for mushroom development. Dif-
fused light and cross ventilation were also provided
during the entire cropping period. Beds were watered twice a
day. Watering was stopped a day before harvesting and again
continued next day after harvesting. Mature sporophores were
harvested just before edges of pilei began to fold or curl
upwards. Harvested sporophores were weighed for their fresh
••ei~ht ar1d processed cts required for further experiments.
The spawned bags were l<ept for 45 days from the date of
spawning where as spawned trays were ke~t for 55 days for
production of successive flushes and their timely harvests.
The beds were also observed for disease and disorders.
Table: Meterological observation recorded during the cropping periods.
Cultivation period Year 1991-92 Year 1992-93
Mean Mean Mean Mean Tempe- Humidity% Tempe- Humidity% rature•c rature°C (Min-Max) (Min-Max) (Min-Max) (Min-Max)
January-February 26-30 50-82 25-31 31-60
February-March 25--34 31-79 26-34 36-61
Jul y·-August 22-31 75-·94 23-30 71-92
August--September 24-32 72-88 24-28 75-88
September-October 21-30 47-70 21-26 62-80
October--November 1 8-28 50-66 19-26 58-80
November-December 11- 26 4 7-92 17-24 52-80
December-Jaruary 1 3-23 41-94 20-26 60-82
---------------------------------------------------------------
1. Studies on Substrdte for Cultivation:
The relative suitability of different straw substrates
viz wheat straw, paddy straw, combinations of paddy straw
with 5, 10 and 25% rice· bran and paddy straw with 5, 10 and
~5% wheat bran were evaluated for cultivation of Pleurotus
spp. Ft·esh and clean rice bran and wheat bran were separately
sterilised in autoclavable polythene bags and mixed with
pasteurised paddy stt·aw just before spawning. Observations
were recorded for time required for complete spawn run, fresh
weight yields and biological efficiency, average size and
number of sporophores and duration of 4 flushes for selected
test species. Four replicates were maintained throughout the
'",.peri ment.
2. oitudies on Factorainfluencing Cultivation :
Selected P7eurotus spp. were cultivated on paddy straw
for the duration of eight months during the year 1991.
sr,"·.:n~n'J was done in every month starting from January,
FebrLnr; a0d JLJly l.o Ooc2mcer under bag method of cultiva
L' :)r~. ·Jbservat.ions wet·<~ ,.,,corded for t~me required for com-
~lete spawn run, Fresh weight yield and biological efficien-
41
cy, average size and number of sporophores and duration of
four flushes for all three P7eurotus spp.
were maintained throughout the study.
3. Studies on Mode of Cultivation :
Four replicates
Three P7eurotus spp. were cultivated by bag <:t'nd tray
method of cultivation during the period of September-October
and October-t<ovember 1992. Observation were recorded for
time required for complete spawn rutl, fresh weight yield and
biological efficiencies and average size and number of spore-
phores for all three P7eurotus spp.
maintained for the study.
PHYSIOLOGICAL STUDIES
1. Cultural Level :
a. In vitro Growth
Four replicates were
The three P7eurotus spp. were cultured on synthetic
and natural infusion growth media. The media taken were :
Malt extract medium (Raper and Thorn, 1949)
Malt extract
Dextrose
20 g
20 g
Peptone 01 9
Distilled Water 1000 ml
Straw medium
Paddy straw powder 10 9
NaN0 3 02 9
K2HP04 01 g
MgS04 0.5 g
KCl 0.5 g
FeS04 0. 01 g
Distilled Water 1000 ml
Actively growing mycelial discs of Smm diameter were
inoculated to 30 ml media in 150 ml conical flasks and
incubated at 25t..i>C for mycelial growth. Mycelial mats were
harvested after 5,10,15,20,25 and 30 days of incubation and
dried at 50°C for constant weight. Dry mycelial weights were
recorded for each species and each incubation period sepa-
rately. Three replicates were maintained throughout the
experiment.
. -..
b. Extra Cellular Cellulase Activity :
Cell free culture filtrates of different incuba-
tion. periods for both the culture media were
enzyme preparation.
(i) Relative enzyme activity :
used as
It was determined by viscometric method according to
described by Hancock et al. (1964). The reaction mixture was
used as described below.
1.2% CMC solution 3.5 ml
Mc-Ilvain's buffer- 1.5 ml
Distilled water 1.5 ml
Enzyme preparation - 1.5 ml
Percent loss in viscosity of carboxy methyl
cellulose solution was calculated by the formula given below:
ET 0 - ETt
= -------------------x100
ET O - ETw
Where ETo - Efflux time at 0 hour
ETt - Efflux time at t hours
ET - Efflux time of water w
The relative enzyme activities were then calculated
from 1000/Time required to reduced viscosity by 25%.
(ii) Enzyme Units
Cellulose units were determined by the DNS method of
Gascoigne and Gascoigne (1960). One unit of cellulase enzyme
was defined as the amount of enzyme required to liberate
reducing sugar equivalent to 10 ug of glucose (Olutiola,
1976).
2. Spawn Level
a. Spawn Substrate Supplementation :
Maize grains were processed to prepare substrate
and then mixed with urea (0.05% W/W); DAP (0.1% W/W);
mixture of NaCl (0.2% W/W), MgS047H 2o (0.1% W/W), K2HP04
( 0. 1% W /W) and KNO~ .:0
(0.2% W/W) and mixture of Dextrose
and Peptone (0.2% W/W) rice bran and wheat bran (5% W/W)
separately, alongwith usual addition of gypsum and
calcium carbonate before sterilization. Inoculation was done
with same amount of master cultures and spawn bottles were
incubated at 25t.z0 C for spawn growth. Observation
recorded for percent colonisation of developing spawn as
compared to control in each case. Three replicates were
maintained for the study.
3. Crop Level :
a. Effect of different grain spawn :
Spawn prepared on jowar, wheat and maize grains
were used for cultivation on paddy straw by bag method.
Observations were recorded for time required for complete
spawn run, total yield and biological Pfficiencies and
avere~ge size and number of spor·ophores by all three P7eurotus
species. Four replicates were mai:1tained for the studies.
b. Effect of supplemented spawn:
Spawn prepared on maize grain, maize grain+ rice bran
vi/W) , maize grain + wheat bran (5% W/W) were used for
cultivation on paddy straw by the bag method. Obser•;ations
were recorded for time required for complete spawn run, total
yield and biological efficiencies and average size and
number of sporophores for three P7eurotus spp. Four repl i-
cates were mai11tained for the study.
c. Effect of certain nutrients on P. f7orida:
Solutions of different amino acids Arginine,
sine, Methionine, Phenylalanine and Tryptophan (0.5%
Vitamins Ascorbic acid, Biotin and Riboflavin (0.02%
and Carbohydrates Glucose and Mannitol ( 1% W/V),
Ly
W/V);
W/V)
Rice
bran extract (20% W/V) and Hoaglands nutrient solution
were studied for cultivation of P. florida. 3upplementation
were done seperately at a rate of 10% (V/W) at the time of
spa1ming. A second d,:,~'" uf nutr·ients were applied after
completion of spawn r·un. Cultivation was done by both the
bag '"nd t;·ay methodc'3. The compost quantity was reduced to
500 g straw/bag and 250 g ctraw/tray (dry weight basis) for
the eAperiment. Four trays and two bags were maintained
throughout the experiment alongwith suitable controls.
Obsr~rvations were recorded fer time rec,~ired for complete
spawn n.n, total yield and biological efficiencies and
average riLimber of sporophores.
BIOCHEMICAL AND NUTRITIONAL PARAMETER STUDIES
Mature sporophores ::>f P. florida, P. ostreatus and
P. sajor-caju were analysed for their biochemical compostion.
Heal thy and mature sporophor·es wer·e se~e~ted and dried 2<·
47
70c'C in a hot ct·i r o,·,;,,, ,;ilU powdered before being used CIS
samples (e•:(ept r .. r ~,,.,' ;:,lt n.c;isture content.
1. Determination of Percent Moisture Content
Percent mo i :; tu re con tent was determined by
metl,ud given i11 A.O.A.C. (1975). Fresh sporophores were
driGd in oven at 110°C for constant weight and percent
moisture content was calculated by the formula given below:
Initial wt. - Final wt.
Percent moi~ture = ------------ --------- x 100
Initial wt.
2. Estimation of Total Protein Content :
The test samples were first digested for nitro-
gen by Microjeldahl method given by Ha~ija (1986) for plant
tissues. The values were then multiplied with a conversion
factor of N x 4.38 as described by Crisan and Sands (1978).
Ammonium chloride _.,
solution of 3.56 x 10 ~M concentration
was used as standard for reference.
3. Estimation of Carbohydrate Content
(a) Soluble Carbohydrates :
48
Test sa1nples were e~tracted in 70% ethanol and
concentrated over water bath to constant volume. Soluble
Carbohydrate contents were estimated by the method of Dubois
et a7. (1956).
· (b) Tot a 1 Carbohydrates :
Total carbohydrate contents were also estimated by the
method of Dubois et a7. (1956). Test samples were hydrolysed
with 72% sulphuric acid and diluted hydrolysates were used
for estimation.
(c) Total Sugars and Reducing Sugars :
Total sugar and reducing sugar contents were esti-
mated by hot titration method. Aqueous extracts of the test
samples were purified with 10% lead acetate solution and
used for estimation of reducing sugars. Purified extracts
were further hydrolysed with concentrated hydro~hloric acid
and neutrilised samples were taken for estimation of total
sugars. Benedict's quantitative reagent was used for estima
tion and D-glucose solution (1 mg/ml) as standard for refer-
ence.
f9
4. Estimation of Lipid Content
(a) Total Lipids :
Test samples were extracted by the method of
Mishra (1965) in Chloroform : Methanol solvent (2:1 V/V).
Lipid extracts were then purified and total lipid con
tetlt was estimated by the method of Folk et a7. (1958).
(b) Total Phospholipids
Total phospholipid contents was estimated from total
lipid extract by digesting the lipid for phosphorus. The
total phosphorus content was then converted to phospholipid
values by a conversion factor of P x 25. The values were
expressed as relative percentage of phospholipid according to
described by Bartlett (1959).
5. Determination of Crude Fibre Content
Crude fibre content was determined by the method given
in A.O.A.C. (1975). Residues left after acid and alkali
digestion were dried at 80°C till constant weight. Dry
residues were heated in a muffle furnace at 600°C for 2
hours. The differences between the two weight were expressed
as weights of fibre contents.
50
6. Determination of Total Ash Content
Total ash contents of the test samples were determined
by the method desc r· i bed by Bano et a 1. ( 1981 ) . Initially
charred test samples were heated in silica crucibles at 600°C
in a muffle furnace ti 11 complete ashing. The final weights
of ash were recorded.
