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Domestication of wild strain of Pleurotus giganteus

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Domestication of wild strain of Pleurotus giganteus

N. Klomklung1,2,3, S. C. Karunarathna1,2,3, E. Chukeatirote1,2 &

K. D. Hyde1,2,3,*

1
Institute of Excellence in Fungal Research, Mae Fah Luang University,
Chiang Rai 57100, Thailand
2
School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand
3
Mushroom Research Foundation, 128 M.3 Ban Pa Deng T. Pa Pae, A. Mae Taeng,
Chiang Mai 50150, Thailand

Klomklung N., Karunarathna S. C., Chukeatirote E., Hyde K. D. (2012) Domestication

of wild strain of Pleurotus giganteus. – Sydowia 64 (1): 39–53.
Pleurotus giganteus has relatively large fruiting bodies and is a saprobe in heavily
rotting underground wood in forests; it is collected and widely consumed in many tropical
countries including Thailand. Although P. giganteus is a popular edible mushroom, it is not
cultivated in Thailand or Sri Lanka as a commercial mushroom. Recently a method for the
cultivation of P. giganteus at the experimental level using saw dust as a substrate has been
developed. The strain was isolated from a fresh fruiting body of P. giganteus (MFLU10
0154) using a piece of cap tissue and cultivated on Potato Dextrose Agar (PDA). Spawn was
grown in sorghum (Sorghum bicolor) seeds. The cultivation method involves two steps, in-
oculating on a saw dust substrate in polypropylene bags as the preliminary step and trans-
ferring to the soil as a second step, which is very important for fruiting. The developed
method of growing P. giganteus is fully described with all necessary steps.
Keywords: mushroom, polypropylene bags, saw dust substrate, tropical countries, un-
derground wood.

Recently Pleurotus giganteus was transferred from Lentinus based on

morphological and molecular evidences (Karunarathna et al. 2012). This
species was previously named as Lentinus giganteus Berk. and was first de-
scribed from Sri Lanka locally referred to as “Uru Paha” and classified in
‘Decades of Fungi’ (Berkeley 1847). Pleurotus giganteus has been treated as
a special food since ancient times as mentioned in Buddhist literature
(Berkeley 1847, Udugama & Wickramaratna 1991). When fully grown, the
basidioma is typically infundibuliform measuring up to 35 cm in diameter
and 28 cm high (Berkeley 1847, Udugama & Wickramaratna 1991). The
mushroom may be solitary but often forms in groups on the ground. Pleuro-
tus giganteus has a thick, radicant stipe and subdistant broad lamellae
which is typical of P. giganteus (Pegler 1983, Karunarathna et al. 2012). It is
a very popular mushroom because of its high protein content, excellent taste,
bioactive components and the health-related functions (Udugama & Wickra-
maratna 1991, Huang 2005).

*  e-mail: kdhyde3@gmail.com
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40 Klomklung et al.: Domestication of Pleurotus giganteus

