Fungal succession associated with the decay of leaves of an evergreen oak, Quercus myrsinaefolia

Abstract

Fungal succession associated with the decay of leaves of an evergreen oak, Quercus myrsinaefolia. Fungal Diversity 34: 87-109. In Japan, the species component of fungal communities and fungal succession on decaying fallen leaves of broadleaf-evergreen trees has not been elucidated. In this study, we investigated the species components and structures of fungal communities inhabiting the decaying leaves of an evergreen oak, Quercus myrsinaefolia. Fungal succession occurs with the progressive decay of Q. myrsinaefolia leaves with the phyllosphere fungi, such as Tubakia sp. and Colletotrichum gloeosporioides giving way to early colonizers of fallen leaves on the ground, such as Subramaniomyces fusisaprophyticus and Rhinocladiella intermedia, and then progressing to later colonizers, such as Trichoderma koningii and T. harzianum. The whole succession pattern at this study site is characterized by the fungal succession associated with the decomposition of fallen leaves during the main leaf-fall seasons.

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Fungal Diversity
87
Fungal succession associated with the decay of leaves of an evergreen oak,
Quercus myrsinaefolia
Shirouzu, T.1*, Hirose, D.1, Fukasawa, Y.2 and Tokumasu, S.1
1Sugadaira Montane Research Center, University of Tsukuba, 1278-294, Ueda, Nagano, 386-2204, Japan
2Laboratory of Forest Ecology, Division of Environmental Science and Technology, Graduate School of Agriculture,
Kyoto University, Kyoto 606-8502, Japan
Shirouzu, T., Hirose, D., Fukasawa, Y. and Tokumasu, S. (2009). Fungal succession associated with the decay of leaves
of an evergreen oak, Quercus myrsinaefolia. Fungal Diversity 34: 87-109.
In Japan, the species component of fungal communities and fungal succession on decaying fallen leaves of broadleaf-
evergreen trees has not been elucidated. In this study, we investigated the species components and structures of fungal
communities inhabiting the decaying leaves of an evergreen oak, Quercus myrsinaefolia. Fungal succession occurs with
the progressive decay of Q. myrsinaefolia leaves with the phyllosphere fungi, such as Tubakia sp. and Colletotrichum
gloeosporioides giving way to early colonizers of fallen leaves on the ground, such as Subramaniomyces
fusisaprophyticus and Rhinocladiella intermedia, and then progressing to later colonizers, such as Trichoderma koningii
and T. harzianum. The whole succession pattern at this study site is characterized by the fungal succession associated
with the decomposition of fallen leaves during the main leaf-fall seasons.
Key words: broadleaf-evergreen forest, fungal community, leaf decomposition, saprobic succession, warm temperature
Article Information
Received 1 May 2008
Accepted 9 July 2008
Published online 5 January 2009
*Corresponding author: Takashi Shirouzu; e-mail: shirouzy@sugadaira.tsukuba.ac.jp
Introduction
Since the first systematic study of sa-
probic fungal succession associated with the
decay of fallen pine needles by Kendrick and
Burges (1962), many studies on fungal suc-
cesssion have been conducted on fallen needles
of conifers, such as Pinus, Abies and Picea, in
subtropical, temperate and sub-arctic zones in
the northern hemisphere (Hayes, 1965a, b;
Tubaki and Saitô, 1969; Widden and Parkin-
son, 1973; Mitchell and Millar, 1978; Mitchell
et al., 1978; Soma and Saitô, 1979; Aoki et al.,
1990; Tokumasu et al., 1994; Tokumasu, 1996,
1998a,b; Tokumasu and Aoki, 2002). Fungal
succession on fallen leaves of broadleaf trees
(Saitô, 1956; Hering, 1965; Hogg and Hudson,
1966; Macauley and Thrower, 1966; Aoki,
1987; Promputtha et al., 2002; Osono, 2002,
2005; Tang et al., 2005; Pasqualetti et al.,
2006; Paulus et al., 2006; Duong et al., 2008)
and monocotyledons (Yanna et al., 2002;
Thongkantha et al., 2008) have also been
studied. These studies have accumulated
information on fungal succession with leaf
decomposition in broadleaf trees, conifers and
monocotyledons. In general, the fungal species
involved in fungal succession on fallen leaves
of deciduous broadleaf trees were different
from those of conifers and monocotyledons,
but the successional patterns were funda-
mentally the same (Hudson, 1968); however,
information about fungal succession is still
limited in the leaf litter of broadleaf-evergreen
forests in warm-temperate regions in the
northern hemisphere.
In southwest Japan, the original vege-
tation is broadleaf-evergreen forest, although
the forest has been almost destroyed by human
activity. It is mainly composed of evergreen
trees of Fagaceae (Quercus and Castanopsis
etc.) and Lauraceae (Machilus and Cinnamo-
mum etc.). A series of pioneering studies of
saprobic fungal succession on evergreen

88
Quercus and Castanopsis leaves was per-
formed by Tubaki and Yokoyama (1971,
1973a, b), Yokoyama and Tsubaki (1973) and
Yokoyama et al. (1977) who inserted auto-
claved leaves into the surface layer of the O
horizon and observed the succession occurring
on the leaves. A similar experimental study
using unsterilized Cameria japonica leaves set
on litter has been attempted by Tsubaki and
Yoshida (1980); however, information about
saprobic fungal succession on decaying leaves
of broadleaf-evergreen trees is still inadequate
for Japanese warm-temperate forest litter.
In this study, we attempted to study
fungal succession associated with the decay of
leaves of Quercus myrsinaefolia, a dominant
evergreen oak in the northern broadleaf-
evergreen forest in Japan. To evaluate the
changing fungal community on decaying
leaves, we surveyed fungi colonizing the leaves
at different decomposition stages by a cultural
method. Obtained data were compared among
decomposition stages, and fungal succession
occurring on decaying leaves of Q. myrsinae-
folia was estimated. The results were discussed
by comparing with similar published data on
other trees.
Materials and methods
Study site
Leaf samples were collected from forest
litter in the precinct of Meiji Jingu Shrine,
Shibuya, Tokyo (35° 40’ N, 139° 41’ E; alt. 35
m). The mean temperature and precipitation in
this area from 1971 to 2004 were 16.3 °C and
1532mm, respectively, according to weather
observation data from Tokyo District
Meteorological Observatory (Chiyoda, Tokyo).
Monthly mean temperatures and precipitations
in the study period, February 2005 to January
2006, are shown in Fig. 1. The study area is
covered with about 80-year-old planted trees
and it is a well developed, closed broadleaf-
evergreen forest mainly composed of Quercus
myrsinaefolia, Castanopsis sieboldii and,
Cinnamomum camphora. We selected a stand
in the forest for leaf sampling where Quercus
myrsinaefolia is predominant. At the sampling
site, the O horizon was 3 to 6 cm in depth and
composed of an intermingling of variously
decayed leaves. Sub-layers corresponding to
the F or H layer found at the mor site were not
developed.
Measurement of litter production
To estimate the seasonal fluctuation of
leaf litter production of Quercus myrsinaefolia,
leaf litter samples were collected and weighed
in April, June, August, October and December,
2005 according to the following procedure.
The O horizon was cut from six plots (10 × 10
cm) at the sampling site and each sample was
placed in a paper bag and brought back to the
laboratory where only the leaves of Q. myrsi-
naefolia were selected from litter samples. The
leaves were divided into three groups, L-type
(elastic, intact and olive colored), OL-type
(between L-type and F-type) and F-type
(inelastic, damaged and ash-grey colored) by
external appearance. These leaf samples were
dried for 4 days at 40 °C, and then weighed.
Mean monthly dry weights of fallen leaves at
each decomposition stage were calculated.
Chemical property of fallen leaves
Litter samples at the same decomposition
stages from each of four sampling seasons
(April, August, October and December) were
mixed to make one sample and used for
chemical analyses. The dried litter samples
were ground in a laboratory mill to pass
through a 0.5 mm screen and used for chemical
analyses. Concentrations of total N and C were
measured by automatic gas chromatography
(NC analyzer SUMIGRAPH NC-900, Sumi-
tomo Chemical Co., Osaka, Japan). Soluble
sugar and polyphenol were extracted with 50%
methanol, and their contents were estimated
with the phenol-sulphuric acid method (Dubois
et al., 1956) and Folin-Ciocalteau method
(Waterman and Mole, 1994), respectively.
The carbon to nitrogen ratio (C/N) is an
useful index of litter chemical properties
(Osono and Takeda, 2001) and is calculated
according to the following equation:
C/N = carbon concentration (%) /
nitrogen concentration (%)
Leaf sampling for fungal isolation
Fallen leaves of Quercus myrsinaefolia
were collected from the O horizon at the

