Analysis of indole compounds in fruiting bodies and in mycelia from 
in vitro cultures of Calocera viscosa (Basidiomycota)

BOŻENA MUSZYŃSKA and KATARZYNA SUŁKOWSKA-ZIAJA

Chair and Department of Pharmaceutical Botany, Collegium Medicum, Jagiellonian University 
Medyczna 9, PL-30-688 Kraków, muchon@poczta.fm

Muszyńska B., Sułkowska-Ziaja K.: Analysis of indole compounds in fruiting bodies and in 
mycelia from in vitro cultures of Calocera viscosa (Basidiomycota). Acta Mycol. 47 (1): 57–64, 
2012.

Calocera viscosa (Pers.: Fr.) Fr. (Basidiomycota) from Dacrymycetaceae family is 
a widespread species of mushroom in Poland. The aim of this study was to investigate the 
content of indole compounds in fruiting bodies and in mycelium cultured in vitro on solid and 
liquid medium of this species.

Fruiting bodies of Calocera viscosa were collected in coniferous forests in south Poland 
and were used to derive in vitro cultures. The optimal medium composition for cultures was 
determined. Fresh material: fruiting bodies and mycelium from culture in vitro was frozen 
and then dried by lyophilization. The crushed dry biomass was extracted with petroleum ether 
to remove oil fraction, material was dried and extracted with methanol. Analysis of indole 
compounds was performed in methanol extracts using chromatographic methods: TLC, 
UV Vis, EIMS and HPLC. This analysis presented in all three extracts the following indole 
compounds: L-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, melatonin and indole 
(contents fluctuated in the range: 0.37 to 11.88 mg/100 g d.w.). 5-hydroxytryptophan contents 
in all extracts were significant and amounted to 11.88 mg/100 g d.w. in fruiting bodies, and 
11.42 in mycelium from liquid cultures and and 10.59 in mycelium from solid cultures. In 
addition, the fruiting bodies and mycelium from cultures on liquid medium revealed the 
presence of serotonin (0.39 and 3.19 mg/100 g d.w. respectively).
Key words: β,β-carotene, 5-hydroxytryptophan, mushroom, mycelial culture, serotonin

INTRODUCTION

Since the beginning of human evolution fruiting bodies of mushrooms belonging to 
the taxon Basidiomycota are appreciated for flavor and texture but especially for 
their chemical and dietary properties. Mushrooms are a rich source of a variety of 
biologically active compounds belonging to both primary and secondary metabolites 

 ACTA MYCOLOGICA
 Vol. 47 (1): 57–64
 2012



58 B. Muszyńska and K. Sułkowska-Ziaja 

(Yang et al. 2001; Wasser 2002; Muszyńska et al. 2010). More than 140,000 species of 
mushrooms exist in natural sites, but less than twenty five species are widely accept-
ed as food (Barros et al. 2007). The mushroom glucans are well known for their im-
munomodulatory properties and are used for anticancer therapy (Sułkowska-Ziaja 
et al. 2005; Zaidman et al. 2005; Muszyńska et al. 2011a). Antitumor and cytostatic 
properties presented numerous terpenoids, especially sesquiterpenes and triterpe-
noids. A number of recent mycochemical papers have reported on the presence of 
many different terpene compounds (Barros et al. 2008; Liu 2005). 

The antioxidants found in edible mushrooms were flawonoids, phenolic com-
pounds, ascorbic acid, tocopherols and carotenoids. Recently, Muszyńska (Muszyńska 
et al. 2007; Muszyńska et al. 2009; Muszyńska et al. 2011b) have hinted at the occur-
rence of numerous non-hallucinogenic indole compounds in edible, conditionally ed-
ible and inedible Basidiomycota species. Calocera viscosa (Pers.:Fr.) Fr. (Yellow Stag-
shorn) is a species common in Polish, European and Asiatic coniferous forests, which 
develops intensely yellow-orange branched, bush-like fruiting bodies. This species oc-
curs from June to November on dead conifer wood in mixed and coniferous forests, 
mainly in mossy spruce and pine stumpsas saprotrophs. C. viscosa is a good source of 
free exo- and endogenous amino acids, indole compounds, sterols (especially ergos-
terol) and unsaturated fatty acids (Muszyńska 1999). Although the carotenoids con-
tent in Calocera viscosa fruiting bodies growing in Poland were estimated spectropho-
tometrically at 2.46 μg/g f.w. by Czeczuga (1980) at the end of the seventies years of the 
20th century. Barros (2008) demonstrated that of six Basidiomycota species collected 
from natural sites in the northwestern Portugal, Cantharellus cibarius (Chantarelle) 
contained the greatest amounts of β,β-carotene (13.56 μg/g d.w.) while the contents of 
this metabolite in the remaining species ranged from 1.95 - 12.77 μg/g d.w.