Mushrooms have been domesticated for cultivation since early times,

however most commonly grown strains are temperate species (e.g. Stamets
2000, Muruke et al. 2002, Fereira 2010). Tropical mushrooms are, however,
numerous and recent studies on various genera have shown them to be spe-
cious with numerous new species being described (Le et al. 2007 a, b; San-
mee et al. 2008; Zhao 2008; Kerekes & Desjardin 2009; Wannathes et al. 2009
a, b; Sysouphanthong et al. 2010; Zhao et al. 2010, 2011 a, b; Karunarathna
et al. 2011, 2012). Domesticating tropical mushrooms therefore provides a
huge opportunity for tropical and subtropical countries and progress has
already been made (Boa 2004, Vostrovský & Jablonská 2007, Marshall &
Nair 2009). Tropical mushrooms grow rapidly and produce fruiting bodies at
25 °C or higher and thus can be produced more quickly than temperate spe-
cies. They can also be produced on readily available waste products such as
saw dust, corn cobs, rice straw, sugarcane bagasse, and other forest and agri-
cultural waste. Growing mushrooms will therefore help recycle agricultural
and forest waste products, provide income to various entrepreneurs, provide
nutritional and medicinal foods and prevent pollution through less dumping
and burning of agricultural waste (Vostrovský & Jablonská 2007, Kwon &
Thatithatgoon 2004).
Only a small number of mushrooms are presently commonly cultivated
in Thailand and include the Oyster Mushroom, Pleurotus ostreatus (Jacq.) P.
Kumm., Hed Nanglom Khao in Thai, Straw mushroom, Volvariella volvacea
(Bull.) Singer, Hed Fang in Thai, and Wood ear, Auricularia polytricha (Mont.)
Sacc., Hed Hoo-Nu (Hanko 2001, Boa 2004, Karunarathna et al. 2011). Other
less commonly produced species are King oyster mushroom, Pleurotus eryn-
gii (DC.) Quél., Heh Nanglom Luang in Thai, Abalone mushroom, Pleurotus
cystidiosus O. K. Mill., Hed Pao-hue in Thai, Golden oyster mushroom, Pleu-
rotus citrinopileatus Singer, Black poplar mushroom, Agrocybe cylindracea
(DC.: Fr.) Maire, Hed yanagi in Thai, Enokitake, Flammulina velutipes (Curt.:
Fr.) Karst., Hed Khemthong in Thai, Reishi, Ganoderma lucidum (Leyss.: Fr.)
Karst., Hed Lin Juer in Thai, Shiitake, Lentinula edodes (Berk.) Pegler, Hed
Hom in Thai, Button mushroom, Agaricus bisporus (J. E. Lange) Imbach, Hed
Kradum in Thai, while Inky cap, Coprinus atramentarius (Bull.) Fr., Hed
Muerk in Thai, Lion’s mane, Hericium erinaceus (Bull.) Pers., Hed Hua Ling
in Thai, Silver ear, Tremella fuciformis Berk., Hed Hu-nu-Khao in Thai, and
Parasol mushroom, Macrolepiota gracilenta (Krombh.) Wasser, Hed Nok
Yoong in Thai (Kwon & Thatithatgoon 2004) may be rarely produced. Most of
the mushrooms presently produced are imported strains and as far as we are
aware little use is made of Thai strains. According to Boa (2004) there are
over 1100 species of edible and medicinal fungi from over 80 countries and
when this list is compared with the edible mushroom species that are pres-
ently commercially harvested in Thailand, many opportunities are clearly
available for other species domestication (Jones et al. 2004, Kwon &
Thatithatgoon 2004, Berch et al. 2007). Studies centered around the Mush-
room Research Centre in Chiang Mai have resulted in records and descrip-
tions of numerous mushroom species (Le et al. 2007 a, b; Sanmee et al. 2008;
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Sydowia 64 (2012) 41

Zhao 2008; Kerekes & Desjardin 2009; Wannathes et al. 2009 a, b; Sysou-
phanthong et al. 2010; Karunarathna et al. 2011, 2012; Zhao et al. 2011 a, b),
many of which are edible and have the potential to be domesticated (Zhao et
al. 2011 a, b; Karunarathna et al. 2011, 2012; Chen et al., unpubl.). Thai peo-
ple like to eat mushrooms and there is extensive cultivation of common
mushrooms in Thailand. Mushrooms are also used in traditional Thai medi-
cines and have been shown to contain various bioactive components and are
used in cosmetics (Kwon & Thatithatgoon 2004, Hyde et al. 2010), cancer
treatments (Wisitrassameewong et al. 2012 a, b) and have anti-diabetic prop-
erties (De Silva et al. 2012). The Thai government and Royal project also en-
courages rural farmers to grow mushrooms because of the large income from
the mushroom growing using low cost agricultural wastes (Kwon &
Thatithatgoon 2004). As Thai’s appear to like eating mushrooms, the poten-
tial for introducing new mushrooms to the Thailand market is great.
During studies of the genus Lentinus in northern Thailand we collected
several species of Lentinus including three species new to science (Karunar-
athna et al. 2011). We also collected Lentinus giganteus and following mo-
lecular study found this taxon to be more closely related to Pleurotus
(Karunarathna et al. 2012) showing how molecular methods have revolu-
tionized the study of taxonomy, systematics, phylogeny, biogeography, popu-
lation and microevolutionary processes in basidiomycetes in the last two
decades (Yang 2011). Species of the genus Pleurotus are the best known of
edible higher basidiomycetes as producers of the pharmacologic agent lovas-
tatin (mevinolin) (Gunde-Cimerman et al. 1993 a, b; Gunde-Cimerman &
Cimerman 1995). The presence of lovastatin was determined in four species:
P. ostreatus, P. cornucopiae, P. eryngii, and P. sapidus (Wasser & Weis 1999).
Pleurotus giganteus is one of the largest edible mushrooms in the world
and can be grown on saw dust medium with supplements (Udugama & Wick-
ramaratna 1991). Saw dust from a mixture of wood species or Jak wood is
preferred as the main substrate for P. giganteus growing (Udugama & Wick-
ramaratna 1991). Pleurotus giganteus is cultivated in China (Huang 2005)
and Taiwan (Peng 2006), but even though it has a very good taste (Udugama
& Wickramaratna 1991), it is not yet cultivated in Thailand or Sri Lanka as
a commercial mushroom (Karunarathna et al. 2012). The present experiment
was undertaken to investigate the best conditions for domestication of wild
P. giganteus using saw dust as a locally available substrate.

