Content uploaded by Mariana Drewinski
Author content
All content in this area was uploaded by Mariana Drewinski on Nov 22, 2024
Content may be subject to copyright.
An Acad Bras Cienc (2024) 96(4): e20230838 DOI 10.1590/0001-3765202420230838
Anais da Academia Brasileira de Ciências | Annals of the Brazilian Academy of Sciences
Printed ISSN 0001-3765 I Online ISSN 1678-2690
www.scielo.br/aabc | www.fb.com/aabcjournal
An Acad Bras Cienc (2024) 96(4)
Running title: DOMESTICATION
OF WILD EDIBLE MUSHROOMS
FROM BRAZIL
Academy Section:
MICROBIOLOGY
e20230838
96
(4)
96(4)
DOI
10.1590/0001-3765202420230838
MICROBIOLOGY
Studies on domestication of two species
of wild edible mushroom from Brazil
MARIANA P. DREWINSKI, MARINA P. CORRÊA-SANTOS, DIEGO C. ZIED & NELSON
MENOLLI JR.
Abstract: There are about 80 species of wild edible mushroom that certainly occur in
Brazil and can be used as a natural source of food and medicine. This study aimed to
evaluate the in vitro mycelial development in culture media at different temperatures
and substrates for cultivation of the edible mushroom species Auricularia fuscosuccinea
and Laetiporus gilbertsonii. Additionally, the cultivation and the nutritional composition
of A. fuscosuccinea mushrooms were evaluated. The two best wild strains of each
species were selected for the in vitro cultivation experiment in two different substrates.
Furthermore, an axenic cultivation on sawdust was conduct and the basidiomata
produced were evaluated on their nutritional composition. The temperatures that best
favored the mycelial growth were 30 °C for A. fuscosuccinea and 25 °C and 30 °C for L.
gilbertsonii. The mycelium of both species developed better in the sterile Eucalyptus
sawdust substrate. Despite the success in cultivating the mycelium of L. gilbertsonii, it
was not possible to obtain basidioma for this species. On the other hand, it was possible
to produce basidiomata of the two tested wild strains of A. fuscosuccinea.
Key words: Atlantic Rainforest, Auricularia fuscosuccinea, Laetiporus gilbertsonii, mush-
room cultivation, nutritional content.
INTRODUCTION
Wild edible mushrooms are an important
natural source of food, medicine, and income
for communities in more than 90 countries (Li et
al. 2021). There are about 2,000 species of wild
edible mushrooms worldwide (Li et al. 2021), but
only approximately 130 have been domesticated
(Thawthong et al. 2014). Despite the diversity
of edible species, five genera [Lentinula Earle,
Pleurotus (Fr.) P. Kumm., Auricularia Bull.,
Agaricus L., and Flammulina P. Karst.] constitute
about 85 % of the world production of edible
mushrooms (Royse et al. 2017). Albertó (2017)
summarized the steps for domestication of
naturally occurring species in 14 points, from the
strain isolation to the nutritional composition.
Considering that all these steps are complex, a
good starting point is to determine if the wild
species to be domesticated can be cultivated
using the techniques common for commercial
species (Albertó 2017). Among the almost 80
species of wild edible mushroom that certainly
occur in Brazil (Drewinski 2023), we focus this
work to test few steps for the domestication of
Brazilian wild strains of the edible mushrooms
Auricularia fuscosuccinea (Mont.) Henn. and
Laetiporus gilbertsonii Burds.
Auricularia fuscosuccinea (Basidiomycota:
Auriculariaceae) was originally described from
Cuba and occurs in tropical and subtropical
regions of the Americas (Wu et al. 2021). Fidalgo
& Hirata (1979) reported the use of this species
as food by Txicão and Txucarramãe indigenous
people from the Xingu National Park in Brazil.
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 2 | 14
Additionally, the use of A. fuscosuccinea as food
has been recorded by other communities in the
Americas (Gamboa-Trujillo et al. 2019, Ruán-Soto
et al. 2021). Auricularia fuscosuccinea is easily
found both in forests and in urban areas, and
it has already been recorded for 13 of the 26
Brazilian states: Acre, Amazonas, Mato Grosso,
Goiás, Pará, Paraíba, Pernambuco, Paraná, Rio
de Janeiro, Rondônia, Rio Grande do Sul, Santa
Catarina, and São Paulo (Alvarenga et al. 2015,
Drewinski 2023).
Laetiporus gilbertsonii (Basidiomycota:
Laetiporaceae) was described based on
collections from the Pacific coast of the United
States of America (Burdsall Jr & Banik 2001) and
has a wide distribution in the Americas, occurring
from temperate to tropical and subtropical
zones (Lindner & Banik 2008, Banik et al. 2012,
Pires et al. 2016, Campi et al. 2022). Recently,
Campi et al. (2022) studied the occurrence of
the genus Laetiporus in South America and
concluded that L. gilbertsonii is the correct
name for the species growing in Southern South
America (Campi et al. 2022). In Brazil, the species
has been reported for the Southeastern region,
in the states of São Paulo (Pires et al. 2016) and
Espírito Santo (Drewinski 2023). The species is
associated with brown rot mainly of Eucalyptus
spp. and Quercus spp., occurring in both living
trees and dead trunks and logs (Burdsall Jr &
Banik 2001). As well as for another species of the
genus, Laetiporus sulphureus (Bull.) Murrill, the
species L. gilbertsonii is also known as “chicken
of the woods” and present a pronounced flavor,
excellent texture and is highly appreciated in
gastronomy, although there are no studies on
its cultivation. Tropical regions have a great
potential to be a rich source of cultivatable
fungal species (Thawthong et al. 2014), and Brazil
is a good country for this kind of investigations.
