1. Ganoderma Diseases of Perennial
Crops
Edited by
J. Flood
CABI Bioscience, Egham, UK
P.D. Bridge
Mycology Section, Royal Botanic Gardens Kew, Richmond, UK
M. Holderness
CABI Bioscience, Egham, UK
CABI Publishing
iii
3. Contents
Contents
Contents
Contributors ix
Preface xi
Part I Ganoderma, Organism and Systematics 1
1 Ganodermataceae: Nomenclature and Classification 3
G.-S. Seo and P.M. Kirk
2 Systematics of Ganoderma 23
J.-M. Moncalvo
Part II Ganoderma, Diseases of Perennial Crops 47
3 Status of Ganoderma in Oil Palm 49
D. Ariffin, A.S. Idris and G. Singh
4 Basal Stem Rot of Oil Palm in Thailand Caused by Ganoderma 69
S. Likhitekaraj and A. Tummakate
5 The Current Status of Root Diseases of Acacia mangium Willd. 71
S.S. Lee
v
4. vi Contents
Part III Disease Control and Management Strategies 81
6 A Control Strategy for Basal Stem Rot (Ganoderma) on Oil Palm 83
H. Soepena, R.Y. Purba and S. Pawirosukarto
7 The Use of Soil Amendments for the Control of Basal Stem Rot of
Oil-Palm Seedlings 89
M. Sariah and H. Zakaria
8 The Spread of Ganoderma from Infective Sources in the Field and
its Implications for Management of the Disease in Oil Palm 101
J. Flood, Y. Hasan, P.D. Turner and E.B. O’Grady
9 Basidiospores: Their Influence on Our Thinking Regarding a
Control Strategy for Basal Stem Rot of Oil Palm 113
F.R. Sanderson, C.A. Pilotti and P.D. Bridge
10 Management of Basal Stem Rot Disease of Coconut Caused by
Ganoderma lucidum 121
R. Bhaskaran
11 In vitro Biodegradation of Oil-palm Stem Using Macroscopic Fungi
from South-East Asia: a Preliminary Investigation 129
R.R.M. Paterson, M. Holderness, J. Kelley, R.N.G. Miller and
E. O’Grady
12 Functional Units in Root Diseases: Lessons from Heterobasidion
annosum 139
Å. Olson and J. Stenlid
Part IV Molecular Variability in Ganoderma 157
13 Molecular and Morphological Characterization of Ganoderma in
Oil-palm Plantings 159
R.N.G. Miller, M. Holderness and P.D. Bridge
14 Spatial and Sequential Mapping of the Incidence of Basal Stem Rot
of Oil Palms (Elaeis guineensis) on a Former Coconut (Cocos nucifera)
Plantation 183
F. Abdullah
15 Genetic Variation in Ganoderma spp. from Papua New Guinea as
Revealed by Molecular (PCR) Methods 195
C.A. Pilotti, F.R. Sanderson, E.A.B. Aitken and P.D. Bridge
5. Contents vii
16 Molecular Variation in Ganoderma Isolates from Oil Palm, Coconut
and Betelnut 205
H. Rolph, R. Wijesekara, R. Lardner, F. Abdullah, P.M. Kirk,
M. Holderness, P.D. Bridge and J. Flood
Part V Development of Diagnostic Tests for Ganoderma 223
17 Development of Molecular Diagnostics for the Detection of
Ganoderma Isolates Pathogenic to Oil Palm 225
P.D. Bridge, E.B. O’Grady, C.A. Pilotti and F.R. Sanderson
18 The Development of Diagnostic Tools for Ganoderma in Oil Palm 235
C. Utomo and F. Niepold
19 Ganoderma in Oil Palm in Indonesia: Current Status and
Prospective Use of Antibodies for the Detection of Infection 249
T.W. Darmono
Index 267
6.
7. Contributors
Contributors
Contributors
F. Abdullah, Department of Biology, Faculty of Science and Environmental
Studies, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
E.A.B. Aitken, Department of Botany, University of Queensland, St Lucia,
Queensland, Australia
D. Ariffin, Palm Oil Research Institute of Malaysia, No. 6, Persiaran Institute,
Bangi, PO Box 10620, 50720 Kuala Lumpur, Malaysia
R. Bhaskaran, Coconut Research Station, Tamil Nadu Agricultural
University, Veppankulam 614 906, Tamil Nadu, India
P.D. Bridge, Mycology Section, Royal Botanic Gardens Kew, Richmond,
Surrey TW9 3AE, UK
T.W. Darmono, Biotechnology Research Unit for Estate Crops, Jl. Taman
Kencana No. 1, Bogor, 16151, Indonesia
J. Flood, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 9TY, UK
Y. Hasan, Bah Lias Research Station, P.T.P.P. London, PO Box 1154, Medan
20011, North Sumatra, Indonesia
M. Holderness, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 9TY,
UK
A.S. Idris, Palm Oil Research Institute of Malaysia, No. 6, Persiaran Institute,
Bangi, PO Box 10620, 50720 Kuala Lumpur, Malaysia
J. Kelley, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 9TY, UK
P.M. Kirk, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 9TY, UK
R. Lardner, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 9TY, UK
S.S. Lee, Forest Research Institute Malaysia, Kepong, 52109 Kuala Lumpur,
Malaysia
ix
8. x Contributors
S. Likhitekaraj, Division of Plant Pathology and Microbiology, Department of
Agriculture, Bangkok 10900, Thailand
R.N.G. Miller, Universidade Católica de Brasília Pró-Reitoria de Pesquisa e
Pós-graduação, Campus II, 916 Asa Norte, Brasília, D.F., Brazil
J.-M. Moncalvo, Department of Botany, Duke University, Durham, NC
27708, USA
F. Niepold, Federal Biological Research Centre for Agriculture and Forestry,
Institute for Plant Protection of Field Crops and Grassland, Messeweg
11–12, 38104 Braunschweig, Germany
E.B. O’Grady, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20 9TY,
UK
Å. Olson, Department of Forest Mycology and Pathology, Swedish University
of Agricultural Sciences, Box 7026, S–750 07 Uppsala, Sweden
R.R.M. Paterson, CABI Bioscience, Bakeham Lane, Egham, Surrey TW20
9TY, UK
S. Pawirosukarto, Indonesian Oil Palm Research Institute (IOPRI), Jl.
Brigjen Katamso 51, Medan 20158, Indonesia
C.A. Pilotti, PNG OPRA, Plant Pathology Laboratory, PO Box 36, Alotau,
Milne Bay Province, Papua New Guinea
R.Y. Purba, Indonesian Oil Palm Research Institute (IOPRI), Jl. Brigjen
Katamso 51, Medan 20158, Indonesia
H. Rolph, Level 9, Glasgow Dental School and Hospital, 378 Sauchiehall St,
Glasgow G2 3JZ, UK
F.R. Sanderson, PNG OPRA, Plant Pathology Laboratory, PO Box 36,
Alotau, Milne Bay Province, Papua New Guinea
M. Sariah, Department of Plant Protection, Universiti Putra Malaysia,
43400UPM, Serdang, Selangor, Malaysia
G.-S. Seo, College of Agriculture, Chungnam National Unviersity, Taejon
305–764, Korea
G. Singh, United Plantations Berhad, Jenderata Estate, 3600 Teluk Intan,
Perak, Malaysia
H. Soepena, Indonesian Oil Palm Research Institute (IOPRI), Jl. Brigjen
Katamso 51, Medan 20158, Indonesia
J. Stenlid, Department of Forest Mycology and Pathology, Swedish University
of Agricultural Sciences, Box 7026, S–750 07 Uppsala, Sweden
A. Tummakate, Division of Plant Pathology and Microbiology, Department of
Agriculture, Bangkok 10900, Thailand
P.D. Turner, PO Box 105, Quilpie, Queensland 4480, Australia
C. Utomo, Indonesian Oil Palm Research Institute (IOPRI), PO Box 1103,
Medan 20001, Indonesia
R. Wijesekara, Coconut Research Institute, Bandirippuwa Estate, Sri Lanka
H. Zakaria, Department of Plant Protection, Universiti Putra Malaysia,
43400UPM, Serdang, Selangor, Malaysia
9. Preface
Preface
Preface
Perennial oilseed crops form a major component of rural economies through-
out the wet lowland tropics of South and South-East Asia and Oceania. Crops
such as oil palm and coconut are grown as both plantation-scale commodity
crops and as smallholder cash and food crops. Perennial oilseed crops contrib-
ute significantly to local livelihoods through not only their husbandry but also
the processing of the crop and crop by-products and their subsequent shipping
and marketing. As export commodities, they form an important component of
national economies and generate valuable foreign exchange.
Species of the basidiomycete fungus Ganoderma occur as pathogens on a
wide range of perennial tropical and sub-tropical crops, including oil palm,
coconut, tea, rubber, Areca and Acacia, as well as various wild palm species.
The effects of Ganoderma infection on productivity decline in palm crops have
been of considerable concern ever since replanting of oil-palm land began in
South-East Asia and recent workshops have identified basal stem rot, caused
by Ganoderma boninense, as the single major disease constraint to oil palm
production in the region. The long-term nature of palm monocultures means
that they are prone to both premature plant death and to the carry-over of
residual inoculum from one planting to the next. This pattern has been clearly
seen in many areas of South-East Asia and creates considerable concern for the
long-term sustainability of palm production from affected land. Basal stem rot
of oil palm is widespread, occurring in the major oil palm growing regions of
the world. By contrast, the disease on coconut appears very restricted; it was
first recorded in India in 1952 and remains confined to South Asia, yet
Ganoderma species occur as saprobes on dead coconut palm tissues in all
palm-growing regions, an anomaly that requires resolution.
xi
10. xii Preface
A crucial factor in developing effective disease management programmes
is the prior understanding of pathogen biology and disease epidemiology.
