ganoderma lucidum from mycelia in anti-cancer in vietnam

172 Pages·2010·7.44 MB·English

Mendel University in Brno
Faculty of Forestry and Wood Technology

DISSERTATION THESIS

Brno 2010 Ing. Vu Thanh Cong
Mendel University in Brno
Faculty of Forestry and Wood Technology

Department of Forest Protection and Wildlife Management

Ganoderma spp – Biology, Species and Culture in
Vietnam and in the Czech Republic

DISSERTATION THESIS

Vu Thanh Cong

Supervisor: Assoc. Prof. Libor Jankovský

Brno, 2010
ACKNOWLEDGEMENT
Many people have inspired me to complete this thesis, and I would
like to take this opportunity to thank some of them.
First, I am extremely grateful to my supervisor, Assoc. Prof. Libor
Jankovský, (Mendel University in Brno) for his patient and generous
assistance throughout the course of my research. His insightful comments
enabled me to avoid wrong turnings and to get the little details right.
I wish to express my sincere gratitude to my mother, Assoc. Prof. Dr.
Nguyen Thi Chinh. (Hanoi University of Natural Sciences) for her
encouragement, helpful and support for me to scientific research in
Vietnam and Czech Republic.
I am grateful to Dr. Lubomír Scháněl and his family, for their help,
teaching and constructive comments.
I would also like to thank Mrs. Miroslava Medková, Mrs. Marie
Bedřichová, Ing. Dagmar Palovčíková and all my friends, for their help for
my working as well as for my studying.
I am also grateful to the Mendel University in Brno, especially The
Faculty of Forestry and Wood Technology, for the good working and
learning conditions.
Above all, there are not enough words to describe how thankful I am
for all that my parents and family has done for me. Without their
unconditional support, love, care and constant encouragement in every step
of the way I could never have come this far.
I hereby declare, that I elaborate this work own on the bases of my
own knowledge, observation and research using cited papers.

Hanoi, February 12, 2010
Ing. Vu Than Cong
Abstract
Wood destroying fungi from genus Ganoderma spp. are distributed worldwide on many
kinds of trees in temperate zone and also in tropics. About 140 species belonging to this genus
have been described. Six species of this genus can be found in the Czech Republic. However
biodiversity of Ganoderma in Vietnam wild is much more higher but there are not any surveys
on it, although Ganoderma lucidum is grown for many centuries. Ganoderma lucidum is the
rarest and valuable bracket fungus in traditional Chinese medicine as Lingzhi or Reishi
mushroom. It has been known in China for over 4,000 years. Is it considered among the most
powerful natural medicines in Far East. Ganoderma lucidum is rich in organic components such
as polysaccharide amino acids, triterpenes, steroids, adenosine alkaloids, lipids, enzyme,
ascorbic acid and trace minerals etc. These compounds are being studied for their effect on the
immune system since they are known to be very useful in health care. The aim of this thesis was
to isolate and characterize new strains of Ganoderma spp. from the Czech Republic and
Vietnam, to characterize genus Ganoderma, find out the possibility of introducing new strains
belonging to Ganoderma lucidum, G. carnosum, G. adspersum, G. applanatum and G.
resinaceum into the culture and also the possibility of applying them in the pharmaceutical
industry.