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0
5
10
15
20
25
30
0
50
100
150
200
250
300
Feb
2005
Mar Apr May Jun Jul Aug Sep Oct Nov Dec Jan
2006
Temperature (°C)
Precipitation (mm)
Precipitatio n
Temper ature
Fig. 1. Monthly average temperatures and precipitation in the study area.
sampling site on April 10, August 2, October 5,
2005 and January 17, 2006. At the same time,
two-year-old symptomless green leaves on the
trees (A-type leaves) were also collected from
lower branches. The collected leaves were
taken back to the laboratory in separate paper
bags and fungi were isolated within 24 hours
after collection.
For fungal isolation, ten representative
leaves were selected from each leaf type and
two discs per leaf were punched out of the
central part of leaves except for the main veins
by a 7 mm diameter corkbore. Twenty discs
per leaf type, a total of 80 discs, were used for
fungal isolations at each investigation time.
The washing method (Tokumasu, 1980) was
used for fungal isolation. Punched leaf discs of
each leaf type were put in a sterile test tube
with a plastic cap, and autoclaved. Then 10 ml
0.005% aero-sol OT solution (Wako, Osaka,
Japan) was poured into the test tube as a
washing solution using a sterile pipette. To
wash the fungal diaspores off the surface of the
leaf discs, the test tube was stirred for 1 min
using a vortex mixer. After stirring, the
washing solution was removed from the test
tube using another sterile pipette, and then new
washing solution was added to the test tube and
stirred again. After five repeats of the washing
operation, the same process was repeated three
times using sterile distilled water. To inhibit
the propagation of bacteria, washed and rinsed
leaf discs were placed on a sterile filter paper in
a 9 cm Petri dish and dried for 24 hours at
room temperature (Widden and Parkinson,
1973). Dried leaf discs were adhered to 0.2%
corn meal agar plates (Nissui, Tokyo, Japan) in
9 mm plastic Petri dishes with four discs per
dish and cultured at room temperature.
Observation and identification of fungi
After 3 days, 10 days, 28 days and 42
days from the start of incubation, the Petri
dishes were observed using both dissecting and
light microscopes, and fungal species were
identified directly based on the morphology of
their fruit bodies formed on agar plates or leaf
discs. Fungi that could not be identified by
direct observation were isolated by their spores
or mycelia using a sterile elgiloy wire and
preserved in slant cultures. These strains were
then plated out on LCA plates (Miura and
Kudo, 1970; glucose 1.0 g, KH2PO4 1.0 g,
MgSO4·7H2O 0.2 g, KCl 0.2 g, NaNO3 2.0 g,
yeast extract 0.2 g, agar 13 g, distilled water
1.0 l) to induce their sporulation. When no
sporulation of fungi occurred on LCA plates,
pieces of sterile Quercus myrsinaefolia leaf
were added to the plates to induce sporulation.
When spores or other propagative structures
were formed on LCA plates or added leaf
pieces, slides were prepared to observe them
under a light microscope for identification. In
this study, sterile strains were treated as
unidentified fungi and excluded from the
fungal species list. Fungal identification was,

0
1
2
3
4
5
6
Apr Jun Aug Oc t Dec
Dry weight (g)/10cm
2
L-type
OL-type
F-type
Fig. 2. Seasonal change of leaf litter production of Q. myrsinaefolia at the sampling site.
10
15
20
25
30
35
L-type OL-type F-type
C/N ratio
Fig. 3. C/N ratio in fallen leaves at each decomposition stage (error bars are standard deviations).
0
5
10
15
20
25
30
35
L-t
yp
eOL-t
yp
eF-t
yp
e
Concentration (mg/g)
Polyphenol
Soluble suger
Fig. 4. Polyphenol and soluble sugar concentrations in fallen leaves at each decomposition stage (error bars are standard
deviations).
90

Fungal Diversity
91
18
52 53
50
0
10
20
30
40
50
60
A-type L-type OL-type F-type
Total number of species
Fig. 5. Total numbers of recorded fungal species for each leaf type.
0
10
20
30
40
50
60
April August October January
To tal numbe r of spe cies
A-type L-type OL-type F-type Total
Fig. 6. Seasonal changes in numbers of recorded fungal species for each leaf type.
mainly according to studies by Ellis (1971,
1976), Matsushima (1971, 1975), Udagawa et
al. (1978a, b), Carmichael et al. (1980),
Domsch et al. (1980a, b), Sutton (1980), Nag
Raj (1993), and Kiffer and Morelet (2000).
Data calculation
The frequency of occurrence and
seasonality index of each fungus at each
decomposition stage, and Sørensen’s similarity
index (Sørensen, 1948) to examine the simila-
rity of fungal species components between leaf
types were calculated according to the
following expressions. Fungi which could be
identified only at a generic level, such as
Fusarium spp. and Penicillium spp., were
calculated as one species.
Frequency of occurrence
Frequency of occurrence (%) = number
of discs on which a certain species occurred /
total number of discs examined (20) × 100
In this study, fungi occurring at ≥50%
frequency were defined as frequent species.
Seasonality index