Ganoderma is a notoriously variable and difficult fungus to characterize and
this has led to much past confusion in disease aetiology and epidemiology.
Such studies have been greatly enhanced through the development and use
of molecular and biochemical markers to discriminate among pathogen
populations and individuals and to diagnose infected palms in advance of
terminal symptoms. These technological tools can form powerful adjuncts to
field observation and experiments in understanding mechanisms of disease
spread and pathogen survival. This new understanding establishes the
fundamental biology of the genus and provides new insight into disease
epidemiology that enables the implementation of appropriate and effective
management strategies.
In perennial crops, infections of woody tissues have the opportunity to
slowly develop further and expand as conditions permit. Infective material can
remain viable in the ground for many months and infect subsequent crops at
replanting. It is therefore very important to manage disease outbreaks in such
a way as to minimize the risks to both existing and future plantings. One
feature of Ganoderma diseases is the persistence of potential pathogens in old
woody tissues and soil-borne debris. Burning of such material is no longer
acceptable and extensive physical clearing is often not feasible due to the input
requirements involved. Alternative treatments are thus required and a
number of approaches are being explored to manage this residual inoculum.
These are centred on the evaluation of biocontrol agents and the rapid
biodegradation of palm woody residues.
This book is a joint effort by 36 authors from 13 countries, each with a
wide expertise in their own fields. In many chapters, joint authors have come
together from different countries, illustrating the collaborative nature of this
initiative. The 19 chapters address many current issues in the development of
sustainable disease management programmes and are grouped into five major
themes. These are, an introduction to the pathogen and its systematics in
Chapters 1 and 2, outlines of the diseases caused by the pathogen (Chapters
3–5), disease management (Chapters 6–12), molecular biological variability
in the pathogen (Chapters 13–16) and the development of diagnostic tools
(Chapters 17–19). The majority of these chapters have been developed from
presentations made at two international workshops on Ganoderma diseases
held in Malaysia in 1994 and 1998 and a technical workshop held in the UK
in 1998. Funding for these workshops was provided by the UK Department
for International Development (DFID Project R6628) Crop Protection
Programme, for the benefit of developing countries and from the European
Community (Stabex fund), the British Council, Governments and institutions
of the countries concerned and numerous private plantation companies. We
are very grateful to the various sponsors of this research for their involvement,
although the book should not be considered to necessarily reflect the views of
our sponsors. We would also wish to acknowledge the pioneering work and
11. Preface xiii
dedication of a number of scientists who have previously advanced knowledge
of this recalcitrant organism and its various diseases and inspired us in our
own labours, notably E.J.H. Corner, P.D. Turner, A. Darus and G. Singh.
This book reflects the sum of knowledge of Ganoderma as a plant pathogen
as at the end of 1998 and we hope will be both useful and informative to a wide
range of readers including scientists in the private and public sectors, students
and growers of perennial crops. Further work continues and we trust that fur-
ther insights will continue to be obtained in the near future to further enhance
the sustainable management of Ganoderma diseases.
J. Flood
P.D. Bridge
M. Holderness
16. 4 G.-S. Seo and P.M. Kirk
Adaskaveg and Ogawa, 1990; Adaskaveg et al., 1991, 1993), or useful
medicinal herbs (Mizuno et al., 1995). Because of these fundamentally
different viewpoints among collectors, the taxonomy of these fungi is very
subjective and confused. Contributions to the morphology and taxonomy of
the Ganodermataceae have been made by many mycologists, including Steyaert
(1972), Furtado (1981), Corner (1983) and Zhao (1989). However, the great
variability in macroscopic and microscopic characters of the basidiocarps
has resulted in a large number of synonyms and in a confused taxonomy,
especially in the genus Ganoderma (Gilbertson and Ryvarden, 1986).
History of Ganoderma Taxonomy and Nomenclature
The genus Ganoderma has been known for a little over 100 years; it was intro-
duced by the Finnish mycologist Peter Adolf Karsten, in 1881. He included
only one species, Polyporus lucidus, in the circumscription of the genus and this
species, therefore, became the holotype species.
P. lucidus was named by William Curtis, the 16th-century British botanist.
Unfortunately, Karsten incorrectly attributed the epithet ‘lucidus’ to von
Leysser and this error has been perpetuated in numerous subsequent
publications. No authentic specimens remain and the type locality, Peckham,
is now very much changed from what it was in the time of Curtis. The area is
now largely developed as residential housing but the type substratum, the
small tree Corylus avellana, is likely to be growing still on Peckham Rye
Common. It is clear, therefore, where any epitype, selected as an interpretive
type, should be sought. The selection of an epitype, in the absence of type or
authentic material, would be important, for any further molecular work will
need to have available a culture of the type species of the genus which has
some nomenclatural standing, i.e. a culture derived from an epitype.
Following Karsten, dozens of species belonging to the genus were reported
by taxonomists (Patouillard, 1889; Boudier and Fischer, 1894; Boudier,
1895; Murrill, 1902, 1908). The identification of Ganoderma in those days
was mainly based on host specificity, geographical distribution, and macro-
morphological features of the fruit body, including the context colour and
the shape of the margin of pileus, and whether the fruit body was stipitate
or sessile. Subsequently, Atkinson (1908), Ames (1913), Haddow (1931),
Overholts (1953), Steyaert (1972, 1975, 1977, 1980), Bazzalo and Wright
(1982), and Corner (1983) conducted the identification of Ganoderma species
by morphological features with geographically restricted specimens. Haddow
(1931) and Steyaert (1980) placed most of their taxonomy on the spore char-
acteristics and the morphology of hyphal elements. However, the basidiocarps
of Ganoderma species have a very similar appearance that has caused confusion
in identification among species (Adaskaveg and Gilbertson, 1986, 1988).
The genus now contains a few hundred names; there are 322 in the CABI
Bioscience fungus names database, but others may have been published that
17. Nomenclature and Classification 5
the major printed indexes, the source of this database, failed to include. The
database of Stalpers and Stegehuis available on the CBS web site lists 316
names in Ganoderma and the recent publication of Moncalvo and Ryvarden
(1997) lists 386 names for the Ganodermataceae as a whole. It has not yet been
possible to compare these three data sets, although such an exercise would
appear to be needed. However, names are only one aspect of this subject
and problems associated with them are, on the whole, easier to resolve than
problems associated with the circumscription of species.
Based on the unique feature of the double-walled basidiospore, the French
mycologist, Patouillard, over a period of some 40 years from 1887, described a
number of new species of Ganoderma and transferred several names from other
genera of the polypores. Patouillard (1889) published a monograph of the
then known 48 species and also distinguished the species with spherical or
subspherical spores as section Amauroderma. Coincidentally, in the same year,
Karsten introduced the genus Elfvingia, based on the name Boletus applanatus
of Persoon, for the non-laccate species. Later, section Amauroderma of
Ganoderma was raised to the rank of genus by Murrill who, in selecting a
species which was not included in section Ganoderma by Patouillard, is
therefore the author of the name, and priority dates from 1905 not 1889.
Subsequent authors have recognized Amauroderma as a distinct genus. The
two genera have been largely accepted, although Corner (1983) and Zhao
(1989) reported species that are intermediate between them. Amauroderma
was revised by Furtado (1981).
Here then we have two important species in the history and the nomen-
clature of the genus, Ganoderma lucidum and Ganoderma applanatum, and these
are probably two of the most poorly understood species of Ganoderma and two
of the most frequently misapplied names.
The late 19th-century and early 20th-century mycologists contributed
significantly, in terms of volume of published information, on the genus,
describing many new species or perhaps, more correctly, introducing many
new names. Many of these names were based on single collections or on only a
few collections from the same locality, and the taxonomic status of the species
to which these names were applied is, therefore, often open to the criticism
of being unsound. Throughout the remainder of the 20th century various
workers, Steyaert, Corner and Zhao perhaps being the more prominent,
contributed to our knowledge of the genus by providing revisions, mono-
graphs, descriptions of new taxa (again, often based on single collections or
on only a few collections from the same locality) and observations on both
anatomy and ontogeny.
Recent workers have used characters other than morphology to deter-
mine relationships within the genus. These have included, in the first instance;
cultural and mating characters, primarily by Adaskaveg and Gilbertson
(1986); followed by isozyme studies by Hseu and Gottlieb (Hseu, 1990;
Gottlieb and Wright, 1999), amongst others; and, finally, Moncalvo and his
co-workers (Moncalvo et al., 1995a, b) have used ribosomal DNA sequences
18. 6 G.-S. Seo and P.M. Kirk
and cladistics methods to infer natural relationships. However, as Moncalvo
and Ryvarden have stated, these recent studies have had little impact on
Ganoderma systematics in total because too few taxa were examined. This was
quite clearly through both a lack of human and financial resources and,
perhaps more importantly, a lack of the very important type or authentic col-
lections which will link the names available to any subsequent taxa identified.
