Bioresource Technology 97 (2006) 10121017

Manganese and other micronutrient additions to improve yield

of Agaricus bisporus
David A. Weil a, Robert B. Beelman

a,*

, David M. Beyer

a
b

Department of Food Science, 116D Borland Laboratory, The Pennsylvania State University, University Park, PA 16802, USA
Department of Plant Pathology, 102 Buckhout Laboratory, The Pennsylvania State University, University Park, PA 16802, USA
Received 30 October 2003; received in revised form 26 April 2005; accepted 29 April 2005
Available online 1 July 2005

Abstract
Previous experiments have demonstrated that signicant increases in yield of Agaricus bisporus mushrooms were achieved by
adding a micronutrient rich fertilizer, Micromax, to the compost. This study was performed to determine the mineral(s) that
are responsible for this yield improvement. An initial experiment determined that manganese was the mineral of primary importance; addition of 184 mg kg 1 Mn increased yield by 10.8%, compared to the control. Subsequent experiments demonstrated that
the addition of manganese to the compost has a stimulatory eect on mushroom yields. Signicant yield increases, ranging from
9.6% to 11.8% (compared to the control), were observed as a result of manganese additions varying between 50 and 300 mg kg 1.
Also, data from the last set of experiments indicated that Micromax additions always resulted in greater yields indicating that micronutrients in Micromax, in addition to manganese, may be responsible for increasing yields.
2005 Elsevier Ltd. All rights reserved.
Keywords: Agaricus bisporus; Micronutrients; Micromax; Manganese; Compost; Yield

1. Introduction
The essential elements and microelements required by
Agaricus bisporus have been well established (Styer, 1928;
Bohus, 1959; Wood and Fermor, 1985). Furthermore,
several studies have been conducted to determine the effect of added nutrient supplements to compost in order
to optimize compost formulations utilized for A. bisporus
production (Schisler and Sinden, 1962; Sinden and Schisler, 1962; Carroll Jr. and Schisler, 1976; Beyer and
Muthersbaugh, 1996). Thus, until recently it was assumed that current mushroom composting formulations
provided sucient levels of micronutrients to obtain
optimum yield potentials. Research conducted in Belgium (Desrumaux et al., 2000) and recent experiments
performed at the Penn State Mushroom Test Demon*

Corresponding author. Tel.: +1 814 863 2964; fax: +1 814 863 6132.
E-mail address: [email protected] (R.B. Beelman).

0960-8524/$ - see front matter 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2005.04.042

stration Facility (MTDF; Weil, 2003) demonstrated that

the supply of micronutrients provided by current composting formulations may sometimes be inadequate to support optimum yields. These experiments demonstrated
that yields increased following additions of varying levels
of Micromax to the compost at casing, but they did not
indicate which micronutrient(s) were responsible for
yield increases. Previously, Racz and Tasnadi (1998) reported statistically signicant yield increases when manganese was added to compost at spawning used for A.
bisporus production. Research performed on other individual micronutrients added to the substrate at spawning, such as copper, boron, and iron, resulted in no
signicant eect on cumulative yields (Hayes, 1972;
Cresswell et al., 1990). Regardless, micronutrient supplementation appears to be a potential opportunity for
mushroom growers to improve eciency and quality of
freshly harvested mushrooms. Therefore, the objective
of this study was to determine the micronutrient(s) that

D.A. Weil et al. / Bioresource Technology 97 (2006) 10121017

are responsible for A. bisporus yield improvement

and the eect of varying the concentration of the yield
increasing micronutrient(s) identied.