The contents of this very important antioxidant were analyzed in extracts from 
fruiting bodies of C. viscosa and in extracts from mycelium of in vitro cultures 
(Muszyńska et al. 2012). These contents are higher than those determined by Czec-
zuga (1980). The cultures were maintained under different conditions to optimize 
biomass growth and to establish the most beneficial conditions for β,β-carotene ac-
cumulation. β,β-Carotene content in biomass from solid cultures was comparable 
with that found in fruiting bodies (7.1 and 7.5 μg/g d.w., respectively). Mycelia from 
liquid cultures contained half of that β,β-carotene amount which equaled 3.5 μ/g 
d.w. Polysaccharides extracted from the mycelial culture of C.viscosa and admin-
istrated intraperitoneally into mice at dosage of 300 mg/kg inhibited the growth of 
Sarcoma 180 and Erlich solid cancers by 90% (Ohtsuka et al. 1973). The aim of the 
present study was to initiate Calocera viscosa culture in vitro, to determine optimal 
conditions for mycelia growth and to evaluate the indole compounds contents in 
extracts of fruiting bodies and mycelia from Calocera viscosa in vitro cultures. In-
dole compounds have attracted much interest recently because in vitro and in vivo 
studies suggest that they have a variety biological properties, which play important 
functions in the maintenance of human health. These compounds are antioxidants, 
antidepressants, anticancer, being tissue hormones and neurotransmitters. Seroto-
nin is a long known compound playing the role of a regulator of sleep, body tem-
perature, mood, maturation and regeneration and an inhibitor of cell aging, thereby 
contributing to general strengthening of the immune system. Its daily dose when 
used, for instance, as an antidepressant drug ranges from 100 to 200 mg (Mosovich 



 Analysis of indole 59

et al. 2008). Recent reports have revealed further important biological aspects of 
serotonin action, including its usefulness in prevention of Alzheimer’s disease and 
the antioxidant action (Ouchi et al. 2009). Indole compounds and their derivatives 
play also important role as analgesic and anti-inflammatory medicines. 

MATERIALS AND METHODS

Origin of fruiting bodies. The studies were conducted on fruiting bodies of Calocera 
viscosa (Pers.: Fr.) Fr. collected at natural sites in mixed and coniferous forests in 
southern Poland (Brodła near Kraków) (deposited in the Department of Pharma-
ceutical Botany, Jagiellonian University, Collegium Medicum, Kraków, Poland). 

Initial culture. Initial cultures were derived from explants originating from the 
top parts of branched, bushlike fruiting bodies of Calocera viscosa. These pieces of 
fruiting bodies were sterilized with 70% ethyl alcohol and placed on Petri dishes with 
solid Oddoux medium (Oddoux 1957). Cultures were incubated at a temperature 25 
+/- 2° C under 12-h light (900 lx)/12 dark cycle and were subcultured every second 
week. For more details see Muszyńska (Muszyńska et al. 2009).

Experimental in vitro cultures. Solid culture was maintained on Petri dishes, on 
Oddoux medium (Oddoux 1957).

Stationary liquid culture was established from the solid culture by transferring 
0.1 g of mycelium into an Erlenmayer flask (500 mL) containing 250 mL of liquid 
Oddoux medium. Both types of experimental cultures were maintained under the 
same conditions as initial culture and were subcultured every two weeks. 

Extraction. Lyophilized and powdered fruiting bodies and mycelia from Calocera 
viscosa in vitro cultures on solid and liquid media were extracted in a percolator with pe-
troleum ether to remove oil fraction according to the procedure developed in our labo-
ratory (Muszyńska et al. 2007). After this extraction, biomass was dried and extracted 
by methanol in percolator for 24 h. Extracts were concentrated to 5 mL by distillation 
in a vacuum evaporator under reduced pressure. For the purification of the extracts we 
used TLC method on aluminum – baked silica 60 plates (Merck, Art. No 1.05554.0001), 
on which 1 mL of the extracts was loaded and chromatograms were developed in mobile 
faze: n-butanol / acetic acid / water (12 : 3: 5 v/v/v). Spots were identified at λ=254 nm. 
Obtained fractions were analyzed by UV-Vis, EIMS and HPLC method. 