MATERIALS AND METHODS
Sampling
Collections were carried out in the Atlantic
Rainforest domain, in the Brazilian states of
Espírito Santo, Paraná, Rio de Janeiro, and São
Paulo (Table I). From the fresh wild basidiomata,
a pure mycelium culture was obtained through
the inoculation of fragments of the pileus
context into Petri dishes containing sterile
PDA (Potato Dextrose Agar) medium. The dried
vouchers are deposited at the herbarium SP
(at the ‘Instituto de Pesquisas Ambientais’)
and at the fungarium FIFUNGI (IFungiLab, at
the ‘Instituto Federal de Educação, Ciência e
Tecnologia de São Paulo’), and the live cultures
are at the ‘Coleção de Culturas de Algas, Fungos
e Cianobactérias – CCIBt’ (at the ‘Instituto de
Pesquisas Ambientais’). This study is according
to the Brazilian legislation on access to
biodiversity and is registered in the ‘Sistema
Nacional de Gestão do Patrimônio Genético e do
Conhecimento Tradicional Associado’ (SisGen #
A4A9200).
Species identity
The identification of the collected specimens
was made through morphological and molecular
characteristics, following specific bibliographies
(Lowy 1952, Burdsall Jr. & Banik 2001, Looney et al.
2013, Pires et al. 2016, Wu et al. 2021). For molecular
studies, DNA extraction was performed from
mycelium obtained in liquid culture following a
modified CTAB extraction method. The intergenic
ribosomal region (nrITS1-5.8S-ITS2) was
amplified by polymerase chain reactions (PCR)
with the primers ITS1-F and ITS4 (White et al.
1990). The amplified products were purified with
QIAquick PCR Purification Kit and sequenced at
MacroGen (South Korea) using the same primer
pair. The generated sequences were manually
checked and edited with Geneious v.8.1 (Kearse
et al. 2012). We used the Basic Local Alignment
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 3 | 14
Search Tool (BLAST) to find similar sequences
to build the matrices. Sequences were aligned
using MAFFT online (Katoh et al. 2019) and were
manually optimized using AliView (Larsson 2014).
The ITS region was partitioned into ITS1, 5.8S and
ITS2, and the evolution model was estimated
for each partition using the BIC (Bayesian
Information Criterion) criterion in jModelTest
v.2.0 (Darriba et al. 2012). The Bayesian inference
(BI) analysis was performed with Mr.Bayes v3.2.7a
(Ronquist et al. 2012), with two independent
runs, four simultaneous independent chains and
20,000,000 generations with a sample frequency
every 1,000 generation. The phylogenetic trees
are available as Supplementary Material (Figures
S1 and S2).
Mycelial growth and dry mycelial biomass at
different temperatures
Twelve wild strains of A. fuscosuccinea and three
of L. gilbertsonii were evaluated for mycelial
growth and dry biomass production in PDA
culture medium at different temperatures. After
the preparation and solidification of the culture
medium (30 mL) in Petri dishes (90 mm diam),
a 9.6 mm fragment of the pure culture matrix
was inoculated in the center of the plate for
each wild strain. The plates were incubated at
temperatures of 20 °C, 25 °C, 30 °C and 35 °C in
a BOD (Bio-Oxygen Demand) incubator (Vargas-
Isla & Ishikawa 2008). The experiment was
carried out in 15 replicates per temperature per
strain. The diameter of the mycelial growth was
measured on the day that one of the replicates
of each evaluated wild strain completed
Table I. Data on the Brazilian wild strains of Auricularia fuscosuccinea and Laetiporus gilbertsonii evaluated in this
research.
Species CCIBt
accession Collector
number Fungarium/
Herbarium accession GenBank
accession Locality
A. fuscosuccinea 2381 - - OP851766 São Paulo, Cananeia
A. fuscosuccinea 2959 - - OP851765 Paraná, Curitiba
A. fuscosuccinea 4745 MPD158 SP513097 OP851758 Paraná, Guarapuava, particular property
A. fuscosuccinea 4747 MPD351 SP513098 OP851770 São Paulo, Cananeia, PEIC
A. fuscosuccinea 4748 MPD455 SP513099/FIFUNGI2 OP851752 São Paulo, São Paulo, PEC
A. fuscosuccinea 4749 MPD497 SP513100 OP851764 São Paulo, Iporanga, PETAR
A. fuscosuccinea 4751 MPD527 SP513101/FIFUNGI3 OP851753 Rio de Janeiro, Teresópolis, PARNASO
A. fuscosuccinea 4752 MPD539 SP513102/FIFUNGI4 OP851755 Rio de Janeiro, Teresópolis, PARNASO
A. fuscosuccinea 4753 MPD576 SP513103/FIFUNGI5 OP851768 São Paulo, Campos do Jordão, PECJ
A. fuscosuccinea 4756 MPD600 SP513104 OP851800 São Paulo, Cananeia, PEIC
A. fuscosuccinea 4757 MPD609 SP513105 OP851757 São Paulo, Iporanga, PETAR
A. fuscosuccinea 4758 MPD614 SP513106 OP851754 São Paulo, Iporanga, PETAR
L.gilbertsonii 4709 MPD300 SP512740 OP851756 Espírito Santo, Santa Teresa, RBAR
L. gilbertsonii 4710 MPD306 SP512741 OP851769 Espírito Santo, Santa Teresa, RBAR
L. gilbertsonii 4718 MPD466 SP513107/FIFUNGI7 OP851767 São Paulo, São Paulo, PFC
CCIBt: Coleção de Culturas de Algas, Cianobactérias e Fungos at the Instituto de Pesquisas Ambientais; PETAR: Parque Estadual
Turístico Alto do Ribeira; PEIC: Parque Estadual da Ilha do Cardoso; RBAR: Reserva Biológica Augusto Ruschi; PEC: Parque Estadual
da Cantareira; PARNASO: Parque Nacional da Serra dos Órgãos; PECJ: Parque Estadual Campos do Jordão; PFC: Parque Dr. Fernando
Costa.
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 4 | 14
the growth on the plate. To evaluate the dry
mycelial biomass, the plates were placed in the
microwave and heated for 20 seconds to melt
the culture medium, then the mycelium was
filtered and washed with distilled water and the
biomass was dehydrated until constant weight
(Vargas-Isla & Ishikawa 2008).
