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Characterization and optimization of fruit body yield in Volvariella volvacea

white strain

Article  in  Indian journal of experimental biology · February 2018

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Indian Journal of Experimental Biology
Vol. 56, February 2018, pp. 112-120

Characterization and optimization of fruit body yield in

Volvariella volvacea white strain
OP Ahlawat* & Harleen Kaur
ICAR-Directorate of Mushroom Research, Chambaghat, Solan-173 213, Himachal Pradesh, India
Received 04 June 2015; revised 03 March 2017
The paddy straw mushroom [Volvariella volvacea (Bull.) Singer] normally forms dark grey or brownish coloured fruit
bodies, and it leads to lesser acceptance in market compared to button mushroom with white or off-white fruit bodies. In the
present study, we attempted morphological and molecular characterization of the white strain of V. volvacea vis-a-vis brown
strains, standardization of its cultivation technology and nutritional profiling of fruit bodies. The white strain (GVv-01)
formed a separate clade in phylogenetic tree deduced from the 5.8S rRNA gene sequences compared to brown strains along
with a 21 nucleotides long deletion in ITS-2 region. It exhibited highest downward mycelial growth on paddy straw and
formed morphologically distinct colony on malt extract agar medium. It gave highest fruit body yield both at 25-28 and
30-35°C conditions with shortest first harvest period. It also yielded well on cotton ginning mill waste, cotton ginning mill
waste + paddy straw (1:1, w/w) and paddy straw based substrates. Its mean fruit body weight (16.89 to 23.31 g) on different
substrates was also higher compared to brown strains. The fruit bodies had higher crude fibre, ash and contents of calcium,
potassium, sodium, zinc, magnesium, copper and iron compared to brown strains.

Keywords: 5.8S rRNA gene, Mycelial growth, Paddy straw mushroom

The paddy straw mushroom, Volvariella volvacea
(Bull.) Singer, is known for its unique aroma and
texture, and grows well between the temperature
range of 28-35°C1. It is a fast growing mushroom1 and
has significant pharmacological properties, including
antitumor polysaccharides and immunomodulatory
lectins2,3. In global production of cultivated
mushrooms, it ranks 6th and accounts for 5-6% of
world production4. The lower biological efficiency
(about 10 to 15 % on rice straw) and lesser shelf life
compared to other popular edible cultivated
mushroom species are the most important limiting
factors for its proliferation at a large scale5.
The productivity in this mushroom is attributed to
the hydrolytic enzyme production potential6,7, quality
of substrate used, method of substrate preparation and
the growing conditions. The hydrolytic enzyme
production potential and quality of substrate used for
cultivation has direct bearing on its fruit body
production potential. However, other characteristics of
superior yielding strains viz., mycelial growth rate,
mycelial density, formation of aerial hyphae and
chlamydospores on growing medium vary under varied
growth conditions8. The colour of the mature fruit body
__________
*Correspondence:
Ph.: +91 1792 230767/230541; Fax: +91 1792 231207
E-mail: ahlawat22op@gmail.com
is one of the important attributes for the acceptability
among the consumers. In V. volvacea, the dark
grayish or brownish fruit bodies, which further
darken upon aging hamper its acceptability among
consumers. In the present study, we selected
Volvariella volvacea strain with whitish fruit bodies
and tried to optimize its productivity compared to
the strains giving traditionally brown coloured
fruit bodies.
Materials and Methods
Genetic characterization of the white strain of V. volvacea
The pure mycelial culture from white coloured
V. volvacea fruit body, collected from Port Blair
region of Andaman and Nicobar Islands, India was
raised using tissue culture raising technique on malt
extract agar (MEA) medium at 32±2°C. The
mycelial culture was molecularly identified by
extracting its DNA and amplification of the ITS
regions of 5.8S rRNA gene using PCR, followed by
sequencing and blasting of the PCR amplified
amplicon9. The improved consensus sequence was
blasted using BLASTn tool of NCBI10 and the
species against which highest similarity exhibited,
was considered as the identity for the target culture.