Ryvarden (1994) has stated that the genus is in taxonomic chaos and
that it is one of the most difficult genera amongst the polypores. However, this
realization has come at the very time when there has been a renewed interest
in Ganoderma from a number of quite unrelated sources. These include the
medicinal uses based on very old Chinese traditions and the requirement to
elucidate the structure of possible active ingredients, coupled with the require-
ment (not least of all for patent purposes to protect intellectual property rights)
to apply names to the species identified in this context. Also of significance here
is the apparent increase in the importance of some species of Ganoderma as
pathogens of plants used by man.
However, with the development of cladistic methods to reconstruct
natural classifications and the application of these methods to both traditional
morphological data and, more importantly, new molecular data, the potential
for the resolution of some of these problems appears close to hand. Recently,
the phylogenetic relationships of some Ganoderma species collected from
various regions were studied by allozyme (Park et al., 1994) and DNA analysis
(Moncalvo et al., 1995a, b). Moncalvo and his co-workers (Moncalvo et al.,
1995a, b; Hseu et al., 1996) adopted ribosomal DNA sequences and randomly
amplified polymorphic DNA (RAPD) as the tools for analysing phylogenic
relationships in the G. lucidum complex. The results suggested that some
strains were misnamed and misidentified, and all isolates belonging to 22
species were disposed in six groups based on nucleotide sequence analysis
from the internal transcribed spacers (ITS) of the ribosomal gene (rDNA).
However, while some isolates had the same ITS sequence, all of them could be
clearly differentiated by genetic fingerprinting using RAPDs. Therefore, RAPD
analysis might be helpful for systematics at the lower taxonomic levels to
distinguish isolates from each other. When the results of molecular taxonomy
are compared with the data of traditional taxonomy, such as morphological,
ecological, cultural and mating characteristics, some isolates remain as
exceptions. Of many studies on Ganoderma taxonomy, Adaskaveg’s research
(Adaskaveg and Gilbertson, 1986) indicates the importance of vegetative
incompatibility tests for accurate identification, concluding that the incompat-
ibility test must be adopted for the identification of the G. lucidum complex.
Because of the problems as described above, Ryvarden (1994) has proposed
that no new species be described in Ganoderma in the decade to 2005.
Donk, in 1933, was the first to unite the taxa within what was then
the very large family Polyporaceae when he proposed the subfamily
Ganodermatoideae; he subsequently raised this taxon to the rank of family
with the introduction of the Ganodermataceae and this classification has
19. Nomenclature and Classification 7
subsequently been accepted by most recent workers. Much later, Julich, in
1981, introduced the ordinal name Ganodermatales and this was accepted by
Pegler in the eighth edition of the Dictionary of the Fungi, although other work-
ers have continued to use the traditional Aphyllophorales in a broad sense.
There has been much speculation on the relationship between Ganod-
ermataceae and other families of polypores. Corner (1983) believed that the
family represented an old lineage from which other groups of polypores have
been derived. Ryvarden (1994), however, proposed that the high phenotypic
plasticity observed in the genus is indicative that the taxon is young and that
strong speciation has not yet been achieved. This hypothesis was supported by
more recent molecular evidence from Moncalvo and his co-workers. The lack
of fossils limits the accuracy to which we can attribute a minimum age to the
genus. Some fossils of corky polypores from the Miocene (25 million years old)
have been tentatively referred to Ganoderma adspersum.
Morphological Features of Ganoderma
Macromorphology
The naturally produced basidiocarps of G. lucidum show various morphologi-
cal characteristics; sessile, stipitate, imbricate and non-imbricate (Shin et al.,
1986; Adaskaveg and Gilbertson, 1988; Fig. 1.1). The colour of the pileus
surface and hymenophore varies from deep red, non-laccate, laccate and light
yellow to white, and the morphology also differs between the isolates (Shin
and Seo, 1988b). The morphological variation appears to be affected by envi-
ronmental conditions during basidiocarp development. Table 1.1 summarizes
the representative results from several descriptions of the macromorphology of
G. lucidum. The size and colour of the basidiocarp shows significant differences
between the specimens, but the pore sizes are similar. The manner of stipe
attachment to pileus and the host range also varies (Ryvarden, 1994; Fig. 1.1).
The pileus of the normal fruit body is laterally attached to the stipe, but eccen-
tric, central, imbricate, and sessile fruit bodies are also produced rarely in
nature (Fig. 1.1). Stipe characters, including attachment type and relative
thickness and length, have been considered useful for species identification,
but their importance has been neglected by some mycologists, who describe
fruit bodies only as stipitate or sessile. Hardwoods are the usual host plants of
G. lucidum, but some specimens have been collected from conifers.
The laccate character of the pileus and stipe has been variously employed
in the taxonomy of this family. According to traditional concepts, the pileus
surface of Ganoderma is laccate, but is not so in Amauroderma. However, a few
species of Amauroderma and Ganoderma have been reported with laccate (A.
austrofujianense and A. leptopus) and non-laccate appearance (G. mongolicum).
The laccate character, while playing no important role in the segregation of
genera and sections in this family, remains available as an identification aid.
20. 8 G.-S. Seo and P.M. Kirk
Context colour of Ganoderma varies from white to deep brown and has
been considered a useful character in classification. However, some mycolo-
gists have considered it useless for identification of species and supraspecific
groups because it may change under different environmental conditions.
Context colour is often changeable, especially in dried specimens, not only in
the same species but within a single specimen (Zhao, 1989). Corner (1983)
Fig. 1.1. Macromorphological characteristics of Ganoderma lucidum complex.
21. Table 1.1. Macromorphological descriptions of Ganoderma lucidum.
Characters Steyaert (1972) Pegler and Young (1973) Bazzalo and Wright (1982) Melo (1986) Ryvarden (1994)
Size
Pileus Up to 20 cm –b 2–8 × 2–4(–5) cm Up to 15 cm 2–16 cm
Stipe(L)a Up to 20 cm – 4–10 cm Up to 12.5 cm 1–3 cm
(D)a – – 0.5–2 cm – 1–3.5 cm
Pore – – 4–7 pore mm−1, 4–6 pore mm−1 4–6 pore mm−1
6–200 µm diameter
Colour of
Pore surface – – White to yellowish or greyish-white White to cream White-cream to pale brown
Stipe Dark brown Shiny, yellowish red to Reddish-black to almost black Purplish, reddish-brown, Deep chestnut to almost black
reddish-black crust reddish black
Pileus Reddish-brown Shiny, yellowish red to Light to dark reddish-brown Purplish-red, reddish and White or cream-reddish to deep
reddish-black crust reddish black reddish-black
Contex Nearly white Yellowish wood Ochraceous brown to dark brown Wood coloured and dark Wood coloured to pale brown
brownish
Attachment of
stipe to pileus
Lateral #c Usually Frequently Frequently 46 specimens
Ecentric # – – – 1 specimen
Central # – – – 3 specimens
Nomenclature and Classification
Imbricate # – – – 1 specimen
Sessile # # # # –
Hyphal system – – Trimitic Trimitic Trimitic
Host
Hardwood – Common Common # 23 specimens
Conifer – Occasionally Rarely no 22 specimens
a
L and D in parentheses indicate length and diameter, respectively.
b
Not determined.
c
#: described by author as presence only.
9
22. 10 G.-S. Seo and P.M. Kirk
emphasized the importance of observing the context colour of fresh and living
specimens in the classification of Ganoderma. The size and shape of pores are
also useful characters for species classification. The number of pores per
millimetre may serve as a specific character.
The morphology of basidiocarps of G. lucidum in artificial cultivation on
wood logs and synthetic substrates is affected by environmental conditions
(Hemmi and Tanaka, 1936). Fruit-body formation in G. lucidum usually
requires 3 months on sawdust medium (Shin and Seo, 1988b; Stamets,
1993b). The development of the basidiocarp is very sensitive to light and venti-
lation. The stipe exhibits tropic growth toward light (Stamets, 1993a). Under
dim light or dark conditions with poor ventilation, the pileus does not expand
and often an abnormal pileus of the ‘stag-horn’ or ‘antler-type’ is produced
(Hemmi and Tanaka, 1936; Shin and Seo, 1988b; Stamets, 1993a). Figure
1.2 and 1.3 show fruit bodies of the G. lucidum complex produced by the
Fig. 1.2. Fruit bodies of Ganoderma lucidum complex generated by sawdust-
bottle cultivation.
23. Nomenclature and Classification 11
sawdust-bottle culture method. They show polymorphic features such as the
kidney-type and antler-type with various colours (Shin and Seo, 1988b). Out
of 22 isolates of the G. lucidum complex observed by one of the authors of this
chapter (Shin and Seo, 1988b), 16 isolates formed typically kidney-shaped
fruit bodies, and the remainder formed antler-type fruit bodies. Kidney-shaped
fruit bodies could be further divided into those with a concentric zone on the
surface of the pilei and those without. Antler-shaped fruit bodies also divide
into typical forms and those with abnormal pilei (Table 1.2, Fig. 1.2).
However, the fruit bodies of some species of Ganoderma are very stable in
morphology when generated by artificial cultivation with sawdust media,
including their pileus colour, pileus zonation, attachment type and context
colour. Fruit bodies of representative species of Ganoderma are shown in Fig.
1.3. The pileus colour of all the fruit bodies of all species that are generated
by sawdust-bottle cultivation is reddish-brown to deep brown. In G. lucidum
(ATCC 64251 and ASI 7004), G. oregonense (ATCC 64487), G. resinaceum and
G. oerstedii (ATCC 52411) the fruit bodies have very similar pileus colour,
Fig. 1.3. Asian collection – fruit bodies of Ganoderma lucidum generated by
sawdust-bottle cultivation.