7. Amino Acid Profile Determination
Amino acid profiles were determined for free and total
amino acids in test mushrooms. Dry powdered mushroom
sporophores were extracted in 70% ethanol for free amino acid
extracts and remaining residues were hydrolysed with 6N
hydrochloric acid for total amino acid extracts. Paper
chromatography was done by the method of Block et a7. (1958).
Butanol: Acetic acid: Water (75:20:20 v/v) was used as
solvent system and 0.3% (W/V) Ninhydrin solution was used for
detection. Individual amino acids were identified by co-
chroma~ography of standard amino acids.
IIIIIIUIIII/1/ll~/11 T 12731
8. Protein Profile Determination
The protein profiles of test mushrooms were determined
by polyacrylamide gel electrophoresis (PAGE). Disc gel
electrophoresis was done to asses the position of proteins of
different molecular weights and net charges. Electrophoresis
was done by the method of Dunn (1989). Crude protein was
extracted from the sporophores of each species in N saline
and its concentration was estimated by the method of Lowry et
al. (1951). Approximately 100JJg protein was loaded in each
gel tube.
9. Estimation of Minerals
Sample ashes were dissolved in 10 ml double distilled
water and analysed for different minerals by
Coupled Plasma Scan (ICPC).
10. Estimation of Energy Values
Induction
The energy values of test mushrooms in K cal/100g were
estimated by the method of Crison and Sands (1978). Sum of
the values of Total protein x 2.62, Total lipid x 8.37 and
Total carbohydrate x 3.48 were expressed as energy values of
52.
corresponding species.
PRESERVATION STUDIES
The harvested, healthy and mature sporophores
were subjected to three preservation techniques namely,
controlled condition storage, Steeping preservation and
Dehydration. Each experiment was carried out as under, main
taining three replicates.
1. Controlled Cdndition Storage
Mushroom samples each weighing 50 g were stored in
different packaging conditions at below o0 c, 5-10°C and 25-
28°C. The different packagings were perforated
bags, unperforated polythene bags and no rackage.
polythene
Observa-
tions were made on changes in fresh weight, colour and tex
ture of the stored product after 96 hours of storage.
2. Steeping Preservation
The method suggested by Dang and Singh (1978)
was employed for steeping preservation. 50 g test mush-
rooms were steeped in a preservative solution containing
53
common salt (2.5% W/V), ascorbic acid (0.1% W/V),
(0.2% potassium metabisulphit~ (0.2% W/V) and citric acid
W/V) and stored at 5-1 0°C. Observat i ens were made for
changes in fresh weight, colour and textur~ of steeped
product after 5,10,15 and 20 days of storage.
3. Dehydration
Mushroom samples each weighing 50 g were dehydrated by
five different processes as under :
(i) Sun drying.
(ii) Mechanical drying at 70°C in hot air oven.
(iii) Blanching in hot water for 10 min. and drying in
hot air oven at 70°C.
(iv) Blanching Followed by dipping in 0.2%
(W/Y) citric acid solution for 30 min.and drying
in hot air oven at 70°C.
(v) Blanching followed by sulphitation in 0.5%
(W/Y) potas~ium meta bi-sulphite solution for
30 min. and drying in hot air oven at 70°C.
Observations were recorded for c~anges in dry weight
and colcur of dehydrated product in each case for all three
test mushrooms.
Dried mushrooms were soaked in warm water for 2 hours.
Obs~rvations W8re made for rehydration capacity of dried
products in each case for all three test mushrooms.
STRAIN IMPROVEMENT STUDIES
1. Plate Experiment and Cultivation Trial
Pure cultures of three test fungi were cross inoculated
in all possible combinations and observed for mycelial
growth. Cultures isolated from mating zones were recultured
and used to prepare master culture and spawn.
Spawn prepared by isolated cultures were used for
cultivation on paddy straw by bag method of cultivation.
Observations were made for time required for complete spawn
run, total yield and biological efficiencies and average size
and number of sporophores under maintaining two replicates.
2. Ultra Violet Irradiation and Cultivation Trial :
Culture isolated from mating zone of P. florida x P.
sajor-caju was used for irradiation experiment. UV irradia-
tion was given to actively growing mycelial mats keeping
55
12.5 ,25 and 50 em distance from UV source for 1,2 and 3
minutes respectively. Irradiated plates were incubated at
25t..t' C in dark condition to prevent photoreaction of UV
treatment. Mycelial suspensions were prepared from taking
mycelial discs of each treated plate. Prepared mycelial
suspensions plated over malt extract agar plates separate-
ly and incubated at 25t..:PC for mycelial growth. Culture
plates differing in mycelial morphology were selected and
used for preparation of master culture and spawn after proper
reculturing.
Spawn prepared by selected cultures were used for
cultivation on paddy straw by bag method. Observations were
recorded for time required for complete spawn run, total
yield and biological efficiencies and average size and number
of sporophores. Untreated culture of P. florida was also
used as control for comparison. Two replicates were main-
tained for the study.
56
R E S U L T S
The cultivation of Pleurotus species represents one of
the major current economically profitable biotechnological
process for the conversion of waste plant residues into food.
There are many species which grow in nature and some of them
have been brought under cultivation and commercially exploit
ed (Table-1). Cultiva~io0 technologies have been standar-
dised for the '""''-·ia:3 t~c.r1el:; P. citrinopileatus, P. eryngii,
P. f7abe77atus, P. f7orida, P. ostreatus, P. sajor·- caju and
P. sa,c:c'dus :vijay, 1990). Among species, P. eryngii and P.
fos:cu7atus have relatively low temperature optima for myceli-
a1 srowth where as other species have temperature optima at
~5-3CloC for- mycel ~al gr-owtt-.. P. eryngii, P. florida and P.
t:e,Jtus have been found low temperature strains among
._,mrrl<crcially cultivated species where as P. sajor-caju and P.
:JpiJus have been found high temperature tolerant species.
'aile- I: Data on the bi:l:gy of P'eJrotJs spec·•s obtained under cultivation conditions (literatu•e data).
8pt j ~ "r:': ~empe:·a
~Jre ror '·!;ce I i a I ;rowH.
I oc I
tempera-
fr~~tinJ
: oC:
::itr;::~pi /eatus '* 25-30 26!_2
25-3j 20t2
ec.;s 25-30 2lt2
18-22 16t2
f)ate::atus :t: 25-30 22!_2
f1c(da * 25-30 20!_2
fossu1atus 18-22 18U
25-30 :2-22
s~jor-;a)u t 25-30 24•:.2
25-30 241;.2
Ra:~g8 Jf te!pera-
to1erar.:e ( .:J~)
20-:C
12-25
16-25
12-24
16-23
!2-22
12-25
7-'? '·
'7-3Q
re•ic~ fJr Period for Optimum c:1ple!! furiting Humidity spaor r~· at :ptimum range a: :pti1~1 tempera- (I) t£~pera~Jre ture
:days; (days)
12-2 4 20-28 85-90
Morphology Average Sporophore pi Ia us eel our diameter I em)
3-8 Cream White
Yield Sporophore parlor-texture mance
(B. E,%)
Soft 30-60
'6-1 B 25-30 80-90 5.1-12.7 Yellowish Tough 40-70
20-22 25-30 75
55-60 70-7 5 85-95
: 8-2 2 70-80
25-30 70-90
50-56 6 5-70 70-95
20-2 5 30-35 80-85
~ 2-14 18-25 80-85
1H8 22-28 70-90
or Grey
3-9 Pinkish Brown Fleshy to Brittle
4-15 Greyish to Rusty
Tough
8-11 White Turning Leathery to Red
8-15 Cream White Fleshy
3-13 Pale White Brittle
30-50
20-30
60-9V
50-90
8-20 Greyish White Soft spc~gy 30-5~
5-14 G~e) to B~own Rigid
!-3 Cream White Fleshy ------ ------------------------------------------------------------------------------------------------------------------------
Ir1 respect to spawr1 run period and period for fruiting, P.
flabel latus and P. sajor-caju have been regarded as fast
growing and early fruiting species. All the species have
been found to require high humidity range for growth and
fruiting. P. florida and P. ostreatus have been found to
recover moderately large sporophores among the cultivated
species. In respect to sporophore colour P. florida is
superior· where as P. ostreatus is superior in respect to
sporophore texture. P. flabellatus and P. florida have been
found as high yielding species among the commercially culti
vated species.
Selection of P. florida, P. ostreatus and P. sajor-caju
(Plate-1) for the present study was mainly based on require
ment of physical environment for growth and fruiting, fruit
morphology and productivity. A systematic study has been
undertaken to compare the selected species with reference to
mushroom biotechnology and results obtained are given below.
SPAWN PREPARATION :
1. Effect of Spawn Substrate
All the three Pleurotus spp. investigated showed similar
response towards the spawn substrate tested (Table-2;
• I
• • 11
••• Ill
Table-2 : Effect of different spawn substrate on colonisation of Pleurotus florida, P. ostreatus and P. sajor-caju.
Extent of growth (% colonisation in days)
Spawn P. florida P. ostreatus P. sajor-caju substrate ------- ------
5 10 15 20 5 10 15 20 5 10 15 20
Cereals
Maize 30 100 30 80 100 30 100
Jowar- 10 80 100 10 50 80 100 10 50 100
Wheat 10 50 80 100 10 50 80 100 10 30 80 100
Bajra 10 30 SG 10 SG 10 30 SG
Rice 10 SG 10 SG 10 50 SG
Pulse
Lathyrus 10 SG 30 SG NG -
Lentil 10 SG 10 SG 10 SG
Gram NG 10 SG 10 SG
Moong NG NG - NG -
Urd NG NG - NG -
Soybean NG NG - NG -
Arhar NG NG - NG -
--------------------------------------------------------------------SG - Static growth NG - No growth Data - Mean of three rep 1 i cates
P 1 ate- 2). A marl.ed preference for a cerea 1 substrate was
found over the pulse substrate. Among cereals, maize grain
was colonised the best in only 10 days of spawn growth by P.
florida and P. sajor-caju but P. ostreatus required 15 days
of growth for 100% colonisation. Pulse substrates were not
colonised at all by any of the selected species. The maize
grain because of its desirable acceptability by the P7eurotus
species for good colonisation and growth, was selected as
spawn substrate for further studies.
CULTIVATION AND YIELD
1. Substrate for Cultivation
Selected Pleurotus spp. were cultivated on eight different
materials. It was found that all the species individually
showed similar response towards the material used (Tables-3a,
b & c; Figs. 1a, b & c). Much difference were not observed
for complete spawn run period among materials used and among
the species. However, P. florida responded well and required
minimum days (17) for complete spawn run among the species on
paddy straw substrate. It was also found that paddy straw+
10% rice bran (w/w) substrate was the best for fresh weight
yields and biological efficiencies of all the three P7eurotus
• I
Table-3(a): Evaluation of different substrates for Spawn run, Total
yield and Biological Efficiency of PTeurotus florida.