The consensus sequence was also submitted to
NCBI database with accession number KC142107.
 AHLAWAT & KAUR: FRUIT BODY YIELD IN PADDY STRAW MUSHROOM WHITE STRAIN
Sequence alignment, phylogeny and evolutionary relationship
with high yielding brown strains of V. volvacea
The improved consensus sequence of the ITS region
of 5.8S rRNA gene of the test culture (GVv-01) along
with consensus sequences of four high yielding brown
strains viz., OE-210, OE-272, OE-274 and BBSR-007
(DMR, Solan Accession Nos. DMRO-185, DMRO-
245, DMRO-247 and DMRO-463) of V. volvacea with
NCBI Accession Nos. JN086670.2, JN086662.1,
JN086663.1 and JN086677.1, respectively11,12 were
studied for variability in their nucleotide sequences by
using ClustalW2 tool of European Bioinformatics
Institute (EBI). Phylogenetic and molecular
evolutionary analyses were conducted using MEGA
version 5.013. The evolutionary history was inferred
using the Neighbor-Joining method14. The optimal tree
was drawn with the sum of branch length =
1.07128948. The tree was drawn to scale, with branch
lengths in the same units as those of the evolutionary
distances used to infer the phylogenetic tree. The
evolutionary distances were computed using the
Maximum Composite Likelihood method15 and were in
the units of the number of base substitutions per site.
Codon positions included were 1st+2nd+3rd+
Noncoding. All positions containing gaps and missing
data were eliminated. There were a total of 591
positions in the final dataset.
Mycelial growth characteristics of the white and brown
strains of V. volvacea
The mycelial growth characteristics of the white
strain of V. volvacea were studied against two high
yielding brown strains (OE-210 and BBSR-007) of
V. volvacea11,12. The strains were studied for radial
mycelial growth (diameter in mm) and colony
morphology on malt extract agar (MEA) in
Petridishes, and the downward mycelial growth (in mm)
along with aerial mycelial growth and chlamydospore
formation on pounded paddy straw filled in wide
mouth test tubes. For each strain, 6 mm uniformly
grown mycelial bit was placed in the center of the
MEA Petridish and the inoculated Petridishes were
incubated at 34±2°C for 7 days. Three replications
were kept for each strain. For downward mycelial
growth the pounded paddy straw was wetted
overnight and the substrate with 70-72% moisture was
filled up to 2/3 length of the wide mouth (40 mm) test
tubes, plugged and sterilized at 20 psi for 1.30 h. The
sterilized paddy straw was inoculated with one
mycelial bit of 6 mm/tube. The inoculated tubes were
incubated at 34±2°C for 9 days. The downward
113
mycelial growth was measured in mm along the
extent and the density of the mycelial growth. Three
replications were kept for each strain.
Enzyme assay
The activity of extracellular lignocellulolytic
enzymes of one white (GVv-01) and two high
yielding brown strains (OE-210 and BBSR-007) was
studied first by growing them on sterilized paddy
straw substrate with 70% moisture in flasks. The
enzymes were extracted from the mycelium-colonized
substrate in 50 mL phosphate buffer (0.1 M), pH 7.0
by keeping the buffer mixed substrate at 40°C for
30 min in an incubator shaker maintained at 100 rpm.
The extract was filtered through glass microfibre filter
(GF-C) and stored at 4°C for further use. The enzyme
assay was carried out in triplicate for all the enzymes
and data were subjected to statistical analysis using
AGRES software.