Materials and methods

Isolation of pure cultures

Pure cultures were isolated from the sterile internal fungal tissues.
About 5 ml of PDA medium was poured into 50 ml culture tubes followed by
tight capping with sterile cotton wool, sterilization, and kept as slants. Fresh
juvenile fruiting bodies of Pleurotus giganteus were collected from the
Mushroom Research Center, Chiang Mai, Thailand (MFLU10 0154) and used
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42 Klomklung et al.: Domestication of Pleurotus giganteus

for tissue culture. Small pieces of the internal tissue of the broken mushroom
was cut and removed with a flamed needle.
The needle with the tissue was then transferred into a culture-tube slant
and the tissue was laid on the agar surface, and incubated at 30 °C. After 3
to 4 days, the agar surface was covered with a white mycelium as pure cul-
ture.

Mycelial growth tests

Effect of raw materials: Vigna angularis (Willd.) Ohwi & Ohashi. (red
bean), Phaseolus vulgaris L. (black bean), Vigna radiata (L.) R. Wilczek
Steve Hurst. (mung bean), and Glycine max (L.) Merr. (soybean) were ob-
tained from Tesco-Lotus (Mae Chan) and Sorghum bicolor (L.) Moench (sor-
ghum) was bought from Chiang Rai local market in Thailand. Malt extract
agar (DIFCO) and potato dextrose agar (CRITERION) were used as synthet-
ic media. Fifty grams of each grain type was soaked in 250 ml of distilled
water (Gbolagade et al. 2006) for 12 h and boiled. Each grain type was then
ground using a pestle and mortar and filtered through a clean white cloth.
Twenty grams of agar (UNION SCIENCE) and distilled water were added to
obtain the final volume of 1000 milliliters and autoclaved. The media were
poured into 10 cm Petri dishes and allowed to solidify. The mycelia were sub-
cultured into semi-synthetic media and incubated at 30±2 °C (Gbolagade et
al. 2006). The radial colony diameter was measured after two days incuba-
tion and daily until mycelia reached the edge of the plates. The Soybean raw
material prepared to support the growth of P. giganteus mycelia and used to
determine the effect of pH and temperature for the growth and reproduction.
Effect of pH: The pH of the medium prepared from soybean, was ad-
justed to 5-8 (5, 5.5, 6, 6.5, 7, 7.5, and 8). The best pH for the growth of P. gi-
ganteus was determined by measuring the colony diameter following the
method described below.
Effect of temperature: The growth of P. giganteus mycelium was com-
pared using different temperatures (20, 25, 30, and 35 °C) to determine the
optimal temperature requirement for this mushroom. The growth of the col-
ony diameter was measured and compared to establish the optimum tem-
perature for mycelia growth.

Grain spawns development and production

Red bean, black bean, soy bean, mung bean and sorghum were used as
substrates for spawn production. Materials were washed and soaked in dis-
tilled water for 12 hours, boiled for 5-10 minutes (almost cooked) and filtered
through a clean white cloth. All materials were dried at room temperature.
Forty grams of each substrate were placed in Erlenmeyer flasks and loosely
covered with a cotton plug and aluminium foil on top and autoclaved. The
mycelia from pure culture of P. giganteus were cut and inoculated into each
grain material and incubated at the optimal temperature of 30 ºC. The myce-
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Sydowia 64 (2012) 43

lial growth of each grain was recorded in order to choose the best raw mate-
rial for spawn production.
Spawn was prepared as above using Sorghum bicolor. After sterilization
the bottles filled with Sorghum were allowed to cool under room tempera-
ture, transferred with the mycelial disks of P. giganteus under aseptic condi-
tions, plugged with cotton plug and incubated at 30 ºC, in the spawn room,
in the dark. Observations were recorded on the diameter of mycelium run-
ning through the substrate. After 15–20 days the bottles became white due to
complete colonizing by mycelia. The spawn was then ready for transferring
to substrate bags.