-------------------------------------- ---- --- --- ---------------------------------------------------------------
Scbstrate
Wheat straw
Paddy st ra'li
Paddy straw + Rice :ran ',. \.Jf- n' / W)
?addy straw + Rice b:ar I I 0% I': I ,-I
f u,
Paddy s trafi + Rice ~ran ( 25% w Jr~)
~addy straw + Wheat ~ran ( 5% w/·~·;
Paddy s~r aw + 'dheat ~,.a~ ( : ~% 'fl/ /1:
~addy stra·~ + Wheat ~~D I "'~ ~·
' ~ w ' '1\/w:
T i :r.e
req~ ~red
for
co~ple~e
spa11r. r ... ~
1 8
; -
<0 ''
..
1C
I?
~ ?
2:
f'esr <e';\t ;:e:: :;/kg straw) Biological --------- --------------- Efficimy ,. P: •: R!; .~verage I%)
~~c ,.. '" ' ~ 0 ~ ~ ;!:.s w.o 42, 7C ~\J ~' ·' .,. "v . ~ ~l.i~ ••
'' ~ . ~ 1r . :3::.0 376.5 421.61 4 2. I c .......
~" e ' 533.: 558.3 ~ 3 ~' 9 5 94. I 53.41 '"'
:32.: ::I g 7 ~' ~ 879.3 8 76 '9 87.69
:89.2 ~ e • ' 515.3 :7 3 '8 5 61. 5 56' 15 "J " '
~30.~5 30?' :: :92 '5 ;ou 61U 61 '06
.. '~ ~ ·'~~·- .. 3~ .35 759.5 "f!"f ..
I~ I 'I 7 46 '0 7 4. 60
!:8.:5 "' ' ~42' ~ 5 ~ ~' 4 4 70 '2 47.02 "f I 't"
Ave. size of Average Duration sporophores Number of 4
---------------- of flushes Stipe Pileus seorophores (days) Length Diameter
(em) I em)
0 '8 7. ~5 62 44
0 '8 6.88 54 43
0 '8 7.35 I 02 44
0.86 7 ' 13 146 41
0.82 7.82 93 38
0' 8 7.35 I 03 45
0.3 7. 13 138 43
0 '8 7 '8 2 82 43
------------------------------------------------------------------------------------------------------------------CC at 5% for su~stra~es- 49.52
spp. P. florida responded the best among the species by recov-
ering 87.69% biological efficiency which was followed by P.
ostreatus (66.4%) and P. sajor-caju (65.0%) respectively.
Table--3(b): Evaluation of differ·ent substrates for Spawn run, Total
~ubstrate
yield and Biological rfficiency of Pleurotus ostreatus.
~cr
cc~p1eta
spa'~n rJn (days;
-- - - -- - - --- -------------------------------------------------------------r-esf we•Jht ;ie1d :;:>g straw) Biological
------- ----------------------- Efficiency ~I R2 R3 R4 Average 1~1
Ave. s:ze of A1erage sporophores Number
---------------- of
Durat:on of 4 flushes
Stipe Pi~eus sporophores (days) Length Diameter
I em) I em) ------------------------------- -----------------------------------------------------------------------------------Whea~ straw 21 318. J :2 ~ . s 3Ce. 9 3!J. I 325.6 32 . 55 0.80 5 . 54 57 44
Paddy straw i ~ = 1 5' Q :~ 5 '~ 27:.0 3 2: '0 305.5 30 . 55 0.81 5.36 52 44
Paddy straw + ., ' " :62.3 ~ 71' l
J I '•', I ~ 9 8 ' 9 587.9 577.5 57. 7 5 0.91 7.38 I 06 43 Rice bran [5% ·•/•1
Paddy straw + 2C 554.6 se·.z ec3.7 .664.0 66' 40 0.92 7' 18 .123 42 Rlce bran (::i% w/11)
Paddy straw + 2 i ce5.2 HU 422.7 4:6.7 447.2 4U2 c . 8 6 6 . 30 82 38 Rice bran (2!1 w/w)
Paddy s:raw + 55C.4 515.7 5:f.J E5B. 1 5ED.8 56.08 0.86 7.38 96 Whea~ bran :E% w/11;
Paddy straw + oL:.2 0128 Cl2.9 65~.2 645.0 64.50 0.88 6.78 117 40 ~hsa~ ~ran :~:% ~.'~:
35~.+ 29C.5 362.7 32~.45 375.5 37.55 0' 81 7.96 67 43
C~ at 5% 'cr s~bstrates - 29.98
Lowest fresh weight yields and biological efficiencies were
obtained on paddy straw substrate for P. florida and P. os-
treatus and on wheat straw substrate for P. sajor-caju. Other
Table-3(c): Evalu,1tion of different substrates for Spawn run, Total
yield a:1d Biobgical Efficiency of P7eurotus sa.jor-caju.
---------------------------·-- --- -- •-----------•-•-·••••w••••••• -•••••••••••••••••• ••••••••••••••••••----•••--
S;~strate
~·r.:e
required
for
spa~"~·~ : .• ·~
( d a,. 3)
~r:s,: we·sl·: ,· ·e~d :;/kg/straw) Biological - ------------ m:c'ency
Ri R2 P: P~ Average (%)
Ave. size of Ama;e Dura~ icr: sporophores Numter of ' ,
---------------- of flushes Stipe Pileus sporo;hms [days) length Diameter
I:!) I em) -------------------------- ------------------- ··-----------------------------------------------------------------
'•'\·- ~ ~ •I ·r<;;:G-. s ~ r a·r~ :o 24 5 '5 I)") r'\ 7 {.{.,,' 2~~.: :32. 3 271.25 27. 12 0.82 5.35 42 45
Pad~/ straw '1 32' . 0 ~ ~0 'c ~r-r. " ... ~. 'w 223. ~ 32 2. 12 32. 21 0.78 5 '7 7 62 43
Pad~ I straw ' ~ s "~ ~ ~ c.;~ . ·: 5C 2 . .: e: 3. : 517. 3 625.9 62.59 0.83 7 '36 116 45
P::e ~ 1· an I tdl ~/1!; ' ~'
p ., ~ ~ \ a_. ... , s t:a't~ ' 2~ C5?.5 c::.: 555.: 565.6 550.8 65.0 0. 81 : . 3 2 130 45 Rjce :ran : '~% ',''o I
Pad: 1 straw ' " ,, 459.7 443.7 ~.:s.: !E9.8 456.4 45.64 0.85 U5 89 37 Rica ~ran I I) ~ 1' Yl,'t~.) '• ~ ~'~
Pad:y s~:aw ' 2? ~27.~ 525.3 533.0 53. 30 0.82 7.60 98 45 ~lf:s~: bran ::% f\'/'f.')
Pa~d;. stra~v ' '1 625.7 62U 623.6 533.0 629.2 62.92 0. 81 7.82 102 43 l~hsat b;an ( ;~% ',\ / '.\ )
Pad:, stra\\ 20 34:.: 347.2 222.9 334.75 338.2 33.82 0.83 7.52 61 45 ·,~h:at Cr an :25% ~· ,'~~)
--------------------------------------------------------------------------------------------------------------------CD at 5% for su~strates - 37.88
substrate were found to recover moderate fresh weight yields
and biological efficiencies. It was also observed that aver-
age size of sporophores were not affected due to different
1000 TOTAL YIELD(g)
!.======= NIE. No. OF SPOROPHORE --------~~~~~~~~~200
- FRESH WEIGHT
800 c=J SPOROPHORE
600
400
200
0
'----------
ws PS
SUBSTRATES
Flg.1a:Evaluation of different substrata for total yield & sporophore
number of P. florida
TOTAL YIELD (g) AVE.No.OF SPOROPHORE
800
600
400
200
0
M FRESH WEIGHT
0 SPOROPHORE
SUBSTRATES
Fig.1b:Evaluation of different substrata for total yield & sporophore
number of P.ostreatus
1150
100
GO
150
100
50
0
TOTAL YIELO(g) AVE.NO.OF SPOROPHORE 1000 ------------------- 160
800
600
400
200
0
- FRESH WEIGHT
0 SPOROPHORE
WS PS PS•RB• PS•RB••PS•RBoo• PS+WB• PS+WB•.PS•WB•oo
SUBSTRATES
Fig.1c:Evaluation of different materials for total yield & sporophore number of
Psajor-caju
"'
140
120
. 100
80
60
40
20
0
substrates used for cultivation. Average number of sporo
phores were found to well suited to their corresponding bio-
logical efficie•1cies. raddy straw + 25% rice bran (w/w)
substrate was fouGd to require minimum duration for complete
harvest for all th:-ee Pleurotus spp.
2. Factor·s Influencir.3 Cultivation:
The three Pleurotus spp. were cultivated on paddy straw
during January to Marc~ and July to January under the prevail-
ing temperatura and relative humidity.
October-November was found the best period for cultiva
tion of P. florida (Table-4a; Fig. 2a). Minimum time of 14
days for complete spawr run was required during December-
January period. However, much variations were not observed
for complete spawn run time among the cultivation periods.
Highest biological efficiency of 52.23% was recorded during
the cultivation period of October-November which was followed
by 52.07% duri:1g November-December. Cultivation in different
per~cds was found to result in variation of average pileus
dia~eter of sporophores. Maximum 8.2 em wide pileus was
reco~dad during October-November period followed by 7.21 and
1.·: em during November-December and September-Octob~r peri-
uds, respecti~Gly. It W3S also observed that average number
<Jf sporophores wGre wa'l suited to their corresponding biolog-
ical efficier1cie.: .. Cultivati:m during October-November period
was found to require minimum time of 39 days for complete 4
f1 ushes of mush ro0m y i e 1 d.
Table-4(a): Effect of di~fo~or1t periods of cultivation on Spawn run, Total ,i<cld and Sio~ogical Efficiency of P7eurotus florida.
------------------------------------- -- -------------------------------------------------------------------
)W)
~ 1891-3:; spwn ~Jn
Ja~.-~~~.
"' . I \ i.<U} ~
" . (
" '.
" ..
:' 15
" (.