Seasonality index (%) = number of
investigation times in which a certain species
occurred / total investigation times (4) × 100
In this study, fungi recorded at 100%
seasonality index were defined as constant
species.
Sørensen’s similarity index
Sørensen’s similarity index = 2c / (a + b)
a: number of species occurring in sample
A.
b: number of species occurring in sample
B.
c: number of species occurring in both
samples.
Results
Leaf litter production of Quercus
myrsinaefolia
Seasonal change of leaf litter production
of Q. myrsinaefolia at the sampling site is
shown in Fig. 2. The maximum total dry
weight of fallen leaves was recorded in June,
and the second in August. Although the leaf
fall of Q. myrsinaefolia was observed through-
out the year, the dry weight of L-type leaves
increased from June to August and peaked in
June.
Chemical property of fallen leaves
The C/N ratio, polyphenol and soluble
sugar concentrations in fallen leaves at each
decomposition stage are shown in Figs. 3 and
4. These values decreased with the progress of
leaf decomposition. Standard deviations of
polyphenol and soluble sugar concentrations in
L-type leaves were large compared with other
leaf types (Fig. 4). Although it is not shown in
Fig. 4, especially in December, these values in
the L-type (polyphenol: 27.7 mg/g; soluble
sugar: 38.8 mg/g) were higher than in other
seasons (polyphenol: 10–15.8 mg/g; soluble
sugar: 18.4–23 mg/g).
Numbers of species and their seasonal
changes
The list of all recorded fungi is shown in
Table 1. In this study, 83 species of 57 genera
were recorded. Eighteen species of 17 genera
were collected from living leaves and 80
species of 55 genera from fallen leaves. In A-
type leaves, three fungi were recorded from the
A-type only (▲ in Table 1), and the remaining
15 species of 14 genera occurred also on fallen
leaves. Seven species were recorded from all
leaf types (* in Table 1), and 22 species were
recorded from all fallen leaf type (♦ in Table 1).
The total numbers of species at each leaf
type are shown in Fig. 5. Eighteen species of
A-type leaves was the smallest for all leaf
types. The numbers recorded from other leaf
types were almost the same, 50 to 53 species.
Seasonal changes in the number of species
recorded from each leaf type are shown in Fig.
6. The total number of species recorded from
all leaf types and the number recorded from A-
type leaves were relatively stable throughout
the year. In L-type leaves, the species number
increased gradually toward October. The
species number on OL-type leaves peaked in
January, with the minimum in October, while
on F-type leaves it peaked in October, with the
minimum in August.
Similarity indexes of species components
between leaf types
The results of calculating Sørensen’s
similarity index for fungal species components
between leaf types are shown in Table 2.
Similarity indexes of L-type and OL-type
(0.667), OL-type and F-type (0.757), and L-
type and F-type (0.588) were higher than other
combinations, and the A-type and L-type
(0.423) also had a relatively high value.
Distribution patterns of fungal species in all
leaf types
To understand the distribution patterns of
individual fungi in all leaf types, the average
occurrence frequencies of each fungus on every
leaf type was calculated (Table 3). In three
fungi recorded from the A-type only (▲ in
Table 3), Stenella sp. showed a high average
frequency of occurrence (84%) on A-type
leaves. The average frequencies of Tubakia sp.,
Tripospermum prolongatum and Colleto-
trichum gloeosporioides were also high in A-
type and decreased after leaf-fall.
In seven species collected from all leaf
types (* in Table 3), Aureobasidium pullulans
and Phoma sp. showed high average
frequencies of occurrence on L-type leaves and
tended to decrease with the progress of leaf
decomposition.
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Fungal Diversity
93
Table 1. Presence or absence of all recorded fungi for each leaf type (+ and - are presence and
absence, respectively).
Fungi A-type L-type OL-type F-type
Zygomycetes
Backusella circina - - + +
Gongronella butleri - - + +
Mortierella alpina - - + -
Mortierella sp. - - - +
Mucor hiemalis - - + +
Mucor sp. - - + -
Umbelopsis isabellina - - - +
Umbelopsis ramanniana - - - +
Anamorphic Fungi
Acremonium sp.1◆ - + + +
Acremonium sp.2 - - + -
Acremonium sp.3 - + - -
Alternaria alternata★ + + + +
Anungitea fragilis - - + -
Arachnophora sp. - - - +
Arthrinium phaeospermum▲ + - - -
Apiospora montagnei + + - -
Aureobasidium pullulans★ + + + +
Beltrania rhombica◆ - + + +
Beltraniella portoricensis◆ - + + +
Blastophorum truncatum - - - +
Camposporium japonicum◆ - + + +
Camposporium sp. - - + +
Centrospora gracilis - - - +
Chaetopsina fulva◆ - + + +
Chaetospermum camelliae - + - -
Chloridium sp. - - + -
Cladosporium cladosporioides★ + + + +
Cladosporium herbarum◆ - + + +
Cladosporium tenuissimum◆ - + + +
Clonostachys compatiuscula - + + -
Clonostachys rosea◆ - + + +
Colletotrichum gloeosporioides + + + -
Cylindrocladium parvum◆ - + + +
Dactylaria sp.1 + + + -
Dactylaria sp.2 - + - -
Dactylaria spp.◆ - + + +
Dictyochaeta simplex◆ - + + +
Diplocladiella scalaroides - - - +
Discosia artocreas◆ - + + +
Epicoccum nigrum + + - -
Fusarium spp.◆ - + + +
Geotrichum sp. - - + -
▲Collected from A-type leaves only. ★Collected from all leaf types. ◆Collected from all fallen leaf types.

94
Table 1 (continued). Presence or absence of all recorded fungi for each leaf type (+ and - are
presence and absence, respectively).
Fungi A-type L-type OL-type F-type
Gliocladium viride - + - -
Gonytrichum macrocladum - - + +
Hyphomycete sp.1 + + + -
Hyphomycete sp.2 - - + +
Idriella sp. - + + -
Leptographium sp. - + - -
Mariannaea elegans - - + +
Monacrosporium sp. - - + +
Parasympodiella longispora - - - +
Penicillifer superimpositus - - - +
Penicillium spp.★ + + + +
Pestalotiopsis sp.★ + + + +
Phoma sp.★ + + + +
Ramichloridium sp.1 - + - -
Ramichloridium sp.2 + + - -
Rhinocladiella intermedia◆ - + + +
Scolecobasidium cateniphorum◆ - + + +
Scolecobasidium fusiforme - + - +
Scolecobasidium humicola - + - -
Scolecobasidium sp. - - + -
Solosympodiella clavata◆ - + + +
Solosympodiella sp. - - + -
Sporidesmium sp. - - + +
Stenella sp.▲ + - - -
Subramaniomyces fusisaprophyticus◆ - + + +
Trichoconis sp.◆ - + + +
Trichoderma aureoviride◆ - + + +
Trichoderma hamatum◆ - + + +
Trichoderma harzianum◆ - + + +
Trichoderma koningii◆ - + + +
Trichoderma viride - - + +
Trichoderma spp.★ + + + +
Tripospermum acerinum - + - -
Tripospermum myrti - + - -
Tripospermum prolongatum + + - -
Tritirachium bulbophorum▲ + - - -
Tubakia sp. + + - -
Ulocladium sp. - - - +
Volutella sp.1 - + + -
Volutella sp.2 - + - -
Volutella sp.3 - + - -
▲Collected from A-type leaves only. ★Collected from all leaf types. ◆Collected from all fallen leaf types.
In 22 species recorded from all fallen
leaf types (♦ in Table 3), Subramaniomyces
fusisaprophyticus, Chaetopsina fulva and
Beltraniella portoricensis appeared with high
average frequencies on L-type leaves and their
frequencies fell on more decayed leaves. The
average frequencies of Rhinocladiella interme-
dia and Acremonium sp.1 increased in OL-type
and F-type leaves, and those of Trichoderma
harzianum and Fusarium spp. increased in F-
type leaves.
Seasonal change of the fungal species
component of each leaf type
To understand the seasonal change of
fungal species composition on individual leaf