The cellulases were measured according to the
method of Mandels et al.16 as modified by Sandhu and
Kalra17. The reaction mixture for exoglucanase (FPase,
EC 3.2.1.91) comprised of eight filter paper (Whatman
No. 1) discs of 0.6 cm diameter in 0.5 mL acetate buffer
of pH 5.0 and 0.25 mL of the enzyme source. The
reaction mixture was incubated at 40°C for 4 h and the
reducing sugars released were measured by Nelson
Somogyi method18. The endoglucanase (CMCase,
EC 3.2.1.4) activity was measured following the above
method, replacing filter paper discs with 0.5 mL of 5 mg
mL-1 carboxy methyl cellulose. Xylanase (EC 3.2.1.8)
was assayed at 40°C following a method described by
Reese and Mandels19.
Laccase (EC 1.10.3.2) was assayed by adding
0.3 mL enzyme source to 2.5 mL of 30 µM Guaiacol
in phosphate buffer (0.1 M) of pH 6.0 and ∆ A was
read at 470 nm after incubating the reaction mixture
for 30 min at 25-28 °C against zero time control.
Polyphenol oxidase (EC 1.10.3.1) was assayed using
chatechol as substrate in place of guaiacol. One unit
of laccase and polyphenol oxidase activity was
calculated as change in absorbance of 0.001 min-1 mL-1
of enzyme source at 25°C, while that of FPase,
CMCase and xylanase as the µ mol glucose released
h-1 mL-1 of enzyme source.
Cultivation trial
The cultivation trials were conducted at
environment controlled mushroom growing unit,
ICAR-Directorate of Mushroom Research, Solan,
India. The first trial was conducted in July, 2011 by
114 INDIAN J EXP BIOL, FEBRUARY 2018
involving four high yielding brown strains and one
white strain at comparatively low temperature
conditions (25-28°C) using composted substrate
prepared from 1:1, w/w combination of paddy straw
(PS) and cotton ginning mill waste (CGMW). The
paddy straw based ready to use spawn and composted
substrate for crop raising were prepared as per the
method of Ahlawat et al.20. Five replications each with
two beds of 15 kg composted substrate were used for
each strain and the experiment was conducted in
Randomized Block Design (RBD). The second trial
was conducted in July, 2012 by using four strains
including three high yielding brown strains also used
in first trial and one white strain by following the
same protocol as for the first trial but at optimum
temperature conditions (32-35°C) recommended for
brown strains of V. volvacea. The last two fruit body
yield optimization trials were conducted during
cultivation seasons of 2013 and 2014 (July-September)
by using only white strain (GVv-01) of V. volvacea.
The substrates prepared with PS, CGMW and 1:1,
w/w combination of PS + CGMW were used for these
trials20. Beds involving three different quantity of
substrate/bed (12, 15 and 18 kg) were prepared from
all three types of substrates. Five replications each
with two beds were kept for all nine treatments (beds
with three different quantity of substrate/bed for all
three types of substrates) in RBD. The temperature
during mycelial colonization of substrate and fruiting
was 34±2 and 30±2°C, respectively. All cropping
conditions and protocols were similar to that for
brown strains of V. volvacea adopted for earlier study
conducted at DMR, Solan (HP)11. Data for time taken
for first harvest (days post-spawning), mushroom
(fruit body) yield (kg/q dry substrate) and mean
weight of fruit bodies (g) were recorded for 15 days
of cropping21. The data was subjected to statistical
analysis by single factorial ANOVA using AGRES
software.
Comparative nutritional analysis of the mushroom fruit
bodies of white and brown strains of V. volvacea
The randomly drawn fifteen to twenty fruit bodies
from one brown strain (BBSR-007) and one white
strain (GVv-01) of V. volvacea were got analyzed for
fifteen different parameters viz., dry matter, ash, crude
fibre, carbohydrates, protein, fat, vitamin-D and
7 different minerals (calcium, sodium, potassium,
iron, zinc, copper, magnesium and selenium) from
Punjab Biotechnology Incubator, Mohali, India
(a NABL accredited Agriculture and Food Testing
Laboratory, Govt. of Punjab, India). Standard
procedures of AOAC were used for the determination
of dry matter, ash, crude fibre, fat, carbohydrate and
protein content22,23. The protein content was
determined first by determination of nitrogen
percentage using Micro-Kjeldahl method, followed by
its multiplication with a factor of 6.25. The
percentage of crude protein, crude fat, minerals and
ash were combined and subtracted from 100 to obtain
the total carbohydrate percentage for each sample.