Bag preparation, cultivation and fructification

Saw dust from a mixture of wood species was used as the main sub-
strate. For 1 kg of clean sawdust, 50 g of rice bran, 10 g of brewer’s waste,
10 g of Leucaena leaf, 10 g of pumice sulfate, 10 g of calcium carbonate,
and 10 g of flour were added to prepare the substrate. The components
were mixed well and water gradually added until the moisture content was
around 65–70 %. Polypropylene bags (25 × 8 cm) were filled with 800 g
prepared substrate and packed tightly. A hole (about 5 cm) was made with
a PVC pipe at the centre for space to place the mycelial plugs. A plastic
ring was used to make a “bottle neck” for easy handling. Plastic rings were
used on the bags end, the bags end was pulled out through the ring, the
pulled out part was folded down, tied with a rubber band and the hole
plugged with cotton plug. The substrate bags were autoclaved at 15 psi for
15 min at 121 °C or by using a steamer at 90–100 °C for 3 h. After sterili-
zation the substrate bags were allowed to cool to room temperature, trans-
ferred with the spawn, normally using about 80 g for a substrate bag which
was about 10 % of the weight of a substrate bag, with P. giganteus under
aseptic conditions. The bags were kept in a dark incubation room at 25±1 °C
under 70–80 % relative humidity and opened when the mycelia had com-
pletely colonized the substrate.
After the mycelia had completely grown in the substrate, the upper por-
tions of the bags were opened. The opened surface of the substrate was
scraped slightly with a sterile teaspoon to remove the thin whitish mycelia.
The substrate bags were then placed on the shelf and covered with black
cloth to give appropriate ventilation. To maintain 80–85 % relative humidity
in the culture house, water was sprinkled on the open end of the growing
bags. Water spraying was carried out daily until pin heads developed.
When the pin heads had started to develop the polypropylene bags were
completely removed and the contents transferred to the soil and buried or
the top part of the growing bags was covered with soil and transferred to the
growing house. The experiment was composed of 30 growing bags, ten bags
were transferred to the soil, ten bags were covered with soil at the top, and
ten bags were kept without soil as controls. Water spraying was carried out
daily until the fruiting bodies had fully developed.
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44 Klomklung et al.: Domestication of Pleurotus giganteus

Data collection and statistical analysis

A Completely Randomized Design (CRD) with five replications was used
in the experiment. Data were collected for mycelium growth rate in culture
plates and growing bags, time required for completion of mycelium running,
optimum temperature and pH for mycelium growth, duration from stimula-
tion to primordia initiation, duration from stimulation to harvest, number of
mature fruiting bodies, stalk length and diameter, pileus diameter and thick-
ness. The data were analyzed statistically in terms of variance and mean
showing statistical significance using Duncan’s multiple range tests by
SPSS-16 program (Brosius 2008).

Results
Mycelial growth tests
Effect of raw materials: All five types of raw materials used enhanced
mycelia growth and radial mycelia extension of P. giganteus at 30 °C, being
the optimal temperature. The best mycelia growth occurred on soybean me-
dia (Tab. 1, Fig. 1).

Tab. 1. – Effect of raw materials on mycelia growth of Pleurotus giganteus at 30 °C. Each
value represents a mean of five replicates. Means followed by the same letters are not sig-
nificantly different by Duncan’s multiple range test (P<0.05)

Days
Raw materials Radial mycelia extension (mm)
2 4 6 8 10
Mung bean 7.6 cd
16.4 c
37c
60.4 c
74.4c
Black bean 7bc 19.4d 43d 66.6d 80.4d
Red bean 8.8e 27.8f 49.4e 70.2e 82.8e
Sorghum 6.8b 20.4d 44.6d 71.8f 85f
Soy bean 7.8d 24.2e 51f 73f 90g
MEA 6.4ab 7.4a 19a 24a 31.2a
PDA 6a 11.8b 22.2b 27b 33.2

Effect of pH: Mycelia growth of P. giganteus was observed at a pH of 5–8.

Mycelia of P. giganteus can grow in acidic, neutral and alkaline conditions
(Tab. 2), but the maximum growth was obtained at pH 5–6.5.
Effect of temperature: Radial mycelia growth at 20, 25, 30 and 35 °C is
shown in Tab. 3, with an optimum at 25 °C followed by 30 °C and 20 °C. No
growth occurred at 35 °C.