Fungal Diversity
95
types, the frequency values of frequent and
constant species were compared between the
investigated seasons.
Table 2. Sorensen's similarity indexes
calculated for fungal species components
between each leaf type.
L-type OL-type F-type
A-type 0.423 0.278 0.203
L-type - 0.667 0.588
OL-type - - 0.757
A-type
Seasonal fluctuation of frequency values
of fungi occurring on A-type leaves is shown in
Table 4. In this leaf type, all frequent species
were also constant species. Among these
species, Tubakia sp., Colletotrichum gloeo-
sporioides and Stenella sp. appeared at high
frequencies of 50% or more in all seasons.
Tripospermum prolongatum was recorded at
high frequency in April (60%), while the
frequency was low in other seasons.
L-type
Seasonal fluctuation of frequencies of
fungi occurring on L-type leaves is shown in
Table 5. In this stage, 9 species were
recognized as constant. In these fungi,
Subramaniomyces fusisaprophyticus was re-
corded at a high frequency of 50% or more in
all seasons other than winter (January).
Rhinocladiella intermedia was recorded at high
frequencies of 50% and 80% in August and
October, respectively, while the frequency of
these fungi was low in April and January. The
frequency of Phoma sp. was high in April
(55%) and lower in other seasons.
Aureobasidium pullulans was recorded at a
high frequency (55%) in August, but fell in
other seasons. The other constant species,
Dictyochaeta simplex, Penicillium spp.,
Discosia artocreas, Beltrania rhombica and
Cladosporium cladosporioides were not
recorded at such high frequency (lower than
50%).
Besides constant species, Chaetopsina
fulva was recorded at high frequencies in
August (70%) and October (65%) but fell
dramatically in January (5%) and disappeared
in April. Tubakia sp. was recorded at a high
frequency (60%) in January, and Beltraniella
portoricensis occurred at a rather high
frequency of 55% in October. Chaetospermum
camelliae appeared only in August at a
frequency of 55%.
OL-type
Seasonal fluctuation of the frequencies of fungi
collected from OL-type leaves is shown in
Table 6. In this stage, 7 species were recorded
as constant. Rhinocladiella intermedia
occurred at a high frequency of 55% or more in
all seasons. Subramaniomyces fusisaprophy-
ticus was frequently recorded in August (60%)
but fell in other seasons. Acremonium sp.1 was
frequently recorded in October (55%). The
frequencies of other constant species, Chaetop-
sina fulva, Phoma sp., Discosia artocreas and
Beltrania rhombica, were comparatively low in
all seasons.
Excluding constant species, Trichoderma
harzianum was recorded frequently in August
(55%) and January (75%). Fusarium spp.
occurred at a high frequency in January (65%),
and Trichoderma koningii frequently appeared
in October (80%).
F-type
Seasonal fluctuation of frequencies of fungi
collected from F-type leaves is shown in Table
7. In this decomposition stage, 10 species were
recorded as constant. In these species,
Rhinocladiella intermedia was recorded at a
high frequency of 60% or more in all seasons.
Trichoderma harzianum was occurred
frequently in April (85%) and January (75%),
and Fusarium spp. was recorded at 55% in
April, October and January. Subramani-omyces
fusisaprophyticus and Trichoderma koningii
appeared at a high frequency of 55% in
August. Alternaria alternata and, Acremo-nium
sp.1, which is not a constant species, was
recorded at 50% in April.
Discussion
Falling leaf seasons and decomposition
processes of fallen leaves
Although the leaf fall of Quercus

96
Table 3. Average frequencies of occurrence of fungi for each leaf type (- indicates absence).
Fungi A-type L-type OL-type F-type
Stenella sp.▲ 84 - - -
Arthrinium phaeospermum▲ 4 - - -
Tritirachium bulbophorum▲ 1 - - -
Tubakia sp. 89 31 - -
Tripospermum prolongatum 30 10 - -
Ramichloridium sp.2 3 3 - -
Apiospora montagnei 1 1 - -
Epicoccum nigrum 1 1 - -
Colletotrichum gloeosporioides 84 13 3 -
Dactylaria sp.1 4 4 1 -
Hyphomycete sp.1 1 8 1 -
Phoma sp.★ 13 33 26 20
Pestalotiopsis sp.★ 10 8 5 5
Alternaria alternata★ 6 10 8 19
Penicillium spp.★ 4 19 14 24
Trichoderma spp.★ 4 18 28 9
Cladosporium cladosporioides★ 3 14 6 3
Aureobasidium pullulans★ 1 26 16 8
Chaetospermum camelliae - 14 - -
Dactylaria sp.2 - 3 - -
Leptographium sp. - 3 - -
Scolecobasidium humicola - 3 - -
Acremonium sp.3 - 1 - -
Gliocladium viride - 1 - -
Ramichloridium sp.1 - 1 - -
Tripospermum acerinum - 1 - -
Tripospermum myrti - 1 - -
Volutella sp.2 - 1 - -
Volutella sp.3 - 1 - -
Clonostachys compatiuscula - 4 1 -
Idriella sp. - 4 8 -
Volutella sp.1 - 3 1 -
Subramaniomyces fusisaprophyticus◆ - 48 41 40
Rhinocladiella intermedia◆ - 44 74 70
Chaetopsina fulva◆ - 35 30 5
Beltraniella portoricensis◆ - 24 4 1
Dictyochaeta simplex◆ - 21 6 8
Discosia artocreas◆ - 16 24 5
Beltrania rhombica◆ - 15 8 1
Trichoderma harzianum◆ - 14 39 56
Dactylaria spp.◆ - 11 21 20
Trichoderma koningii◆ - 9 29 21
Fusarium spp.◆ - 6 29 50
Acremonium sp.1◆ - 5 30 21
Trichoderma aureoviride◆ - 5 6 5
Cladosporium herbarum◆ - 4 5 8
▲Collected from A-type leaves only. ★Collected from all leaf types. ◆Collected from all fallen leaf types.