Abstrakt
Dřevní houby z rodu lesklokorka Ganoderma jsou rozšířeny po celém světě na širokém
spektru hostitelských dřevin v temperátní zóně a v tropech. Celosvětově bylo popsáno zhruba
140 druhů. V České republice bylo popsáno šest druhů lesklokorek. Významně vyšší je
biodiverzita lesklokorek ve Vietnamu, stále však není toto spektrum dostatečně prozkoumáno,
přestože jsou zde lesklokorky pěstovány po mnoho staletí. Ganoderma lucidum je vzácnou a
zvláště ceněnou chorošovitou houbou. V tradiční čínské medicíně je známa pod jménem
Lingzhi nebo Reishi mushroom. V Číně je známa přes 4000 let. Na Dálném východě je
považována za jednu z nejsilnějších a nejúčinnějších přírodních drog. Ve velkém množství
obsahuje polysacharidy, aminokyseliny, triterpeny, steroidy, adenosin alkaloidy, lipidy,
enzymy, kyseliny askorbovou a stopové prvky. Tyto sloučeniny a prvky byly zkoumány
v medicíně pro jejich příznivé léčebné účinky. Cílem této práce je izolovat a charakterizovat
nové kmeny lesklokorek z České republiky a Vietnamu, charakterizovat rod Ganoderma,
hledat možnosti introdukce nových kmenů Ganoderma lucidum, G. carnosum, G. adspersum,
G. applanatum a G. resinaceum a možnosti jejich uplatnění ve farmaceutickém průmyslu.
iii. Content
CONTENT
I: INTRODUCTION..........................................................................................................3
1. Characterization of Ganoderma spp................................................................................3
1.1 Classification of Ganoderma spp..................................................................................3
1.1.2. Biological characteristics of Ganoderma spp..........................................................24
1.1.2.1 Ecology of Ganoderma spp...................................................................................24
a) Mycelia of Ganoderma.spp.....................................................................................24
b) Fruiting body of Ganoderma spp............................................................................26
c) Spore of Ganoderma spp..........................................................................................27
d) Molecular systematic of Ganoderma spp..............................................................29
1.2.2 Characterictics of Ganoderma species......................................................................42
1.2.2.1 Ganoderma applanatum (Pers.) Pat. .....................................................................42
1.2.2.2 Ganoderma adspersum (Schulz.) Donk ................................................................43
1.2.2.3 Ganoderma resinaceum Boud. in Pat....................................................................44
1.2.2.4 Ganoderma pfeifferi Bres......................................................................................45
1.2.2.5 Ganoderma carnosum Pat.....................................................................................46
1.2.2.4 Ganoderma lucidum (W, Crutis) P. Karst.............................................................48
1.2.3 Effect of environment factors to Ganoderma lucidum.............................................51
1.2.4. Components of Ganoderma and their medical value..............................................51
1.2.4.1 Important components Ganoderma ......................................................................51
a- Polysaccharides (β-glucan)....................................................................................51
b- Bitter Triterpenoids ...............................................................................................52
c- Organic Germanium (Ge) .....................................................................................52
1.1.4.2 The medicinal effects of Ganoderma spp..............................................................55
a - Effect on tumor and cancer.............................................................................62
b- Liver protection ..............................................................................................63
c- Effect on cardiovascular.................................................................................64
d- Ageing prevention...........................................................................................64
e- Effect on neurasthenia....................................................................................65
f- Effect on hypertension ..................................................................................65
g- Treatment of diabetes......................................................................................66
h- Effect on chronic bronchitis and bronchus asthma........................................ 66
i- Effect on hyper susceptibility ........................................................................66
j- Effect on beauty care......................................................................................67
1.2.5 Commercial value of Lingzhi ..................................................................................67
iii. Content
1.3. Culture of Ganoderma spp.........................................................................................68

1.3.1 Cultivate Ganoderma spp in the World . .................................................................68

1.3.2 Ganoderma in Vietnam wild....................................................................................68
1.3.3 Ganoderma cultures in Vietnam ..............................................................................69

II: MATERIAL AND METHODS....................................................................................70
2. Material and Methods....................................................................................................70
2.1 Biological material.......................................................................................................70
2.1.1 Standard Ganoderma lucidum..................................................................................70
2.1.2 Ganoderma strains isolated in the Czech Republic and Vietnam.............................70
2.2 Material........................................................................................................................71
2.3 Study methods .............................................................................................................72
2.3.1 Cultivation methods..................................................................................................72
2.3.1.1 Isolating procedure of Lingzhi mushroom in the Czech republic and Vietnam....74
2.3.2 Identify properties of isolated Ganoderma strains....................................................74
2.3.2.1 Antibiotic features..................................................................................................74
2.3.2.2 Identification of enzymatic of Ganoderma strain.................................................75
2.3.2.3 Identification of some major bioactive compound of Ganoderma........................77
2.4 Cultivation of Lingzhi mushroom................................................................................78
2.4.1 Produce of the first strains........................................................................................78
2.4.2 Produce of the second strains....................................................................................78
2.5.1 Cultivation of fruiting body Lingzhi mushroom.......................................................70
2.5.2 Technology of Lingzhi mycelia products.................................................................80

III: RESULTS.................................................................................................................83
3. Results........................................................................................................................... 83
3.1 Isolation of Ganoderma spp....................................................................................... 83
3.2 Species of some isolated Ganoderma strains............................................................. 83
3.2.1 Ganoderma strains in the Czech Republic...............................................................83
3.2.2 Ganoderma strains in Vietnam.................................................................................85
3.3 Biological feature of Vietnam and CR Ganoderma strains.........................................90
3.3.1 Effect of temperature on mycelia growth of Ganoderma spp..................................90
3.3.2 Effect of pH on growth and development mycelia Ganoderma spp........................92
3.3.3 Components and bioactive compounds of some Ganoderma strain……………….95
3.3.3.1 Enzymatic activities of isolated and cultivated G. lucidum...................................96
iii. Content
3.3.4 Productivity calculation............................................................................................99
3.4 Cultivation of Lingzhi mushroom in pilot project.................................................…110
3.4.1 Cultivation of some Lingzhi strains in wood......................................................…110
3.4.2 Cultivation of Lingzhi mushroom in bioreactor.................................................…110

IV.DISCUSSION…….… …………………………………………………………...120
V: CONCLUSION.........................................................................................................125