Mycelial development on different substrates
From the results of the mycelial biomass and
growth tests at different temperatures, two
wild strains of each species were selected
for the experiment of mycelial development
on two substrates: i) autoclaved, based on
eucalyptus sawdust; ii) pasteurized, based on
sugarcane bagasse and grass (Brachiaria sp.).
The sawdust-based substrate was donated by
the company Yuri Cogumelos (Sorocaba, São
Paulo state, Brazil), which sells blocks for the
shiitake production, and is composed of 80 %
eucalyptus sawdust and 20 % grass bran, with
a moisture content of 68 %. The substrate was
distributed in glass jars (600 mL), closed with a
metal lid with a cotton filter, and then sterilized
in an autoclave at 121 °C for two hours. The
substrate JunCao (Jun = mushroom and Cao =
grass), based on sugarcane bagasse and grass
(Brachiaria sp.), was donated by a producer of
Pleurotus ostreatus from Bragança Paulista city
(São Paulo state, Brazil), and is composed of 60
% sugarcane bagasse, 35 % Brachiaria sp., and
5 % wheat bran. The substrate was pasteurized
with fluent steam for seven days and distributed
in sterile glass jars (600 mL). The substrates were
inoculated with 1 % of spawn (myceliated wheat
grains), which was divided into three fractions
and inoculated in three portions on the surface
of substrates. The jars were maintained in BOD
incubator at 30 °C and the daily growth was
measured from the three inoculation points
with a pachymeter. The experiment was carried
out with 15 replicates per substrate per strain.
Axenic cultivation experiment
The substrate based on eucalyptus sawdust was
used for the cultivation in blocks. The substrate
(2.5 kg) was packed in polypropylene bags with
filters, which were sterilized in an industrial
autoclave for 3h and 40min at 121 °C. After
sterilization and cooling of the substrate, the
packages were inoculated with 2 % of spawn.
The packages were incubated in the dark, in a
culture chamber, with the temperature set to
30 °C. After complete mycelial growth, the
packages were submitted to a rustic cultivation
environment, without temperature control,
with average temperature of 24 °C (ranging
from 7.4 °C to 35.5 °C), average humidity of 64
% (ranging from 34 % to 99 %), and average
CO2 concentration of 659 ppm (ranging from
585 ppm to 745 ppm). To induce primordia, the
packages of A. fuscosuccinea were cut at the top.
For the cultivation of L. gilbertsonii, four forms
of induction of primordia were tested, following
Pleszczyńska et al. (2013): i) incubation of the
blocks in a refrigerator (7–8 °C) for 24 hours
and transfer to the grow environment without
opening the package; ii) injection of 300 mL of
cold sterile distilled water through the package
filter; iii) incubation of the blocks in a refrigerator
(7–8 °C) for 24 hours, cutting on the surface of
the package and removal of mycelium from the
surface of grow block (scratching technique); and
iv) cutting on the package surface and scratching
technique. The A. fuscosuccinea experiment
was carried out with 12 replicates and the L.
gilbertsonii experiment was carried out with
eight replicates per treatment. The blocks were
monitored for 60 days after primordia induction.
Bromotalogical analyses of A. fuscosuccinea
The basidiomata obtained were analyzed for
moisture content, ash, crude protein, crude
fat, and crude fiber (Zenebon et al. 2008). The
bromatological analyzes were carried out at the
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 5 | 14
Bromatology Laboratory of the Animal Production
Department at ‘Universidade Estadual Paulista
Júlio de Mesquita Filho - UNESP’, in Dracena,
São Paulo, Brazil. The crude protein content was
determined indirectly from the total nitrogen
value using Kjeldahl method (Zenebon et al.
2008), using the conversion factor of 4.38 (Crisan
& Sands 1978). All analyzes were performed in
triplicate and the results express the arithmetic
mean.
Statistical analyses
For statistical evaluation, the data obtained
were submitted to the Shapiro-Wilk normality
test and then analyzed using Two-way ANOVA
test. The averages were compared by the Tukey
test, using level of significance of 0.05 (Vieira
1980). Statistical analyzes were performed using
the software GraphPad Prism version 9 (http://
www.graphpad.com).
RESULTS
Auricularia fuscosuccinea
The effects of different temperatures on 12 wild
strains of A. fuscosuccinea growth in PDA are
shown in Figure 1. The mycelial growth and dry
mycelial biomass were higher at 30 °C (p ≤ 0.05)
with the first wild strains completing the Petri
dish colonization in seven days (Figure 3a-d).
Figure 1. Effects of different
temperatures on growth of
12 Brazilian wild strains of
Auricularia fuscosuccinea
on the seventh day. (a)
Mycelium diameter (mm); (b)
Dry mycelial biomass (mg).
Capital letters compare the
means of all wild strains at
different temperatures. The
asterisk indicates statistical
significance by Tukey’s test
at 0.05 probability of the
best values obtained by
the wild strain at the best
temperature.
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 6 | 14
Comparing the mycelial growth diameter among
all wild strains at 30 °C, there was no difference
(p > 0.05) between the wild strains CCIBt2381,
CCIBt2959, CCIBt4747, CCIBt4748, CCIBt4751,
CCIBt4753, CCIBt4756, and CCIBt4758 (Figure 1a).
However, in relation to dry mycelial biomass
(Figure 1b), on the seventh day of colonization at
30 °C, the wild strain CCIBt2381 had the highest
biomass production (171.20 mg ± 10.01 mg),
followed by the wild strain CCIBt4753 (151.11 mg ±
6.77 mg). Based on these results, the wild strains
CCIBt2381 and CCIBt4753 were selected for the
substrates experiments.