Mineral constituents (iron, copper, zinc, selenium and
magnesium) were determined by using ICP-MS
method 999.1022, while sodium and potassium by
using flame photometer. Calcium was determined by
Titrimetric Macro method, 910.0122. Vitamin D as
Vitamin D2 was determined using HPLC as per the
protocol standardized at PBTI, Mohali. It was
determined at a wavelength of 266 mm by using
Eclipse XCB-C18 column, DAD (UV-VIS) detector
and Methanol/Acetonitrile/Water as the mobile phase.
From 10 g mushroom sample, saponofied extract was
prepared and out of it 1/10th extract was loaded on
silica gel column, followed by elution with n-heptane,
drying and reconstitution using n-hexane/isopropanol.
The reconstituted sample was run on HPLC along
with requisite standard.
Results
Molecular and morphological characterization of the white
strain
The five strains used in the study including four
high yielding brown strains and one white strain,
formed three different groups in phylogenetic tree
deduced from the sequences of 5.8S rRNA gene. The
first group included all three high yielding brown
strains (OE-272, OE-274 and BBSR-007), while the
second group was having only one white strain and
the third group was with one high yielding brown
strain, OE-210 (Fig. 1). The study proved the
distinctness of the white strain from rest 4 brown
strains of V. volvacea. In nucleotides sequence
Fig. 1 — Phylogenetic tree derived from the sequences of the 5.8S
rRNA gene of the white and high yielding brown strains of
V. volvacea. The NJ-tree was constructed using neighbors joining
algorithm in MEGA 5.0 software.
 AHLAWAT & KAUR: FRUIT BODY YIELD IN PADDY STRAW MUSHROOM WHITE STRAIN 115

variability, the PCR amplicon of 5.8S rRNA gene of while it was brownish and white in rest two brown
white strain, GVv-01 was shortest in length strains (Fig. 3A & Table 1). On paddy straw, the strains
(617 nucleotides). It was short by 21 nucleotides varied in downward mycelial growth and it was highest
compared with three high yielding brown strains of 68 mm in white strain, compared to 60.11 and 43.11
(OE-272, OE-274 and BBSR-007) and 19 nucleotides mm in brown strain BBSR-007 and OE-210,
compared with brown strain OE-210 (Fig. 2). respectively. The extent and density of aerial hyphae
Compared with the four high yielding brown strains was highest in white strain, which formed light
of V. volvacea, the white strain, GVv-01 exhibited a brownish chlamydospores (Table 1). The strains did
deletion of 21 nucleotides in ITS-2 region of the 5.8S show variation at the level of the activities of
rRNA gene. This is an information of high extracellular lignocellulolytic enzymes and the white
importance, as along with other morphological growth strain exhibited highest activity of laccase, while
features, this long stretch of 21 nucleotides deletion lowest of exoglucanase and polyphenol oxidase
makes this strain an entirely a new strain. Such types compared with two high yielding brown strains of
of information are missing in mushrooms and V. volvacea.
specially in V. volvacea.
One strain each from three different groups as
depicted in phylogenetic tree was used to study the
variability in their mycelial growth characteristics on
MEA and pounded paddy straw. All the three strains
attained same level of radial growth (90 mm) on MEA
after 7 days of incubation at 34±1°C. However, the
strains varied with respect to extent and density of
aerial hyphae, and the colour of chlamydospores
formed. The white strain (GVv-01) formed highest and
dense level of aerial hyphae of creamy white colour.