VI: SOUHRN ..............… ……………………………………………………………. 127

VII: REFERENCES
7. References………………………………………………………………..…............. 130

Annexes

Papers
1 . Introduction
Introduction
The Lingzhi mushroom or Reishi mushroom (Ganoderma lucidum Curtis P.
Karst.) is one of the most valued nature products in traditional Chinese medicine and
has been widely used all over the World for centuries and is described with Ginseng as
one of the two most important elixirs (Chang 2006). This medicinal mushroom has been
shown to boost heart health, lower the risk of cancer promote immune function, ward of
viruses, bacteria and fungi, reduce inflammation, combat allergies, help balance blood
sugar levels and support the body's detoxification mechanisms (Willard 1990).
In historic times the use of Lingzhi was highly restricted by the limitations of a
wild-crafted supply. Today however, modern cultivation methods are used for the
commercial production of Lingzhi mushrooms and mycelium. Contemporary inquiry
and analytical techniques have also greatly increased our knowledge of the biochemistry
of Lingzhi and confirmed it is profound benefits to human health. Lingzhi has been
grown in many countries in the world, such as China, Korea, Singapore, Thailand,
Taiwan, and Vietnam. These countries get a large amount of money from this
mushroom (Tham 1996).
Vietnam is a tropical country which has good condition for Lingzhi production.
Since Vietnam is aware of the medical and commercial benefit of Lingzhi, this
mushroom has been cultivated on farms in natural condition and family scale. Besides,
there are so many wild strains which are not isolated and identified in many provinces
in the North and the South like Thai Binh, Thai Nguyen, Lang Son, Quang Binh, Quang
Ninh, Hoa Binh and Lam Dong. The biggest output is 20-25 t per year (Chinh 2000). In
artificial cultivation, we can’t achieve the strains that have high productivity as well as
suitability for each locality. Furthermore, Lingzhi is a kind of wood destroying
mushroom so it is necessary to isolate Ganoderma strains in Vietnam and to investigate
the cultivation procedure in order to control the production, reduce price and help many
people to use this medicinal mushroom (Chinh 2000).
According to research works of Moncalvo and Ryvarden (1998) announced also
in 1998, actually, there were about 150-200 described species of Ganoderma all over
the world. Whereas in Vietnam approximately 60 of these species were found (Tham
2005).
Within the genus Ganoderma, over 250 taxonomic names have been reported
worldwide by Moncalvo et al.v(1995b) and Ryvarden (1994), few of them are :
3
1 . Introduction
G. adspersum, G. applanatum, G. australe, G. boninense, G. cupreum, G. lipsiense,
G. lobatum, G. oerstedii, G. oregonense, G. pfeifferi, G. platense, G. resinaceum,
G. sessile, G. sinense, G. trornatum, G. tsugae and G. weberianum. However, the
majority of reports in the literature appear to be on the one species G. lucidum.
The most frequently cited species in research publications on the cultivation,
chemical analysis, pharmacology, and a medicinal effect of Ganoderma is the type
species G. lucidum or Lingzhi. It is most commonly used and commercially cultivated
in North America, China, Taiwan, Japan, Korea, Thailand, Vietnam and other
countries. Other common names are Lucky Fungus, Ganoderma, Happy Fungus,
Lingzhi, in Vietnam “Ling Chi”, in Japanese “Reishi, Mannentake or Sachitake”, in
Korean “Youngzhi” or herb of immurtality. Chinese tradition proclaims that
Ganoderma is also called “mariculous zhi”, or “auspicious herb” and usually considered
to “Symbolize happy augury, and to bespeak good fortune, good health and longevity,
even immortality” (Wasson 1968). The Latin word “lucidum” means "shiny" or
"brilliant" and refers to the varnished surface of Lingzhi cap, which is reddish orange to
black. Also Czech name “lesklokorka” has similar meaning. Recent taxonomic studies
indicate that several species are probably represented in medicinal studies, and possibly
none of them is G. lucidum sense. Rather, the name G. lucidum often refers
inadvertently to a complex of species, including at least seven species (Hseu et al.,
1996). Taxonomic research is only now, through molecular methods such as ITS
sequences of rDNA, able to begin description of the component taxa. In many
publications, reference is omitted to data of strain numbers or origin of the Ganoderma
species used, thus limiting evaluation and usefulness of the reported findings.
This situation forces us set up present project to research “Biology characteristics
of some Ganoderma strains isolated in Vietnam, in the Czech Republic and their
cultivation” for the purposes as follows:
1. To isolate and identify some Ganoderma strains.
2. To investigate biological characteristics of isolated Ganoderma strains.
3. To compare biological characteristics of Ganoderma strains of Vietnam and in
the Czech Republic to G. lucidum
4. To identify component and some active compound in Ganoderma.
5. To cultivate Ganoderma lucidum and some other isolated species of
Ganoderma.
4