Table 1.2. Classification of stocks in Ganoderma lucidum according to the
morphology of fruit bodies generated by sawdust-bottle cultivation.
1. Typically kidney-shaped fruit body-------------------------------(A and B)
A. Concentric zones on the surface of the pileus ---------------------------10 isolates
B. No concentric zones on the pileus ----------------------------------------- 6 isolates
2. Antler-shaped fruit body --------------------------------------------(a and b)
a. Typically antlered--------------------------------------------------------------- 2 isolates
b. Antler-shaped with abnormal pileus --------------------------------------- 4 isolates
24. 12 G.-S. Seo and P.M. Kirk
zonation and pattern of stipe attachment. Although one isolate (ASI 7024)
of G. lucidum produced typical antler-shaped fruit bodies, isolates ASI 7024
and ASI 7004 were confirmed as conspecific by mating tests with mono-
karyotic mycelia. Another isolate (MRI 5005) of G. lucidum showed a very
specific pileus pattern with well-developed concentric zones. The species G.
applanatum, G. microsporum, G. subamboinense and G. pfeifferi have unique
morphological characters. The fruit body of G. meredithae (ATCC 64490) has a
long stipe attached parallel to the pileus and no concentric zones on the surface
of the pileus. In G. applanatum (ATCC 44053) the fruit body is reddish-brown
and has no distinct stipe; the surface and margin of the pileus are rough. The
pileus of G. microsporum (ATCC 6024) has a yellowish-brown margin and
the stipe is black; the surface of the pileus is smooth and has many narrow
concentric zones. In G. subamboinense (ATCC 52420) the pileus is deep brown,
although the growing margin is white, and it has a typical stipe; the surface
of the pileus has many concentric zones. An abnormal pileus was produced
in G. pfeifferi (CBS 747.84), with an upturned margin; the pileus is also
comparatively very thick (up to 30 mm).
Micromorphology
The structure of the pileal crust and cortex are useful characters in the
taxonomy of the Ganodermataceae. The former character occurs mainly in
Ganoderma and Amauroderma, but the latter also occurs rarely in Amauroderma.
Fruit bodies of Ganoderma mostly have an hymenioderm or characoderm and
anamixoderm (Steyaert, 1980). In Elfvingia, the pileal crust is a trichoderm
or an irregular tissue; it is also an irregular tissue in Trachyderma (Zhao,
1989). This character is considered to be very useful for identification by some
taxonomists. However, it often differs in different specimens of a single species
and may show various structural forms.
In Ganodermataceae, the hyphal system is usually trimitic, occasionally
dimitic, the generative hyphae are hyaline, thin walled, branched, septate or
not, and clamped. Clamp connections may often be difficult to observe in
dried specimens. However, they are easily observed in the youngest parts of
the hymenium and context of fresh specimens. Skeletal hyphae are always
pigmented, thick walled, and arboriform or aciculiform; skeletal stalks may
end in flagelliform, branched binding processes. Binding hyphae are usually
colourless with terminal branching. Some species of Ganoderma, such as G.
lucidum and G. ungulatum, show Bovista-type binding hyphae which are
produced from the generative or skeletal hyphae. G. mirabile and G. oregonense
have a pallid context and exhibit intercalary skeletals, which are derived from
a transformed and elongated generative cell. On the other hand, Amauroderma
has no Bovista-type binding hyphae and many species have intercalary
skeletals. Hyphal characters are also influenced by environmental factors.
Zhao (1989) observed great variation in hyphal diameter and in frequency of
25. Nomenclature and Classification 13
septation due to differences in age as well as in nutrition. For species identifica-
tion, however, hyphal characters are often useful (Zhao, 1989).
Basidia and basidiospores are considered as the most important characters
for species identification in basidiomycetes. Basidia in Ganodermataceae attain a
relatively large size and range from typically clavate to pyriform. Intermediate
forms are often seen in the same specimen. Basidiospores show several
dependable characters for identification. Ganodermataceae have a unique
double-walled basidiospore; Donk’s (1964) concept for the Ganodermataceae is
based on characters of the basidiospores. Basidiospores of Ganoderma are ovoid
or ellipsoid–ovoid, occasionally cylindric–ovoid, and always truncate at the
apex. The wall is not uniformly thickened, with the apex always thicker than
the base. It is very distinctly double-walled, with the outer wall hyaline and
thinner, and the inner one usually coloured and thicker and echinulate or
not. In Amauroderma the basidiospores are globose to subglobose, occasionally
cylindrical, and form a uniformly thickened wall. In Haddowia the basidio-
spores are longitudinally double-crested, with small, transverse connecting
elements.
Microscopic observations, such as the size and morphology of basidio-
spores, have been adopted as the criteria for the taxonomy of Ganoderma. The
basidiospores, which commonly have double walls and are ellipsoid and
brownish, vary in size (based on descriptions in the literature; Table 1.3). A
basidium of G. lucidum has four sterigma with a hilar appendix (Fig. 1.4) and
1–2 vacuoles. Basidiospores have an eccentric hilar appendix on a rounded
spore base, and vacuoles. The surface of basidiospores is smooth or wrinkled,
and most of them have numerous small and shallow holes (Fig. 1.4). The sizes
of basidiospores of naturally grown specimens from Japan and Korea were
8.5–11 × 6.5–8.5 µm (average 10.1 × 7.5 µm), and 8.5–13 × 5.5–7 µm
(average of 10.4 × 6.6 µm), respectively. The mean spore indexes (the ratio of
spore length to width) were 1.62 and 1.58, respectively.
Cultural Characteristics
Critical studies on cultural characteristics are very important in species identi-
fication of some groups of higher basidiomycetes. However, useful studies of
cultural characteristics of Ganoderma for species identification are rare. In vitro
morphogenesis and cultural characteristics of basidiomycetes are affected by
various environmental factors, such as light, aeration, temperature, humidity
and nutritional condition (Schwalb, 1978; Suzuki, 1979; Manachère, 1980;
Kitamoto and Suzuki, 1992). Among these, light is an essential factor for
fruiting and pileus differentiation (Plunkett, 1961; Kitamoto et al., 1968,
1974; Perkins, 1969; Perkins and Gordon, 1969; Morimoto and Oda, 1973;
Schwalb and Shanler, 1974; Raudaskoski and Yli-Mattila, 1985; Yli-Mattila,
1990). Primordium formation, pileus differentiation and tropic growth of the
stipe of G. lucidum were affected positively by light (Hemmi and Tanaka, 1936;
26. 14 G.-S. Seo and P.M. Kirk
Table 1.3. Morphological comparison of basidiospores of Ganoderma lucidum.
Basidiospore Size Spore
Reference sources (µm) indexa Microscopical feature
Ito (1955) Wild fruit 9.5–11 × 5.5–7 – Deep yellowish brown,
body ovoid and double wall
Steyaert (1972) Wild fruit 8.5–13 × 5.5–8.5 – Ovoid, chamois
body
Pegler and Wild fruit 9.0–13 × 6–8 1.64 Ovoid to ellipsoid
Young (1973) body (av. 11.5 × 7)
Bazzalo and Wild fruit 9–13 × 5–6.9 – Subovoid with the apex
Wright (1982) body truncate, perisporum
hyaline, smooth and
thin endosporic pillars
Melo (1986) Wild fruit 8.2–13.5 × 6.8–8.1 – Truncate, ovoid,
body brownish to brown
Adaskaveg Wild fruit 10.6–11.8 × 6.3–7.8 1.50 Brown, ovoid with
and Gilbertson body (av. 11.5 × 7.4) holes and eccentric
(1986) hilar appendix, double
wall and vacuole
Mims and Wild fruit 9–12 × 6–7 – Ellipsoid with holes and
Seabury (1989) body eccentric hilar appendix
Seo et al. Wild fruit 8.6–10.9 × 6.6–8.3 1.62 Brown, ovoid with
(1995a)b body (av. 10.1 × 7.5) holes and eccentric
hilar appendix, double
wall and vacuole
Seo et al. Wild fruit 8.3–12.8 × 5.6–7.2 1.58 Brown, ovoid with
(1995a)c body (av. 10.4 × 6.6) holes and eccentric
hilar appendix, double
wall and vacuole
Seo et al. Atypical 6.4–9.6 × 3.2–5.1 1.74 Brown, ellipsoid with
(1995a) fruiting (av. 7.3 × 4.2) holes and eccentric
structures hilar appendix, double
wall and vacuole
aSpore index = ratio of spore length to width; –, not determined.
bBasidiospores from a Korean specimen.
cBasidiospores from a Japanese specimen.
Stamets, 1993a, b). On the contrary, the growth of mycelium was suppressed
by light (Shin and Seo, 1988a, 1989a; Seo et al., 1995a, b). However, critical
studies on the effects of light on mycelial growth and basidiocarp formation of
Ganodermataceae have not been reported.
In vitro, cultures of Ganoderma species produce various hyphal structures,
such as generative hyphae with clamp connections, fibre or skeletal hyphae,
‘stag-horn’ hyphae, cuticular cells and vesicles, and hyphal rosettes (Adaska-
veg and Gilbertson, 1989; Seo, 1995). The colony is white to pale yellow and
even, felty to floccose at the optimum temperature on potato dextrose agar
27. Nomenclature and Classification 15
Fig. 1.4. Basidiospores (a and b) and basidia (c and d) of Ganoderma lucidum,
generated from fruit body (left) and atypical fruiting structures (right). Scale bars:
2 µm (basidiospores) and 3 µm (basidia).