UV VIS analysis of indole compounds. Purified by preparative TLC method ex-
tracts of fruiting bodies and mycelia from cultures in vitro on solid and liquid me-
dium were analyzed for the presence of indole compounds by spectrophotometry 
using UV-Vis apart: UV-Vis Cary - Varian Spectrophotometer. Absorption measure-
ments were carried out in the λ = 200-500 nm; solvent: methanol AR; standards: 
indole compounds - manufactured by Sigma-Aldrich. In all three extracts was found 
an increase of the growth of an absorption maximum (for example at λmax = 227 and 
λmax = 254) characteristic for indole compounds.

EIMS analysis of indole compounds. Electron Impact Mass Spectrometry Anal-
ysis (EIMS) was performed at the Regional Laboratory of Physicochemical Analy-
ses and Structural Research. Apparatus: High Resolution Mass Spectrometer with 



60 B. Muszyńska and K. Sułkowska-Ziaja 

options: EI, ESI, GC-M, Finnigan MAT 95S. In the results of these studies were 
obtained the spectra for the methanol extracts from the fruiting bodies and mycelia 
from cultures on solid and liquid medium. The EIMS spectra for the methanol ex-
tracts from the fruiting bodies and mycelia from in vitro cultures on solid and liquid 
medium containing peaks characteristic for indole compounds (m/e=115, m/e=129, 
m/e=130, m/e=135, m/e=143, m/e=157).

Estimation of indole compounds by the HPLC method. Contents of indole com-
pounds in extracts from fruiting bodies and in mycelia maintained in in vitro cultures 
on solid and liquid medium were determined after preliminary separation with pre-
parative TLC method. Contents of L-tryptophan, 5-hydroxytryptophan, 5-methyl-
tryptophan, serotonin, melatonin, tryptamine, kynurenic acid, kynurenine sulfate, 
indoleacetic acid, β-indoleacetonitrile, indole and kynurenine were determined ac-
cording to the procedure developed by Kysilka and Wurst (1985) with our modifi-
cations (Muszyńska et al. 2009). Briefly, the analytical conditions were as follows: 
HPLC apparatus: Hitachi; pump: L-7100; column: Purospher® RP-18 (4 x 200 mm, 
5 μm) thermostated at 25°C. The solvent system used were: methanol/water/ammo-
nium acetate (15:14:1 v/v/v); flow rate:1ml/min. Detection was carried out in a UV 
detector, using λ=280 nm; standards: manufactured by Sigma-Aldrich. Standards 
solutions were prepared in HPLC grade methanol. 

The identification of the indole compounds was made by comparing the retention 
times of samples peaks with standards. The results are expressed in mg/100 g of dry 
weight, calculated by internal normalization of the of the chromatographic peak area. 

RESULTS

We established that good mycelial mass growth of Calocera viscosa could be ob-
tained in agitating liquid cultures and solid cultures on modified Oddoux (1957) 
medium at 25 +/- 2°C under 16 h photoperiod (900 lx/8 h dark). A 20-fold fresh 
biomass growth in cultures on solid medium and a 15-fold growth in liquid cultures 
were obtained within a 14-day growth cycle. The biomass growth in the initiated 
cultures averaged 8.3 g d.w./1 L of medium. The obtained biomass increments and 
dynamics of mycelium growth did not differ from the results that we obtained for 
Sarcodon imbricatus L. (Sułkowska-Ziaja 2010), Xerocomus badius (Fr.) Kühn. ex 
Gilb., Tricholoma equestre (L.: Fr.) Kumm. (Muszyńska et al. 2009) and Cantharellus 
cibarius Fr. cultures studied earlier. The in vitro cultures of these species were used 
for evaluation of qualitative and quantitative composition of non-hallucinogenic in-
dole compounds and proved to be a valuable model for investigation of their me-
tabolism. The present study is an extension of the previous studies on accumulation 
of indole compounds and is the first report about their presence in: fruiting bodies 
and in mycelia of Calocera viscosa cultured in vitro. 