! 4
c~22~ 'l•t;Jht i':,d :g/:; stra·~) B~clogical
----------------------------------- Effi:ie~cy
Rl .,2 "3 ~~ ~verage (%~
212.C 3~e.o 31;.J 3C?.O 330.5 33.05
2!2.3 25?.0 2EE.e 272.0 263.2 2€.32
n:.55 HU n:.3 383.25 404.4 40.44
~ • .., 11 't ' ' ''w 363.0 494.0 446.0 44.60
424.~ ~57.0 423.0 434.5 43.45
5;;. 0 487.0 51C.O 555.0 522.3 52.23
HC.75 5:6.0 582.75 543.5 52U 52.07
5~:.0 522.3 527.2 47i.O 509.0 50.90
C8 at 5% 7cr ::'eriods - 66.23
Ave. size of sporophores
!·mage rlumber
of
Duratien of 4
flushes Stipe Pileus sporoptores (days) length D~ameter
I em) I em)
0.90 5. I 0 42 45
0.35 4.30 45
0.82 6.32 55 42
0.80 6.45 58 45
0. 81 7. 15 60 4S
0.80 8.2C 67 39
0.81 7. 21 72 43
0.80 6.30 68 45
0ctober-November was found the best period of cultiva-
tion fot- P. ostreatus (Table-4b; Fig. 2b). Minimum time of 15
Table-4(b): Effect of different periods of cultivation on Spawn run,
Total yield and Biological Efficiency of Pleurotus ostreatus.
---------------- --
:l<:tl 1atiJn ?er i od :dur~n~ ~~e
)edr)
: 1991-n;
r2~J 1 rs~
7:Jr :a~:ple~e
s;awn rcr
-------------··---- -- --------------------------------------------------------------rr~s~ ne;;~t )lE:d :;/~;, 1 strawl Biological
··------- ----------------Efficiency
PI 12 r: R4 A·1erage (X)
A;e, size of Average Duration sporophores Nu~ber of 4
---------------- of flushes Stipe Pileus sporophores (days) Length Diameter I em) I em)
-------------- -- ------- -----------·----------------------------------------------------------------------
:an.-reb. :59.~ : c 1' 6 16.26 0 '93 us 12 45
Feb.-March ~ 05 '~ ~28.0 15~.5 131.0 13. 10 0.90 3.85 18 45
Ju1y-A~g. 30U 2G 5. 2 30 2. 2 288.2 30.82 0.80 6,10 76 44
., ... ,., 31 g' 0 31 '90 0.76 5' 30 82 44
Sept.-8:~. '. "' I' 7 ~
.,. It '" m.75 350.3 35 '0 3 0.81 5.40 94 43
. 3 ~29.: ~~9.C 4C ~ • : m.: 429.2 42.92 0.80 6.25 98 42
'ici.-Dc:. '' 285.5 252.5 353.0 218.8 29' 80 0.80 6 I 15 67 45
Jec. -Ja~. 258.25 2?4.5 2::.:5 165.: 167.5 26.75 0 '81 5.82 53 45
:8 at ~% fJr Par:ods - 87.05
days was required for complete spawn run during the periods of
November-December and December-January while a maximum of 21
days was required during September-October. It was found that
Gklximum biol~;Ji<."ll ,_;ff"kic:I<Y (42.92%) was recovered during
October-~ovemb~r per icd which was followed during September-
October (SS.03't) c:ttd Augu::.t September (31.90%) periods respec--
Lively. Muderate size cporophores ranged from 3.85 em to 6.25
em 1n averaGe pileus diameter during different periods of
cultivation. Tl1e average number of sporophores were found
well suited to t: .. ,oir cor:·esr;,Jtlding biological efficiencies
Cultivatio11 dt1ring Octc~er-November period resulted in minimum
duration (42 days) for complete harvest.
September-October was observed the best period of
cu1tivaticr1 D ' . sajcr--caju (Table-4c; Fig. 2c).
Cultivation during July-August and August-September required
minimuM time (~7 days) for complete spawn run while maximum
time (22 dayc) was required during February-March period.
Highest biological efficiency of 39.83% was recovered during
Septembsr-Ootcber period which was followed by 38.97% and
33.11% during November-December and December-January periods,
respect~vely. It was also found that different periods of
culti ation were found to result in moderate size (4.3 to 6.82
em p'leus diameter) sporophores. Average number of sporophores
w2re a~so found to suit well to their corresponding biological
effi c.: er:c: i es. Cultivation during January-February and
October-November wcr€ fcund to require less time (43 days)
for complete 4 flushes of mushroom yield.
Table-4(c): Effect of different periods of cultivation on Spawn run,
Total yield and Biological Efficiency of PTeurotus sajor-caju. --------------------------------- ----- --- -----· --------------------------------------------------------
:~1t:.,·atj~·~
re;~' rec f: r
,.;ar) co~t~~e~e
::;31-92) 3P~~~~ r~~
~a~.-Feb.
ceb. -March
~~1y-Aug.
A~8. -Sept.
Ss~~.-Oct.
:ca;s;
'' ' . 22
17
1;
" ,,
Bio1:gical ------ Efficiency
Image I%)
224.0 22~.5 2:2.E 222.~ 224.0 22.40
~5'.25 14:.25 :~8.8 15~.~ 15:!.7 15.37
24:. 7 2 7 3. 5 27.35
:s~.c 2::.0 ~?:.c 277.c 2~s.2 24.82
2~E.~ 3~3.0 355.0 225.5 33~.1 33.11
4:7.5~ 3S?.95 ~31.0 38~.5 389.7 38.97
2~C.J 253.0 2E~.O 2:S.5 256.5 25.65
3. Mode of Cultivation:
Ave. si:e of sporophores
Average Number
of
Duration of 4 flushes
Stipe Pileus sporophares (days) Length Diamete~
I em) I em I
0.80 I '' ..... loj
0 .so uo
0 '81 4.45
0.?8 5.20
0.30 6' 82
0.80 5 ·'' '"
0 .EJ 5.90
0.12 5.20
46
32
50
43
61
56
67
48
45
45
42
'' "
~hree species of Pleurotus were cultivated by Bag and
Tra~ method of cultivation during the periods of September-
600 TOTAL YIELD (J)
TEMP lc RH
500
400
:300
200
100
0
.... 100 ... 80
60
40
20
JAN-FEB FEB-»AR JUL-AUG AUG-SBP SIP-OCT OCT-NOV NOV-DBC DEC-IAN O
PERIOD
--<>- TEMP (min) _,._ TEMP (mu) -~- RH (min) ·~>· RH (mu)
Fig.2a:Effect of different periods of cultivation on total yield of ?.florida
' .
TOTAL YIELO(g) TEMP &. RH · 600.-------~~------------------------------.100
400
300
200
100
0
/,>--·-···-I> '
/),.. ___ ---e;.·
~- ..
JAN-FEBFEB-MARJUL-AUGAUG-SEPSEP-OClOCT-NO\fiOV-OECOEC-JAN
PERIOD
-&- TEMP (min) _,._ TEMP (max) ·+- RH (min) ·A· RH (max)
Fig.2b:Effect of different periods of cultivation on total yield of P.ostreats
80
60
40
20
0
TOTAL YIELD(g) TEMP & RH 500.-----------~------------------------~-----.100
400
300
200
100
0
-"- 1!.-~------l:o
'
JAN·FEBFEB·MARJUL·AUGAUG·SE1'6EP-OC10CT·NO\ti0V·DECDEC·JAN
PERIODS
----TEMP (min) --TEMP (max) -·0 · RH (min) - ,._ RH (max)
Fig.2c:Effect of different periods of cultivation on total yield of
P.sajor-caju
73
80
80
40
20
0
Data on Bag nrethod of cultivation (Table·-5a; Fig. 3· •
Plate-3A)r-e.;ealcd that P. flo.·ida and P. sajor-caju required
17 and 18 da;s respect~vely for complete spawn run during
Table-5(a):Evaluation of Bag rr;ethod of cultivation for Spawn run, Total yield and Biologi:al EFficiency of rleurotus spp.
Cultivat~o,-~ T~me Req,Jir·ed
period for complete rresh weight
weight yield*
Biological Ave. size of Efficiency sporophores (%) -------------
Average number
of s;:awn run (days) (g/kg straw) Stipe Pileus
Lenght Diameter sporophores
(em) (em)
P. florida Sep.-Oct. 17 428.8 42.88 0.81 7. 30 86
Oct. ·Nov. ~7 492.3 49.23 0. 85 7. 42 92
P.ostreatus Sep.~-oct. 374.25 37.42 0.80 6.32 53
Oct. -NO\/. 18 379.0 37.90 0.86 6.25 50
P.sajor-caju Sep.-Cct. 13 430.9 43.09 0.86 5.40 62
Oct.-Nov. 10 .u 324.4 32.44 0.82 5.61 47
CC at 5% for Periods - 10. 68
for Species - 13.08 for Periods x Species- 18.5
t - yie1d given is sum of 4 flushes
both the cultivat~on periods where as P. ostreatus required
21 daiS dur-i:1g Stptuclccr Octvber· and 18 days during October-
l~cvernbe r-. r. Fl,,, id,l r c~ponded the best among the species
and r~covordd higheot (49.2%) biological efficier1cy during
OctvLer-- Novelll 1'•:>r, fol 'o ' b p · · (4" 09%) d · u~ r \'leu y -. Sajor-CaJU ~. ur1ng
September·Octuber. P. ostreatus was found to recover moderate
cio~ogical efficiency ctmun::J the species. Average size of
sporophorus ~ere found correlated with cultivation periods.
Average nctrrrbar of sporophores were also found to suit their
corresponding biological efficiencies.
nata on Tray method of cultivation (Table-5b; Fig. 3• ,
Plate-·38) re'lealed that all the three Pleurotus species re-
quired almost similar time for complete spawn run during their
cultivation periods. P. ostreatus responded the best among
the species and recovered 84.10% biological efficiency during
September-~ctober period. P. sajor-caju was found second best
(65.66% biolosical efficiency) where as P. florida (65%) was
found third among species. However, P. ostreatus was found to
recover highest biological efficiency during both the cultiva-
~ion periods. P. florida was found to recover large size
sporophores (9.83-10.5 em average pileus diameter) during
both the cultivation periods, respectively. The average number
Table-5(b):Cvalua~inl1 CJf Tray rnetl.ud of cultivation for Spawn run,
Specios
Total ;ield .<nd 3ic.lugiql Efficiency of Pleurotus spp.
Culti;ati0tl Time Required period for- con,pl·3tc
spawn ~-un
Biological Ave. size of Efficiency sporophores
(%) -- ----------
Average number
of ( ,.J '":'I""" '\ \ U<.l].:;)
Fresh weight weight yield*
(g/kg straw) Stipe Pileus sporoLenght Diameter phores
P. florida 2.;;;p.- Oct.
Oct. -Nov.
P.ostreatus Sep.-Oct.
Oct. --Nov.
P. sajor-caju Sep. -Oct.
Oct.-Nov.
