Bull. Natn. Sci. Mus., Tokyo, Ser. B, 23(1), pp. 25- 34, March 24, 1997

Host Range of Aciculosporium take, the Causal Agentof Witches' Broom of Bamboo Plants
Mitsuya Tsuda\ Kiminori Shimizu\ Keishi Matsumura\
Eiji Tanaka\ Chihiro Tanaka1
and Yoshimichi Doi 2
1
Faculty of Agriculture, Kyoto University,
Kitashirakawa, Sakyo-ku, Kyoto, 60(H)l Japan2
Department of Botany, National Science Museum,4- 1- 1 Amakubo, Tsukuba, Ibaraki, 305 Japan

Abstract The host range of Aciculosporium take, the causal agent of witches'broom of bamboos, was surveyed in Japan, and 17 species, 8 varieties, 9 forms
and 2 horticultural cultivars in 5 genera of bamboo plants were confirmed as hostplants. Of these, 11 species, 6 varieties, 6 forms, and 2 horticultural cultivars in
4 genera were new host plants. Here, in addition to the study of host range, themorphology and characteristics of conidia and conidial germination of thefungus are discussed.
Key words:
new host.

Albomyces take, bamboo taxonomy, conidial morphology, fungi,

Introduction
Bamboo plants with densely ramified young twigs without normal branchlets
and normal leaves have recently been observed in many Madake (Phyllostachys
bambusoides Sieb. et Zucc.) forests in the Kansai area of Japan. This is a typicalsymptom of witches' broom of Madake. The disease is one of the most destructive
causes of the deterioration of bamboo forests and has been well known in Japan
since the Meiji era (Hara, 1907; Tsuboi, 1907; Shinohara, 1965). The disease has
also been reported in China (Zhu and Huang, 1988) and Taiwan (Chen, 1970) on
some Phyllostachys bamboo species. The causal fungus, Aciculosporium takeMiyake (anamorphic stage = Albomyces take Miyake), is considered to be anallied species of Balansia (Hino, 1962) and therefore treated as a member of theclavicipitaceous fungi (Diel, 1950; Kobayashi, 1992). However, few studies on
this fungus and the disease have been reported. Generally, the fungus is believed
to attack only bamboo plants belonging to the genus Phyllostachys (Kobayashi,1992), although bamboo species other than Phyllostachys have been recorded as
host plants in Japan (Hara, 1938). The pathogenicity of the fungus to Shibataea
kumasaca (Zoll.) Nakai (Okame-zasa) has also been proved by artificial inocula-26

Mitsuya Tsuda et al.

tion (Chen, 1970; Shinohara, 1970). Accordingly, the host range of the fungus
has not been fully explored. In this study, we describe the host plants of thefungus on the basis of our survey in several bamboo gardens. Furthermore, weinvestigated some morphological characteristics of the conidia and their germination process of the fungus on artificial culture.
Materials and Methods
The survey of host plants. The study was principally made during 19941996 on bamboo forests around Kyoto-shi and the bamboo sample stands in thefollowing botanical gardens: The Kyoto Botanical Garden, Shimogamo Nakaragi-cho, Sakyo-ku, Kyoto-shi; Rakusai Bamboo Park, Ohe Kitafukunishi-cho,Nishikyo-ku, Kyoto-shi; Kamigamo Experimental Forest, Kyoto University,
Kamigamo Motoyama, Kita-ku, Kyoto-shi; Kansai Research Center, Forestry
and Forest Products Research Institute, Momoyama-cho, Fushimi-ku, Kyoto-shi;
Yoro Bamboo Garden in Yoro-Koen, Yoro-cho, Gifu-ken. Other sampling sites
from which voucher collections were obtained are indicated as necessary.
Determination of the fungus. Because the anamorphic stage of the fungus
was detected throughout the year and the period of the teleomorphic stage islimited to summer, our survey was made on the basis of the anamorphic stage.
When the witches' broom symptom was observed, the presence of white spindlelike stromatic pseudoparenchymatal tissues was first confirmed. Then a portion of
twigs with symptoms was collected and the presence of conidia or other fungal
structures was examined. If conidia were detected, conidial measurements weremade on conidia from conidial masses naturally produced on stromatic tissues by
fixation with lactophenol. Only the total length was determined because the width
was less than 2 f.lm and precise measurement was difficult.
Observation of some cultural characteristics. The shape of conidia produced on artificial culture was compared with that of naturally produced conidia.
Some cultural characteristics were also investigated. In this study, the fungus wascultured on PDA (Difco) and PDB (Difco) media at 25 °C in the dark.
Deposition of voucher collections. All of the sample specimens collected in
the field were deposited at the National Science Museum, Tokyo with TNS
accession numbers TNS F-100,001-100,088. The names of collected bamboo
plants with the fungus and their accession numbers are indicated in Tables 1 and2. Names of the host plants follow Suzuki ( 1978) or the indication by eachbamboo garden.
Results and Discussion
Disease symptoms.

