Mycobiology 37(2) : 109-113 (2009)
© The Korean Society of Mycology
Determination of Mineral Components in the Cultivation Substrates of Edible
Mushrooms and Their Uptake into Fruiting Bodies
1, Jeong-Eun Park
2, Bo-Bae Kim
2, Sun-Mi Kim
2 and Hyeon-Su Ro
1Greenpeace Mushroom Co.,
2Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University, Chinju 660-701, Korea
(Received April 21, 2009. Accepted June 5, 2009)
The mineral contents of the cultivation substrates, fruiting bodies of the mushrooms, and the postharvest cultivation sub-
strates were determined in cultivated edible mushrooms Pleurotus eryngii, Flammulina velutipes, and Hypsizigus marmoreus.
The major mineral elements both in the cultivation substrates and in the fruiting bodies were K, Mg, Ca, and Na. Potassium
was particularly abundant ranging 10~13 g/kg in the cultivation substrates and 26~30 g/kg in the fruiting bodies. On the
contrary, the calcium content in the fruiting bodies was very low despite high concentrations in the cultivation substrates,
indicating Ca in the cultivation substrates is in a less bio-available form or the mushrooms do not have efficient Ca uptake
channels. Among the minor mineral elements determined in this experiment, Cu, Zn, and Ni showed high percentage of
transfer from the cultivation substrates to the fruiting bodies. It is noteworthy that the mineral contents in the postharvest
cultivation substrates were not changed significantly which implies that the spent cultivation substrates are nutritionally intact
in terms of mineral contents and thus can be recycled as mineral sources and animal feeds.
KEYWORDS: Edible mushroom, Flammulina, Hypsizigus, Mineral, Pleurotus, Substrate
Mushrooms belong to fungal groups that form characteris-
tic fruiting bodies in which spores reside. They not only
play an important role in the recycling of plant materials
in an ecosystem but also are appreciated as good sources
of food and medicine. Edible mushrooms, Agaricus
bisporus and Pleurotus ostreatus, are popular mushrooms
with high commercial values and are thus cultivated
world wide. Pleurotus eryngii, Flammulina velutipes, and
Hypsizigus marmoreus are edible mushrooms particularly
popular in East Asia, cultivation of which are facilitated
using specially formulated substrates in semi-automated
cultivation facilities. The former two are generally culti-
vated on a culture bed consisting of dairy manure-wheat
straw or dairy manure-rice straw composts. The latter
three are cultivated in wide-mouth polypropylene bottles
which contain a substrate mixture constituted of various
agricultural wastes including rice bran, corncob, soybean
hull, and sawdust. In either case, proper composition of
the cultivation substrate with good preparation practice is
crucial for the reliable production of mushrooms. For
example, basal substrate supplemented with Mn and
ground soybean resulted in significant enhancement in the
production yield of P. eryngii (Rodriguez Estrada and
Royse, 2006). Addition of limiting mineral components
promoted mycelia growth rate of P. ostreatus up to 25%
(Curvetto et al., 2002). Because mycelia propagation
within the cultivation substrate takes at least a month to
cultivate these mushrooms, an enhancement in the myce-
lia growth rate by 25% means a huge reduction in pro-
In this report, we determined the mineral contents of the
cultivation substrates for the mushrooms P. eryngii, F.
velutipes, and H. marmoreus. The percentage of mineral
transfer from the cultivation substrate to the mushroom
fruiting body was assessed by comparing the mineral con-
tents in the fruiting bodies with those in the input sub-
strates. We also determined the mineral contents in the
postharvest cultivation substrates which are often recycled
for an additional round of cultivation and are utilized as
animal feed sources (Kim et al., 2007b; Kwak et al., 2008).