(PDA) (Seo, 1987; Adaskaveg and Gilbertson, 1989). The colony becomes
more yellowish under exposure to light.
The different optimum temperatures and growth rates among various
species and strains of the G. lucidum complex have been described (Table 1.4).
Hyphal growth of most isolates was 2–4 mm day−1 on PDA but chlamydo-
spore (CHL) forming isolates grew faster than those that did not form
chlamydospores. In vitro, colonies showed various features, such as sectoring,
pigmentation, formation of fruit-body primordia (FBP) and atypical fruiting
structures (AFSs) which formed basidia and basidiospores without basidiocarp
formation (Shin and Seo, 1988a). AFSs were induced by light with ventilation
from the white mycelial colony stage (Shin and Seo, 1989b). Some isolates
28. 16
Table 1.4. Cultural characteristics of the Ganoderma lucidum complex.
Temperature (°C)
Growth rate
Species Reference Colour Growth habit Opt. Max. (mm day−1) Chlamydosporeb Fruitingb
G. lucidum Adaskaveg and White Even, felty 30–34 37 7–8 − +
Gilbertson (1989)
Seo (1995) White to pale yellow Even, felty to floccose 25–30 33–35 2–7 ± +
G. tsugae Adaskaveg and White to pale yellow Even, felty to floccose 25–25 30 2–3 − −
Gilbertson (1989)
Seo (1995) White to pale yellow Even, felty 25–30 33 1–2 − −
G. oregonense Adaskaveg and White to pale yellow Even, felty to floccose 20–25 30 2–4 − +
Gilbertson (1989)
G.-S. Seo and P.M. Kirk
Seo (1995) White Even, felty to floccose 25–30 a#a
2–3 − −
G. resinaceum Seo (1995) White Even, felty to floccose # # 3–4 − −
G. valesiacum Seo (1995) Grey Even # # 1–2 − −
a#: not determined.
bFormation of chlamydospore, vesicle, atypical fruiting structures and fruit-body primordia on agar media (+), or not (−).
29. Nomenclature and Classification 17
produced FBP on agar medium, but these did not develop into mature fruit
bodies during the 30 days of cultivation (Seo et al., 1995a). In vitro, higher rate
of ventilation was required for AFS formation, but FBP could be formed under
conditions of lower ventilation. This fact suggests that FBP and AFSs may
be initiated by a common morphogenetic control system, but that subsequent
development to either FBP or AFSs may be determined by environmental con-
ditions in addition to the genetic characteristic of the strains. The formation of
AFSs and FBP on agar media was noted particularly in the G. lucidum complex,
especially the Korean and Japanese collections, and in G. oerstedii (ATCC
52411, Argentina).
A few reports have described the formation of aberrant fruit bodies of
G. lucidum in vitro (Bose, 1929; Banerjee and Sarkar, 1956; Adaskaveg and
Gilbertson, 1986). Adaskaveg and Gilbertson (1989) reported that G. lucidum
occasionally produced aberrant fruit bodies with basidiospores on agar media.
The basidiospores were formed on red, laccate, coral-like fruit bodies. These
fruit bodies might be AFSs because of similarity in their appearance and in
their ability to form basidiospores. In this case, chlamydospore formation was
observed on the same colony, although the AFS- and FBP-forming isolates
examined by Seo et al. (1995a) did not produce chlamydospores. Furthermore,
chlamydospore-forming isolates formed neither AFSs nor FBP under any of the
conditions examined (Seo et al., 1995a).
Among 30 isolates of G. lucidum collected from Japan, Korea, Papua
New Guinea, Taiwan and the USA, 20 isolates (about 66% of the isolates
tested), none of which was from the USA, formed AFSs with basidiospores, and
another five isolates (about 17% of the isolates tested), none of them from
Papua New Guinea, induced FBP. Of the remaining five isolates, one isolate
from Korea formed a callus-like structure without producing basidiospores,
this structure differing from AFSs and FBP in form, and the other four isolates
from Korea, Papua New Guinea and the USA formed neither AFSs nor FBP.
Among the latter, three strains formed chlamydospores. One isolate did not
form any fruiting structure under standard conditions, but it could produce
AFSs in dual culture with a species of Penicillium known to produce a fruit-
body-inducing substance (Kawai et al., 1985).
Taxonomy of the Ganoderma lucidum Complex
The Ganodermataceae Donk was created to include polypore fungi characterized
by double-walled basidiospores. Large morphological variations in the family
resulted in the description of about 400 species, of which about two-thirds
classify in the genus Ganoderma Karst, many of them belonging to the
G. lucidum complex.
The variable morphological features of the G. lucidum complex, such as the
size, colour and shape of fruit bodies, may be caused by different environmental
conditions during development. Because of the morphological variation in
30. 18 G.-S. Seo and P.M. Kirk
Norwegian laccate specimens of G. lucidum, Ryvarden (1994) commented that
‘Macro-morphology is of limited value for criterion of species in the G. lucidum
group and at least 3–5 collections with consistent microscopical characters
should be examined before new species are described in this group’.
Cultural characteristics of Ganoderma species have been studied and
employed to determine taxonomic arrangement (Nobles, 1948, 1958;
Stalpers, 1978; Bazzalo and Wright, 1982; Adaskaveg and Gilbertson, 1986,
1989), but these attempts caused more confusion as they were often quite
different from classical identifications based on morphological features. For
example, Nobles (1948, 1958) described the differences in the cultural charac-
teristics of G. lucidum, G. tsugae and G. oregonense. Later, the isolates previously
listed as G. lucidum were changed to G. sessile (Nobles, 1965). However,
Steyaert (1972) and Stalpers (1978) classified it as G. resinaceum. The cultural
characteristics of G. resinaceum given by Bazzalo and Wright (1982) agree with
the description of Nobles (1965) and Stalpers (1978) and the description of
G. lucidum cultures given by Bazzalo and Wright (1982) is very similar to that
of G. tsugae as described by Nobles (1948). Furthermore, Stalpers (1978)
considered that the cultural characteristics of the European G. valesiacum were
identical to those of G. tsugae from North America, and listed it as a synonym of
G. valesiacum. Nobles (1958) suggested that the use of cultural characters in
the taxonomy of the Polyporaceae reflects natural relationships and phylogeny.
References
Adaskaveg, J.E. and Gilbertson, R.L. (1986) Cultural studies and genetics of sexuality
of Ganoderma lucidum and G. tsugae in relation to the taxonomy of the G. lucidum
complex. Mycologia 78, 694–705.
Adaskaveg, J.E. and Gilbertson, R.L. (1988) Basidiospores, pilocystidia, and other
basidiocarp characters in several species of the Ganoderma lucidum complex.
Mycologia 80, 493–507.
Adaskaveg, J.E. and Gilbertson, R.L. (1989) Cultural studies of four North American
species in the Ganoderma lucidum complex with comparisons to G. lucidum and
G. tsugae. Mycological Research 92, 182–191.
Adaskaveg, J.E. and Ogawa, J.M. (1990) Wood decay pathology of fruit and nut trees in
California. Plant Disease 74, 341–352.
Adaskaveg, J.E., Blanchette, R.A. and Gilbertson, R.L. (1991) Decay of date palm wood
by white-rot and brown-rot fungi. Canadian Journal of Botany 69, 615–629.
Adaskaveg, J.E., Miller, R.W. and Gilbertson, R.L. (1993) Wood decay, lignicolous
fungi, and decline of peach trees in South Carolina. Plant Disease 77, 707–711.
Ames, A. (1913) A consideration of structure in relation to genera of the Polyporaceae.
Annals of Mycology 11, 211–253.
Atkinson, G.F. (1908) Observations on Polyporus lucidus Leys., and some of its allies
from Europe and North America. Botanical Gazette (Crawfordsville) 46, 321–338.
Banerjee, S. and Sarkar, A. (1956) Formation of sporophores of Ganoderma lucidum
(Leyss.) Karst. and Ganoderma applanatum (Pers.) Pat. in culture. Indian Journal of
Mycological Research 2, 80–82.
31. Nomenclature and Classification 19
Bazzalo, M.E. and Wright, J.E. (1982) Survey of the Argentine species of the Ganoderma
lucidum complex. Mycotaxon 16, 293–325.
Blanchette, R.A. (1984) Screening wood decayed by white rot fungi for preferential
lignin degradation. Applied Environmental Microbiology 48, 647–653.
Bose, S.R. (1929) Artificial culture of Ganoderma lucidus Leyss from spore to spore.
Botanical Gazette (Crawfordsville) 87, 665–667.
Boudier, E. (1895) Description de quelques nouvelles espèces de Champignons recoltées
dans les regions elevées des Alpes du Valais, en aout 1894. Bulletin de la Société
Mycologique de France 11, 27–30.
Boudier, E. and Fischer, E. (1894) Rapport sur les espèces de Champignons trouvées
pendant l’assemblée a Geneve et les excursions faites en Valais. Bulletin de la Société
Mycologique de France 41, 237–249.
Corner, E.J.H. (1983) Ad Polyporaceas I. Amauroderma and Ganoderma. Nova Hedwigia
75, 1–182.
Donk, M.A. (1964) A conspectus of families of Aphyllophorales. Persoonia 3, 199–324.
Furtado, J.S. (1981) Taxonomy of Amauroderma. Memoirs of the New York Botanical
Garden 34, 1–109.