The wild strains CCIBt4753 and CCIBt2381
completed the colonization of the eucalyptus
sawdust substrate in 21 and 22 days, respectively
(Figure 2). The wild strain CCIBt4753, although
it took longer to start growing in the sawdust
(sixth day), completed the substrate colonization
before the other wild strain (CCIBt2381) and
presented an average daily growth of 5.21
mm ± 2.99 mm, against 4.88 mm ± 0.96 mm
of the wild strain CCIBt2381. In the JunCao
substrate, the wild strain CCIBt2381 showed a
good development at the beginning of the
experiment (only until the tenth day), with an
average daily growth of 1.00 mm ± 1.45 mm, but
without surpassing the development in sawdust.
The wild strain CCIBt4753 developed little in the
JunCao substrate, showing an average growth of
0.26 mm ± 0.36 mm. For both wild strains, the
substrate based on eucalyptus sawdust showed
better mycelial growth results (p ≤ 0.05).
The wild strain CCIBt2381 took 25 to 27 days
to fully colonize the 2.5 kg sawdust substrate
block, while the wild strain CCIBt4753 took 28 to
34 days. However, in the blocks with CCIBt4753,
primordia were observed from 14 to 17 days
after induction, not on the surface but in the
lower half of the blocks (Figure 3e-f), and the
harvest were performed 35 days after induction
of primordia. The primordia of CCIBt2381
developed 21 days after induction, and the first
harvest was performed 45 days after primordia
induction (Figure 3g-h). Nutritional composition
of basidiomata produced is shown in Table II.
Laetiporus gilbertsonii
For the three wild strains of L. gilbertsonii
evaluated, the temperature that best favored
the growth of the mycelium was at 30 °C (p ≤
0.05), with all the wild strains completing the
mycelial growth in the Petri dish in seven days
(Figures 4a and 6a-d). However, in relation to
dry mycelial biomass (Figure 4b), the values
were higher at both 25 °C and 30 °C (p ≤ 0.05).
The wild strain CCIBt4710, despite not showing
differences in growth diameter against the
other two wild strains evaluated, was the one
that obtained the best values (p ≤ 0.05) of dry
mycelial biomass, both at 25 °C (78.71 mg ± 5.10
mg) and 30 °C (78.00 mg ± 4.56 mg).
The wild strains CCIBt4710 and CCIBt4718
were evaluated in the substrates experiment
and completed the colonization of the sawdust-
based substrate in 11 and 12 days, respectively
(Figure 5). The mycelium of L. gilbertsonii is very
Figure 2. Cumulative mycelial growth of two Brazilian
wild strains of Auricularia fuscosuccinea in substrates
JunCao and based on eucalyptus sawdust. The asterisk
indicates statistical significance by Tukey’s test at 0.05
probability.
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 7 | 14
aerial, fragile, and powdered, but it grew fast
in the substrate based on eucalyptus sawdust
after the second day from the inoculation. The
average daily growth in this substrate was 9.60
mm ± 2.85 mm for the wild strain CCIBt4710 and
9.39 mm ± 2.88 mm for the wild strain CCIBt4718.
On the JunCao substrate, the mycelium started
to grow on the fourth day after inoculation,
and it developed little, with an average daily
growth of 0.27 mm ± 0.36 mm for the wild
strain CCIBt4710 and 0.16 mm ± 0.24 mm for the
wild strain CCIBt4718. The substrate based on
eucalyptus sawdust was better for the mycelial
development (p ≤ 0.05) of both wild strains of
L. gilbertsonii when compared to the JunCao
substrate.
Figure 3. Cultivation of Brazilian wild strains of Auricularia fuscosuccinea. (a-d) Mycelial growth of the wild strain
CCIBt4753 in PDA medium on the seventh day; (a) Temperature at 20 °C; (b) Temperature at 25 °C; (c) Temperature
at 30 °C; (d) Temperature at 35 °C; (e) Primordia of the wild strain CCIBt4753; (f) Beginning of the basidiomata
development of the wild strain CCIBt4753; (g) Basidiomata of the wild strain CCIBt4753 at harvest point; (h)
Basidiomata of the wild strain CCIBt2381 at harvest point. Scale bars = 3cm. Photos by M.P. Drewinski.
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 8 | 14
The wild strains CCIBt4710 and CCIBt4718
were evaluated in the experiment of cultivation
in blocks (2.5 kg) with substrate based on
eucalyptus sawdust. The wild strains colonized
the block quickly (Figure 6f), taking from 9 to 10
days (CCIBt4710) and from 10 to 11 days (CCIBt4718)
to complete the blocks (Figure 6g). After 30 days
of inoculation, the blocks were induced to form
primordia. In the blocks in which the bag was
opened and/or the surface layer of the mycelium
was removed from substrate, there was new
mycelial growth with an intensification of the
orange color of the mycelium, but without the
formation of primordia. In the blocks where the
bag was not opened, it was possible to observe
the mycelium growing out of the filter (Figure
6h-k) in: four bags of the wild strain CCIBt4710
incubated in the refrigerator; one bag of the wild
strain CCIBt4710 that was injected cold water
through the filter; and one bag of the wild strain
CCIBt4718 that was incubated in the refrigerator.
Despite the formation of this external mycelial
mass, over time other contaminating fungi
began to grow on the surface of the mycelium of
L. gilbertsonii and it was not possible to obtain
basidiomata for this species.
DISCUSSION
The growth experiments on PDA culture medium
at different temperatures showed a better
mycelial development of the A. fuscosuccinea
wild strains at 30 °C. Coniglio et al. (2021)
studied five wild strains of A. fuscosuccinea
from Argentinian Paranaense Rainforest and
also indicates 30 °C as the optimal temperature
for mycelial development in PDA medium. The
evaluated wild strains of L. gilbertsonii developed
well at both 25 °C and 30 °C. Hitherto, there are
no studies on cultivation of L. gilbertsonii, only
a few studies on another species of the genus, L.
sulphureus, for which the suitable temperatures
for mycelial growth also ranged between 25 °C
and 30 °C (Okamura et al. 2000, Luangharn et
al. 2014). Normally, tropical mushroom species
grow rapidly at 25 °C or higher temperatures,
and thus they can be produced in tropical areas
more quickly than species from temperate
climate areas (Klomklung et al. 2012).