The brown strain OE-210 formed least extent and
dense aerial hyphae compared to rest two strains. The
colony formed by white strain was creamy in colour,

Fig. 2 — ClustalW analysis of the ITS-2 region of 5.8S rRNA Fig. 3 — Characteristics of the white and brown strains of V.
gene of four high yielding brown and one white strain of V. volvacea [(A) mycelial growth characteristics; and (B) fruit body
volvacea characteristics]

Table 1 — Mycelial growth characteristics of selected brown and white strains of Volvariella volvacea on malt extract agar and paddy straw media
Strains Mycelial Growth Characteristics
MEA medium in Petridishes Paddy straw in Test tubes
Radial Aerial mycelial Chlamydospores Colony colour Downward Aerial mycelial Chlamydospores
growth growth mycelial growth
(mm) Density Growth growth (mm) Density Growth
GVv-01 90 mm 5+ 5+ Light orange Creamy white 68.00 2+ 3+ Light brownish
BBSR-007 90 mm 4+ 4+ Brownish circle Brownish white 60.11 2+ 2+ Brownish
OE-210 90 mm 4+ 3+ Brownish circle White 43.11 2+ 2+ Light Brownish
circle
[+ Scare; 5+ highest; - absent]
116 INDIAN J EXP BIOL, FEBRUARY 2018

Fruit body yield potential of the white strain of V. volvacea ability to give superior fruit body yield at lower
The initial evaluation trial conducted at temperature conditions, where the high yielding
comparatively lower temperature conditions, revealed brown strains failed.
lowest first harvest time of 11.93 days in white strain, The white strain evaluated on beds with three
GVv-01. It also gave highest fruit body yield (20.79 kg/q different quantities of substrate/bed prepared from PS,
dry substrate) and the mean fruit body weight (20.74 g) CGMW and 1:1, w/w combination of PS + CGMW,
compared with four high yielding brown strains of revealed lowest first harvest time of 9.0 days in 15 kg
V. volvacea (Fig. 3B &Table 2). In evaluation trial substrate beds of CGMW, followed by 9.67 days in
conducted at optimum temperature conditions (32-35°C) 18 kg substrate beds of CGMW. In trial-I, the beds
for V. volvacea cultivation, lowest time for first prepared from CGMW took 1-3 days less time for
harvest (11 days) was again recorded in white strain first harvest than beds of PS + CGMW and PS alone.
compared with brown strains. This strain also gave The difference in first harvest time was not so
significantly higher fruit body yield (41.12 kg/q dry significant in trial-II (Table 3). However, lowest time
substrate) and mean fruit body weight (20.40 g) was again recorded in beds of CGMW. In trial-I, the
compared with brown strains. The fruit body yield effect of the quantity of substrate used for bed making
and mean fruit body wt. in white strain was higher by was more visible compared to trial-II. In trial-I, beds
27.66 and 36.91%, respectively compared with the prepared with 18 kg substrate/bed of CGMW gave
next best performing brown strain (BBSR-007). The higher fruit body yield compared with beds prepared
study proved the white strain as better strain in all with 12 and 15 kg substrate/bed. The trend was just
respect, giving first harvest nearly two days earlier, reverse with beds prepared with PS and beds with
and fruit body yield and mean fruit body wt. higher by smaller quantity of substrate/bed gave higher fruit
nearly 28 and 37%, respectively than the best body yield compared to beds with higher
performing brown strain. This strain also revealed its substrate/bed. The difference in fruit body yield in
Table 2 — Fruit body yield of brown and white strains of V. volvacea at different temperature conditions on composted
substrate of paddy straw and cotton ginning mill waste
Strain Time taken for first harvest Mushroom yield Average fruit body wt.