Materials and Methods
Substrate composition and mushroom cultivation con-
The substrate for H. marmoreus consisted of
pine sawdust (23%), concob (32%), rice bran (32%), and
soybean hull (22%). The cultivation of H. marmoreus was
carried out at 15
4000 ppm CO2 and 95% relative humidity (RH). The sub-
strate for P. eryngii was rather complex. It contained pine
sawdust (23%), concob (29%), rice bran (18%), beet pulp
(4%), wheat bran (14%), cottonseed hull (4%), and shell
powder (4%), and dehydrated beverage by-product from
soybean (14%). The cultivation conditions for P. eryngii
were similar to H. marmoreus with slight differences:
500~2000 ppm CO2 and 87% RH. F. velutipes was cul-
tured at 5
substrate consisted of concob (31%), rice bran (40%), beet
oC in an incubating room with 3000~
oC with 3000~5000 ppm CO2 and 90% RH. Its
*Corresponding author<E-mail: firstname.lastname@example.org>
110Lee et al.
pulp (16%), wheat bran (5%), cottonseed hull (5%), and
shell powder (4%). The total growth period for H. mar-
moreus, P. eryngii, and F. velutipes was 23 days, 18 days,
and 30 days, respectively. Each individual component in the
cultivation substrate was measured as weight percent (wt%).
substrates, and postharvest cultivation substrates of H.
marmoreus, F. velutipes, and P. eryngii were collected
from mushroom farms in southern Korea. The samples
(50.0 g) were dried at 100
weights were determined. The dried sample was ground
using a mortar and pestle. For complete digestion, the
ground sample was digested by a solution containing a
10 : 1 mixture of 50% HNO3 and 30% H2O2 solutions
(digestion solution) as previously described (Gergely et
al., 2006). Briefly, the sample solution containing 0.5 g of
the ground powder in 100 ml of the digestion solution was
The fruiting bodies, cultivation
oC for 48 hrs and their dry
Table 1. The determination of dry weight of mushroom fruiting bodies and cultivation substrates
Fruiting body (50 g)Substrate (50 g)Post-harvest substrate (50g)
6.1 ± 0.1
8.3 ± 0.2
6.8 ± 0.2
18.1 ± 0.5
16.6 ± 0.7
17.1 ± 0.4
18.7 ± 0.8
20.1 ± 0.5
16.1 ± 0.5
aThe measurement was triplicated. Data are expressed as mean ± SEM (standard error of the mean).
Table 2. The production yield of mushroom fruiting bodies
Substrate input (g)Harvested fruiting body (g)
Dry weight (A)Wet weight Dry weight (B)
556.0 ± 1.2
748.3 ± 4.1
840.4 ± 3.4
180.2 ± 2.5
289.6 ± 2.7
199.8 ± 2.8
aThe wet weight of 5 samples per each mushroom was measured separately and expressed as mean ± SEM.
Table 3. The mineral concentrations (mg/kg) of dried mushroom fruiting bodies and cultivation substrates
H. marmoreus F. velutipesP. eryngii
Substrate Fruiting bodySubstrate Fruiting body Substrate Fruiting body
aThe measurement was carried out 5 times. Data in the table are mean values of the measured data. Standard error of the mean (SEM) of each
data was less than 1% of the mean value. SEM values are omitted for the clarity in this table.
heated for 24 hrs. When the solution became clear with
slightly yellowish color, the digestion was stopped and the
volume of the sample solution was adjusted to 40 ml with
Quantification of mineral elements in the cultivation
substrates and fruiting bodies.
the distribution of minerals and trace elements in mush-
room production, we determined the concentration of
mineral components in the fruiting bodies of mushrooms
and their cultivation substrates. The digested sample solu-
tions were subjected to inductively coupled plasma spec-
trometry (ICP spectrometer Optima 5300DV, Perkin
Elmer, CT, USA) or ICP mass spectrometry (ICP-MS,
Elan DRC II, Perkin Elmer). Mineral elements including
Na, K, Ca, Mg, Al, Mn, and Fe were quantified by ICP.