Fig. 1. Effect of raw materials on mycelia growth of Pleurotus giganteus. A, B Soybean

medium (best growth), C Sorghum medium, D red bean medium, E black bean medium,
F mung bean medium, G PDA, H MEA (poorest growth).
Author’s personal copy

Sydowia 64 (2012) 45
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46 Klomklung et al.: Domestication of Pleurotus giganteus

Tab. 2. – Effect of pH on mycelia growth of Pleurotus giganteus. Each value represents a

mean of five replicates. Means followed by the same letters are not significantly different by
Duncan’s multiple range test (P<0.05)

Days
pH Radial mycelia extension (mm)
2 4 6 8 10 12
5 11.4 c
21.6 b
33.6 b
52 b
76.4 c
90b
5.5 12.2c 24.4d 40.6d 58.8c 78.6d 90b
6 11.6c 22.8c 35.8c 52.4b 79.2d 90b
6.5 10.2c 21b 35c 51.8b 76c 90b
7  9.8b 19.8a 35.2c 49.2a 70.6b 81a
7.5  8.2a 18.6a 31.4a 48.6a 67.6a 80.8a
8  8.6a 19a 33.6b 48.4a 67a 80.4a

Tab. 3. – Effect of temperature on mycelia growth of Pleurotus giganteus. Each value rep-
resents a mean of five replicates. Means followed by the same letters are not significantly
different by Duncan’s multiple range test (P<0.05)

Days
Temperature (°C) Radial mycelia extension (mm)
2 4 6 8 10
20 10 a
18.2 a
27.8 a
41 a
53.8a
25 13.4 b
35.8 c
35.8 c
77.8 c
90c
30 12.6b 25.8b 25.8b 52.8b 72.2b
35 5 5 5 5 5

Grain spawn development and production

Mycelia of P. giganteus could grow in all five grain types to a varying
extent. The best growth in terms of mycelial extension was obtained on soy-
bean (12 days after inoculation) followed by black bean, red bean, mung
bean and the poorest mycelia growth occurred in Sorghum (Fig. 2).

Fructification
We observed that the mycelia of P. giganteus took 30-32 days to run from
the top to the bottom of the substrate bags until pin heads developed. After
31 days of encasing, P. giganteus produced fruiting bodies in two bags and
after 53 days in eight bags. In control bags, the fruiting was observed only in
two bags after 33 days and until 53 days it was still only in two bags. The
results of fruiting body development are shown in Tab. 4 and Fig. 3.

Fig. 2. Pleurotus giganteus mycelial growth on different grain types. A Soy bean (best
growth), B black bean, C red bean, D mung bean, and E Sorghum (poorest growth).
Author’s personal copy

Sydowia 64 (2012) 47
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48 Klomklung et al.: Domestication of Pleurotus giganteus

Tab. 4. – Fruiting body production of Pleurotus giganteus in substrate bags. * Total number
of fruiting bodies after 53 days. **Total number of bags completed with mycelia running
from the top to the bottom of the substrate bags after 32 days

Mycelia running Fruiting bodies development

Fruiting bodies No.
Days Bags Days
Casing Non casing
30  5 31  2 0
31 11 33  1 2
32  4 36  1 2
20** 42  2 2
48  1 2
53  3 2
10* 2*

Discussion
Huge quantities of waste are freely available from the agro-forest and
timber industries in Thailand (Kwon & Thatithatgoon 2004). Mushroom
yields of 317 million metric tons (317 billion kg) of fresh mushrooms per year
could be achieved only using 25 % of the yearly volume of burned cereal
straws in the world (Chang & Miles 1989). In fact, we could simply grow 360
billion kg of fresh mushrooms on the total of 600 billion kg of dry waste, us-
ing the annual available world waste in agriculture (500 billion kg) and for-
estry (100 billion kg). A yearly mushroom yield of 60 kg per head per year
could be achieved, all containing the 4 % protein of fresh mushrooms. Recent
analysis has shown that 200 g of mushrooms can efficiently replace 100 g of
meat as a protein source which could solve 30 % protein deficiency in their
diet of the world population (Souci et al. 1975–1989). The fast mycelial
growth and multilateral enzyme system of Pleurotus ostreatus (oyster mush-
room) which is very special among mushrooms could be used to biodegrade
nearly all types of different available wastes (Kwon & Thatithatgoon 2004).
Usually, sawdust is used as a substrate for mushroom cultivation (Stamets
2000). The sawdust used in composting does not have sufficient nitrogen and
other components required for the fermentation process therefore, the com-
pounding mixture is supplemented with nitrogen and carbohydrate sources,
in our case rice bran and meal concentrate, to enhance this process (Pathak
et al. 1998).
The comparative mycelia growth rate of P. giganteus on culture media of
different substrates, pH and temperature showed varying responses. The my-
celia growth on saw dust substrate was best at 25 °C (Tab. 3) with a pH of
5–6.5 (Tab. 2) and the superior raw material for mycelia growth in culture at

Fig. 3. Fruiting bodies of Pleurotus giganteus. A, B Developing young fruiting bodies.