(days post-spawning) (kg/q dry substrate) (g)
Low Optimum Low Optimum Low Optimum
temperature temperature temperature temperature temperature temperature
OE-210 (brown) 13.25 13.22 7.51 18.17 7.83 6.40
OE-272 (brown) 15.00 ND 1.75 ND 10.29 ND
OE-274 (brown) 13.88 13.94 7.75 31.38 8.12 15.18
BBSR-007 (brown) 12.62 13.11 8.93 32.21 14.83 14.90
GVv-01 (white) 11.93 11.00 20.79 41.12 20.74 20.40
CD 0.05 1.87 1.32 3.06 2.94 3.21 2.85
[Low temperature – 25 to 28 °C, Optimum temperature – 32 to 35 °C, ND – not done]

Table 3 — Yield potential of white strain of V. volvacea on beds with different quantities and types of substrate
Substrate and substrate Time taken for first harvest Mushroom yield Average fruit body wt.
quantity (kg/bed) (days post-spawning) (kg/q dry substrate) (g)
Trial - I Trial - II Average Trial - I Trial - II Average Trial - I Trial - II Average
CGMW - 12 10.17 12.75 11.46 30.49 25.25 27.87 19.66 19.33 19.50
CGMW - 15 9.0 12.12 10.56 30.43 23.63 27.03 19.87 20.91 20.39
CGMW - 18 9.67 12.75 11.21 33.17 18.08 25.63 19.12 16.89 18.01
CGMW + PS - 12 11.17 13.37 12.27 35.04 22.2 28.62 20.96 20.51 20.74
CGMW + PS - 15 10.83 12.62 11.73 36.21 16.02 26.12 22.23 19.34 20.79
CGMW + PS - 18 11.33 12.37 11.85 35.86 22.29 29.08 20.98 18.33 19.66
PS - 12 12.17 12.75 12.46 31.78 26.64 29.21 20.24 19.37 19.81
PS - 15 12.33 12.87 12.60 23.02 28.71 25.87 20.66 19.75 20.21
PS - 18 12.67 13.00 12.84 19.57 26.33 22.95 23.31 17.55 20.43
CD0.05 1.19 0.98 - 2.67 2.83 - 2.38 2.64 -
[CGMW – cotton ginning mill waste, PS – paddy straw, 12, 15, 18 – quantity of composted substrate in kg]
 AHLAWAT & KAUR: FRUIT BODY YIELD IN PADDY STRAW MUSHROOM WHITE STRAIN 117

Table 4 — Proximate composition of the fruit bodies of the brown and white strain of V. volvacea
Nutritional parameters Brown strain of White strain of Difference over
V. volvacea V. volvacea brown strain (± %)
Dry matter (%) 10.10 9.0 -10.89
Ash (%) 9.01 10.30 +14.32
Fat (%) 0.97 0.79 -18.56
Carbohydrates (%) 42.30 42.83 +1.25
Protein (F=6.25) (%) 38.10 36.88 -3.20
Crude fibre (%) 4.40 6.02 +36.82
Vitamin D (IU/g) 462.04 268.55 -41.88
Potassium (%) 4.16 5.34 +28.37
Sodium (mg/kg) 345.34 417.08 +20.77
Potassium: Sodium 120.46 128.03 +6.28
Calcium (mg/100 g) 39.74 59.88 +50.68
Iron (mg/kg) 72.51 84.59 +16.66
Copper (mg/kg) 42.55 56.13 +31.92
Zinc (mg/kg) 94.28 100.52 +6.62
Magnesium (%) 0.11 0.15 +36.36

relation to quantity of substrate used/bed was not
evident in case of substrate prepared with 1:1, w/w
combination of PS + CGMW both in trial-I and II.
Not much relationship could be established with the
type of substrate and the quantity of substrate/bed
with the mean fruit body wt. in all the three substrate
types (Table 3).
Proximate composition of the fruit body of the white strain
vis-à-vis brown strain of V. volvacea
The proximate nutritional analysis of the fruit
bodies of white and brown strains of V. volvacea
revealed higher contents of ash, crude fibre and all
seven minerals (potassium, sodium, calcium, iron,
copper, zinc and magnesium) in fruit bodies of the
white strain compared with the fruit bodies of brown
strain. The enhancement ranged between lowest of
6.62% for zinc to highest of 50.68% for calcium. The
fruit bodies of white strain exhibited lesser levels of
dry matter, fat, protein and vitamin D compared to
fruit bodies of brown strain (Table 4). For these four
parameters, except of vitamin D, the difference was
insignificant between two strains. The two strains
differ marginally with respect to protein content and
the difference was only 3.20%. Selenium was not
detected in the fruit bodies of both types of strains. In
nut shell, the white strain was superior in minerals
and crude fibre contents, while the brown strain in
vitamin D content, rest other parameters were by and
large at similar levels.