The disease symptoms closely matched those reportedHost Range of Aciculosporium take
Table I.

27

List of voucher bamboo plants with Aciculosporium take.
Collection site*2

Japanese common
nameLatin name*'

Ma-dake
Shibo-chiku
Ginmei-shibo-chiku
Kashiro-dake
Kon-shima-dake
Mutsuore-dakeKinmei-chiku
Ginmei-chiku
Ougon-chiku
Usan-chiku
Hotei-chiku
Shima-hotei-chiku
Ginmei-hachiku
Ougon-hotei
Kuro-chiku
Ha-chiku
Unmon-chiku
Shima-hachiku
Tosa-torafu-dake
Meguro-chiku
Taiwan-ma-dakelnyo-chiku
Hakuho-kanchiku
Hime-hachiku
Okame-zasa
Narihira-dakeAo-narihira
Nikko-narihira
Bizen-narihira
Kenashi-Yasha-dake
Rikuchu-dake
Taimin-chiku
Ne-zasa
Chimaki-zasa
Nambu-suzu

Phyllostachys bambusoides
var. marliacea
cv. ginmeishibochiku*
f. kashirodake
f. subvariegata
f. geniculata
var. castillonis
var. castilloni-inversa
var. holochrysa
cv. usanchiku*
P. aurea
f. albo-variegata
var. jiavescens-inversa
f. holochrysa*
P. nigravar. henonis
f. boryana
f. albo-variegata
var. tosaensis
f. meguorchiku
P. makinoi*
P. tranquillans
P. dulcis*
P. humiris*
Shibataea kumasaca
Semiarundinaria fastuosavar. viridis
S. yoshi-matsumurae
S. okuboi
S. tatebeana
S. kagamiana
Pleioblastus gramineus
Pleioblastus Sect. Nezasa sp.
Sasa Sect. Sasa sp.
Sasa Sect. Lasioderma sp.

A

B

c

D

E

F

010
011

sym
031

040

001

055
056
057
058

071*a

012

002

041
042

013
014
015

003
004
sym
no
no

no
no
no
no
005
006
no
007
008

009
no
no
no

016
017
018
no
no
no
019
020
no
021
022
023
no
024
025
026
027
028
029
030
no
no
no
no

032

033
034
035
036
no
sym
no

037
038

no
no
039

sym
043
sym
044
no
no
045
no
no
no
046
047
048
no
049
050
051
052
053
054
no
no
no
no

059
060
061
062
sym
063
064
no
no
no

072*b

073*a

065
066
no
067
no
no
068
069
no
070
no
no
no
no
no

074*c
075*d

*' The bamboo names are after Suzuki (1978): Index to Japanese Bambusaceae. The nameswith (*) are followed by the indication of each bamboo garden.
*2 Collection sites: A, Kansai Research Center, Forestry & Forest Products Research Institute;B, The Kyoto Botanical Garden; C, Yoro Bamboo Garden; D, Kamigamo Experimental
Forest, Kyoto University; E, Rakusai Bamboo Park; F, Others (*a, Takeda Pharmaceutical
Plant Garden at Kyoto-shi; *b, The Fukiage Gardens in the Imperial Palace Grounds;*c, Kohata, Uji-shi, Kyoto-fu, same as 079 in the Table 2; *d, Kyoto University Forest in
Ashiu) .
- , Not planted; no, Not confirmed; sym, Symptom was confirmed.

28

Mitsuya Tsuda et al.
Table 2. List of vouchers of P. bambusoides with Aciculosporium take other than those in
Table 1.
076; Isonokami, Tenri-shi, Nara-ken
077; Isonokami, Tenri-shi, Nara-ken
078; Sumiyama, Uji-shi, Kyoto-fu
079; Kohata Kanakusahara, Uji-shi, Kyoto-fu
080; Kohata Kanakusahara, Uji-shi, Kyoto-fu
081; Shirakawa, Uji-shi, Kyoto-fu
082; Shirakawa, Uji-shi, Kyoto-fu