Fungal Diversity
97
Table 3 (continued). Average frequencies of occurrence of fungi for each leaf type (- indicates
absence).
Fungi A-type L-type OL-type F-type
Clonostachys rosea◆ - 4 6 19
Cylindrocladium parvum◆ - 4 1 5
Trichoderma hamatum◆ - 3 11 11
Camposporium japonicum◆ - 1 4 13
Cladosporium tenuissimum◆ - 1 10 4
Scolecobasidium cateniphorum◆ - 1 8 4
Solosympodiella clavata◆ - 1 4 9
Trichoconis sp.◆ - 1 11 5
Scolecobasidium fusiforme - 1 - 5
Geotrichum sp. - - 6 -
Scolecobasidium sp. - - 4 -
Acremonium sp.2 - - 1 -
Anungitea fragilis - - 1 -
Chloridium sp. - - 1 -
Mortierella alpina - - 1 -
Mucor sp. - - 1 -
Solosympodiella sp. - - 1 -
Trichoderma viride - - 8 18
Backusella circina - - 5 6
Camposporium sp. - - 4 3
Hyphomycete sp.2 - - 4 3
Gongronella butleri - - 3 3
Mariannaea elegans - - 3 1
Mucor hiemalis - - 3 8
Gonytrichum macrocladum - - 1 3
Monaclosporium sp. - - 1 9
Sporidesmium sp. - - 1 5
Arachnophora sp. - - - 6
Blastophorum truncatum - - - 5
Centrospora gracilis - - - 3
Diplocladiella scalaroides - - - 3
Parasympodiella longispora - - - 3
Umbelopsis ramanniana - - - 3
Mortierella sp. - - - 1
Penicillifer superimpositus - - - 1
Ulocladium sp. - - - 1
Umbelopsis isabellina - - - 1
◆Collected from all fallen leaf types.
myrsinaefolia was observed throughout the
year, fallen leaves were abundant from June
to August (Fig. 2). In Fig. 2, the main fallen
leaf season of this evergreen oak is probably
around June in the forest, because the dry
weight of L-type leaves in this period was
heaviest. This result almost agrees with other
studies on litter production of Q.
myrsinaefolia (Nagao and Harada, 1996;
Kuramoto, 1997).
The leaves in the maximum falling leaf
period appeared to decay at almost the same
speed. The decay stage of these leaves at a
certain sampling time can be assessed from
the fluctuation of the dry weight of more
decomposed leaf types; therefore, we estimate
that L-type leaves that fell in June
decomposed to the OL-type stage in August,
the F-type in October, and the F-type or more
decomposed to the OL-type stage in August,
the F-type in October, and the F-type or more
decomposed stages in December (Fig. 2). L-
type leaves that fell in August, a later leaf-fall
season, might decompose to the OL-type

98
Table 4. Seasonal changes in frequencies of occurrence, average frequencies and seasonal indexes
of fungi occurring from A-type leaves (- indicates absence).
Fungi Apr Aug Oct Jan Average
Seasonal
index
Tubakia sp. 100 100 100 55 89 100
Colletotrichum gloeosporioides 85 95 95 60 84 100
Stenella sp. 80 95 90 70 84 100
Tripospermum prolongatum 60 30 10 20 30 100
Phoma sp. 10 25 - 15 13 75
Pestalotiopsis sp. - 30 5 5 10 75
Penicillium sp. - 5 5 5 4 75
Alternaria alternata 15 10 - - 6 50
Dactylaria sp.1 5 - - 10 4 50
Cladosporium cladosporioides 5 - - 5 3 50
Ramichloridium sp.2 - - 5 5 3 50
Trichoderma sp. 15 - - - 4 25
Arthrinium phaeospermum - - 15 - 4 25
Apiospora montagnei - 5 - - 1 25
Aureobasidium pullulans - 5 - - 1 25
Epicoccum nigrum - - - 5 1 25
Hyphomycete sp.1 - - - 5 1 25
Tritirachium bulbophorum - - - 5 1 25
stage in October and the F-type in December
(Fig. 2).
Chemical components of each leaf type
The C/N ratio, polyphenol and soluble
sugar concentration in the fallen leaves
decreased with progress of leaf decomposition
(Figs 3, 4). This change of chemical
components is generally seen during the
decomposition process of fallen leaves (Berg
and McClaugherty, 2003). These results
therefore support the validity of the leaf
decomposition stages classified in this study.
Successional patterns of fungal species with
the decay of leaves
Comparison of typical fungi characterized by
each leaf type
In earlier studies, fungal succession with
the decay of fallen leaves has been presumed
by comparing the species components of
typical fungi characterizing each decompo-
sition stage (Kendrick and Burges, 1962; Aoki
et al., 1990, 1992; Heredia, 1993; Tokumasu et
al., 1994; Tokumasu, 1996). In this study,
frequent and constant species were selected as
typical fungi in each leaf type and fungal
successsion with the decay of leaves was
estimated by comparing these fungi.
The list of frequent and constant species
is shown in Table 8. The frequent species in A-
type leaves, Stenella sp., Tubakia sp.,
Tripospermum prolongatum and Colletotri-
chum gloeosporioides, were recorded constant-
tly from only A-type leaves (Table 8). In
particular, Stenella sp. did not occur on other
type leaves (Table 1). On the other hand, two
frequent and constant species in all fallen
leaves types, Rhinocladiella intermedia and
Subramaniomyces fusisaprophyticus, frequent
species in the L-type such as Beltraniella
portoricensis, Chaetopsina fulva and Chaeto-
spermum camelliae, and frequent species in the
OL-type and F-type, such as Trichoderma
harzianum and T. koningii, were not recorded
from A-type leaves (Table 1, 8). These results
suggested the conspicuous difference of fungal
species components between living leaves on
the tree and fallen leaves on the ground.
Indexes of similarity according to Sørensen
also showed that the species composition of A-
type leaves was characteristic compared with
leaf types on the ground (Table 2). In contrast,