The identity of indole compounds was confirmed on the basis of their param-
eters TLC, UV-Vis, EIMS and HPLC methods. The HPLC method was used for 
quantitation of indole compounds and optimum conditions were established by this 
method for separation: L-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, 



 Analysis of indole 61

serotonin, melatonin, tryptamine, kynurenic acid, kynurenine sulfate, indoleacetic 
acid, β-indoleacetonitrile, indole and kynurenine in extracts from fruiting bodies and 
in mycelia from in vitro cultures on solid and liquid medium. This analysis presented in 
all three extracts the following five indole compounds: L-tryptophan, 5-hydroxytrypto-
phan, 5-methyltryptophan, melatonin and indole. In addition, the fruiting bodies and 
mycelium from cultures on liquid medium revealed the presence of serotonin (0.39 
and 3.19 mg/100 g d.w. respectively). Results are presented in Table 1.

Contents of indole compounds in fruiting bodies ranged from 0.39 to 11.88 
mg/100 g d.w., in mycelium from liquid cultures from 0.37 to 11.42 mg/100 g d.w., 
and in mycelium from solid cultures from 0.39 to 10.59 mg/100 g d.w. 5-hydroxytry-
pophan contents in all extracts were significant and amounted 11.88 mg/100 g d.w. 
in fruiting bodies, and 11.42 in mycelium from liquid cultures and 10.59 in mycelium 
from solid cultures. The second metabolites because of their amounts was 5-meth-
yltryptophan and presented: 3.39 mg/100 g d.w. in fruiting bodies, 3.34 in mycelium 
from solid medium and 3.36 in mycelium from liquid medium. The contents of L-
tryptophan and indole were very comparable and ranged from 1.19 mg/100 g d.w. 
to 1.79. Melatonin was found in comparable but small amounts in all three extracts: 
0.47 mg/100 g d.w. in fruiting bodies, 0.39 in mycelium from solid medium and 0.37 
in mycelium from liquid medium. In none of the extracts: tryptamine, kynurenic 
acid, kynurenine sulfate, kynurenine, indoleacetic acid, β-indoleacetonitrile and 
tryptamine were found.

DISCUSSION

Among indole compounds, indole alkaloids and hallucinogenic compounds origi-
nating from mushrooms have been the main focus of interest. Studies of nonhal-
lucinogenic indole compounds have concentrated on tryptophan, among other com-
pounds. Tryptophan was identified in many Basidiomycota species (Kohlmünzer 
et al. 2000). Tryptamine, serotonin, 5-hydroxytryptophan, indoleacetic acid, β –in-
doleacetonitrile, melatonin, kynurenic acid and kynurenine sulfate have also been 
quite frequently identified (Muszyńska et al. 2009). 

Table 1 
Contents of indole compounds (mg/100 g d.w.) in fruiting bodies of Calocera viscosa  

and in mycelia from cultures in vitro

 Fruiting bodies  
of C.viscosa

Mycelium of C.viscosa
 from solid medium

Mycelium of C. viscosa 
from liquid medium

Inndole compounds mg/ 100 g d.w.
melatonin 0.47 ± 0.02 0.39 ± 0.01 0.37 ± 0.01
L-tryptophan 1.26 ± 0.04 1.19 ± 0.01 1.79 ± 0.05
5-hydroxytryptophan 11.88 ± 0.19 10.59 ± 0.11 11.42 ± 0.20
5-methyltryptophan 3.39 ± 0.02 3.34 ± 0.03 3.36 ± 0.02
serotonin 0.39 ± 0.01 – a 3.19 ± 0.07
indole 1.26 ± 0.04 1.21 ± 0.02 1.25 ± 0.03

Data presented as mean of three series ± SE; a - content lower than 0.001 mg/100 g d. w.