1013

500 mL paper bag by repetitively inverting the bag for

30 s, which was then added to the compost at spawning.
The amount of each micronutrient added to the compost was equivalent to the amount contained in
7350 mg kg 1 Micromax.
The second set of experiments consisted of three
crops of o-white hybrid mushrooms (A. bisporus)
grown at the MRC to determine the eect of adding
varying levels of manganese to the compost at spawning
on yield and quality of fresh mushroom production.
The rst two crops (# 3010 and 3011) of the second
set of experiments consisted of 30 trays, with six trays/
treatment, incorporated the addition of 92, 184, and
276 mg kg 1 manganese (Sigma), 7350 mg kg 1 Micromax (which contained 184 mg kg 1 manganese), and a
control (no Micromax or manganese added) to the
compost. The third crop (# 3101) consisted of 48 trays,
with six trays/treatment, incorporated additions of 50,
100, 150, 200, 250 and 300 mg kg 1 Mn (Sigma),
6000 mg kg 1 Micromax (which contained 150 mg kg 1
manganese), and a control (no Micromax or manganese
added) to the compost. All treatments were thoroughly
mixed with the supplement before being added to the
compost. The spawn for each crop of the second set of
experiments was treated with benomyl (Benlate; DuPont
Co., Wilmington, DE) to minimize the occurrence of
Trichoderma green mold. For every 1000 g of spawn,
1.34 g of benomyl was thoroughly mixed with 66.7 g
of food-grade gypsum (United States Gypsum Co.,
Southard, OK) in a gallon size Ziploc storage container (S.C. Johnson & Son, Inc., Racine, WI). The
spawn was subsequently added and thoroughly coated
with the gypsum/benomyl mixture.
Market mature mushrooms, with intact veils, from
each crop were harvested over three ushing periods.
Yields for each ush and cumulative yield (overall three
ushes), for each treatment, were expressed as kg m 2,
the weight of the mushrooms harvested per unit area
of bed space. Solids determination and mineral analysis
of the mushrooms were performed for all the crops,
except crop 3101. Initial whiteness was analyzed only
on mushrooms from crops 3010 and 3011.

2. Methods
This study was divided into two sets of experiments.
The rst set involved the addition of six of the individual
elements contained in Micromax (Scotts-Sierra Horticultural Products Co., Marysville, OH) to the compost
at spawning for one crop. The second set of experiments
involved the addition of varying levels of manganese to
the compost at spawning for three crops.
2.1. Mushroom production
For both sets of experiments, the crops were grown
using standard MTDF cultural practices including traditional horse manure compost for Phase I and Phase II
composting (Beyer and Beelman, 1995). Mushroom
Research Center (MRC, University Park, PA) cultural
methods similar to those described by Spolar (1997) were
utilized for spawning, casing, and harvesting. Spawn and
supplement (Campbells S-41 Full-house, Napoleon,
OH) were added to the crop at a rate of approximately
2% and 4% (of the compost dry weight), respectively.
Details on all the crops are presented in Table 1.
In the rst set of experiments, one crop (# 3006) of
o-white hybrid mushrooms (A. bisporus) was grown
at the MRC to determine which micronutrient(s) in
Micromax were active in improving crop yield. This
crop consisted of 48 trays, with six trays/treatment.
The eight treatments consisted of 3.7 mg kg 1 molybdenum, 7 mg kg 1 boron, 74 mg kg 1 copper, 74 mg
kg 1 zinc, 184 mg kg 1 manganese, 1176 mg kg 1 iron
(Sigma, St. Louis, MO) and 7350 mg kg 1 Micromax,
expressed on a compost dry weight basis. Each micronutrient was added as sodium molybdate, sodium borate,
copper sulfate, zinc sulfate, manganese sulfate, and ferrous sulfate, respectively. Each micronutrient compound was eectively mixed with the supplement in a

Table 1
Spawn strains and production information for experimental mushroom (Agaricus bisporus) crops grown at the Mushroom Research Center
(University Park, PA) to evaluate the eect of manganese sulfate added to the compost at spawning on yield and quality
Crop number

Spawn manufacturer
and strain

Compost moisture
at spawning (%)

Compost dry matter

(kg m 2)

Total N (%)

pH

Compost Mn
content (mg kg 1)a

3006
3010
3011
3101

Ital F58b
Sylvan 130c
Sylvan 140c
Le Lion X-25d

71.9
72.8
70.6
72.4

17.21
16.66
18.13
16.91

1.8
2.2
2.3
2.6

7.5
7.9
8.1
7.3

283
297
234
226

a
b
c
d

Prior to the addition of MnSO4.

Ital Spawn Co., Italy.
Sylvan America, Kittanning, PA.
Le Lion Plus, Varrains, France.