Fungal Diversity
99
Table 5. Seasonal changes in frequencies of occurrence, average frequencies and seasonal indexes
of fungi occurring from L-type leaves (- indicates absence).
Fungi Apr Aug Oct Jan Average Seasonal
index
Subramaniomyces fusisaprophyticus 50 60 50 30 48 100
Rhinocladiella intermedia 25 50 80 20 44 100
Phoma sp. 55 30 25 20 33 100
Aureobasidium pullulans 15 55 20 15 26 100
Dictyochaeta simplex 20 35 10 20 21 100
Penicillium spp. 5 25 25 20 19 100
Discosia artocreas 40 5 10 10 16 100
Beltrania rhombica 5 20 25 10 15 100
Cladosporium cladosporioides 10 10 25 10 14 100
Chaetopsina fulva - 70 65 5 35 75
Tubakia sp. 35 30 - 60 31 75
Beltraniella portoricensis - 30 55 10 24 75
Dactylaria spp. - 30 10 5 11 75
Alternaria alternata 20 10 10 - 10 75
Fusarium spp. 5 15 5 - 6 75
Trichoderma spp. - - 35 35 18 50
Trichoderma harzianum 25 - 30 - 14 50
Colletotrichum gloeosporioides - 30 - 20 13 50
Tripospermum prolongatum 5 - - 35 10 50
Pestalotiopsis sp. - - 20 10 8 50
Trichoderma aureoviride 5 15 - - 5 50
Cladosporium herbarum 5 - 10 - 4 50
Cylindrocladium parvum - - 5 10 4 50
Scolecobasidium humicola - 5 5 - 3 50
Chaetospermum camelliae - 55 - - 14 25
Hyphomycete sp.1 - - - 30 8 25
Trichoderma koningii - - 35 - 9 25
Acremonium sp.1 - - 20 - 5 25
Dactylaria sp.1 15 - - - 4 25
Clonostachys rosea - 15 - - 4 25
Idriella sp. - 15 - - 4 25
Clonostachys compatiuscula - - 15 - 4 25
Volutella sp.1 - 10 - - 3 25
Leptographium sp. - - 10 - 3 25
Dactylaria sp.2 - - - 10 3 25
Ramichloridium sp.2 - - - 10 3 25
Trichoderma hamatum - - 10 - 3 25
Acremonium sp.3 5 - - - 1 25
Apiospora montagnei 5 - - - 1 25
Camposporium japonicum 5 - - - 1 25
Ramichloridium sp.1 5 - - - 1 25
Scolecobasidium fusiforme - 5 - - 1 25
Solosympodiella clavata - 5 - - 1 25
Volutella sp.2 - 5 - - 1 25
Volutella sp.3 - 5 - - 1 25
Cladosporium tenuissimum - - 5 - 1 25
Epicoccum nigrum - - 5 - 1 25
Scolecobasidium cateniphorum - - 5 - 1 25
Trichoconis sp. - - - 5 1 25
Tripospermum acerinum - - - 5 1 25
Tripospermum myrti - - - 5 1 25
Gliocladium viride - - - 5 1 25

100
they indicated that species composition was
similar among leaf types found on the ground.
Moreover, the species composition of a certain
decomposition stage was most similar to that of
the next decomposition stage. All frequent
species found on the ground, except Chaeto-
spermum camelliae, were recorded from all
fallen leaf types (Tables 1, 8). For example,
Subramaniomyces fusisaprophyticus and Rhi-
nocladiella intermedia appeared as frequent
and constant species from all fallen leaf types
(Table 8). These results suggest no remarkable
differences among fallen leaf types in the
composition of frequent and constant species;
however, some fungi showed a tendency to
occur with a rather higher frequency value in
early decomposition stages such as Aureobasi-
dium pullulans, Phoma sp., Chaetospermum
camelliae, Chaetopsina fulva and Beltraniella
portoricensis, and some showed an opposite
tendency, viz Acremonium sp.1, Fusarium spp.,
Trichoderma harzianum and T. koningii (Table
8), suggesting the species alternation of fungi
on fallen leaves of this oak. An outline of
fungal succession associated with the decay of
Quercus myrsinaefolia leaves was estimated as
follows. Fungal colonizing leaves change from
living leaf inhabitants, such as Stenella sp.,
Tubakia sp., Tripospermum prolongatum and
Colletotrichum gloeosporioides, through early
fallen leaf colonizers, such as Chaetospermum
camelliae, Chaetopsina fulva and Phoma sp.,
to later colonizers, such as Fusarium spp. and
Trichoderma spp.
Ecological features of frequent species
In four frequent species colonizing living
leaves, Stenella sp. was a frequent and constant
species on A-type leaves, and was not recorded
from leaves on the ground (Tables 1, 8).
Tubakia sp. has been isolated from living and
fallen leaves of Quercus and Castanopsis (Yo-
koyama and Tsubaki, 1971; as Actinopelte).
In this study, it also occurred on living
and fallen leaves at an early decomposition
stage (Table 1). Tripospermum prolongatum is
known as a phylloplane fungus often seen on
leaves infected with sooty mold, and Col-
letotrichum gloeosporioides is known as a
multiple-host pathogen of various plants (Sato,
1996). These four species are probably phyllo-
sphere fungi invading green leaves on the tree.
According to research on Pinus densiflora
(Tokumasu, 1996), Abies firma (Aoki et al.,
1990) and Fagus crenata (Osono, 2002),
Phoma spp. has been mainly reported from
living and fallen leaves at early decomposition
stages. Our results showed the same tendency
(Tables 3, 8). Phoma sp. might be a phyllo-
sphere fungus colonizing green leaves on the
tree. Aureobasidium pullulans is known as a
common primary saprophytes recorded from
senescent leaves on trees and fallen leaves at an
early decomposition stage (Hudson, 1968). In
this study, A. pullulans also appeared from
living and L-type leaves at high frequency and
also declined with the progression of leaf decay
(Tables 3, 8).
The five species mentioned below have
ecological features as they quickly infect new
fallen leaves. Beltraniella portoricensis in-
fected newly fallen leaves and its frequency of
occurrence decreased with decay (Table 3).
According to Aoki et al. (1990) and Tokumasu
(1996, 1998a), Chaetopsina fulva is a fungus
that quickly infected freshly fallen leaves of
Pinus densiflora and Abies firma in summer
and autumn, and its frequency of occurrence
declined sharply with leaf decomposition. Our
results also showed that this fungus is an early
fallen leaf colonizer in summer and autumn
(Tables 3, 5). Chaetospermum camelliae
showed a distribution pattern similar to Chae-
topsina fulva, but occurred only in summer
(Tables 3, 5). Subramaniomyces fusisaprophy-
ticus was mainly found on fallen leaves in the
early decomposition stage at high frequency,
and then declined slowly (Table 3). Rhino-
cladiella intermedia quickly colonized freshly
fallen leaves and the frequency of occurrence
increased gradually as leaf decay progressed
(Table 3).
Acremonium sp. 1 first occurred on
freshly fallen leaves and the frequency
increased in OL-type leaves (Table 3). Tricho-
derma spp. has been recorded frequently on
well-decomposed leaves in other studies
(Kendrick and Burges, 1962; Domsch et al.,
1980a, b; Aoki et al., 1990; Tokumasu, 1996;
Osono, 2002; Sadaka and Ponge, 2003). In this
study, T. harzianum and T. koningii also
occurred frequently and became constant
species on F-type leaves (Table 8). The