62 B. Muszyńska and K. Sułkowska-Ziaja 

Our earlier studies analyzed nonhallucynogenic indole compounds in fruiting bodies 
of the following edible species from natural habitats: Boletus edulis Bull.: Fr , Cantharel-
lus cibarius Fr., Lactarius deliciosus (L.: Fr.) Gray, Leccinum rufum (Schaeff.) Kreisel, 
Suillus luteus (L.: Fr.) Roussel, Xerocomus badius (Fr.:Fr.) Kühner ex Gillbert, and of 
commercial origin: Agaricus bisporus (J.E. Lange) Imbach, Pleurotus ostreatus (Jacq.: 
Fr.) P. Kumm , and the species considered as conditionally edible: Armillaria mellea 
(Vahl:) P. Kumm. ss. lato., Lactarius deterrimus Gröger and Tricholoma equestre (L.: Fr.) 
P. Kumm. ss. lato. The contents of indole compounds in all these species were diverse 
and had a very wide variability range from 0.01 to 39.20 mg/100 g d. w. similar to that 
of C. viscosa (Muszyńska et al. 2007; Muszyńska et al. 2009; Muszyńska et al. 2011a, b). 
Mycelial cultures of three species: Cantharellus cibarius, Tricholoma equestre, Xerocomus 
badius were established in our laboratory and the contents of compounds under study 
range from 0.01 to 20.49 mg/100 g d. w. Found melatonin content in the extracts from 
fruiting bodies of Calocera viscosa was comparable to the content of these compounds 
in the previously studied species (ranged from 0.08 to 1.29 mg/100 g d.w.). In mycelia 
of cultures in vitro melatonin was established only in the extract from mycelium of T. 
equestre (0.60 mg/100 g d.w.) and was comparable with the contents of this compound in 
the mycelia of C. viscosa from solid and liquid medium (0.39 mg/100 g d.w. and 0.37, re-
spectively). In the case of tryptophan, the highest content of this compound was found in 
the fruiting bodies of A. mellea (4.47 mg/100 g d. w.), while the other examined fruiting 
bodies and mycelia from cultures in vitro (X. badius - 0.83 mg/100 g d.w. and T. equestre 
- 1.04) content were identical for all extracts obtained from C. viscosa. The content of 
5-hydroxytryptophan was previously the highest in the currently studied extracts from 
fruiting bodies and mycelium C. viscosa. In other species the highest content of this com-
pound was in P. ostreatus (2.08 mg /100 g d.w.). In the present analysis for first time we es-
tablished in extracts from fruiting bodies and mycelium from culture in vitro of C. viscosa 
5-methyltryptophan and indole. Compared with previously studied species the content 
of serotonin in the fruiting bodies of C. viscosa was low - 0.39 mg/ 100 g d.w., while for 
example in the fruiting bodies of S. luteus, L. rufum and C. cibarius ranged from 29 to 
almost 38 mg/100 g d.w. However, in the mycelium of C. viscosa from cultures on liquid 
medium found content of serotonin was higher than had previously been detected in 
mycelium of T. equestre (3.19 mg/100 g d.w. and 0.59, respectively). Tryptamine was la-
beled compound in most of the earlier fruiting bodies and mycelium of T. equestre and X. 
badius, but it was not detected in any extracts from C. viscosa. Greater variety of indole 
compounds than in the fruiting bodies of C. viscosa was showed only in previously ana-
lyzed fruiting bodies of S. luteus, while mycelia from in vitro culture on solid and liquid 
medium of C. viscosa presented that amount of these metabolites was comparable with 
the amount indicated in the mycelia of T. equestre and X. badius (Muszyńska et al. 2009).

CONCLUSIONS

The obtained results indicate that Calocera viscosa in vitro cultures can be a good 
model for the studies on accumulation and metabolism of indole compounds in 
mushrooms. These similar contents indicates also that it is possible to use in vitro 



 Analysis of indole 63

cultures as a model for studies on the physiological activity of above compounds. 
The comparable quantity of indole compounds obtained in the present study in C. 
viscosa mycelium with fruiting bodies collected from natural condition indicate that 
in vitro cultures are a good source of these compounds. High serotonin precursor 
5-hydroxytrypophan contents in this material prove also a potential for the use of the 
mycelium cultured in vitro as a source of this physiologically important compound 
for humans. In vitro studies demonstrated that serotonin, melatonin and their indole 
derivatives (N-acetylserotonin, 6-methoxytryptamine) dose-dependently decreased 
lipid peroxidation (Sewerynek et al. 2005). 

REFERENCES

Barros L., Baptista P., Correira D.M., Casal S., Oliveira B., Ferreira C.F. R. I. 2007. Fatty acid and sugar 
compositions, and nutrional value of five wild edible mushrooms from Northeast Portugal. Food 
Chem. 105:140–145.