Most studies on the domestication of A.
fuscosuccinea were published by researchers
from Mexico (Castillejos-Puón et al. 1996,
Calvo-Bado et al. 1996, Carreño-Ruiz et al. 2014,
Morales & Sánchez 2017), although there are
studies with specimens from other locations,
such as the United States of America, Brazil,
Peru, Colombia, and Ecuador (Wong 1993,
Vargas et al. 2015, Niño et al. 2017, Rodríguez et
al. 2018). Wong (1993) studied two strains of A.
fuscosuccinea from Brazil with success on the
basidiomata production, although the authors
did not describe in detail the substrate used
neither the cultivation conditions. Here,A we
demonstrate that it is possible to use eucalyptus
sawdust as substrate to produce wild strains of
A. fuscosuccinea. The use of substrates based on
eucalyptus sawdust has already been reported
to produce other wild mushrooms such as A.
auricula-judae (Chen et al. 2013), Oudemansiella
Table II. Nutritional composition of two Brazilian wild
strains of Auricularia fuscosuccinea produced on
sawdust-based substrate.
CCIBt2381 CCIBt4753
Moisture 12.79 % 12.17 %
Ash 4.79 % 4.49 %
Crude Protein 10.73 % 12.11 %
Crude Fat 0.91 % 0.82 %
Crude Fiber 3.85 % 3.73 %
Carbohydrate 70.78 % 70.41 %
Values are expressed on dry matter. Protein conversion factor
N × 4.38
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 9 | 14
Figure 4. Effects of different
temperatures on growth of
three Brazilian wild strains
of Laetiporus gilbertsonii
on the seventh day. (a)
Mycelium diameter (mm); (b)
Dry mycelial biomass (mg).
Capital letters compare the
means of all wild strains at
different temperatures. The
asterisk indicates statistical
significance by Tukey’s
test at 0.05 probability of
the best values obtained
by the wild strains at each
temperature.
Figure 5. Cumulative mycelial
growth of two Brazilian
wild strains of Laetiporus
gilbertsonii in substrates
JunCao and based on
eucalyptus sawdust. The
asterisk indicates statistical
significance by Tukey’s test
at 0.05 probability.
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 10 | 14
Figure 6. Cultivation of Brazilian wild strains of Laetiporus gilbertsonii. (a-d) Mycelial growth of the wild strain
CCIBt4710 in PDA medium on the seventh day; (a) Temperature at 20 °C; (b) Temperature at 25 °C; (c) Temperature
at 30 °C; (d) Temperature at 35 °C; (e) Specimen MPD306 (= CCIBt4710) in the field; (f) Fully colonized block
CCIBt4710; (g) Fully colonized block CCIBt4718; (h-k) Mycelial growth out of the filter of a block of the wild strain
CCIBt4710. Scale bars = 3cm. Photos by M.P. Drewinski.
canarii (Jungh.) Höhn. (Ruegger et al. 2001),
and Gymnopilus pampeanus (Speg.) Singer
(Colavolpe & Albertó 2014).
Regarding the composition of A.
fuscosuccinea produced in this study, the
crude protein values found (10.73–12.11 %) were
higher than those obtained by Mau et al. (1998)
but lower than those obtained by Sánchez et
al. (2018), which found 8.62 % and 13.5 % of
crude protein, respectively. For crude fiber, the
values obtained here (3.73–3.85 %) were higher
than those found by Sánchez et al. (2018) but
lower than those recorded by Mau et al. (1998),
which found 5.8 % and 11.69 % of crude fiber,
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 11 | 14
respectively. The value of the crude fat in the
samples studied here (0.82–0.91 %) was much
lower than that found by Mau et al. (1998) and
Sánchez et al. (2018) for A. fuscosuccinea, which
were 4.48 % and 13.5 %, respectively.
This is the first study on cultivation of
L. gilbertsonii. Regarding on the evaluated
substrates, it was possible to observe a fast
colonization of the mycelium of L. gilbertsonii in
the substrate based on eucalyptus sawdust. This
result was already expected because the species
occurs naturally in Eucalyptus spp. wood (Burdsall
Jr. & Banik 2001). Unlike other commercially
produced mushroom species, L. gilbertsonii is a
brown rot fungus and does not produce lignin-
degrading enzymes (Burdsall Jr. & Banik 2001).
Pleszczyńska et al. 2013 were the first to report
the successful initiation and development of a
Laetiporus species mushrooms in large scale
experiments. They studied the development
of twelve wild strains of L. sulphureus from
Poland in a substrate based on a mixture of
sawdust. Among the twelve studied strains, only
two produced basidiomata. It was found that
shocking the fungus mycelium with cold water
or at low temperature was the most suitable
method for forcing L. sulphureus basidioma to
grow (Pleszczyńska et al. 2013). In this study, we
tested the same induction methods used by
Pleszczyńska et al. 2013 for L. gilbertsonii, but we
were not successful in obtaining primordia and
neither producing basidiomata, unfortunately.
Besides the mushrooms, mycelia and the
culture media used in the fungi cultivation also
have been explored as potential sources of food
and bioactive compounds (Cheung 1996, Gan et
al. 2012, Ma et al. 2016, Souilem et al. 2017, Stoffel
2019). Compared to mushrooms production,
mycelium cultivation has potential advantages
for higher production of biomass in a more-
compact space over a shorter incubation time
(Gan et al. 2012).