Fungal Diversity
101
Table 6. Seasonal changes in frequencies of occurrence, average frequencies and seasonal indexes
of fungi occurring from OL-type leaves (- indicates absence).
Fungi Apr Aug Oct Jan Average Seasonal
index
Rhinocladiella intermedia 55 75 100 65 74 100
Subramaniomyces fusisaprophyticus 30 60 30 45 41 100
Acremonium sp.1 5 20 55 40 30 100
Chaetopsina fulva 35 15 25 45 30 100
Phoma sp. 40 10 25 30 26 100
Discosia artocreas 20 40 20 15 24 100
Beltrania rhombica 5 5 15 5 8 100
Trichoderma harzianum - 55 25 75 39 75
Fusarium spp. 35 - 15 65 29 75
Trichoderma koningii - 25 80 10 29 75
Trichoderma spp. 45 - 30 35 28 75
Dactylaria spp. 15 45 - 25 21 75
Penicillium spp. - 15 20 20 14 75
Trichoconis sp. 10 15 20 - 11 75
Scolecobasidium cateniphorum 10 10 - 10 8 75
Cladosporium cladosporioides 5 - 15 5 6 75
Dictyochaeta simplex 10 10 - 5 6 75
Aureobasidium pullulans 25 - - 40 16 50
Trichoderma hamatum - 25 20 - 11 50
Cladosporium tenuissimum 15 - - 25 10 50
Alternaria alternata 15 - - 15 8 50
Trichoderma viride - 25 5 - 8 50
Idriella sp. - 10 - 20 8 50
Clonostachys rosea - 5 - 20 6 50
Pestalotiopsis sp. 10 - 10 - 5 50
Backusella circina - - 15 5 5 50
Camposporium japonicum 10 - 5 - 4 50
Camposporium sp. 5 10 - - 4 50
Hyphomycete sp.2 5 10 - - 4 50
Scolecobasidium sp. - 5 - 10 4 50
Solosympodiella clavata - 5 - 10 4 50
Colletotrichum gloeosporioides 5 - - 5 3 50
Trichoderma aureoviride 25 - - - 6 25
Geotrichum sp. - - 25 - 6 25
Cladosporium herbarum 20 - - - 5 25
Beltraniella portoricensis - - - 15 4 25
Gongronella butleri - 10 - - 3 25
Mariannaea elegans - 10 - - 3 25
Mucor hiemalis - - - 10 3 25
Acremonium sp.2 - 5 - - 1 25
Anungitea fragilis 5 - - - 1 25
Chloridium sp. 5 - - - 1 25
Clonostachys compatiuscula 5 - - - 1 25
Dactylaria sp.1 5 - - - 1 25

102
Table 6 (continued). Seasonal changes in frequencies of occurrence, average frequencies and
seasonal indexes of fungi occurring from OL-type leaves (- indicates absence).
Fungi Apr Aug Oct Jan Average Seasonal
index
Hyphomycete sp.1 5 - - - 1 25
Solosympodiella sp. 5 - - - 1 25
Gonytrichum macrocladum - 5 - - 1 25
Sporidesmium sp. - 5 - - 1 25
Volutella sp.1 - 5 - - 1 25
Cylindrocladium parvum - - - 5 1 25
Monacrosporium sp. - - - 5 1 25
Mortierella alpina - - - 5 1 25
Mucor sp. - - - 5 1 25
frequencies of these Trichoderma species
fluctuated every season (Tables 5, 6, 7).
Because these fungi have rapid growth rates
and various temperature preferences (Widden
and Hsu, 1987), their growth rate and activity
are influenced by temperature and humidity
just before sampling time.
Alternaria alternata is a common
primary saprophyte invading senescent leaves
on trees or fallen leaves at early decomposition
stages (Hudson, 1968); however, in our results,
the frequency of this fungus increased in F-
type leaves. Namely, the tendency of
appearance was different from other studies
(Table 3). The appearance directionality of this
fungus was not clear (Table 3), so it is not easy
to interpret the ecological features only from
the data obtained in this study.
Estimation of fungal succession on Quercus
myrsinaefolia leaves
The fungal succession illustrated in Fig.
7 was estimated from comparing the
appearance and disappearance patterns of
frequently occurring fungi with leaf decay. On
leaves with phyllosphere fungi on trees, fungi
such as Tubakia sp. and Colletotrichum gloeo-
sporioides change to early colonizers of fallen
leaves on the ground, such as Subrama-
niomyces fusisaprophyticus and Rhinocladiella
intermedia, and then progress to later
colonizers, such as Trichoderma koningii and
T. harzianum.
Tokumasu (1998a,b) reported the
seasonal change of fungi on decaying pine
needles. Similarly, on fallen leaves of Quercus
myrsinaefolia, Chaetospermum cameliae and
Chaetopsina fulva are involved in fungal
succession only in limited seasons of summer
and autumn (Table 5, Fig. 7). The seasonal
fluctuation of chemical components in fallen
leaves, for example, increased polyphenol and
soluble sugar in December, might be a factor in
the seasonal alternation of members of the
fungal community (see Results).
Furthermore, in this research, we tried to
estimate fungal succession by tracing frequent
species on leaves fallen in the main leaf-fall
seasons throughout a whole year. Leaves of
Quercus myrsinaefolia fell in August, later in
the main leaf-fall season, were decomposed to
the OL-type stage in October and reached the
F-type stage in January (Fig. 2). The frequent
species on each leaf type in each season are
shown in Table 9. Fungal succession associated
with the decomposition of fallen leaves in this
period characterizes the whole succession
pattern at this study site (Fig. 7, Table 9).
Characteristics of the fungal community
inhabiting Quercus myrsinaefolia leaves
Although fungal succession accompa-
nying the decay of Q. myrsinaefolia leaves was
estimated as above, it is difficult to compare
fungal succession directly with that on other
tree leaves, especially conifers, firstly because
differences in the structure of the O horizon
may prevent a comparison. Although the leaf-
fall of Pinus densiflora is seen throughout the
year, the decomposition rate of P. densiflora
leaf litter is usually very slow on mor or moder
sites where a thick O horizon is formed and the
horizon divides into sub-layers corresponding
to different decomposition stages. Species

Fungal Diversity
103
Table 7. Seasonal changes in frequencies of occurrence, average frequencies and seasonal indexes
of fungi occurring from F-type leaves (- indicates absence).
Fungi Apr Aug Oct Jan Average
Seasonal
index
Rhinocladiella intermedia 85 65 70 60 70 100
Trichoderma harzianum 85 45 20 75 56 100
Fusarium spp. 55 35 55 55 50 100
Subramaniomyces fusisaprophyticus 40 55 45 20 40 100
Penicillium spp. 5 30 45 15 24 100
Trichoderma koningii 5 55 15 10 21 100
Dactylaria spp. 45 15 15 5 20 100
Alternaria alternata 50 10 10 5 19 100
Clonostachys rosea 5 30 25 15 19 100
Camposporium japonicum 15 20 10 5 13 100
Acremonium sp.1 50 - 30 5 21 75
Phoma sp. 45 - 25 10 20 75
Trichoderma hamatum 5 10 30 - 11 75
Solosympodiella clavata 20 - 5 10 9 75
Dictyochaeta simplex 5 - 15 10 8 75
Arachnophora sp. 10 10 5 - 6 75
Backusella circina 5 5 15 - 6 75
Chaetopsina fulva 10 5 5 - 5 75
Discosia artocreas 5 - 10 5 5 75
Trichoconis sp. 5 - 5 10 5 75
Trichoderma viride - 30 40 - 18 50
Monaclosporium sp. - - 20 15 9 50
Cladosporium herbarum 10 - 20 - 8 50
Aureobasidium pullulans 5 - - 25 8 50
Mucor hiemalis - 25 - 5 8 50
Pestalotiopsis sp. 5 - 15 - 5 50
Scolecobasidium fusiforme 15 - 5 - 5 50
Sporidesmium sp. - 5 15 - 5 50
Cylindrocladium parvum - 5 - 15 5 50
Cladosporium tenuissimum 10 - - 5 4 50
Scolecobasidium cateniphorum 10 - - 5 4 50
Gongronella butleri 5 5 - - 3 50
Diplocladiella scalaroides 5 - - 5 3 50
Centrospora gracilis - 5 5 - 3 50
Hyphomycete sp.2 - 5 5 - 3 50
Trichoderma spp. - - 35 - 9 25
Blastophorum truncatum - - 20 - 5 25
Trichoderma aureoviride - - - 20 5 25
Camposporium sp. - - 10 - 3 25
Cladosporium cladosporioides 10
- - - 3 25
Umbelopsis ramanniana - 10 - - 3 25
Gonytrichum mactocladum - - 10 - 3 25
Parasympodiella longispora - - - 10 3 25
Beltraniella portoricensis 5 - - - 1 25
Ulocladium sp. 5 - - - 1 25
Umbelopsis isabellina 5 - - - 1 25
Beltrania rhombica - 5 - - 1 25
Mariannaea elegans - 5 - - 1 25
Penicillifer superimpositus - - 5 - 1 25
Mortierella sp. - - - 5 1 25