Discussion
The straw mushroom V. volvacea grows optimally
on paddy straw at 30 to 35°C, which gives it
advantage over other mushrooms, especially while
growing under tropical or subtropical agroclimatic
conditions. However, if specifically compared to
white button mushroom, A. bisporus, the attributes
like brown or off-brown colour of V. volvacea fruit
bodies along with their short shelf life and tendency
of liquefaction on storage at refrigerated conditions
have put it at disadvantageous position for its
cultivation and marketing at a wider scale. Some of
these limitations can be sorted out through selection
or identification of strains with white colour of the
fruit bodies and productivity at par or superior to that
of brown strains of V. volvacea.
In present study, out of five strains used for
studying the variability in sequences of their 5.8S
rRNA gene and mycelial growth characteristics, one
was the white strain, while the rest four were the
proven high yielding brown strains11,12. The white
strain (GVv-01) giving consistently superior fruit
body yield both at lower as well as optimum
temperature conditions exhibited a deletion of 21
nucleotides in ITS-2 region of the 5.8S rRNA gene
compared with four high yielding brown strains,
which is a rare occurrence in V. volvacea24. Along
with the white colour of fruit bodies, the deletion in
ITS-2 region also proves the distinctness of the white
strain from high yielding brown strains. Along with
these characteristics, formation of a separate clade in
phylogenetic tree by the white strain and one
combined clade by the three high yielding brown
strains also proves the distinctness of white strain
from rest brown strains. A similar finding with respect
to this white strain and several brown strains has also
118 INDIAN J EXP BIOL, FEBRUARY 2018
been reported earlier by the same group24, which
reflects its genetical distinctness from brown strains.
The variability in the ITS region of 5.8S rRNA gene
is widely used for species identification in case of
fungi9,24 and minimum variability with in species is
expected in this region. In present context the level of
variability recorded in white strain compared to
brown strains is worth to be acknowledged. In respect
to genetical variability, Volvariella is a poorly studied
genus and variability based upon the sequences of the
ITS region of 5.8S rRNA gene has not been exploited
much, except limited study carried out for few
selected strains of V. volvacea24. The variability in
sequences of the laccase 3 gene has also revealed
heterozygocity in one of these brown strains
(OE-210)25. The reports on phylogenetic and
evolutionary relationship between strains of
V. volvacea are again quite scanty and in few studies
carried out using RAPD, it has been reported that
strains originated from same region exhibit nearly
85-95% similarity, while strains originated from
diverse regions were reported as quite dissimilar7. The
single spore isolates from two different strains have
again been reported to exhibit intra-specific variations
of 52 and 65%26.
In the present study, strains with higher downward
mycelial growth and forming higher aerial hyphae as
well as chlamydospores have also been recorded to
exhibit higher fruit body yield potential compared
with strains with lower downward mycelial growth
and forming lesser aerial hyphae/chlamydospores.
Relationship between mycelial growth characteristics
and variability at molecular level as well as fruit body
yield potential in V. volvacea strains has also been
studied earlier7,11,21 and has proved its worthiness in
selecting high yielding strains. Reports specifically on
relationship between downward mycelial growth and
the fruit body yielding potential in V. volvacea are
quite scanty, except one where in the strain showing
higher downward mycelial growth also exhibited
higher fruit body yielding potential27. However,
similar types of findings in button mushroom,
A. bisporus, have proved worthiness of this approach
for selection of high yielding strains28.