C, D fully grown fruiting bodies. Bar A = 20 cm.
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Sydowia 64 (2012) 49
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50 Klomklung et al.: Domestication of Pleurotus giganteus

30 °C was soybean which could easily be available in local markets (Tab. 1,

Fig. 1). On the other hand the best material for spawn run was also soybean
whereas black bean and red bean are also good according to the results ob-
tained (Fig. 2). In Thailand, mushroom growers normally use Sorghum seeds
as the spawn carrier (Kwon 2004). Sorghum was the least suitable substrate
according to our results (Fig. 2), when compared to soy bean, black bean, red
bean and mung bean. The reason why mushroom growers use Sorghum, could
be its readily availability and low cost.
There was a significant difference of biological yield between the control
group and the experimental group. Biological yield of P. giganteus grown on
control bags was poor. According to Moorthy (1993), 25–28 °C were found to
be the optimum for P. sajor-caju in vitro studies. Cartwright & Findlay (1934)
had observed that most of the fungi prefer a temperature range of 25–30 °C
for mushroom production.
The total biological yield of the experimental group was 10 fruiting
bodies/10 growing bags after 53 days (Tab. 4). The control group showed a
very poor yield of only two fruiting bodies/10 bags even after 53 days (Tab. 4).
Saw dust bags are mainly used for Pleurotus growing especially with P. os-
treatus, whereas for P. giganteus growing it is essential to use soil casing in
order to obtain a better yield.
Pleurotus giganteus has been shown to have medicinal properties
(Huang 2005), and the protein content of P. giganteus in dry weight basis
reported as 37.8 %, which was the highest compared to most other cultivated
popular mushrooms (Udugama & Wickramaratna 1991). Our effort is to in-
troduce P. giganteus to Thai markets as a new member, like Yanagi matsutake
in Thailand (Agrocybe cylindracea) growing on saw dust substrate, which
has a high demand and brings a handsome income to mushroom farmers
(Kwon & Thatithatgoon 2004).
The low yield and long time for Pleurotus giganteus to produce fruiting
bodies would possibly make it an expensive mushroom. It is now important
to develop better protocols for growing it at high yields which are produced
quickly. It is also important to isolate other strains and find better strains for
mushroom production. It may also be possible to breed hybrid strains using
the methods described in Callac (1995).
The study showed that it is possible to domesticate local strains of P.
giganteus that can grow at a temperature consistent with Thailand farm pro-
ductions. In China it was successfully domesticated in the 1980’s and strains
are now extensively grown there (Chen & Hu 2002; Wang-Qiu et al. 2006).

Acknowledgements

We are grateful to Jian Kui Liu, Komsit Wisitrassameewong, Bencha-

rong Thonbai and Phongeun Sysouphanthong for their help in collecting,
growing experiments and suggestions. A special thank goes to Else Vellinga
(Department of Plant and Microbial Biology, University of California, Berke-
ley, USA), Rui Lin Zhao (Southwest Forestry University, Kunming 650224,
 Author’s personal copy

Sydowia 64 (2012) 51

China) and Philippe Callac (INRA, MYCSA (Mycologie et sécurité des ali-
ments), Villenave d’Ornon cedex, France) for their valuable suggestions.
This study was financially supported by the project “Value added products
from basidiomycetes: Putting Thailand’s biodiversity to use” (BRN049/2553),
by the French-Thai cooperation PHC SIAM 2011 (project 25587RA), by the
National Research Council of Thailand (NRCT) with the project “Taxonomy,
Phylogeny and cultivation of Lentinus species in northern Thailand”
(NRCT/55201020007), and by the Mae Fah Luang University research divi-
sion with the project “Taxonomy, Phylogeny and cultivation of Lentinus spe-
cies in northern Thailand” (MFU/54 1 01 02 00 48).

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(Manuscript accepted 11 Jun 2012; Corresponding Editor: I. Krisai-Greilhuber)

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54 Klomklung et al.: Domestication of Pleurotus giganteus

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