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D.A. Weil et al. / Bioresource Technology 97 (2006) 10121017

2.2. Initial whiteness

Immediately after the mushrooms were harvested, the
mushrooms from each treatment were analyzed for their
initial whiteness (L-value; Solomon et al., 1991) using
a hand-held Chromameter (Model CR-200, Minolta
Crop., Ramsey, NJ). Four replicates consisting of eight
mushrooms each were randomly selected for each treatment and placed into 8-ounce linear polystyrene tills
(Tray-Pak Corp., Reading PA). Three readings were
taken from the top of each individual mushroom cap
giving a total of 24 color measurements for each replication. The means of the four replications were averaged
to provide the L-value for each treatment for the peak
day of production of each ush.
The Chromameter was calibrated prior to each use on
a standard white calibration plate that was included with
the instrument. L * a * b color coordinates were used for
all color measurements. The standard white tile with
coordinates of L = 97.00, a = 2.00, and b = 0.00 were
used as an arbitrary standard. Increasing L-value (ranging from 0 to 100) indicates a higher degree of whiteness.
2.3. Solids determination
Six replicates of four randomly selected quarter
wedges from four dierent mushrooms were combined,
placed into aluminum weigh dishes, weighed, and frozen
at 20 C for each treatment. The frozen samples were
then freeze-dried (VirTis model Genesis 25XL, VirTis
Inc., Gardiner, NY) until constant weight was obtained.
Dried samples were immediately weighed, placed in
sterile sample bags (Fisher) and then stored for further
analysis. The means of the replicates were averaged to
provide solids values for each treatment for each ush.
Solids content was reported as a percentage of dry
matter.

(replication) and treatment. Analysis of variance (ANOVA) and mean separation analysis using Fishers least
signicant dierence test were used to determine signicant dierences between treatments for yield, initial
whiteness, solids content and mineral content of the
mushrooms.

3. Results and discussion

3.1. Addition of individual micronutrients to compost
The eect of 7350 mg kg 1 of Micromax and six of
the individual elements present in that level of Micromax added to the compost at spawning are presented
in Table 2. Manganese (184 mg kg 1) was the only element added to the compost at spawning that produced
a statistically signicant increase (p < 0.05) in cumulative yields compared to the control. The predominant increase as a result of adding manganese occurred during
the rst ush and the cumulative yield increase greater
than the control was 1.56 kg m 2 (+10.8%). None of
the added elements had any signicant eect on the solids content of the harvested mushrooms (Table 2).
Addition of boron and copper were the only individual treatments to signicantly increase (p < 0.05) the
amounts in the mushrooms as a result of their additions
(data not shown). The tendency for more boron and
copper to be taken up by the mushrooms as more is
added to the compost was observed in previous experiments performed at the MTDF (Weil, 2003) and by
Cresswell et al. (1990). These increases do not appear
to be correlated to yields or solids content of the harvested mushrooms. The concentration of Zn, Fe, Mo
and Mn in the mushrooms were not signicantly impacted by any of the respective treatments (data not
shown).

2.4. Mineral analysis

Three dried mushroom samples for each treatment
and ush for each crop were sent to the Agricultural
Analytical Services Laboratory at University Park, PA
for mineral analysis. Mineral values determined by standard procedure using inductively coupled plasma atomic
emission spectroscopy (Miller, 1998) included phosphorus (P), potassium (K), calcium (Ca), magnesium
(Mg), manganese (Mn), iron (Fe), copper (Cu), boron
(B), aluminum (Al), zinc (Zn) and sodium (Na).
2.5. Statistical analysis
All statistical analyses were performed using Statistical Analysis System software, version 8.2 (SAS Institute
Inc., Cary, NC). The experiments were completely randomized with the sources of variation consisting of tray

Table 2
Inuence of individual micronutrients and Micromax (Scott-Sierra
Horticultural Products Co., Marysville, OH) on cumulative yield,
percent yield dierence compared to the control, and overall solids
content of Agaricus bisporus mushrooms for crop 3006 grown at the
Mushroom Research Center (University Park, PA)
Treatment

Yield
(kg m 2)a

Dierence
(%)

Solids
(%)a

Control
Micromax (7350 mg kg 1)
Mn (184 mg kg 1)
Mo (3.7 mg kg 1)
B (7 mg kg 1)
Cu (74 mg kg 1)
Zn (74 mg kg 1)
Fe (1176 mg kg 1)

14.46BC
15.29AB
16.02A
14.06C
14.06C
14.11C
14.01C
14.36BC

0.0
+5.8
+10.8
2.7
5.8
2.4
3.1
0.7

7.23A
7.43A
7.26A
7.16A
7.27A
7.46A
7.30A
7.31A

a
Means within a column followed by the same letter are not signicantly dierent at p < 0.05 according to Fishers LSD; values are the
means of six replications.