20
19
21
on LU
22
21
609.5
650.0
841.0
716.0
656.6
594.0
CD at 5% for Periods for Species
(em) (em)
60.95 0.83 9.83
65.00 0.93 10.50
84.10 0. 98 8. 30
71.60 1.05 8.0
65.66 0.96 6.10
59.40 0. 96 7. 82
- 27.102 - 33.19
For Periods x Species- 46.94 * - yield given is sum of 5 flushes.
of sporophores were found to suit their corresponding biolog-
~cal efficiencies and average size of sporophores.
118
121
128
125
106
98
1.1
0.9
TOTAL YIELD(a) (Thousands)
c:J SBPT-OCT BAG
1111111111111 OCT-NOV BAG
P.florida
- SEPT-OCT TRAY
lllill OCT-NOV TRAY
P.ostreatus P .sajor-caju
Fig.3:Evaluation of mode of cultivation for Pleurotus spp.
~HYSIOLOGICAL STUDIES :
1. Cultural Level
a. In vitro growth
It was found that all the three species of P7eurotus
responded well to strav1 medium (Table-6; Fig. 4). However,
the growth pattern was found similar on both the culture
Tab1e-E J,o: v·i~-: ;~: .. : 1 ~ ct ''f~·~·::~:;~ 3~~. :~ ~/nt 11et1:: and natura~ hfus1cn growth med1a at different ~e~:cjs :f i~: b.~~:::·,
r~cubatjc~ ----------
P. florida P. ostreatus P. sajar-caju
" "0 "1: ' I' 1'\r\. '\ • ''>. ~".:.. ' .!\I ',:;
1'. r~~~:" 1'11'1'~ l•, ... ,~_ •• l·~ ~ C .C232i).OC:j4 0.1203±_0.00014 0.1032±_0.001:7 0.0991±.0. 000?0
" ~.1s1e:,:.~n: .. : c. :sc1:_8.oo::~.~ :.~~52-t.~.JC~15 o. 2Co5t.o. oo390 0 .1522±_0.00052 0. 1373t.O. C0046 " H J.2!54:_~.:J~·::: I' 1'1 ~ C ?.:. ~'> II f"" •!) : .2~.,J!_C .:!C2C27 u::s:c.ooos4 0. 2307!.0. 00023 0, 3253-t_O, OOOH ' \.>,~.Jv_ •• v~. 'J
:~ c.13:·:_:.~cc:: c.=~~~::.8c~~') .25~:-t_C.CGC 1 7 1. W7 t.0.003C5 ~.4219!,0.000072 0.4792-t_?.C~~\4
. , • 1/fiC:,;.."' "1\ fl ~- r •· '~ ~ r.: " r'l r "" 7 'l ,., 'fl~ 1: .j. fl 1'11\ ~:'\ 0.5€12!_1.00030 U307t.0. 00021 C.S0'2:_:.~~02' ' ...... ,. - ' ' •'' ~' ~·~·-· ... ·.~~ '- ·• ',..,.'"" :...~ '~ J .... - -
:G " 117".,_' I'H\(1~1:: .. -~ ~--·-~··-· C.?iS~:_: .2C2:~ :·.3~05±_2.~C~22 U5~0t.0.0003i G. 4328:_0. 000118 o. sn~:.o. oooss
med i ~-- A steady increase in dry biomass was found upto 25
days cf growth. Among species, P. florida produced higher
dry biomass during different incubation periods followed by
P. ostreatus at<d P. sajor· caju.
b. Extracellular cellulase activity
Determination for eYtra cellular cellulase production
in malt extract broth and straw broth media at different
incubation periods showed that it was constitutive in all the
Pleurotus spp. (Tables-7a,b & c; Figs. 5a,b & c).
Table-7(a): In vitro cellulasE activity of Pleurotus florida.
Incubation Per1 od (days)
Malt e~tract medium Straw medium
5
1 0
15
20
25
30
REA* Eu** REA*
< 1 25.88 6. 11
10.582 46.44 10.92
7. 168 32. 18 37.03 . c; 26.0 25.31
< 1 25.32 21.27
<1 23.20 13.98
* REA - Relative enzyme activity (Viscometric) ** EU - Enzyme units (DNS Colorimetry)
Eu**
26.92
48.73
136.04
84.47
52. 14
47.53
Malt extract broth medium did not support cellulase produc-
t~on. Ail Pleurotus spp. appeared to posses the cellulolytic activ-
0.8
0.6
0.4
0.2
0
DRY WT. {g)
P. florida P. ostreatue
MIDIUM
R MALT IX TRACT
CJ 8TR- MI!DIUM
',: .. : .. . '· .. ·:·
P. eajor-caju
Fig.4:/n vitro growth of Pfeurotus spp. after 30 days of Incubation
ENZYME UNITS 160~~~~---------------~========~
140
120
100
80
60
MIDI A
-ll<- Malt extract
--l- Straw mltllum
40 ./' ~-# ~----~----~~----*
20
oL-----L-----J_----~-----L-----l----~ s ro ~ w u ao ~
DAYS
Flg.5a:/n vitro cellulase activity of P.florida
80
ity of c't lower nt deer and t 1rat too during the late growth phases.
Higher enzyme ,rn its >1-HC· r-ecovered after 10 days of growth by all
Pleurotus c;r·p.
Table-7(t). In vitrcJ cGllul,,coc activity of Pleurotus ostreatus.
Inccbat i o", Period (days)
Malt extr·oct medium Straw medium
5
10
15
20
25
30
------
Eu** REA* EU**
<1 21.86 <1 27.45
7. 017 31.92 8.73 32. 16
< 1 28.17 9.25 34.58
< 1 25. 16 7.84 30.63
<1 26.03 7.01 31.36
< 1 23.94 5. 57 29.74
------------------------------------------------------------± REA - Relative enzyme activity (Viscometric)
** EU - Enzyme units (DNS Colorimetry)
'
Table-7(c): In vitm cellulase activity of P7eurotus sajor-caju.
Incubation Period (days)
5
15
20
30
*
Malt e>:tract medium Straw medium
nrA* 1\C
i 1
6. 11 G
: 1
( 1
( < ''
< 1
cu** REA*
13. S7 < 1
30.86 6.17
28.26 18.34
27.03 11. 56
25.46 7. 01
25.91 7.66
REA - Relative enzyme activity (Viscometric) ** EU - En:yme units (DNS Colorimetry)
Eu**
26.53
28.08
73.41
53.20
39.73
46.52
St~aw mediu~ was found best to support cellulase pro-
duction by P7eurc~us spp. All species were invariably found
to pn,duce high amount of cellulase beyond 10 days and up to
15 :Jays. The ma;,imum enzyme units were recovered after 15
days of growth a11d then decreasing trend was found up to 30
days of growth.
Tfle cellulolytic potential of Pleurotus species varied
a:1d suggested superiority of o florida among them.
ENZYME UNITS 40 ·---- ·-----·-··--- ------- --
NIDI A
35 _.... Malt extract
-+- atraw mlcllum
20L--------L _________ lL--------~------~------~ 5 ro ~ 20 25
DAYS
Fig.5b:/n vitro cellulase activity of Postreatus
ENZYME UNITS
30
80~----------------------~==========~
60
40
20 -
o+---5
MEDIA
--*"" Malt extract
-+ Straw medium
10 15 20 25
DAYS
Fig.5c:/n vitro cellulase activity of P sajor-caju
30
2. Spawn Level
a. Effect of spawn substrate supplementation
Suprolernentatiur-, of spa1m base with rice bran and wheat
bran (5% w/w) as woll a~ un~upplemented controls were only
found ·~u result -jr~ cumrolete ;;pawn growth (Tables-8a,b & c).
Table-B(a): ~ffect of spaw~ substrate supplementation on growth oF Pleutuius florida.
E~ta~t of growth (% colonisation in days) Substrate Supplement
2
U;·ea (0.05% whi) 20
DAP (0.01% w/w) 20
NaCl (0.2~~ w/w) + MgS047H20 10 (0.1% w/w) + K2HP0 4 (0. H;
W/W) + KNOo J
(0.2% w/w)
Peptone + Dextrose (0.2% w/w)
Rice bran (5% w/w)
Wheat bran ( 5% w/w)
Control (1-~a~ze grain)
10
10
10
10
4 6 8 10 12
30 SG
30 SG
30 50 SG
30 SG
50 80 100
60 90 100
40 70 80 100 -----------·-----------------------------------------------------------
SG - Static growth. Data given is Mean of three replicates.
able-3(b): Effect of spnwn substrate s:1pplementation on growth of P!eurotu;; o,;t/ e.;tus .
. .. - ------- - --------- -----------------------------E~te~t of growth (% colonisation in days)
:.bstrate Supplement 2
Jrea (0.05% w/w)
20
~aCl (0.2% w/w + MgS047H2o 10 (0.1% w/w) + 1: 2HP0 4 (0.1% w/w) + KN0 3 (0.2% w/w)
Peptone + Dextrose (0.2% w/w)
Rice bran (5% w/w)
WhEat bran (5% w/w)
Control (Maize grain)
10
10
30
10
4 6 8 10
30 50 SG
SG
30 SG
~0 SG
50 80 100
60 90 100
40 70 80 100
SG - Static growth. Do.ta given is Mean of three replicates.
12
P. florida and P. ostreatus showed similar growth and
colonised best in only 8 days on both supplemented spawn base
where as P. sajor-caju required 10 days for rice bran
supplemented base and 8 days for wheat bran
supplemented spawn base. Unsupplemented maize grains were
colonised -: n 10 days by all Pleurotus spp. Other
supple,.,2ntat~ons ·.-~en: f .. .:1d to result in static growth after
4-6 d:ty:: of iJ"Ov.tl, by all three P7eurotus spp.
Table-8(c): [fFcct of spawn substt c~te supplementation on gt·owth ;,,f P7eur,Jtus :;ajor-caju.
Cxtet:t of growth (% colonisation in days) Substrate S~pplement
Un::a (0. 05~~ ·w/w) 1 0
DAP (O.Oi?~ ~,.;;'w) :'0
NaCl (0.2~£ v;/v;· -r t(;S'J 11 7H('\C 10 hj L.
(0.1?~ '1'1/W) + l(,...t-lF'C,~ (C.~?~ " " w/w) + KN0 0 (C.2c; ·yJv~)
~
Peptone + Dex~rose (0.2~~ ~/w)
Rice ~ran (5% w/w)
Wheat bran (5~ w/w)
Control (Maize grain)
1 0
10
20
20
4 6
30 50
:;G
30 SG
20 SG
50 60
50 80
30 50
SG Static growth.
8
SG
90
100
70
Cat a given is t~ean of three replicates.
3. Crop Level
10 12
100
100
a. Effect of spawn prepared with different grains
Data of effect on cu1tivation (Table-9; Fig. 6) showed
tl~at ~a~ze grain spawn was required minimum time for complete
. Table-9: Effect of spawn prepared by different substrate on Spawn run, Total yield and Biological Efficiency of PTeurotus spp, by Bag method of cultivation.