Barros L., Cruz T., Baptista P., Estevinho L.M., Ferreira I. 2008. Wild and commercial mushrooms as 
source of utrients and nutraceuticals. Food Chem. Toxicol. 46: 2742–2747 

Czeczuga B. 1980. Badania nad karotenoidami u grzybów IX Dacrymycetaceae. Acta Mycol. 16 (1): 115–120.
Kysilka R., Wurst M. 1985. High Performance Liquid Chromatographic Determination of Hallucino-

genic Indoleamins with Simultaneous UV Photometric and Voltametric Detection. J. Chrom. 320: 
414–420. 

Liu J. 2005. N-Containing Compounds of Macromycetes. Chem. Rev. 105: 2723–2744. 
Kohlmünzer S., Węgiel J., Muszyńska B., Sułkowska-Ziaja K, Gouillot J. 2000. Metabolity indolowe w ho-

dowli mycelialnej grzybów wielkoowocnikowych. Herba Polonica 66: 98–104. 
Mosovich S.A., Boone R.T., Reichenberg A., Bansilal S., Shaffer J., Dahlman K., Harvey P.D., Farkouh 

M.E. 2008. A new perspective on the relation between cardiovascular diseases and depression. Folia 
Cardiologica Excerpta 3: 296–307.

Muszyńska B. 1999. Chemical and biological studies of fruiting bodies and mycelium from solid and 
liquid cultures of Calocera viscosa (Pers.: Fr.) Fr. PhD Thesis. Jagiellonian University, Collegium 
Medicum, Kraków, 38–46.

Muszyńska B., Maślanka A., Sułkowska-Ziaja K., Krzek J. 2007. Analysis of indole compounds and nitric 
bases in fruiting bodies in Lactarius deterrimus by TLC-UV. J. Plan. Chrom. Modern TLC. 20: 55–58.

Muszyńska B., Sułkowska-Ziaja K., Ekiert H. 2009. Indole compounds in fruiting bodies of some selected 
Macromycetes species and their cultured in vitro. Pharmazie 64: 479–480. 

Muszyńska B., Sułkowska-Ziaja K., Ekiert H. 2010. Główne grupy związków i pierwiastki z aktywnością 
biologiczną w wybranych gatunkach grzybów jadalnych z taksonu Basidiomycota. Farmacja Polska 
66: 804–814.

Muszyńska B., Sułkowska-Ziaja K., Ekiert H. 2011a. Indole compounds in fruiting bodies of some edible 
Basidiomycota species. Food Chem. 125: 1306–1308.

Muszyńska B., Sułkowska-Ziaja K., Ekiert H. 2011b. Analysis of indole copounds in fruiting bodies of 
Armillaria mellea. Acta Polon. Pharm. Drug Res. 168: 93–97.

Muszyńska B., Sułkowska-Ziaja K., Ekiert H. 2012. An antioxidant in fruiting bodies and in mycelia from 
in vitro cultures of Calocera viscosa (Basidiomycota) – preliminary results. Acta Polon. Pharm. Drug 
Res. 169: 10.

Oddoux L. (ed.). 1957. Recherches sur les mycéliums secondaires des Homobasidiés en culture pure. 
Imprimerie de Trevoux, Lyon.

Ohstuka S., Ueno S., Yoshikumi C., Hirose F., Ohmura Y., Wada T., Fujii T., Takahashi E. 1973. Polysac-
charides having an anticarcinogenic effect and a method of producing them from species of Basidi-
omycetes. UK Patent 1331513.

Ouchi Y., Yoshikawa E., Futatsubachi M., Yagi S., Ueki T., Nakamura K. 2009. Altered brain serotonin 
transporter and associated glucose metabolism in Alzheimere disease. J. Nucl. Med. 50: 1260–1266.

Sewerynek E , Stuss M., Oszczygieł K., Kułak J , Lewiński A. 2005. Protective effect of indole compounds 
on lipopolysccharide induced lipid peroxidation in in vitro conditions. Endokrynol. Polska 4: 508–511.



64 B. Muszyńska and K. Sułkowska-Ziaja 

Sułkowska-Ziaja K., Muszyńska B., Końska G. 2005. Biologically Active Compounds of Fungal Origin 
Displaying Antitumor Activity. Acta Polon. Pharm. Drug Res. 6: 153–160.