D.A. Weil et al. / Bioresource Technology 97 (2006) 10121017

Despite signicant increases in the amount of manganese in the compost as a result of the manganese additions (data not shown), no signicant increases taken
up by the mushrooms were observed. This tendency
was previously observed by Racz and Tasnadi (1998).
The results seem to indicate that the yield simulating
eect produced by adding manganese occurred in the
compost, since it was not taken up by the mushrooms.
Since previous research has established manganese
peroxidase as a crucial enzyme in lignin degradation of
wood rotting fungi (Kirk and Farrell, 1987; Tien and
Cai, 1987), it appears that manganese additions may
possibly facilitate enzymatic reactions in compost for
A. bisporus production.

1015

Table 4
Cumulative yield and percent yield dierence compared to the control
of Agaricus bisporus mushrooms as a result of manganese sulfate
(MnSO4) and Micromax (Scott-Sierra Horticultural Products Co.,
Marysville, OH) additions to the compost at spawning for crop 3101
grown at the Mushroom Research Center (University Park, PA)
Treatment

Yield (kg m 2)a

Dierence (%)

Control
50 mg kg 1 Mn
100 mg kg 1 Mn
150 mg kg 1 Mn
200 mg kg 1 Mn
250 mg kg 1 Mn
300 mg kg 1 Mn
6000 mg kg 1 Micromax

13.72B
15.04A
15.34A
14.85AB
15.09A
14.85AB
15.33A
15.63A

0.0
+9.6
+11.8
+8.2
+10.0
+8.2
+11.8
+14.0

3.2. Addition of manganese to compost

a
Means within a column followed by the same letter are not signicantly dierent at p < 0.05 according to Fishers LSD; values are the
means of six replicates.

In one crop (# 3010), a statistically signicant increase (p < 0.05) in cumulative yield was observed as
a result of adding 184 and 276 mg kg 1 manganese
and 7350 mg kg 1 Micromax (containing approximately
184 mg kg 1 Mn) to the compost at spawning compared
to the control (Table 3). The yield increases compared to
the control, were 10.0%, 11.7%, and 18.3% when 186
and 276 mg kg 1 manganese and 7350 mg kg 1 Micromax were added to the compost, respectively (Table 3).
The yield increases as a result of adding 92 mg kg 1 manganese was 2.5% and was not signicantly dierent
(p < 0.05) when compared to the control.
A similar experiment was conducted in a second crop
(# 3011) but there were no statistically signicant dierences (p < 0.05) in cumulative yields as a result of adding
manganese or Micromax treatments compared to the
control despite yield increases of 5.8%, 12.1%, 2.7%,
and 16.8% greater than the control when 92, 184,
276 mg kg 1 manganese and 7350 mg kg 1 Micromax

were added to the compost, respectively (Table 3).

Greater variability in response to the treatments resulted
in no statistically signicant dierences observed in this
crop as a result of adding micronutrient treatments to
the compost. However, the yield increases were similar
to that of crop 3010, except for the highest level
(276 mg kg 1) of manganese added where an unexplained lower yield was observed.
A third crop (# 3101) was grown to examine a wider
range of manganese additions to the compost at spawning. Yield increases, compared to the control, ranging
between 8.2% and 11.8% were observed (Table 4).
Yields as a result of 50, 100, 200 and 300 mg kg 1 manganese and 6000 mg kg 1 Micromax were statistically
signicant (p < 0.05) compared to the control. Also, observed yield increases occurring due to added manganese
resulted in an interesting pattern (Fig. 1). Yield increased signicantly with 50100 mg kg 1 manganese

Table 3
Cumulative yield, percent yield dierence compared to the control, overall solids content, initial whiteness (L-value) and manganese concentration
(expressed on a dry weight basis) of Agaricus bisporus mushrooms as a result of manganese sulfate (MnSO4) and Micromax (Scott-Sierra
Horticultural Products Co., Marysville, OH) additions to the compost at spawning for crops 3010 and 3011 grown at the Mushroom Research
Center (University Park, PA)
Crop number

Treatment

Yield (kg m 2)a

Dierence (%)