~c i es Spawn T'me rresh Wqht BiJiorcal Average size Average substrate required y:eld (g/kg EFicie~cy cf sporophores number of
for com-- stra•) (I --------------- sporophores p1ete Stipe Pileus spa•n rcr Lenght Diameter (:ays) (ern) (em)
J owar g r a i n 20 ~: 0. s ~ 5. c 0.80 6.23 86
r:orida ~1 h~at g r a~ ~ ! 9 :87.5 38. ., 0. 81 6' 50 73
Ha i ze g r a i ~ 17 6:15.2 63.:2 0.80 6. 41 92 -----------------------------------------------------------------------------------
J 81'iar g r a i n 2S ~:a.: 57. s "! 0. 82 7,13 89
ostreatus Wheat gra;n 19 404' 8 40. 48 0.76 6.92 65
M ' .. a , ze g r a i n : 5 035.0 53. 50 0.85 7.20 85
----------------------------------------------------------------------------------~owar g ra i r 22 468.: 46.87 0.80 6.81 72
sajor:.caju Wheat g r a; ~ 1 s 455 'c 45.50 0.83 6.50 76
Mai:e grain .. 51 i. 2 51. ! 2 0.80 6.67 86 :,'
·----------------------------------------------------------------------------------C2 at 5% fer Substrate - 20. 14
for Species - 20.14 for Substrate x Species - 34.88
spawn run among spawn used. This was recorded in 17, 15 and
17 days for P. florida, P. ostreatus and P. sajor-caju,
respectively. Maximum biological efficiencies were also found
by maize grain spawn for P. florida (63.62%) and P. sajor-
caju (51 .21%) but P. ostreatus recovered maximum biological
efficiency (57. 17%) by jowar grain spawn. Use of wheat grain
spawn was found to result in lower fresh weight yields and
biological efficiencies for all three P7eurotus spp. It was
also observed that average size of sporophores was not af-
fected by different grain spawn used. Average number of
sporophores were found to suit their corresponding biological
efficiencies.
b. Effect of spawn substrate supplementation
Data on effect of supplemented spawn on cultivation
showed that supplementation of spawn base could not result in
superior effects over control (Table-10; Fig. 7). Almost
similar time was required for complete spawn run by all three
P7eurotus spp. Highest fresh weight yields and biological
efficiencies were recorded by unsupplemented spawn. P.
florida performed the best in respect to biological efficien
cy (64.80%) which was followed by P. ostreatus (45.71%) and
P. sajor-caju (51.48%), respectively. The average size of
sporophores were not
-·~
Table-10: Effect of supplemented spawn on Spawn run, Total yield a11d Biological Efficiency Pleurotus spp. by Bag method of cultivation.
;ec 1 es
f!or:'da
, ~ ~: e bra c
~st•eat~s .!.;\ 11; 3. t 1-r;:.,.. c' _.,
... ' .... ~ ~" 1 "~ '• c'
~ '):
' '" ~ ~,-a·~
S&w:;J1<5>1;J ._,,: :3. t br3.~
::·~~~:·
. '
"" i.
2'
" ,,
13
• s
" ,,
'.A 1 ,J I~ 'I,~ ,• , c u , :l· r. :;1
'" w ~ v'.::.
! ')
:~~.2
Jf'lfl II
-:".::..~
!2 t.:
~5 i' 1
., '' -.
~ 1. 2 2
51 .~:3
:.,rage size of sporophores
Average number of
-------------- sprophore Stipe Pileus Lenght Diameter
[em) I em)
0.80 7.12 102
C.SI 7.55 88
0.80 6.52 :22
0.81 8. I 2 75
0 '82 8.05 54
0.84 3.20 32
-------------------------------------
~3.22 0.78 7 " ' • w.,) C7
•'l !:!: .,.k.~~ 0.80 8. 21 59
J ~ 7. .,. V, > I 0.80 8.03 69
--------------------------------------------------- -------------------------------
-'.'e·:e s··~~-~ ,.,.~h s~~~·s,~e::t
a~ ~~ tcrS~~~·s~eG~ - 41.50
- 41 . 50
S~e:i e3 - NS
800
600
400
200
0
700
600
1500
400
300
200
100
0
TOTAL YIELD (g) ------------------------ ------- ------------,
P .florida P. otreatua
OAAJNI
8 JOMA
CJ WHI!AT
- MAIZI!
P. aajor-caju
Flg.6:Effect of different grain spawn on total yield of Pleurotus spp. ·
TOTAL YIELD (g)
-CONTROL
-RICE BRAN
P. florida P. oetreatue P. aajor-caJu
Fig.7:Effect of spawn substrate supplementation on total yield of Pleurotus
spp.
affected by supplementation where as average number of
sporophores were found to s~it their corresponding biological
efficiencies.
c. Effect of certain nutrients on P. florida
Cer·tai,l nutrie ts were studied for cultivation of P.
florida ur.Jer both the Bag and Tray methods of cultivation.
It was found that all the amino acid treatments respond
ed wel.l over ccmtrol under both the cultivation methods
(Table-11; It was observed that time required
for complete spawn run varied with different amino acids.
Minimum time of 7 days was reequired on methionine treatment
under both the cultivation methods and on arginine treatment
by tray method. The result showed preference of P. florida
towards utilization of tryptophan and lysine. Application of
tryptophan produced highest biological efficiency of 58.2% and
71.14% under Bag and Tray methods, respectively. Average
number of sporophores were not·found correlated with their
corresponding biological efficiencies. Application of methio
nine produced maximum sporophores (94) followed by phenylala
nine (89) and tryptophan (70) under Bag method whereas appli-
cation of phenylalanine produced maximum sporophores (36)
followed by tryptophan (35) and methionine (30) under Tray
method of cultivation.
Table-·:: offect of Amh ac';s 'Pn S>m run, Tc~al reid and Biological Efficiency of P!eurotus florida.
~------------------------------------------------------------------------------------------------------------------Bag met~cd of cu~t'vatlon Tray method of cultivation
reatment C.51 w/1)
------------------------- --- ------------- - ----------------------------------------------------------Time required Fresh \eight Biclag'cal Average Time required Fresh weight Biological Average for :omp1::~e; ; :aid : ;! Ef~icisncr nl.r~er of for complete yield Is/ Efficiency Number of spawc ('U~ 5JC g stra\\ ~ :%: &>:'ophores spawn run 250 g straw) (I) sporophroes
, 'a • ) j ·' Y~ (days)
--------------------------------------------------------------------------------------------------------------------rgirine l')~'r '
L J I , ' 48. n 138' 15 55.98 25
)Sine 13 270.~ 54'(: s I 0 148 .I? 65.38 29
,., "" 't! ,1,..:, 137.25 5UO 30
1eny:alar:~re 12 2 39' 3 4 7' 2 6 89 9 159.75 53.90 36
-yptophan 12 2 g 1 '~ 58.28 70 I 0 177.85 11.14 35
! 91. ! 5 55 10 I 02, 75 41' 10 21
CD at 5% ~cr ~reatment - I o.?O for ~·~ath:d o~ cJ~~ivation- 18.54 f:;r Treatment x i~ethod - NS
Data on vitamin treatment revealed that P. f7orida
responded well over control for application of ascorbic acid
and biotin under both the cultivation method (Table-12; Figs.
9a & b). Minimum time of 10 days was recorded for complete
400 TOl:_AL_YI_E L_D ~-g) ___ ·-· ~-· ~~ _____ , __ 141_E_._N_o • ..;:.O_F..,:8.:_P~ORO.::.P:...:H..:..O.::.R..:..E::;.. 200
300
200
100
0
190
170
150
130
110
90
70
50
[ f,\\\Wl VIE~~- --8POROPHORI I
CONTROL ARG LYS MET PHE
Fig.Sa:Effect of amino acids upon total yield & sporophore number of P. florida
under bag cultivation
1110
100
150
0 TRY
TOTAL YIELD (g) AVE.No. OF SPOROPHORE .~-~~·~:~~---------~~~~~~~--.100
I HYIELD -SPOROPHORE I
CONTROL ARG LYS MET PHE
Fig.Sb:Effect of amino acids upon total yield & sporophore number of P. florida
under tray cultivation
80
60
40
20
0 TRY
spawn r~n wl10n trc~tcd with ascorbic acid under both the
cultivation melit)CS, as well as in control under Tray method.
Applicat'otl of biotin produced maximum biological efficiency
of 51. ~8% and 74.6S% u11der Bag and Tray methods, respective-
ly. Avet-age number of sporophores were found to suit their
correc;~0ndi1t9 biological efficiencies under both the cultiva-
Tablc-12: Effect of vitamins upon Spawn run, Total yield and Biological
. '
!' n" ~,, l H\ "v : • ~ {_
Efci c ~ .sncy of f' 7 curotus f7orida.
" ~~ "7
'· 'J w -,o,;;:
255.~ 5; '; 8
~; ~ . ~3.22 .;, "v' 0
2~?.3~ j; .,.., 't1, ,'I
Tray method of cJltivation
Average Time required Fresh weight number of for complate tield (g/ sproplmes spa;n run 250 g stm I
I days I
H 10 157.72
64 12 186.65
43 12 142. 81
44 10 152.95
Biological Efficiency
(%)
63.08
7 4. 65
57.12
6C . 98
Average Number of sporophroes
28
'? '"
" ,,
22
TOTAL YIELD (g) AVE.No.OP' SPOROPHORE 360 ,-----------=:__ _________ ___.,:_.:;:_;_:~..:..;_:.;_;,;,.:.:...:....:..:.:.::.:; 100
300
250
200
CONTROL ASCORBIC ACID BIOTIN RIBOLNIN
Fig.9a:Effect of vitamins upon total yield & sporophore number of PJ/orlda under bag cultivation
80
40
20
0
TOTAL YIELD (g) AVE.No. OF SPOROPHORE 21SOr-------~=-----------------------------,100
200 I CJ Y;ELD -SPOROPHORE I
150
100
ISO
0 L__J, __
CONTROL ASCORBIC ACID BIOTIN RIBOFLNIN
Fig.9b:Effect of vitamins upon total yield & sporophore number of P. florida under tray cultivation
80
60
40
20
,,.,
A clear differat1ce was observed on crop level due to
application of differen~ chemical nutrients (Table-13; Fig.
1 Oa & b) • Mir.im~m tirr,e of 8 days was recorded for complete
spawn run on applic~tion of glucose under both the
1118 thods. 7able-13: EffE:ct JF chemkal nu~"rients upan Spawn run, Total yield and
3i.)logical :ff;cicncy of Pleurotus florida.
... -.,.~
~!: " 1.' "~ ... '-'"' oi \'::0.