Gilbertson, R.L. and Ryvarden, L. (1986) North Americal Polypores. Part 1. Fungiflora,
Oslo, Norway.
Gottlieb, A.M. and Wright, J.E. (1999) Taxonomy of Ganoderma from southern South
America: subgenus Ganoderma. Mycological Research 103, 661–673.
Haddow, W.R. (1931) Studies in Ganoderma. Journal of the Arnold Arboretum 12,
25–46.
Hemmi, T. and Tanaka, I. (1936) Experiments for developing sporophores of Ganoderma
japonicum. Botany and Zoology 4, 13–23 (in Japanese).
Hepting, G.H. (1971) Diseases of forest and Shade Trees of the United States. US
Department of Agriculture, Agricultural Handbook, 386, 1–658.
Hseu, R.S. (1990) An identification system for cultures of Ganoderma species. PhD
thesis, National Taiwan University, Taipei (in Chinese).
Hseu, R.S., Wang, H.H., Wang, H.F. and Moncalvo, J.M. (1996) Differentiation and
grouping of isolates of the Ganoderma lucidum complex by random amplified
polymorphic DNA-PCR compared with grouping on the basis of internal
transcribed spacer sequences. Applied and Environmental Microbiology 62,
1354–1363.
Ito, S. (1955) Mycological Flora of Japan 2(4), 450 pp Yôkendo.
Julich, W. (1981) Higher Taxa of Basidiomycetes, J. Cramer, 485 pp.
Karsten, P. (1881) Enumeratio Boletinearum et Polyporearum Fennicarum, Systemate
novo dispositarum. Revue Mycologie (Toulouse) 3, 1–19.
Kawai, G., Ikeda, Y. and Tubaki, K. (1985) Fruiting of Schizophyllum induced by certain
ceramides and cerebrosides from Penicillium funiculosum. Agricultural and Biological
Chemistry 49, 2137–2146.
Kitamoto, Y. and Suzuki, A. (1992) Seiri. In: Hurukawa, H. (ed.) Kinokogaku.
KyouritsuSyuppan, Tokyo, pp. 79–115.
Kitamoto, Y., Takahashi, M. and Kasai, Z. (1968) Light induced formation of fruit
bodies in a basidiomycete, Favolus arcularius (Fr.) Ames. Plant and Cell Physiology 9,
797–805.
Kitamoto, Y., Suzuki, A. and Furukawa, S. (1972) An action spectrum for light induced
primordium formation in a basidiomycetes, Favolus arcularius (Fr.) Ames. Plant
Physiology 49, 338–340.
32. 20 G.-S. Seo and P.M. Kirk
Kitamoto, Y., Horikoshi, T. and Suzuki, A. (1974) An action spectrum for photo-
induction of pileus formation in a basidiomycete, Favolus arcularius. Planta 119,
81–84.
Manachere, G. (1980) Conditions essential for controlled fruiting of macromycetes.
A review. Transactions of the British Mycological Society 75, 255–270.
Melo, I. (1986) Studies on the Aphyllophorales of Portugal: The genus Ganoderma.
International Journal of Mycology and Lichenology 2, 183–204.
Mims, C.W. and Seabury, F. (1989) Ultrastructure of tube formation and basidiospore
development in Ganoderma lucidum. Mycologia 81, 754–764.
Mizuno, T., Wang, G.Y., Zhang, J., Kawagishi, H., Nishitoba, T. and Li, J.X. (1995)
Reishi, Ganoderma lucidum and Ganoderma tsugae: Bioactive substances and
medicinal effects. Food Reviews International 11, 151–166.
Moncalvo, J.M., Wang, H.F. and Hseu, R.S. (1995a) Gene phylogeny of the Ganoderma
lucidum complex based on ribosomal DNA sequences. Comparison with traditional
taxonomic characters. Mycological Research 99, 1489–1499.
Moncalvo, J.M., Wang, H.H. and Hseu, R.S. (1995b) Phylogenetic relationships in
Ganoderma inferred from the internal transcribed spacers and 25S ribosomal DNA
sequences. Mycologia 87, 223–238.
Moncalvo, J.-M. and Ryvarden, L. (1997) A nomenclatural study of the Gano-
dermataceae Donk. Synopsis Fungorum 11.
Morimoto, N. and Oda, Y. (1973) Effect of light on fruit body formation in a
basidiomycete, Coprinus macrorhizus. Plant and Cell Physiology 14, 217–225.
Murrill, W.A. (1902) The polyporaceae of North America. I. The genus Ganoderma.
Bulletin of the Torrey Botanical Club 29, 599–608.
Murrill, W.A. (1908) Family 5. Polyporaceae. North American Flora 9, 73–132.
Nobles, M.K. (1948) Studies in forest pathology. VI. Identification of cultures of wood-
rotting fungi. Canadian Journal of Research C 26, 281–431.
Nobles, M.K. (1958) Cultural characters as a guide to the taxonomy and phylogeny of
the Polyporaceae. Canadian Journal of Botany 36, 883–926.
Nobles, M.K. (1965) Identification of cultures of wood-inhabiting Hymenomycetes.
Canadian Journal of Botany 43, 1097–1139.
Overholts, L.O. (1953) Polyporaceae of the United States, Alaska, and Canada. University of
Michigan Press, Ann Arbor.
Park, D.S., Sung, J.M., Kim, Y.S., Yoo, Y.B., Ryu, Y.J. and Cha, D.Y. (1994) Analysis of
interspecific allozyme variation within the genus Ganoderma by polyacrylamide gel
isoelectric focusing. RDA Journal of Agricultural Science 36, 212–221.
Patouillard, N. (1889) Le genre Ganoderma. Bulletin de la Société Mycologique de France
5, 64–80.
Pegler, D.N. and Young, T.W.K. (1973) Basidiospore form in the British species of
Ganoderma Karst. Kew Bulletin 28, 351–364.
Perkins, J.H. (1969) Morphogenesis in Schizophyllum commune I. Effect of white light.
Plant Physiology 44, 1706–1711.
Perkins, J.H. and Gordon, S.A. (1969) Morphogenesis in Schizophyllum commune. II.
Effects of monochromatic light. Plant Physiology 44, 1712–1716.
Plunkett, B.E. (1961) The change of tropism of Polyporus brumalis stipes and the effect
of directional stimuli on pileus differentiation. Annals Botany, N. S. (London) 25,
206–222.
33. Nomenclature and Classification 21
Raudaskoski, M. and Yli-Mattila, T. (1985) Capacity for photoinduced fruiting in a
dikaryon of Schizophyllum commune. Transactions of the British Mycological Society
85, 145–151.
Ryvarden, L. (1994) Can we trust morphology in Ganoderma? In: Buchanan, P.K.,
Hseu, R.S. and Moncalvo, J.M. (eds) Ganoderma – Systematics, Phytopathology and
Phamacology. Proceedings of contributed symposia 59A, B, Fifth International
Mycological Congress, Vancouver, August 14–21, 1994, pp. 19–24.
Schwalb, M.N. (1978) Regulation of fruiting. In: Schwalb, M.N. and Miles, P.G. (eds)
Genetics and Morphogenesis in the Basidiomycetes. Academic Press, New York,
pp. 135–165.
Schwalb, M.N. and Shanler, A. (1974) Phototropic and geotropic responses during
the development of normal and mutant fruit bodies of the basidiomycetes
Schizophyllum commune. Journal of Genetic Microbiology 82, 209–212.
Seo, G.S. (1987) Studies on cultural characteristics of Ganoderma lucidum (Fr.) Karst.
MSc Thesis (in Korean).
Seo, G.S. (1995) In vitro photomorphogenesis and genetic diversity in the basidio-
mycete, Ganoderma lucidum. PhD dissertation, Tottori University, Tottori, Japan.
Seo, G.S., Shin, L.G.C., Otani, H., Komada, M. and Kohmoto, K. (1995a) Formation
of atypical fruiting structures in Ganoderma lucidum isolates on a nutrition agar
media. Mycoscience 36, 1–7.
Seo, G.S., Otani, H. and Kohmoto, K. (1995b) Effect of light on the formation of atypical
fruiting structures in Ganoderma lucidum. Mycoscience 36, 227–233.
Shin, G.C. and Seo, G.S. (1988a) Formation of the nonbasidiocarpous basidiospore of
Ganoderma lucidum. Korean Journal of Mycology 16, 230–234 (in Korean).
Shin, G.C. and Seo, G.S. (1988b) Classification of strains of Ganoderma lucidum. Korean
Journal of Mycology 16, 235–241 (in Korean).
Shin, G.C. and Seo, G.S. (1989a) Effect of light on the formation of non-basidiocarpous
basidiospores of Ganoderma lucidum. Korean Journal of Mycology 17, 189–193
(in Korean).
Shin, G.C. and Seo, G.S. (1989b) Effect of temperature and aeration on the formation
of non-basidiocarpous basidiospores of Ganoderma lucidum. Korean Journal of
Mycology 17, 194–196 (in Korean).
Shin, G.C., Park, Y.H., Seo, G.S. and Cha, D.Y. (1986) Morphological characters
of Ganoderma lucidum (Fr) Karsten grown naturally in Korea. Research Reports of
Institute of Agricultural Science and Technology 13, 44–51 (in Korean).
Stalpers, J.A. (1978) Identification of wood-inhibiting Aphyllophorales in pure culture.
Centraalbureau Voor Schimmelcultures, Baarn. Studies in Mycology, pp. 1–248.