104
components on individual needles are highly
homogeneous in each layer and clearly
different from other layers (Tokumasu, 1980);
therefore, we were able to estimate fungal
succession from the differences of species
composition among sub-layers. However, the
O horizon under the Quercus myrsinaefolia
forest was thin because of the fast progress of
leaf decomposition. As a result, the
development of sub-layers corresponding to
each decomposition stage was very poor and
distinction of the decay stages of individual
leaves was often very difficult. As another
reason, because leaves of Quercus have a large
area compared with conifer needles, one
Quercus leaf recognized as one decomposition
stage may be composed of various parts with
miscellaneous decay conditions.
As a result of comparing with earlier
studies using other tree species, there are some
Table 8. Frequent and constant species for each leaf type.
A-type
Colletotrichum gloeosporioides
Stenella sp.
Tripospermum prolongatum
Tubakia sp.
L-type
Aureobasidium pullulans Beltraniella portoricensis◆ Beltrania rhombica▲
Phoma sp. Chaetospermum camelliae◆Cladosporium cladosporioides▲
Rhinocladiella intermedia Chaetopsina fulva◆ Dictyochaeta simplex▲
Subramaniomyces fusisaprophyticus Tubakia sp.◆ Discosia artocreas▲
Penicillium spp.▲
OL-type
Acremonium sp.1 Fusarium spp.◆ Beltrania rhombica▲
Rhinocladiella intermedia Trichoderma harzianum◆ Chaetopsina fulva▲
Subramaniomyces fusisaprophyticus Trichoderma koningii◆ Discosia artocreas▲
Phoma sp.▲
F-type
Alternaria alternata Acremonium sp.1◆ Camposporium japonicum▲
Fusarium spp. Clonostachys rosea▲
Rhinocladiella intermedia Dactylaria spp.▲
Subramaniomyces fusisaprophyticus Penicillium spp.▲
Trichoderma harzianum
Trichoderma koningii
bold type are frequent and constant species at the same time. ◆frequent species. ▲constant species.
differences in the member of frequently
recorded fungi. For example, Cladosporium
cladosporioides, which was recorded
frequently from fallen leaves of Pinus
densiflora, Abies firma and Fagus crenata
(Aoki et al., 1990; Tokumasu, 1996; Osono,
2002), occurred only infrequently on Quercus
myrsinaefolia leaves (Tables 5, 6, 7). In
contrast, the following species have not been
recorded frequently from other tree species,
viz Tubakia, Subramaniomyces fusisapro-
phyticus and Rhinocladiella intermedia,
which appeared on Quercus myrsinaefolia
leaves at high frequentcies (Tables 5, 6, 7).
One primary factor of this phenomenon may
be the host specificity or selectivity of
individual species of fungi. Subramaniomyces
fusisaprophyticus has been reported as a
fallen leaf colonizer of broadleaf-evergreen
trees, especially oak trees (Matsushima, 1975;
Kirk, 1982, 1983; Ellis and Ellis, 1985;
Cooper, 2005). On the other hand, a sapro-
trophic pine leaf litter fungus has been
reported, the distribution pattern of which is
strongly affected by climatic factors such as
annual mean air temperature or annual range
(Tokumasu, 2001). The fungal species
component colonizing on decaying Quercus

Fungal Diversity
105
Fig. 7. Appearance and disappearance pattern of frequently occurred fungi with the decay of Q. myrsinaefolia leaves.

106
Table 9. Tracing the succession of frequent fungi with decomposition of fallen leaves in August
(figures after fungal names are frequency of each fungus).
A-type in April L-type in August OL-type in October F-type in January
Tubakia sp. (100) Tubakia sp. (30)
Colletotrichum
gloeosporioides (85) C. gloeosporioides (30)
Stenella sp. (80)
Tripospermum
prolongatum (60)
Chaetopsina fulva (70) C. fulva (25)
Subramaniomyces
fusisaprophyticus (60) S. fusisaprophyticus (30) S. fusisaprophyticus
(20)
Aureobasidium
pullulans (55)
A. pullulans (25)
Chaetospermum
camelliae (55)
Rhinocladiella intermedia (50) R. intermedia (100) R. intermedia (60)
Trichoderma koningii (80) T. koningii (10)
Acremonium sp.1 (55) Acremonium sp.1 (5)
Trichoderma harzianum (25) T. harzianum (75)
Fusarium spp. (15) Fusarium spp. (15) Fusarium spp. (55)
myrsinaefolia leaves is probably decided by
both the host specificity/selectivity of indivi-
dual species of fungi and the climate in the
study area.
Future work
In this study, fungi from leaves were
isolated using traditional methodology which
incorporated media. It is therefore unlikely that
all of the fungi involved in leaf decay would
have been detected (Hyde and Soytong, 2007).
The origin of fungal saprobes on Castanopsis
diversifolia leaf litter has discussed by Duong
et al. (2008) that it may derive from aerially
dispersed spores or ground soil. In addition,
endophytes present within living leaves may
participate in the decomposition process of
fallen leaves (Osono, 2003; Hyde and Soytong,
2008). However, an entire fungal species
component inhabiting decaying leaves has not
yet been established. In future it might be wise
to incorporate molecular techniques (e.g.
DGGE, T-RFLP, construction of clone libra-
ries) to detect more taxa (Duong et al., 2006;
Seena et al., 2008). The fungi occurring on
decaying leaves have also been shown to differ
from those occurring on wood (Kodsueb et al.,
2008a,b; Küffer et al., 2008) and it would be
interesting to establish if this is true for
Japanese broadleaf trees.
Acknowledgements
We wish to thank Meiji Jingu Shrine for kindly
offering the investigation site for this research. We also
thank Dr. Takashi Osono, Graduate School of
Agriculture, Kyoto University, for his support with the
chemical analyses of the fallen leaves. We want to
acknowledge reviewer’s works
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