CONCLUSIONS
We obtained twelve wild strains of A. fuscosuccinea
and three wild strains of L. gilbertsonii from the
Brazilian Atlantic Rainforest. The temperature
that best favored the mycelial growth of A.
fuscosuccinea was 30 °C. For L. gilbertsonii, the
temperatures of 25 °C and 30 °C were suitable
for the mycelium development. Mycelia of
both species better developed in the sterile
sawdust substrate, in which it was possible to
produce basidiomata of the two wild strains
of A. fuscosuccinea evaluated. The nutritional
values of A. fuscosuccinea studied here are
similar to those described for other species of
the genus. This is the first study on cultivation
of L. gilbertsonii. Although the mycelia of the
two evaluated wild strains of L. gilbertsonii
colonized the eucalyptus-based substrate
quickly, they do not produced primordia neither
basidiomata. However, it was possible to observe
the mycelium of L. gilbertsonii growing out of
the filter in some bags in which methods based
on low temperature induction were performed.
There is a huge potential associated to the
collecting, identification, and maintenance
of mushroom strains and further studies are
needed for domestication and optimization of
wild species production.
Acknowledgments
The authors are grateful for the financial support
received from the ‘Fundação de Amparo à Pesquisa
do Estado de São Paulo – FAPESP’ (grants #2017/25754-
9 and #2018/15677-0). MPD thanks Jorge Arruda Neto
for donating the pasteurized substrate. NMJ thanks
the ‘Conselho Nacional de Desenvolvimento Científico
e Tecnológico–CNPq’ (Research Productivity grant
314236/2021-0).
REFERENCES
ALBERTÓ E. 2017. Naturally occurring strains of edible
mushrooms: a source to improve the mushroom industry.
In: Zied DC & Pardo-Giménez A (Eds), Edible and Medicinal
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 12 | 14
Mushrooms: Technology and Applications, John Wiley &
Sons, p. 415-425. https://doi.org/10.1002/9781119149446.
ch19.
ALVARENGA RLM, NAVES LRR & XAVIER-SANTOS S. 2015. The
Genus Auricularia Bull. ex Juss. (Basidiomycota) in
Cerrado (Brazilian Savanna) areas of Goiás state and the
Federal District, Brazil. Mycosphere 6: 532-541. https://
doi.org/10.5943/mycosphere/6/5/3.
BANIK MT, LINDNER DL, ORTIZ-SANTANA B & LODGE DJ. 2012. A
new species of Laetiporus (Basidiomycota, Polyporales)
from the Caribbean basin. Kurtziana 37: 15-21.
BURDSALL JR. HH & BANIK MT. 2001. The genus Laetiporus
in North America. HPB 6: 43-55.
CALVO-BADO LA, SÁNCHEZ-VÁZQUEZ JE & HUERTA-PALACIOS G.
1996. Cultivation of Auricularia fuscosuccinea (Mont.)
Farlow on agricultural substrates in Soconusco region,
Chiapas, Mexico. Micología Neotropical Aplicada 9:
95-106.
CAMPI MG, AZEVEDO-OLIVEIRA C, COSTA-REZENDE D, CANO YM,
MORERA G, URCELAY C, DRECHSLER-SANTOS ER & ROBLEDO
GL. 2022. What are the Laetiporus species presente in
Southern South America? Lilloa 59: 193-218. https://doi.
org/10.30550/j.lil/2022.59.S/2022.09.19.
CARREÑO-RUIZ SD, CAPPELLO-GARCÍA S, GAITÁN-HERNÁNDEZ
R, CIFUENTES-BLANCO J & ROSIQUE-GIL E. 2014. Crecimiento
de tres hongos comestibles tropicales en medios de
cultivo y residues agrícolas. Rev Mexicana Cienc Agric 5:
1447-1458.
CASTILLEJOS-PUÓN V, SÁNCHEZ-VÁZQUEZ JE & PALACIOS
GH. 1996. Evaluación de cepas del hongo comestible
Auricularia fuscosuccinea nativas del Soconusco,
Chiapas, México. S F 12: 23-30. https://doi.org/10.33885/
sf.1996.3.845.
CHEN L, CHEN Z, WANG Q, SHAO L & HUANG Z. 2013. Application
of Eucalyptus sawdust on strain production of edible
fungi. South J Agric Econ 44: 644-648. https://doi.
org/10.3969/j:issn.2095-1191.2013.4.644.
CHEUNG PCK. 1996. Dietary fiber content and composition
of some cultivated edible mushroom fruiting bodies
and mycelia. J Agr Food Chem 44: 468-471. https://doi.
org/10.1021/jf950455l.
COLAVOLPE MB & ALBERTÓ E. 2014. Cultivation requirements
and substrate degradation of the edible mushroom
Gymnopilus pampeanus—A novel species for mushroom
cultivation. Sci Hortic 180: 161-166. https://doi.
org/10.1016/j.scienta.2014.10.011.
CONIGLIO R, DÍAZ G, LÓPEZ C, RESTELLI M, GRASSI E, ALBERTÓ E
& ZAPATA P. 2021. Solid-state bioprocessing of sugarcane
bagasse with Auricularia fuscosuccinea for phenolic
compounds extraction. Prep Biochem Biotech 52: 701-
710. https://doi.org/10.1080/10826068.2021.1986722.
CRISAN EV & SANDS A. 1978. Nutritional value. In: Chang ST
& Hayes WW (Eds), The Biology and Cultivation of Edible
Mushrooms, Academic Press, New York, p. 137-168.
DARRIBA D, TABOADA G, DOALLO R & POSADA D. 2012.
jModelTest 2: more models, new heuristics and parallel
computing. Nat Methods 9: 772. https://doi.org/10.1038/
nmeth.2109.
DREWINSKI MP. 2023. Cogumelos comestíveis do Brasil:
diversidade e viabilidade de cultivo [thesis]. Instituto de
Pesquisas Ambientais, Sâo Paulo, SP.
FIDALGO O & HIRATA JM. 1979. Etnomicologia Caiabi, Txicão
e Txucarramãe. Rickia 8: 1-5.
GAMBOA-TRUJILLO P ET AL. 2019. Edible Mushrooms of
Ecuador: consumption, myths and implications for
conservation. Ethnobot Res Appl 18: 1-15. http://dx.doi.
org/10.32859/era.18.38.1-15.