The high temperature requirement for cultivation of
V. volvacea, is providing competitive advantage to
this mushroom while growing in tropical and
subtropical regions of the world, however, the same
advantage becomes disadvantage when it has to be
grown under subtropical or temperate conditions. In
present study, the initial evaluation trial conducted
under suboptimal temperature (25 to 28°C) conditions
revealed significantly higher fruit body yield in the
white strain GVv-01 compared to high yielding brown
strains of V. volvacea. The yield enhancement was
nearly 2.33 folds higher than second best performing
brown strain. Higher fruit body yield compared to
brown strains was again recorded in white strain on
cultivation at optimum temperature conditions for
V. volvacea. The brown strains used in the study were
the proven high yielder, confirmed by the earlier
studies carried out at different scales of
cultivation11,12,21. However, present study has revealed
the white strain as the superior yielder compared with
the brown strains. It provides a better alternative of
brown strains both in respect of fruit body yield as
well as white colour of the fruit bodies. Further, the
superior fruit body yield of the white strain both at
sub-optimal and optimal temperature conditions,
makes it a better choice for the mushroom growers to
grow it under varied temperature conditions. Selection
of a superior yielding strain has remained a
continuous endeavour in V. volvacea11,21,29,30,
however, only limited attempts have been made on
evaluation of strains for fruiting at low temperature24
and white colour of the fruit bodies. In an earlier
study, two strains generated through EMS
mutagenesis and screened at 0°C have been reported
to give 46.1 and 40.5% higher mushroom yield,
respectively than the control strain on cultivation at
27°C. The strains were also reported to have longer
shelf life of their fruit bodies on storage at 16°C than
control strain31.
In trials to optimize the fruit body yield of white
strain, lowest time for first harvest was in CGMW,
followed by 1:1, w/w combination of CGMW + PS
and PS alone. In one earlier study the substrate
prepared with CGMW, CGMW + PS and PS alone
was evaluated for the fruit body yield potential of a
brown strain of V. volvacea and CGMW was reported
as the best substrate, followed by CGMW + PS and
PS alone20. However, in present study, the white
strain performed well on all the three substrates,
which proved its worthiness of growing on different
types of substrates. Alike present study, positive
correlation between average fruit body weight and
early first harvest with yielding potential of a strain is
well documented in literature11,21. However, in these
studies, the higher yield is also attributed to higher
hydrolytic enzymes activity.
 AHLAWAT & KAUR: FRUIT BODY YIELD IN PADDY STRAW MUSHROOM WHITE STRAIN
Mushrooms are well known for their nutraceutical
properties and straw mushroom in not the exception.
There are several studies where comparative nutritional
analyses of commercially grown mushrooms have been
attempted32,33. However, studies involving the
comparative proximate nutritional analysis of the
strains of V. volvacea giving fruit bodies with varied
colours are quite scanty, except of one study where the
brown coloured fruit bodies of V. volvacea have been
compared with grayish fruit bodies of V. bombycina23.
The current study has presented a comparative
proximate composition of one high yielding brown
strain and one white strain with potential of
acceptability due to giving white coloured fruit
bodies. The study has proved that the fruit bodies of
white strain of V. volvacea contain higher level of
crude fibres and ash leading to higher values of
calcium, zinc, magnesium, copper, iron, sodium and
potassium. It contained lesser level of dry matter,
protein, fat and vitamin D compared to fruit bodies of
high yielding brown strain of V. volvacea. However,
except vitamin D content, the difference in rest three
parameters was insignificant making it nutritionally a
very important strain. The values of parameters like
crude fiber, protein, fat, total ash and carbohydrates
presented in an earlier study34 vary from the present
study and the values of most of the parameters in the
present study are higher than the values reported
earlier, and this can be attributed to the variation in
protocols adopted in two studies.
Acknowledgement
We thank SERB, DST, Govt. of India for funding the
project under which the study was carried out and the
Director, Directorate of Mushroom Research (ICAR),
Solan for constant encouragement and providing the
requisite facilities for conducting the study.
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