,, ; "1 ,I I " I j ' ~ I \• , :;\1
" ' ' ~ >-. J
C!: 0 1 , •: A" 1 owlv:l ,_,(;0,
';:'
vier of ~ime req~ired Fresh ~ejght
for complete yield (g/ :~:-·ophJrss spa;.;~ rJn 2EO g s~raw)
(days:
'. 3 151.42
s:o1ogica~
Effici-&~GY
rx:
64.54
sporophroes
l·;; •·, '·
11 :;,1 :z 9 125.59 54' 2 3 '•:• J I
10 ~27.52 :4.62
5: . 3} 12 125' 90 54. 1 e J,
z::.:: .. I, 15 2. C E 61. c 6
TOTAL YIELD (g) AaiE.No. 011 SPOROPHORE 350~---------~--------------~--~~~~~~200
300
250
200
150
100
50
@YIELD - &POROPHORe j
Flg.10a:Effect of chemical nutrients upon total yield & sporophore number
of P. florida under bag cultivation
100
50
· TOTAL YIELD (g) AVE.No. OF SPOROPHORE 250r---------~-----------------------------,100
200
150
100
50
I E:J YIELD -SPOROPHORE I
Flg.10b:Effect of chemical nutrients upon total yield & sporophore number
of P. florida under tray cultivation
97
60
40
20
0
J i Ffen::n:.. :lL-.~r i8l.l._ ' . ,'_• ·~ :'·Jc,i\d to well support to biolog--
_p '1'~0 L,,,_,, e·L·-._,,t and glucose were found to
hir lc:sical cJ;;iciencies (74.62% and 64.54%
. 1.::0:..•ect i '/;;; l ·.~ ~ ·Y·c,r - "'' r' ., r·c~~~r Tray method of • • •• - -'·•''- -' -•<C. cultivation.
1. Gross Chemical and Biochemical Characteristics
IL
..
1\ ._ .. ·.:>
,r ~·
:. I ;;JiiS:'" _.;.,.I'-" -•' ' ~ -'
. .
~ ,.. ~ ~ ...... "' ' \- ·-- ' . ...:..,:;; "
s.1._;._ ~--c ....... _,
by u3ing dry powedered mushroom
The contained considerably
:F IJr~c:eitio wi.ich was found highest in P.
fc~~cwe~ ~i 0 ostreatus (22.33%) and P.
Carbohydrates were found
higher
.,j··~ ... ; ···:...~,-:-·,
• •_( '-. , \' f,:_• I J
Pieurntu.~ :3,:;1. r. ostreatus contained high
It was
_, ~' high crude fibre
,,., f"\70/\ I ~· • ~' I '~ 1 ::~~c:tc than other two species.
c.b1e-"!: G:···)"'· .;:,e11r'::c' .:rJ t:io:h,ei':i.;al cr.aracteristics (%dry weight) of P7eurotus sp~
,.... ' '- .J.. ~ ~ ,. : '\.-.~Ill
Lipid
~2~g~L; (N~4.:3) S:~J~18 Total Total Reduc- Total Relative sugar i ng phospho
sugar 1 ipid
Crude Fibre
Total Ash
19.9J 50.97 11.61 2.125 2.598 65.14 3.20 9.07
89.-17 ~0.57 58.04 16.89 1.17 2.076 68.15 2.975 6.125
00 ')/"" ·_.u • '-)
,.., -1 1 f'l .;... '. '~
Amino acid Profiles
3Z.30 61.55 15.25 0.59 2.520 66.07 2.05
~hE paper chromatograp:~i: analysis of free and bound amino
~id coo1ros~ti:c·-, '1 d:-y sporophore ext·racts showed that all the
:ec:Jrot.,:o spe:.-ies possessed a'.nost similar amino acid compostion
.,...?..b"'s-;5; The bound amino acid composition was of
7.325
Cystine At-~ inir1e '31/c.ine GL .. L.·iWic r~c:d
Al2.11ine Aspartic Isoleucinat Serine Pr-ol~ne
Metilinonine* Tryptophan' :yt·os i ne* 'v'a l i ne:t
· · 'd·. it ~luding 7 essential amino ,1cids.
1, 2~ -- :u l•.Jur.c ,Jtlit'" _,,_ id profile of Pleurotus spp.
F': '..J':t:: j II
/,r~i nr)
Jc i c!
+
+ +
+ +
+
~ ee ' An, i tv:
~ ... : ~ d
+
+
+
+ +
+ t-
+
Jl1u~ein
Am;no ·)C i d
+
+
t
+ + +
+ +
+ + + + +
+ + + + + +
Free Amino acid
P. sajor Cdju
Protein
+ + + + + + + + + +
Amino acid
Free
+ t
+ + t-
+ + + + + +
Amino acid
Phenylala~+ne:t:
+ + + + +
Leucine* Lysine*
+ + + + +
+ for presence; - for absence * Esssnt1a1 amino acids
+
The P7e~rotus spp. ware found to possess 11 common amino
acids ''. their free amino acid compostion which included 5
amino acids. Proline was found absent in free
a.rn; ·:, c;:~::l corr;postion of P. florida. Isoleucine, methionine,
ty,·csine and leucine were found common in all Pleurotus spp.
Valine, phen;lc•hr~ine a.:1d lysine were additionally present in
protein ami~o acid e/tracts and tryptophan was additionally
present jr: free am·ino acid extracts of all three Pleurotus
spp.
3. Pr·otei n Profi 1 e of f'leurotus spp.
An electrophoretis protein profile analysis was done in
fresh cellular homogenate to educidate genetic relationship
among them. The electrophorograms are shwon in ~~ate 4& ~d
Fig. 11. It shwoed that P. sajor-caju had 13 clear protein
bands lvhereas P. florida and P. ostreatus had 16 and 12 bands
respectively. On comparison it appeared that P. sajor-caju
and P. florida have 3 common bands (2,3,4,8,9, 11, 12, 13)
whereas P. ostreatus was similar only in respect of 3 bands
(3,11 and 12). Pleurotus florida and P. ostreatus resembled
in 2 bands only i.e. a and b. Protein specific only to P.
florida are marked as 'y' and those of P. ostreatus as 'x'.
In this respect P. florida has only 2 specific proteins and
P.ostreatus has 3 of them. This protein profile established
distjnguishing differences in their genetic rrrakeup to accord
thEm separate specific status. Genetic nea~ness of P. flori
da ~>.ith P. sajor-caju become evident.
101
13 13.---1
1 2 3 Fig.11: Protein ·profile ·of Pleurotus
spp.
1. P. sajor-caju
2. R ftorida
3. P. ost.reatus
____ ___)
4. Mineral Nutrient Status
Analysis for the various macro and micro-nutrients
present in the mushroom tissue was made. The results ob-
tained from ash analysis on Plasma scan have been given in
table-16.
Table-16: Analysis of mineral nutrient* in Pleurotus spp.
Mineral P. florida P. ostreatus P. sajor-caju
I r-on 7 5 6 Manganese BDL BDL Magnesium 220 195 220 Zinc 14 1 1 1 3 Cadmium BDL BDL BDL Cobalt BDL BDL BDL Copper 2 1 1 Calcium 85 72 50 Phosphorus 845 343 339 Sodium 39 30 26 Nickel BDL BDL BDL Molybdenium BDL BDL BDL Vanadium BDL BDL BDL Potassium 5025 3430 2955 Aluminium 10 5 1 Boron 6 1 1 -------------------------------------------------------------
* - Values given are in ppm. BDL- Below detection level.
J 1-: =
It was found that P. florida possessed a much superior
level of both the micro and macro nutrients in its tissue as
compared to P. ostreatus and P. sajor-caju. The amounts of
iron,zinc, copper and boron as well as of magnesium, calcium,
potassium, sodium and phosphorus were significantly much
higher in P. florida. Some of the micro nutrients 1 ike the
manganese, cadmium, nicl,el and molybdenum were found only in
negligible amounts (BDL-below detection level). However, the
quantit·ies of iron, magnesium, zinc, calcium, phosphorus,
sodium and potassium in all the three species appeared to
have a good dietry value besides the protein and carbohydrate
r· i chness.
PRESERVATION STUDIES
1. Effect of Storage Condition and Temperature
The fresh sporophores of three P7eurotus spp. were stored
under three different storing conditions and temperatures.
It was found that all three species showed somewhat similar
pattern for weight loss, colour change and change in texture
(Tables-17 a,b & c). Minimum weight loss and cf·,ange in
' ," '
al: three F'l<-t<~ ,,tu:~. c·PP ~ P~ Florida showed minimum weight
loss cmr:l '\,,_\1\Qe in t'2> tur-e under unperforated pacLage and
A 11 storing
cc1nditi<J11::' were found ;;ill,ilar·l; affect the sporophore colour
chc!llge and ·~to:~ .. 1ge at 2S 23°C sho'.ved rna>, imum colour change
Table-17(n): Effec:t cf un~e··forateJ polythene package on shelf 1 ife of sp:;r~ophor·es * of Pleurotus spp.
S~:ra;e
% wel;~t 1""" ,vC>;:,
-------------------------------------
Cc~c-.r Tex~~'"e % V!·el;r.t Colour T2.xt.~rc
:oss
3;~t:1e :5.64t)."'~ Pa~e
1 e: 1 Vii
--------------------------------------------------------------------
s~:rage.
Table-17(b): Effect of per·forated polythene package on shelf 1 ife of sporophor·es* Pleurotus spp.
------------------- ------------------·------------- - -------------------------------------------------P. cs~rea~us P. sajor-caju
~e~perature - ------------------------------------------------------------------------------------------I we:ght CcloJr ~e;t~re % wei;ht Cc1~ur Te,o;t~re %weight Co1ot.~r Text~re
loss less loss
8e~o·• OoC 13. :0~0.26 Pate :e!'e< Lrgid 5.5C!_Q.07 3reyis~ Flaccid 9.21'.:_0.25 Grey Flaccid w h ~ te
5-: OoC ~~.?0:_0.8€ Pa1e ya:1o~ Fiacc~d S.5i.,O.C6 Pa'e Flaccid 12.51!_0.73 Pale Flaccid 1 e 1 I o; ye 11 OW
25-!So: 23.8±_2.41 ve~low'sl: Brittle '9 1c:_U5 Yellowish Brittle 19.90';.0.30 Yellowish Brittle Brown Brown Brown
~- Data ~eccrded after 95 hcurs cf storage.
2. Effect of Steeping Preservation :--
It was found that all species of P7eurotus showed
similar response towards steeping preservation (Table- 18).
The fresh weight of sporophores were found to increase after
storage. Maximum weight increase was recorded in P. florida
followed by P. ostreatus and P.sajor-caju respectively.