GAN D, MA LP, JIANG CX, WANG MC & ZENG XX. 2012. Medium
optimization and potential hepatoprotective effect of
mycelial polysaccharides from Pholiota dinghuensis Bi
against carbon tetrachloride-induced acute liver injury
in mice. Food Chem Toxicol 50: 681-2688. https://doi.
org/10.1016/j.fct.2012.05.003.
KATOH K, ROZEWICKI J & YAMADA KD. 2019. MAFFT online
service: multiple sequence alignment, interactive
sequence choice and visualization. Briefings in Bioinf 20:
1160-1166. https://doi.org/10.1093/bib/bbx108.
KEARSE M ET AL. 2012. Geneious Basic: an integrated
and extendable desktop software platform for
the organization and analysis of sequence data.
Bioinformatics 28: 1647-1649. https://doi.org/10.1093/
bioinformatics/bts199.
KLOMKLUNG N, KARUNARATHNA SC, CHUKEATIROTE E & HYDE KD.
2012. Domestication of wild strain of Pleurotus giganteus.
Sydowia 64: 39-53.
LARSSON A. 2014. AliView: a fast and lightweight alignment
viewer and editor for large datasets. Bioinformatics
30: 3276-3278. https://doi.org/10.1093/bioinformatics/
btu531.
LI H ET AL. 2021. Reviewing the world’s edible mushroom
species: A new evidence‐based classification system.
Compr Rev Food Sci Food Saf 20: 1982-2014. https://doi.
org/10.1111/1541-4337.12708.
LINDNER DL & BANIK MT. 2008. Molecular phylogeny
of Laetiporus and other brown rot polypore genera
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 13 | 14
in North America. Mycologia 100: 417-430. https://doi.
org/10.3852/07-124r2.
LOONEY BP, BIRKEBAK JM & MATHENY PB. 2013. Systematics of
the genus Auricularia with an emphasis on species from
the southeastern United States. North American Fungi 8:
1-25. https://doi.org/10.2509/naf2013.008.006.
LOWY B. 1952. The genus Auricularia. Mycologia 44:
656-692.
LUANGHARN T, KARUNARATHNA SC, HYDE KD & CHUKEATIROTE E.
2014. Optimal conditions of mycelia growth of Laetiporus
sulphureus sensu lato. Mycology 5: 221-227. https://doi.or
g/10.1080/21501203.2014.957361.
MA G, YANG W, FANG Y, MA N, PEI F, ZHAO L & HU Q. 2016.
Antioxidant and cytotoxicites of Pleurotus eryngii
residue polysaccharides obtained by ultrafiltration. LWT
Food Sci Technol 73: 108-116. https://doi.org/10.1016/j.
lwt.2016.05.049.
MAU JL, WU KT, WU YH & LIN YP. 1998. Nonvolatile taste
components of ear mushrooms. J Agric Food Chem 46:
4583-4586. https://doi.org/10.1021/jf9805606.
MORALES V & SÁNCHEZ JE. 2017. Self-heating pasteurization
of substrates for culinary-medicinal mushrooms
cultivation in Mexico. Int J Med Mushrooms 19: 477-484.
https://doi.org/10.1615/IntJMedMushrooms.v19.i5.90.
NIÑO YM, PEÑA ER & ENAO LG. 2017. Aislamiento y producción
de semilla de Auricularia fuscosuccinea (Mont.) Henn. y
Crepidotus palmarum Sing. Usados tradicionalmente en
Pauna (Boyacá, Colombia). Rev Colomb Cienc Hortic 11:
151-158. https://doi.org/10.17584/rcch.2017v11i1.5616.
OKAMURA T, TAKENO T, FUKUDA S, MOHRI A, NODA H, IEMOTO
A, HORIE N & OSHUGI M. 2000. Laetiporus sulphureus,
producing an anti-thrombin substance. Mushroom
Sci Biotechnol 8: 121-125. https://doi.org/10.24465/
apmsb.8.3_121.
PIRES RM, MOTATO-VÁSQUEZ V & GUGLIOTTA AM. 2016. A new
species of Laetiporus (Basidiomycota) and occurrence
of L. gilbertsonii Burds. in Brazil. Nova Hedwigia 102: 477-
490. https://doi.org/10.1127/nova_hedwigia/2016/0320.
PLESZCZYŃSKA M, WIATER A, SIWULSKI M & SZCZODRAK J. 2013.
Successful large-scale production of fruiting bodies of
Laetiporus sulphureus (Bull.: Fr.) Murrill on an artificial
substrate. World J Microbiol Biotechnol 29: 753-758.
https://doi.org/10.1007/s11274-012-1230-z.
RODRÍGUEZ EJO, INSUASTI JAP, TRUJILO ASD, ARROYAVE CPS, SOTO
CAP & DEL AMBIENTE CEDB. 2018. Auricularia fuscosuccinea:
una cepa native ecuatoriana para hacer frente a la crisis
alimentaria. Rev Biorrefinería 1: 32-39.
RONQUIST F, TESLENKO M, VAN DER MARK P, AYRES DL, DARLING
A, HÖHNA S, LARGET B, LIU L, SUCHARD MA & HUELSENBECK
JP. 2012. MrBayes 3.2: efficient Bayesian phylogenetic
inference and model choice across a large model space.
Syst Biol 61: 539-542. https://doi.org/10.1093/sysbio/
sys029.
ROYSE DJ, BAARS J & TAN Q. 2017. Current overview of
mushroom production in the world. In: Zied DC & Pardo-
Giménez A (Eds), Edible and Medicinal Mushrooms:
Technology and Applications. John Wiley & Sons, p. 5-13.
https://doi.org/10.1002/9781119149446.ch2.
RUÁN-SOTO F, DOMÍNGUEZ-GUTIÉRREZ M, PÉREZ-RAMÍREZ L &
CIFUENTES J. 2021. Etnomicología de los lacandones de
Nahá, Metzabok y Lacanjá-Chansayab, Chiapas, México.