Solids (%)a

L-valuea

Mn content (mg kg 1)a

3010

Control
92 mg kg 1 Mn
184 mg kg 1 Mn
276 mg kg 1 Mn
7350 mg kg 1 Micromax
(contained 184 mg kg 1 Mn)

16.32C
16.71BC
17.93AB
18.23A
19.31A

0.0
+2.5
+10.0
+11.7
+18.3

8.67A
8.52AB
8.52AB
8.36B
8.03C

91.78B
92.15AB
92.21A
92.41A
92.04AB

5.1A
4.9A
5.2A
5.3A
5.3A

3011

Control
92 mg kg 1 Mn
184 mg kg 1 Mn
276 mg kg 1 Mn
7350 mg kg 1 Micromax
(contained 184 mg kg 1 Mn)

13.92A
14.75A
15.63A
14.31A
16.27A

0.0
+5.8
+12.1
+2.7
+16.8

7.56B
7.20C
7.84A
7.41BC
7.60AB

92.79A
92.33A
92.70A
92.72A
92.05A

5.8A
5.8A
5.8A
6.3A
6.1A

a
Means within a column followed by the same letter are not signicantly dierent at p < 0.05 according to Fishers LSD; values are the means of six
replicates.

Yield (kg/m2)

1016

D.A. Weil et al. / Bioresource Technology 97 (2006) 10121017

16
15.5
15
14.5
14
13.5
13
12.5
12
0

50

100

150

200

250

300

Mn Added (mg/kg)

Fig. 1. Inuence of varying levels of manganese added to compost at

spawning on yield of mushrooms produced. The values depicted in the
gure are from crop 3101 grown at the Mushroom Research Center
(University Park, PA) and are mean yields SEM.

appears that other micronutrients and/or interactions

between other micronutrients present in Micromax
may also stimulate mushroom yields. This suggests that
further studies are needed to determine which micronutrient(s) may be involved. Furthermore, results of these
experiments conrm earlier work that demonstrated the
potential to improve yields of A. bisporus mushrooms by
adding micronutrients (especially manganese) to compost. However, due to the complex and variable nature
of compost used in mushroom cultivation, further
experiments will be necessary to determine how growers
can best utilize this practice at the commercial level.

Acknowledgements
added and then leveled o indicating a possible saturation eect at higher levels of manganese.
The results from this study were consistent with
previous research performed in Hungary (Racz and
Tasnadi, 1998). They observed yield increases of 14%,
8% and 12% (for three separate trials) when a level of
100 mg kg 1 manganese was added to the compost at
spawning. They also observed no signicant increases
(p < 0.05) in manganese content of the mushrooms as
a result of adding manganese to the compost.
Since the uptake of manganese by the mushrooms
was not aected (Table 3), and since the role of manganese peroxidase in lignin degradation of wood rotting
fungi has been well established (Kirk and Farrell,
1987; Tien and Cai, 1987), it was further speculated that
the addition of manganese increased the decomposition
of lignin in compost ingredients such as straw, providing
an increase in carbon and nitrogen sources to the
mushrooms.
The initial whiteness (L-value) of the mushrooms
(Table 3) was signicantly improved as a result of adding manganese (184 and 276 mg kg 1) to the compost
in one crop (# 3010), but no eect was observed with
mushrooms from another crop (# 3011). In general,
changes in color were not expected, since the manganese
added apparently facilitated enzymatic reactions occurring in the compost and manganese levels in the mushrooms were not aected (Table 3).
However, the additions of micronutrients to the compost had a signicant eect (p < 0.05) on the solids content of the harvested mushrooms (Table 3). Signicant
decreases (p < 0.05) in solids occurred as a result of adding 276 mg kg 1 manganese (crop 3010) and 92 mg kg 1
manganese (crop 3011) to the compost at spawning. A
negative impact on solids as a result of adding Micromax
has been observed during previous research (Weil, 2003).
It was speculated that cation imbalances in the compost
or casing may have produced this eect (Reid, 2001).
In general, yield increases observed by adding Micromax were generally greater than those with equivalent
levels of manganese added (Tables 3 and 4). Hence, it

The authors would like to acknowledge the nancial

support of Mushroom Industry Farmer Based Applied
Research (MIFBAR). The technical assistance of the
MDTF and MRC sta was also greatly appreciated.

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