Sułkowska-Ziaja K. 2010. Chemical analysis of Sarcodon imbricatus (L.) P. Karst fruiting bodies and my-
celia from in vitro cultures and biological activity of their polysaccharide fractions. PhD Thesis. Jag-
iellonian University Collegium Medicum, Kraków, 97-107.

Wasser S.P. 2002. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccha-
rides. Appl. Microbiol. Biotechnol. 60: 258–274.

Yang J-H., Lin H-C., Mau J-L. 2001. Non-volatile taste components of several commercial mushrooms. 
Food Chem. 72: 465–471.

Zaidman B.Z., Yassin M., Mahajana J., Wasser S.P. 2005. Medicinal mushroom modulators of molecular 
targets as cancer therapeutics. Appl. Microbiol. Biotechnol. 67: 453–468.

Analiza zawartości związków indolowych w owocnikach i mycelium  
z kultur in vitro Calocera viscosa (Basidiomycota)

Streszczenie

Od tysięcy lat owocniki grzybów wyższych były wykorzystywane jako źródło pożywienia. 
Obecnie mogą być pozyskiwane nie tylko ze stanu naturalnego i hodowli komercyjnych lecz 
również z kultur in vitro prowadzonych w odpowiednich warunkach. Grzyby jadalne są coraz 
intensywniej badane ze względu na produkcję biologicznie aktywnych metabolitów wtórnych. 
Biologicznie i leczniczo aktywne metabolity grzybów są używane w terapii tak poważnych 
schorzeń jak np.: choroby krążenia, cukrzyca, miażdżyca oraz choroby nowotworowe. Niektó-
re z metabolitów wykazują działanie: przeciwwirusowe, przeciwbakteryjne, przeciwrobacze. 
W konwencjonalnej medycynie, w leczeniu onkologicznym najdłużej stosowane są polisa-
charydy grzybowe, stąd też są ich najlepiej poznanymi metabolitami. Innymi grupami szerzej 
badanych związków są również liczne związki fenolowe, terpenowe, indolowe, witaminy, bio-
pierwiastki (np. selen) o działaniu antyoksydacyjnym. 

Obiektem badań w ramach niniejszej pracy był gatunek grzyba wielkoowocnikowego 
Calocera viscosa (Pers.: Fr.) Fr. (Basidiomycota). Owocniki tego gatunku występują pospolicie 
w lasach iglastych Polski południowej. Materiał do badań stanowiły owocniki zebrane w No-
wej Białej (powiat Nowy Targ) oraz mycelium otrzymane z kultur in vitro, które wyprowadzo-
no z owocników tego gatunku. Kultury prowadzono na pożywce stałej i płynnej wytrząsanej wg 
Oddoux w temperaturze 25 ± 2°C przez 14 dni w warunkach sztucznego oświetlenia o inten-
sywności 900 lx. Owocniki oraz zebrane biomasy z kultur in vitro wysuszono metodą liofilizacji, 
ekstrahowano eterem naftowym w celu usunięcia frakcji lipidowej, a następnie prowadzono 
ekstrakcję metanolem. Uzyskane wyciągi metanolowe po zagęszczeniu, oczyszczono metodą 
preparatywnej chromatografii TLC na płytkach (DC Alufolien Kiesel gel F-254), a następnie 
analizowano na zawartość związków indolowych metodą HPLC (identyfikacje związków indo-
lowych dokonano metodami spektralnymi UV-VIS i EIMS). Na podstawie przeprowadzonych 
analiz stwierdzono obecność we wszystkich trzech ekstraktach następujących związków indo-
lowych: tryptofanu, 5-hydroksytryptofanu, 5-metylotryptofanu, melatoniny i indolu (zawar-
tości wahały się w przedziale: 0.37 do 11.88 mg/100g s.m.). Związkiem indolowym występują-
cym w największych ilościach we wszystkich ekstraktach był 5-hydroksytryptofan. Dodatkowo 
w owocnikach i w mycelium z kultur na podłożu płynnym stwierdzono obecność serotoniny 
(odpowiednio: 0.39 i 3.19 mg/100g s.m).


		2014-01-02T12:03:01+0100
	Polish Botanical Society