Ciencias Sociales y Humanidades 8: 24-42. https://doi.
org/10.36829/63CHS.v8i1.1112.
RUEGGER MJS, TOURNISIELO SMT, BONONI VLR & CAPELARI M.
2001. Cultivation of edible mushroom Oudemansiella
canarii (Jungh.) Höhn. in lignocellulosic substrates.
Braz J Microbiol 32: 211-214. https://doi.org/10.1590/
S1517-83822001000300009.
SÁNCHEZ JE, MORENO L & ANDRADE RH. 2018. Auricularia
spp. In: Sánchez JE, Mata G & Royse DJ (Eds), Updates
on Tropical Mushrooms. Basic and Applied Research,
ECOSUR, Mexico, p. 137-150.
SOUILEM F, FERNANDES Â, CALHELHA RC, BARREIRA JC,
BARROS L, SKHIRI F & FERREIRA IC. 2017. Wild mushrooms
and their mycelia as sources of bioactive compounds:
Antioxidant, anti-inflammatory and cytotoxic properties.
Food Chem 230: 40-48. https://doi.org/10.1016/j.
foodchem.2017.03.026.
STOFFEL F, OLIVEIRA-SANTANA W, GREGOLON JGN, KIST TBL,
FONTANA RC & CAMASSOLA M. 2019. Production of edible
mycoprotein using agroindustrial wastes: Influence
on nutritional, chemical and biological properties.
Innovative Food Sci Emerg Technol 58: 102227. https://
doi.org/10.1016/j.ifset.2019.102227.
THAWTHONG A ET AL. 2014. Discovering and Domesticating
Wild Tropical Cultivatable Mushrooms. Chiang Mai J Sci
41: 731-764.
VARGAS LAF, AGURTO MEP & FLORES MA. 2015. Identificación
de las técnicas de propagación y cultivo en residues
agroindustriales de hongos comestibles originarios de
San Juan de Cacazúen la Selva central de la Amazonía
peruana. Producción Agropecuaria y Desarrollo
Sostenible 4: 47-63. https://doi.org/10.5377/payds.
v4i0.3963.
VARGAS-ISLA R & ISHIKAWA NK. 2008. Optimal conditions of
in vitro mycelial growth of Lentinus strigosus, an edible
MARIANA P. DREWINSKI et al. DOMESTICATION OF WILD EDIBLE MUSHROOMS FROM BRAZIL
An Acad Bras Cienc (2024) 96(4) e20230838 14 | 14
mushroom isolated in the Brazilian Amazon. Mycoscience
49: 215-219. https://doi.org/10.1007/S10267-007-0404-2.
VIEIRA S. 1980. Introdução à bioestatística. Editora
Campus, Rio de Janeiro.
WHITE TJ, BRUNS T, LEE SJWT & TAYLOR J. 1990. Amplification
and direct sequencing of fungal ribosomal RNA genes
for phylogenetics. PCR protocols: a guide to methods
and applications 18: 315-322.
WONG GJ. 1993. Mating and fruiting studies of Auricularia
delicata and A. fuscosuccinea. Mycologia 85: 187-194.
https://doi.org/10.1080/00275514.1992.12026266.
WU F, TOHTIRJAP A, FAN LF, ZHOU LW, ALVARENGA RL, GIBERTONI
TB & DAI YC. 2021. Global diversity and updated phylogeny
of Auricularia (Auriculariales, Basidiomycota). J Fungi 7:
933. https://doi.org/10.3390/jof7110933.
ZENEBON O, PASCUET NS & TIGLEA P. 2008. Métodos físico-
químicos para análise de alimentos. Instituto Adolfo
Lutz, São Paulo.
SUPPLEMENTARY MATERIAL
Figures S1 and S2.
How to cite
DREWINSKI MP, CORRÊA-SANTOS MP, ZIED DC & MENOLLI JR N. 2024.
Studies on domestication of two species of wild edible mushroom
from Brazil. An Acad Bras Cienc 96: e20230838. DOI 10.1590/0001-
3765202420230838.
Manuscript received on July 26, 2023;
accepted to publication on July 24, 2024
MARIANA P. DREWINSKI1,2
https://orcid.org/0000-0002-7299-8477
MARINA P. CORRÊA-SANTOS1,2
https://orcid.org/0000-0002-4329-9861
DIEGO C. ZIED3
https://orcid.org/0000-0003-2279-4158
NELSON MENOLLI JR.1,2
https://orcid.org/0000-0002-1841-8179
1Instituto de Pesquisas Ambientais, Núcleo de Pós-
graduação Stricto Sensu, Pós-graduação em Biodiversidade
Vegetal e Meio Ambiente, Av. Miguel Stefano 3687,
Água Funda, 04301-012 São Paulo, SP, Brazil
2Instituto Federal de Educação, Ciência e Tecnologia de São
Paulo, Departamento de Ciências da Natureza e Matemática,
Subárea de Biologia, IFungiLab, Câmpus São Paulo, Rua
Pedro Vicente 625, 01109-010 São Paulo, SP, Brazil
3Universidade Estadual Paulista Júlio de Mesquita Filho
(UNESP), Faculdade de Ciências Agrárias e Tecnológicas,
Câmpus Dracena, Rod. Comandante João Ribeiro de
Barros, km 651, 17900-000 Dracena, SP, Brazil
Correspondence to: Mariana de Paula Drewinski
E-mail: maridrewinski@gmail.com
Author contributions
MARIANA P. DREWINSKI: Data curation, Formal analysis,
Investigation, Methodology, Resources, Software, Validation,
and Writing – original draft; MARINA P. CORRÊA-SANTOS: Data
curation, Formal analysis, and Investigation; DIEGO C. ZIED:
Methodology, Funding acquisition, Resources, Supervision,
and Writing – review & editing; NELSON MENOLLI JR.:
Conceptualization, Funding acquisition, Methodology, Project
administration, Resources, Supervision, Visualization, and
Writing – review & editing.