Sporophores of P. florida were found to show colour change
after 15 days of storage onwards while sporophores ofP.ostreatu§ard
P.sajor-caju showed colour change after 10 days onwards.
Table-17(c): Effect of unpacked storage on shelf 1 ife of sporophores * of Pleurotus spp.
------------------------------------------- ------------------------------------
st,rage te1peratm
P. ostreatus
------------------------
P. sajcr-caj'
%weigh: ;ass
~olcur Te:.;ture % v;elght Co,:Jr Texture % wei;ht Colour Text~re
1 oss
Below 0o: 15.~7:_0.73 r~:: 1e:low Flaccjd ·~.3Q-t_O.i3 ~aie Turaid 10.26t.1.3~ Pale '!ac:id
re 11 ow ye: lo~•
s-::oc 19.C:_: .52 Pale 1e:'c·; 3r'ttle 19.4Qt0.21 Pale Brittle 12. ~2!_0. 18 Ye~1c·~is~ F~a~cid
y e :1 o,r Brown
25-2SoC
* - Data rac:r~ed after 95 ~ours of stostorage.
Sporophores of P. florida and P. sajor-caju were found to
turn soggi after 15 days of storage while sporophores of P.
ostreatus were turn soggi after 20 days of storage.
Tab~e 18: Effect of steeping preservation on sporophores of rleurotus spp.
Species Period of Fr·esh weight Colour Texture storage (g)
(days)
0 50.00 White Turgid 5 52. 82-t_O. 0012 White Turgid
P. florida 10 53. 77-t_0.003 White Turgid 15 53. 75-t_O. 041 Pale Yellow Soggi 20 54.10-t_0.036 Pale Yellow Soggi
0 50.00 Greyish White Turgid 5 51.31-t_0.006 Greyish White Turgid
P.Ostreatus10 51. 82t_O. 008 Pale Yellow Turgid 15 52. 635-t..O. 0035 Pale Yellow Turgid 20 53.10!.._0.0021 Pale Yellow Soggi
0 50.00 Grey Turgid P. sajor- 5 50.360-t_0.001 Grey Turgid
caju 10 51.315-t_0.00015 Pale Yellow Turgid 15 52. 050-t_O. 0052 Pale Yellow Soggi 20 52. 300-t_O. 0066 Pale Yellow Soggi
3. Evaluation of dehydration process
It was found that different processes of dehydration
showed variable effects over dry weight, colour and rehydra-
tion capacity of Pleurotus spp. (Table-19; Fig. 12a, b & c).
Maximum dry weights were obtained by sun drying similarly
maximum weight recovery after rehydration was found
Table-19: Effect of different processes of dehydration on weight loss, Colour and Rehydration of sporophores of Pleurotus spp.
Weight loss
I I)
- --------------------------------------------------------------------------------------------
WeiJht rega; ~.ed after Rehydrati8n
Weight 1055
I I I (%I
P. ostreatfus o, sajor-cajl
- -· ------------------------------------- -----------:clour Weight
regained after Rehydration
'ie i ght loss
[I) [I)
Cclcur
------------Weight regaine: after Rehydratlo:·
(%)
---------------------------------------------------------------------------------------------------------------------------------Sun Or]ing
Me:hanical Drying
Blanching 1
Drying
Bla~ching 1
Citirc acid treatment • Drying
Blanching t
Sulphitat'on • Drying
ss.6nuo5 Light Brawn 96.4t_O.C5 36±_0.81 Light Brown 96.9t_D.031 88.88t_8.13 L:ght Brawn 95.7;_~.1:2
B8.98t_0.007 ligH Bro·.n 9Ut_O.c:3 SU3t_0.06 Light Brown 97.62t_O.C41 89.4t_C.C04l'gb~ Brown 95.6±_Ui2
93. 596±_U42 Bra'n 8S.24t).~E 3U1!_D.67 Brown 90.6!_0.13
92.11!_0.16 Dark Brow~ 88.6±_0. 23 88.9?2!_1.05 Brown 34.2!_0.29
92.99±_0.68 Pale "e!low 82.i:_O.C47 89.8:.U05 Pale Ye'low 83.9t_O.II5 90.94±_0.16 Pale Yellow s:.o,_:.::"
--------------------------------------------------------------------------------------------------------------------------------
by sundrying in P. sajor-caju whereas P. florida and P.
ostreatus recovered maximum weights by mechanical drying.
Blanching + citric acid tretment before drying showed maximum
weight loss and minimum rehydration for P. florida whereas
WEICIHT L088( .. ) 100
I -DRY WIIGHT
WEIGHT RECIAINED('It) 150
D RIHYDRATION WIIGHT I
90
88
SO MD B•D B•C•D B+S+D DEHYDRATION PROCESS
Flg.12a:Evaluatlon of different processes of dehydration for P. florida
130
110
90
WEIGHT LOSS('Io) WEIGHT REGAINED('It) 100.-------------------------------------~120
95
90
80
76
TO
I -DRY WEIGHT EJ REHYDRATION WEIGHT I 110
100
90
80
"--..._,70 SO MD B+O B•C•D B•S•D
DEHYDRATION PROCESS
Flg.12b:Evaluatlon of different processes of dehydration for P. ostreatus
: . c
WEIGHT LOSS('!tt) WEIGHT REGAINED(-.) lOOr----------------------------------------•110
[•DRY WEIQHT W~EHYDRATION WEIQHT I 106
96 100
95 90
90
86 86
80
80 76 SO MD B+D . B+C+D B+S+O
DEHYDRATION PROCESS
Fig.12c:Evaluation of different processes of dehydration for P.sajor-caju
111
Blanching + Sulphitation before drying showed maximum weight
loss and minimum rehydration for P. ostreatus and P. sajor-
caju. Blanching + Citric acid treatments were founds to
result in maximum colour change for all P7eurotus spp. sporo
phore& were found to show minimum colour change when they
were given Blanching + Sulphitation treatment prior to dry
ing.
Strain Improvement Studies
Attempts were made to improve the strains of the selected
mushrooms with a view to obtain higher yield/biological efficiency
and to reduce sporulation which is known to have some toxic ef
fects. This was carried out by following 1) Somatic hybridization
in cross inocultions and 2) UV-irradiation to cultures.
sults have been given in tables-20 & 21 and Figs. 13 &
14 and Plates-5,6 & 7.
The re-
In cross inoculations the species were not found completely
compatible for somatic hybridization, but there was an evidence at
the borders of the colonies where they fused. This synnematic
mycelial mass was used to develop spawn for mushroom production.
The data of cultivation was not promising in any respect (Table,
\ \~
20). The yield and biological efficiency was rather of a low order
as compared to the parent strains.
able-20: Evaluation ot dJftere~t cross inoculated cultures for spawn ru", yota )ie'd and
Biological Efficiency and Average ~umber and size of Spcrophores.
--------·--------------------------------------------------------------------------------------------------
Culture Time Total Yield * Biological Size of Sporophores Average Remark
Requ 1 red (fresh weight, g) Effic·ency ------------------- Nu'!lber of for ---------------------- (%) Average Average sporophroes compelete R-I R-II Average stipe Pileus spawn run e11gth Diameter
(days) (em) (em)
P.florida x P.ostreatus 18 198.2 200 .5 199 .3 39 .8' 0.8 5.34 42 UP
P.florida x P.sajor-caju 19 212.5 210.6 21' .7 42.34 0.82 6.87 43 UP
P.ostreatus x P.sajor-caju 21 146.7 162.3 154.5 30.90 0.75 6.18 61 UP
-----------------------------------------------------------------------------------------------------------UP - Uncolonized patches * - Yield given is sum of two flushes CO at 5% for cu 1tures - 20.69
Ultra violet irradiation to produced 8-9 distinct types
of co l oni es in pure cultures (Plate- ~ ). All of them were
grown on straw compost to obtain data on sporophore produc-
t ion and the associated characteristics. In this case PFS-6
was a sterile mutant and did not form any sporophores; PFS-?
113
•
was obtained as a sporeless mutant which had a very low yield
potentia l and the consequential
r ..
ble-21: Evaluation of different isolates of UV irradiated cultures for spawn run, Total Yield and Bio log ical Efficiency and Average sizeand Number of Sporophores.
-----------------------------------------------------------------------------------------------------Culture Time Total Yie ld * Biological Stze of Sporophores Average Remark isolates Required (Fresh ~~eight , g) Effic iency ------------------- Number of
for ~--------------------- (') Average Average sporophroes compelete R-I R·II Average stipe Pi 1 eus spawn run length Diameter (days) (em) (em)
-----------------------------------------------------------------------------------------------------
PFS-1 11 262 .8 24 7. I 254 .5* 50.99 0.82 7.03 23
PFS-2** It 96.0 100 .5 98 .2 19.65 0.80 12. 17 3 SD
PFS-3 11 205.6 224 .75 21 5.2 43 .03 0.80 6. 19 34
PFS-4 9 246.4 237 .5 241. 95* 48.39 0.80 8.1 5 22
PFS-5 9 227 .5 244 .6 236.05* 47.21 1.0 7.6 20
PFS-6 10 NF
PFS-7 10 174.3 184.25 179 .20 35.85 0.8 6.55 16
PFS-8 .
10 209 .8 224 .8 217 .30 43 .46 0.8 6. 34 15
HC/F 11 284 .4 272 .3 278.35* 55 .67 0.8 7. 7 4 23
-----------------------------------------------------------------------------------------------------SO- Sporulation deficient sporophore NF - Non fruitin g
* - Yield given is sum of 4 flushes t* - Yield given is of s;ngle f us CD at 5% for cultures - 20.62
114
T _ O~t~~=L_Y~I~E_LD_(=g) ________________ N __ E._N_O_.O_F __ SP_O_R_O_P_H_O_R-,E 250 r-
200
150
275
225
175
125
I - YIELD D SPOROPHORE I
P.flo.XP.oat. P.flo.XP.aaj. P.oat.XP.aaj. CULTURES
Flg.13:Evaluatlon of different cross Inoculated cultures for totai yield
& sporophore number
60
40
20
TOTAL YIELD(g) AVE.NO.OF SPOROPHORES ~----------------------------------------.&0
!•YIELD c:J SPOROPHORE I 40
30
20
10
75 0 PF81 PF82 PF83 PFS4 PF86 PF88 PF87 PF88 CONTROL
ISOLATES
Flg.14:Evaluatlon of different Isolates of UV Irradiated cultures for total ·
yield & sporophore number
115
biological efficiency under the cultivation conditions; PFS-
1, PFS- 4 , PFS- 5 and MC/F were characterized by good yield and
biological efficiency; and PFS-3 , PFS-7 and PFS-8 were only
average or below average then the parent strain. Further
studies are needed on PFS-2 for yie ld and other
considerations.