083; Daigo, Fushimi-ku, Kyoto-fu
084; Rokujizo, Uji-shi, Kyoto-fu
085; Bicchu-Matsuyama, Okayama-ken
086; Shizu, Sakura-shi, Chiba-ken
087 : Sakura, Sakura-shi, Chiba-ken
088; Amakubo, Tsukuba, lbaraki-ken

in the literature (Tsuboi, 1907; Miyake, 1908; Hara, 1938; Hino and Katsumoto,
1961; Shinohara, 1965). Namely, the bamboo twigs attacked by the fungus wererepeatedly ramified with deformations. Plants often drooped from the canopy
and gave a dirty brownish impression due to poor leaf growth. When the fungus
attacked ornamental bamboo plants, the beauty of the bamboo stand was spoiled
by the drooping of brownish-parts diseased by witches' broom. This spoilage was
widely distributed in bamboo culms at the margins of Madake forests near Kyoto
and other areas of the Japanese archipelago. The diseased parts graduallydefoliated and the whole plant died, probably because loss of leaves led todecreased photosynthesis. Production of bamboo culms and bamboo sprout arealso decreased (Tsuboi, 1907). New culms with elongated or shortened nodes andwith irregularly proliferated branches occasionally emerged from aerial parts ofunderground rhizomes. These symptoms were recognized on Semiarundinaria
fastuosa (Mitf.) Makino (Narihira-dake) (Fig. 3), Pleioblastus gramineus (Bean)
Nakai (Taimin-chiku) (Fig. 1), and Sasa Sect. Lasioderma sp. (Nambu-suzu)(Fig. 4 ), respectively.
Host range of the fungus. Conidial sizes were similar among the different
host bamboo plants examined, falling within the range of variation of thespecimens on P. bambusoides collected from stands in different localities (Tables
3, 4). This indicates that the causal agent of witches' broom of these bamboo
plants is one species, the Albomyces stage of Aciculosporium take.
In the field, we found the disease mainly on P. bambusoides stands. In a rarecase, the disease was recognized on Pleioblastus Sect. Nezasa sp. (Ne-zasa) (Fig.
4) growing under heavily diseased Madake-forest. We also detected the disease
on Sasa Sect. Sasa sp. (Chimaki-zasa) (Fig. 5) at the Kyoto University Forest in
Ashiu, in Miyama-cho, Kyoto-fu. In this case, the host plant is formed a purestand with no other bamboo plants around it. Then we surveyed the disease in
several bamboo gardens. As a result, the bamboo plants listed in Table 1 wereconfirmed as host plants of the fungus. The results of conidial measurements ofthe fungus from the above listed host plants are given in Table 3.
In the literature, few species are recorded as the host plants of the fungus. In

Host Range of Aciculosporium takeFig. 1. The disease symptom on the new twigs of Pleioblastus gramineus (Taimin-chiku)
grown from the aerial part of underground rhizomes.
Fig. 2. The disease symptom on Pleioblastus Sect. Nezasa sp. (Ne-zasa) .
Fig. 3. The disease symptom on new twigs of Semiarundinaria fastuosa (Narihira-dake)grown from the aerial part of underground rhizomes.
Fig. 4. The disease symptom on the new twigs of Sasa Sect. Lasioderma sp. (Nambu-suzu)
grown from the aerial part of underground rhizomes.
Fig. 5. The disease symptom on Sasa Sect. Sasa sp. (Chimaki-zasa) .

29

Mitsuya Tsuda et al.

30
Table 3.

Conidial lengths of Aciculosporium take from different bamboo plants.
Collection site

Bamboo species
A

Phyllostachys bambusoides
var. marliacea
cv. ginmeishibochiku
f. kashirodake
f. subvariegata
f. geniculata
var. castillonis
var. castilloni-inversa
var. holochrysa
cv. usanchiku
P. aurea
f. a/bo-variegata
var. jlavescens-inversa
f. holochrysa
P. nigravar. henosis
f. boryana
f. albo-variegata
var. tosaensis
f. megurochiku
P. makinoi
P. tranquillans
P. dulcis
P. humiris
Shibataea kumasaca
S emiarundinaria fastuosavar. viridis
S. yoshi-matsumurae
S. okuboi
S. tatebeana
S. kagamiana
Pleioblastus gramineus
Sect. Nezasa sp.
Sasa Sect. Sasa sp.
Sect. Lasioderma sp.