Other trace elements including Cu, Zn, Pb, Ni, and Se
were quantified by ICP-MS. The results are summarized
In order to investigate
Mineral Components in the Cultivation Substrates of Edible Mushrooms and Their Uptake111
in Tables 3~5. The concentrations of mineral components
in the dried samples in mg/kg (part per million, ppm) unit
are shown in Table 3. The transfer rate of minerals from
the cultivation substrate to the fruiting body is shown in
Table 4. This rate was calculated based upon the concen-
tration of a mineral (mg/kg) per cultivated fruiting bodies
(kg) divided by the concentration of the mineral (mg/kg)
per cultivation substrate input (kg). For example, the rate
of transferred Na for H. marmoreus was (61.3 mg/
kg × 0.024 kg)/(220.3 mg/kg × 0.199 kg) × 100 =
The amounts for the cultivation substrate inputs and the
cultivated fruiting bodies are shown in Table 2. The con-
centrations of mineral components in wet samples are
reconstituted using the water contents in Table 1 and the
mineral contents in dried samples shown in Table 3.
Results and Discussion
Determination of mushroom production yields in dry
The edible mushrooms H. marmoreus, F. veluti-
pes, and P. eryngii are cultivated through formulated sub-
strates which are contained in wide-mouth polypropylene
bottles with a volume capacity× diameter of 850 ml × 58
mm for H. marmoreus, 1030 ml × 75 mm for F. velutipes,
and 1280 ml × 80 mm for P. eryngii in commercial farms.
In order to investigate the mushroom yield per cultivation
substrate input, we measured the dry weights of the fruit-
ing bodies and the cultivation substrates. As shown in
Table 1, the water concentration of the fruiting bodies was
87.8, 83.4, and 86.4% for H. marmoreus, F. velutipes, and
P. eryngii, respectively, while the water concentration of
the cultivation substrates was 63.8, 66.8, and 65.8%,
respectively. The mushroom farms collected 180 g (22.0 g
dry weight), 290 g (48.1 g dry weight), and 200 g (27.2 g
dry weight) of the fruiting bodies for H. marmoreus, F.
velutipes, and P. eryngii, respectively, from 556 g (201.3 g
dry weight), 748g (248.2g dry weight), and 840g (287.3g
dry weight) of the cultivation substrate for each corre-
sponding mushroom. From these figures, the production
yields in dry weight are 10.9% for H. marmoreus, 19.4%
for F. velutipes, and 9.6 % for P. eryngii (Table 2). There-
fore, it is likely that the cultivation substrate for F. veluti-
pes contains a better formulation compared with that for
H. marmoreus and P. eryngii in terms of efficient utiliza-
tion of resources.
Major mineral elements in the cultivation substrates
and the fruiting bodies.
K, Ca, Mg, and Na were the
major mineral elements found in the cultivation sub-
strates and in the fruiting bodies regardless of mushroom
species (Table 3). Potassium, which is commonly found in
other cultivated or wild mushrooms (Mattila et al., 2001;
La Guardia et al., 2005), was particularly abundant in the
cultivation substrates and fruiting bodies for all 3 mush-
rooms. The concentration of K in the mushroom fruiting
bodies was as much as 26~30 g/kg dry weight. K in the
cultivation substrate transferred very efficiently to the
fruiting body. The transfer rate of K was 31.7, 50.5, and
Table 4. The transfer rate of minerals from the cultivation
substrate to the fruiting bodies
H. marmoreusF. velutipesP. eryngii
Table 5. The mineral concentration (mg/kg) of the postharvest cultivation substrates
aThe measurement was carried out 5 times. Data in the table are mean values of the measured data. Standard error of the mean (SEM) of each
data was less than 1% of the mean value. SEM values are omitted for the clarity.
bStandard table of feed composition in Korea, National Institute of Animal Science, Korea (http://www.nias.go.kr/saryo/eng/animal.asp)
112Lee et al.
25.7% for H. marmoreus, F. velutipes, and P. eryngii,
respectively (Table 4). On the contrary, mushrooms were
not good at Ca uptake. Less than 0.4% of Ca in the culti-
vation substrate was taken by the fruiting bodies resulting
in Ca concentrations of 159.8, 324.3, and 162.5 mg/kg dry
weight in the fruiting bodies of H. marmoreus, F. veluti-
pes, and P. eryngii, respectively.