Stamets, P. (1993a) Evaluating a mushroom strain: Photosensitivity. In: Growing Gour-
met and Medical Mushrooms. Ten Speed Press, Berkely, California, pp. 117–126.
Stamets, P. (1993b) The polypore mushrooms of the genera Ganoderma, Grifola and
Polyporus. In: Growing Gourmet and Medical Mushrooms. Ten Speed Press, Berkely,
California, pp. 351–369.
Steyaert, R.L. (1972) Species of Ganoderma and related genera mainly of the Bogor and
Lieden herbaria. Persoonia 7, 55–118.
Steyaert, R.L. (1975) The concept and circumscription of Ganoderma tornatum. Trans-
actions of the British Mycological Society 65, 451–467.
Steyaert, R.L. (1977) Basidiospores of two Ganoderma species and others of two related
genera under the scanning electron microscope. Kew Bulletin 31, 437–442.
34. 22 G.-S. Seo and P.M. Kirk
Steyaert, R.L. (1980) Study of some Ganoderma species. Bulletin du Jardin Botanique
National de Belgique 50, 135–186.
Suzuki, A. (1979) General review on environmental factors affecting primordium
formation in Homobasidiae. Transaction of the Mycological Society of Japan 20,
253–265 (in Japanese).
Willard, T. (1990) Reishi Mushroom, ‘Herb of spiritual potency and medical wonder’. Sylval
Press, Issaquah, Washington, p. 167.
Yli-Mattila, T. (1990) Photobiology of fruit body formation in the basidiomycete
Schizophyllum commune. Reports from the Department of Biology, University of Turku,
No. 27, pp. 1–67. Turku.
Zhao, J.D. (1989) The Ganodermataceae in China. Bibliotheca Mycologica 132.
J. Cramer, Berlin, Stuttgart.
Zhao, J.D. and Zhang, X.Q. (1994) Importance, distribution and taxonomy of
Ganodermataceae in China. In: Buchanan, P.K., Hseu, R.S. and Moncalvo, J.M.
(eds) Ganoderma – Systematics, Phytopathology and Pharmacology. Proceedings
of Contributed Symposia 59A, B, Fifth International Mycological Congress,
Vancouver, August 14–21, 1994, pp. 1–2.
36. 24 J.-M. Moncalvo
should be abandoned for various reasons (Moncalvo and Ryvarden, 1997).
Most species were described in the genus Ganoderma (219 species), mainly
from laccate collections (166 species). Many species are known only from a
single collection or locality. Several names have been considered synonyms
(reviewed in Moncalvo and Ryvarden, 1997), but I believe that more
taxonomic synonyms still exist because a large number of species were
Fig. 2.1. Morphological characters traditionally used in Ganoderma systematics.
(a) Typical basidiospore of Ganoderma. (b) Basidiospore of G. boninense. (c)
Basidiospore of G. formosanum (longitudinal crests are barely seen in light micro-
scopy). (d) Typical basidiospore of Amauroderma. (e) Various types of pilocystidia
found in Ganoderma. (f) Stipitate versus dimidiate basidiocarps: relationships
between stipe formation and location of basidiocarp development on wood.
37. Table 2.1. A summary of the traditional taxonomy in Ganodermaa.
Number of species
Number of Estimated
Known from a names proposed number of
Genera Subgenera Distinctive features Described single locality as synonyms known species
Ganoderma Spore wall enlarged at the apex
Ganoderma Pilear surface laccate (presence of pilocystidia) 168 124 48 60–80
Elfvingia Pilear surface dull (absence of pilocystidia) 51 31 21 10–30
Amauroderma Spore wall uniformly large 96 60 41 30–50
Haddowiab Spore wall uniformly large and spore surface 5 1 2 3
longitudinally crested
Humphreyac Spore wall enlarged at the apex and spore 7 3 3 4
Systematics of Ganoderma
surface reticulate
aDatafrom Moncalvo and Ryvarden (1997).
bSynonym of Amauroderma in Furtado (1981) and Corner (1983).
cSynonym of Ganoderma in Furtado (1981) and Corner (1983).
25
38. 26 J.-M. Moncalvo
distinguished from characters that depend on growing conditions and develop-
mental stage. For instance, careful observation in vivo shows that young,
actively growing fruiting bodies generally have lighter and brighter surface
colours than basidiocarps that are several weeks or months old: the latter have
been exposed to repeated periods of rain and dryness, covered with dust,
attacked by insects, or even colonized by algae. Presence, absence, size and
insertion of the stipe have also been used to circumscribe species (e.g. G.
gibbosum, G. dorsale, etc.), but it has been shown that stipe development can be
controlled in vitro by the duration and intensity of exposure to light and by car-
bon dioxide concentration (Hseu, 1990). In vivo, stipe development also
depends on the location in the host: a basidiocarp that develops from a buried
root is more likely to develop a stipe than a basidiocarp that develops higher in
the trunk (Fig. 2.1). Ryvarden (1995) examined the variability of 53 Norwe-
gian specimens of G. lucidum, and concluded that macromorphological charac-
ters are of very limited value for the identification of Ganoderma species.
Reliable morphological characters for Ganoderma systematics appear to be
spore shape and size, context colour and consistency, and microanatomy of the
pilear crust. However, the typical spore of G. lucidum is similar for dozens of
different species. Scanning electron microscopy (SEM) has been useful in dis-
tinguishing between spores that appear similar under light microscopy (Pegler
and Young, 1973; Gottlieb and Wright, 1999), and has revealed the existence
of distinctive, slightly longitudinally crested basidiospores in the G. australe and
G. sinense species complexes (Hseu, 1990; Buchanan and Wilkie, 1995; Tham,
1998). Context colour and consistency may change slightly with the age of the
fruit body or upon drying, and are also somewhat subjective characters, but it
is still possible to distinguish at least three very distinctive types: (i) light col-
oured and/or duplex context in G. lucidum and its allies; (ii) uniformly brown to
dark brown context as in the G. sinense and G. australe complexes; and (iii) very
soft, cream to pale ochraceous context in G. colossum. Relationships between
the microstructure of the pilear crust, the age of the basidiocarp, and the
exposure to environment are not well known, but different types of pilocystidia
and hyphal arrangement can be distinguished among both laccate and non-
laccate taxa (Steyaert, 1980; Fig. 2.1). The laccate appearance of Ganoderma
basidiocarps is associated with the presence of thick-walled pilocystidia
(Fig. 2.1) that are embedded in an extracellular melanin matrix. The exact
origin and chemical composition of this matrix remain to be elucidated.
High phenotypic plasticity at the macroscopic level, uniformity of micro-
scopic characters, and subjective interpretation of various features such as
colour or consistency have resulted in the creation of numerous unnecessary
names (synonyms), and a lack of handy identification keys. The absence of
a world monograph has also contributed to problems with species circum-
scriptions and identifications in Ganoderma.
Culture and enzymatic studies have produced additional and useful
taxonomic characters in Ganoderma systematics (Adaskaveg and Gilbertson,
1986, 1989; Hseu, 1990; Wang and Hua, 1991; Gottlieb et al., 1995; Gottlieb
39. Systematics of Ganoderma 27
and Wright, 1999). It appears that chlamydospore production and shape, and
to a lesser extent the range and optima of growth temperatures, are extremely
useful culture characters for distinguishing between morphologically similar
species. Mating studies have also been conducted to circumscribe biological
species within species complexes (Adaskaveg and Gilbertson, 1986; Hseu,
1990; Yeh, 1990; Buchanan and Wilkie, 1995). However, all these studies
were restricted in scope, and the techniques employed, although useful at the
species level, have limitations for addressing phylogenetic relationships
between taxa and the development of a natural classification system.
Molecular Systematics of Ganoderma
With recent advances in both sequencing techniques to produce taxonomic
characters and cladistic methods to infer natural relationships between
organisms, molecular systematics has become a paradigm in biology. To date,
the most widely used molecules in fungal molecular systematics have been
the ribosomal genes (rDNA). Hibbett and co-workers (Hibbett and Donoghue,
1995; Hibbett et al., 1997) produced molecular phylogenies for hymenomyce-
tous fungi using sequence data from the nuclear small subunit (18S, or nSSU)
and mitochondrial small subunit (12S, or mtSSU) rDNA, and showed that
Ganoderma belongs to a larger group of white-rot fungi that also includes the
genera Trametes, Fomes, Polyporus, Lentinus, Datronia, Pycnoporus, Cryptoporus,
Daedalopsis, Lenzites and Dentocorticium. Additional phylogenetic studies using
sequence data from the nuclear large ribosomal subunit (25–28S, or nLSU)
rDNA showed that genera Amauroderma, Irpex, Loweporus and Perenniporia
also belong to this group (Moncalvo et al., 2000; Thorn et al., 2000; Moncalvo,
unpublished). Combined evidence of nLSU and mtSSU-rDNA data support the
placement of Amauroderma as a sister genus to Ganoderma (Moncalvo and
Hibbett, unpublished). However, nucleotide sequence data from nuclear and
mitochondrial rDNA encoding sequences do not offer enough variation to infer
phylogenetic relationships between Ganoderma species.
Appropriate nucleotide sequence variation for systematics of Ganoderma
was found in the internal transcribed spacers (ITS) of the nuclear rDNA gene
(Moncalvo et al., 1995a, b, c). The ITS phylogenies produced in these studies
indicated that many names were commonly misapplied (e.g. G. lucidum and
G. tsugae), and that the proposed subgenera and sections in Steyaert (1972,
1980) and Zhao (1989) were not monophyletic and should be abandoned.