Minor mineral elements in the cultivation substrates
and the fruiting bodies.
mineral elements for humans. The mushrooms in this
study could take-up Fe to some extent (Table 3). F. veluti-
pes was the best and accumulated 108.8 mg/kg in its fruit-
ing body with a transfer ratio of 6.4%. Zn was present in
fairly high amounts in all three mushrooms when com-
pared with other trace elements. P. eryngii had the best
accumulation of Cu. The concentration of Ni was intrigu-
ingly high in H. marmoreus even though Ni concentra-
tion in its cultivation substrate was lower than the others.
H. marmoreus may have a special mechanism by which it
can accumulate high amounts of Ni in its fruiting bodies.
Recent evidence has shown that mushrooms can absorb
metal ions in high concentrations (Bystrzejewska-Piotrowska
et al., 2008; Gonen Tasdemir et al., 2008) and the metal
absorption capability appeared to be species-specific
(Alonso et al., 2003). Therefore, it is very natural for dif-
ferent mushrooms to exhibit a preferential difference in
absorbing mineral components. It is notable that metal
binding peptides, namely phytochelatins which are found
in most eukaryotic cells and some prokaryotes, preferen-
tially form complex with transition metals such as Zn, Cu,
and Ni (Clemens, 2006). Therefore, the phytochelatins in
the fruiting bodies can be one explanation to address the
high uptake capacity of mushrooms for Zn, Cu, and Ni.
Se is recognized as an important micronutrient by exhibit-
ing antioxidative protection effects against oxidative dam-
age caused by free radicals and against development of
certain types of cancer (Falandysz, 2008). The recom-
mended daily intake of Se is 75 and 60 µg for men and
women respectively (Barelay et al., 1995). H. marmoreus
and P. eryngii appeared to contain nutritionally significant
but yet insufficient amounts of Se. Selenium enrichment
in the cultivation substrate can be an approach to increase
the Se concentration in fruiting bodies of mushrooms. The
mycelia of P. ostreatus were enriched with Se when they
were grown in Se rich medium (Serafin Muñoz et al.,
2006). Pb is toxic to humans. The legal limit concentra-
tion in food is 0.2 mg/kg (Korea Food and Drug Adminis-
tration). Among three mushrooms, only P. eryngii contained
insignificant amounts of Pb (0.002 mg/kg wet weight)
while others were devoid of any.
Fe is one of the important
Minerals in postharvest substrates.
cultivation substrates are mixtures of materials with high
nutritional values including mushroom mycelia, degraded
cellulosic fibers, degraded lignins, proteins, minerals etc.,
which make them a recognized valuable biological
resource particularly as animal feed (Kim et al., 2007a, b;
Kwak et al., 2008). In this regard, we assessed the micro-
nutritional value of the spent substrates of H. marmoreus,
F. velutipes, and P. eryngii. As shown in Table 5, the con-
centrations of the mineral components in the postharvest
cultivation substrates are largely higher than those in the
input cultivation substrates. This is probably due to the
supply of mineral elements through moisture during the
cultivation. In general, mushroom cultivation consists of
two steps. The first is to propagate mycelia in the cultiva-
tion substrate which requires constant relative humidity
(RH) around 65%. This step takes 25 days for F. veluti-
pes, 35~40 days for P. eryngii, and 85~120 days for H.
marmoreus. The second step is to grow fruiting bodies
from the fully developed mycelia. It takes about a month
of incubation with a relative humidity more than 85%.
Therefore, it is most than likely that the elevated mineral
contents in the postharvest cultivation substrates origi-
nated from the supplied water. In terms of micro-nutri-
tional value, the mineral contents in the postharvest
substrates were highly comparable with those in represen-
tative grains such as corn, barley, and wheat (Table 5).
This study was carried out with the support of the Mush-
room Export Research Program and Technology Develop-
ment Program for Agriculture and Forestry, Ministry of
Agriculture and Forestry, Republic of Korea.
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