Gene trees and species trees
A gene tree is not necessarily equivalent to a species tree, and phylogenetic
trees inferred from the sequences of different genes can be contradictory for
several reasons, including differences in their power or level of phylogenetic
40. 28 J.-M. Moncalvo
resolution, incorrect recovery of evolutionary relationships by phylogenetic
reconstruction methods (e.g. ‘long branch attraction’, Felsenstein, 1978),
discordance in rates and modes of sequence evolution (Bull et al., 1993), differ-
ent phylogenetic histories due to lineage sorting or difference in coalescence
time (Doyle, 1992, 1997; Maddison, 1997), or horizontal gene transfer.
Incongruences between gene trees are more likely to occur at lower taxonomic
levels (species, populations). In fact, it is expected that gene trees are
incongruent among interbreeding individuals because these individuals are
connected by gene flow and recombination: their relationships are therefore
tokogenetic (reticulate) rather than phylogenetic (divergent) (Hennig, 1966;
Doyle, 1997). Overall, a phylogenetic hypothesis is more likely to be correct if it
is supported from multiple, independent data sets rather than from a single
gene tree.
ITS phylogeny versus manganese-superoxide dismutase (Mn-SOD)
phylogeny
Thirty-three Ganoderma taxa were used to conduct separate phylogenetic
analyses of sequence data from ITS and Mn-SOD genes. The incongruence
length difference (ILD) test of Farris et al. (1994), also known as the partition-
homogeneity test, indicated absence of statistically significant conflict
(P = 0.08) in phylogenetic signals between the two data sets. Results of the
analyses are shown in Fig. 2.2. Tree topologies are fully congruent for all
nodes having bootstrap statistical support (BS) greater than 50%, with two
exceptions:
1. the type specimen of G. microsporum clusters with G. weberianum
CBS219.36 in the ITS analysis (88% BS), but clusters with a strain labelled
G. cf. capense ACCC5.71 in the Mn-SOD analysis (98% BS); and
2. the cultivar G. cf. curtisii RSH.J2 nests with strain RSH-BLC in the ITS
analysis (58% BS) but with RSH-J1 (83% BS) in the Mn-SOD analysis.
The latter three collections are known to be intercompatible (i.e. belong to
the same biological species; Hseu, 1990), therefore conflicting gene phylo-
genies for these strains are not surprising. Strains labelled G. microsporum,
G. weberianum and G. cf. capense are probably also conspecific: the synonymy of
the first two names was already suggested by Peng (1990).
Both data sets strongly support similar terminal clades, and do not fully
resolve basal relationships among Ganoderma taxa. The ITS data set offers
slightly more resolution for deeper branches (Fig. 2.2), whereas higher
sequence divergence between closely related taxa was found in the Mn-SOD
gene (in particular in two introns that were excluded from the analyses
because nucleotide sequences could not be unambiguously aligned across all
the taxa sampled). Ongoing sequencing and analyses of β-tubulin genes also
41. Systematics of Ganoderma 29
support similar terminal clades to those from ITS and Mn-SOD data (Moncalvo
and Szedlay, unpublished).
Therefore, preliminary data suggest that phylogenies derived from
ITS sequences are congruent with those from other genes, and that ITS
phylogenies may accurately reflect natural relationships between Ganoderma
species.
Fig. 2.2. Comparison between internal transcribed spacer (ITS) and manganese-
superoxide dismutase (Mn-SOD) nucleotide sequence phylogenies for 33 Gano-
derma taxa. Sequences from one species of genus Amauroderma were used to
root the trees. Trees depicted are strict consensus trees produced from maximum
parsimony searches. Bootstrap statistical supports greater than 50% are shown
above branches. Mn-SOD data were from Wang (1996; GenBank accession
numbers U56106-U56137), and Moncalvo and Szedlay (unpublished). Analyses
were conducted in PAUP* (Swofford, 1998) and employed maximum parsimony
with heuristic searches using 50 replicates of random addition sequences with
TBR branch swapping. Bootstrap statistical supports were evaluated with 100
bootstrap replicates of random addition sequence with TBR branch swapping.
Regions with ambiguous alignment were removed from the alignment, and
unambiguously aligned gaps were scored as ‘fifth character state’. The ITS
data set used 81 parsimony-informative characters and produced 24 equally
parsimonious trees of length 232, with a consistency index of 0.703. The SOD
data set used 105 parsimony-informative characters and produced 58 equally
parsimonious trees of length 329, with a consistency index of 0.623.
42. 30 J.-M. Moncalvo
ITS phylogeny
The current ITS sequence database for Ganoderma and Amauroderma species
includes about 300 taxa. Numerous small nucleotide insertions and deletions
make sequence alignment problematic in several regions, but at least 380
characters can be aligned unambiguously across the entire data set, yielding
about 200 parsimony-informative characters. Phylogenetic analysis of large
molecular data sets is still a controversial field (Lecointre et al., 1993; Hillis,
1996; Graybeal, 1998; Poe, 1998). One commonly encountered problem with
large data sets concerns the applicability and/or accuracy of standard
descriptors commonly used to assess branch robustness. For instance, the use
of branch decay indices (Bremer, 1994) is not practical for large data sets
because of the large number of trees that cannot be sampled; and consistency
indices (Sanderson and Donhogue, 1989), bootstrap (Felsenstein, 1985) and
jackknife (Farris et al., 1996) statistical supports are sensitive to sample size.
However, evidence from various studies (Hillis, 1996, 1998; Moncalvo et al.,
2000) suggests that increasing taxon sampling generally increases phylo-
genetic accuracy, and that bootstrapping or jackknifing methods are still
useful tools to determine the robustness of clades.
Parsimony analyses of ITS data for 248 Ganoderma taxa reveal about
50 clades with bootstrap statistical support greater than 50% (Fig. 2.3 and
Table 2.2), that are also consistent with morphological and/or geographical
data. Terminal clades in this phylogeny represent either a population, a
species, a species complex, or a group of closely related species. In Table 2.2,
tentative names for the most well-supported clades are proposed, although
16 clades have not been named (the original data set included 36 species
names and many unnamed taxa). Basal relationships are either poorly
supported or unresolved, but phylogenetic analyses of various data sets
using maximum parsimony and maximum likelihood consistently reveal
three larger groups: these are labelled Groups 1–3 in Fig. 2.3 and Table
2.2.
ITS phylogeny suggests that the laccate habit has been derived more
than once (or lost several times), making the laccate Ganoderma taxa
polyphyletic. This conflicts with traditional systems of classification that
accommodate laccate and non-laccate Ganoderma taxa in subgenera
Ganoderma (laccate) and Elfvingia (non-laccate), respectively (see Table 2.1).
However, within the Ganodermataceae, there is already evidence for
non-monophyly of laccate taxa because at least three laccate species have
been traditionally classified in genus Amauroderma (Furtado, 1981). A revised
classification for subgenera and sections in Ganoderma seems therefore
necessary, and will be formally proposed elsewhere. For now discussion is
limited to some taxonomic groupings revealed by ITS sequence data, as
summarized in Table 2.2.
43. Systematics of Ganoderma 31
Fig. 2.3. Internal transcribed spacer (ITS) phylogeny for 248 taxa of Gano-
dermataceae (sequences from several Amauroderma species were used to root the
tree). The tree depicted is one of 100 equally parsimonious trees produced using
maximum parsimony in PAUP* (Swofford, 1998) with heuristic searches, random
addition sequences (100 replicates), TBR branch swapping, and MAXTREES set
to 100. Statistical supports for branch robustness were evaluated in PAUP* with
100 bootstrap replicates, random addition sequence, TBR branch swapping, and
MAXTREES set to 10. Bootstrap values are only given for branches in bold that
refer to groups or clades that are presented in Table 2.2. Groups 1 and 1.4 are not
monophyletic in the figure they were retained as such to facilitate the discussion.
Details about Groups 1–3 and unclassified taxa are given in the text and Table 2.2.
44. 32
Table 2.2. Groupings of Ganoderma taxa based on a phylogenetic analysis of ITS nucleotide sequence data (Fig. 2.3), with geographic
origin and host relationships of the strains examined.
Geographic categories Hosts
India, Pakistan
Indo, PNG
S. America
Neotropics
N. America
Palms
S. Africa
Europe
China, Korea
Japan
Taiwan
S.E. Asia
Australia
New Zealand
Florida
Woody dicots
Conifers
Group 1
1.1 G. lucidum complex sensu stricto (84% BS)
G. lucidum • • •
G. valesiacum • •
G. carnosum • •
G. ahmadii • •
G. tsugae • • •
J.-M. Moncalvo
G. oregonense • •
G. praelongum, G. oerstedii • •
1.2 G. resinaceum complex sensu lato (86% BS)
G. resinaceum complex sensu stricto:
G. resinaceum (’G. pfeifferi’) (90% BS) • • •
G. cf. resinaceum (’G. lucidum’) (64% BS) • • •
G. cf. resinaceum (G. sessile, G. platense) (59% BS) • •
G. weberianum complex (59% BS):
G. weberianum (= G. microsporum) (89% BS) • • • •
G. cf. capense (56% BS) • • •
Ganoderma sp. (99% BS) • •
Ganoderma sp. (’G. subamboinense’) (97% BS) • • •
G. trengganuense (87% BS) • •