B

c

D

E

F

48.0 ± 4.1
46.2 ± 3.7 47 .7 ± 3.2
44.6 ± 5.6 48.2 ± 3.5 44.3 ± 7.2
45.8 ± 3.8
47.3 ± 4.6
48.2 ± 4.3 47.1± 3.6 50. 1± 3.9*a
48.8± 3.942.5± 3.7
49.2 ± 3.5
44.6± 4.4 46.2 ± 3.5
47.5± 3.0
48.0 ± 3.8 48.2 ± 3.7*b
45 .5± 3.6
47.3 ± 4. 1
47.0 ± 3.1
49.6± 2.9 46.4 ± 4.4 45.6 ± 4.6
46.8± 4.7 47.3± 4.5
47 .7 ± 3.4
45.1± 3.6
48 .0 ± 3.8 48.4 ± 4.3*a44.7 ± 5.3
48.7 ± 3.4
45 .0 ± 3.4 47.8± 3.2
46.6 ± 4.0
43.2 ± 3.6
49.4 ± 5.1
46.8 ± 2.8
46.5 ± 4.2
46.8 ± 3.5
44.4 ± 3.5
48.4 ± 4.1
44.6 ± 4.1 47.8± 3.4
48.5± 3.7
46.6 ± 3.9
47.0 ± 3.5
46.6 ± 3.5
48.5 ± 3.9 47.2 ± 3.0 48 .1± 3.4
47.2 ± 3.4
43 .6± 4. 1 41.8 ± 3.9 43 .6 ± 4.3 45.9 ± 3.4 46.5± 4.0
44.9 ± 3.4
39.8± 4.1
42.7 ± 5.5 44.5± 3.945.5± 2.6
45.0 ± 4.4 48.8 ± 3.2
46.3 ± 3.5 48.5 ± 3.7
48 .6 ± 3.7
48.4 ± 5.2
46.0 ± 4.1
45.6 ± 3.1
41.9 ± 4.4
43.0 ± 3.9*c42.7 ± 2.8*d48.3± 3.8

The bamboo names and collection sites are the same as in Table 1.
Conidial lengths (in,um) are recorded as average value ± S.D. (n = 50).

this study, however, the fungus was recognized on 17 species, 8 varieties, 9 forms
and 2 horticultural cultivars in 5 genera of bamboo plants in the bamboo gardens
surveyed. Consequently, we can add 11 species, 6 varieties, 6 forms, and 2
horticultural cultivars in 4 bamboo genera as new host plants. The newly
discovered host plants are: Phyllostachys bambusoides cv. ginmeishibochiku (Gin-Host Range of Aciculosporium take

31

Table 4. Conidial lengths of Aciculosporium take on Phyllostachys bambusoides collectedfrom differnt locations.
Voucher no.

Conidial length

Voucher No.

Conidial length010 (B)
040 (C)055 (E)076 (Isonokami)
077 (Isonokami)
078 (Sumiyama)079 (Kohata)
080 (Kohata)

48.0 ± 4.1
46.2 ± 3.7
47.7 ± 3.2
40.5 ± 3.2
48.5 ± 3.7
50.2 ± 3.9
43.5 ± 3.2
46.0 ± 3.3

081 (Shirakawa)
082 (Shirakawa)
083 (Daigo)
084 (Rokujizo)
085 (Takahashi)
086 (Shizu)
087 (Sakura)
088 (Amakubo)

43.3± 4.644.3± 3.444.8± 3.7
45 .7 ± 3.6
44.6 ± 3. 1
44.4 ± 4.3
42.7 ± 4.7
46.4 ± 4.7

Locations are indicated in Tables I and 2.
Conidial lengths (in J.tm) are recorded as average value ± S.D . (n = 50).

mei-shibo-chiku) , P. bambusoides f. subvariegata Makino ex Tsuboi (Kon-shimadake), P. bambusoides f. geniculata (Nakai) Muroi (Mutsuore-dake), P. bambusoides var. castillonis (Marliac ex Carr.) Makino (Kinmei-chiku), P. bambusoides
var. castilloni-inversa Houz. de Leh. (Ginmei-chiku) , P. bambusoides var. holochrysa Pfitzer ex Houz. de Leh. (Ougon-chiku), P. bambusoides cv. usanchiku(Usan-chiku), P. aurea f. albo-variegata Makino (Shima-hachiku), P. aurea var.
fiavescens-inversa (Houz. de Leh.) Nakai (Ginmei-hachiku) , P. aurea f. holochrysa (Ougon-hotei), P. nigra var. tosaensis Makino ex Tsuboi (Tosa-torafudake), P. nigra f. megurochiku (Makino) Nakai (Meguro-chiku), P. tranquil/ans(Koidz.) Muroi (Inyo-chiku), P. dulcis McClure (Hakuho-kanchiku), P. humiris
Muroi (Hime-hachiku), Semiarundinaria fastuosa var. viridis Makino (Aonarihira), S. yoshi-matsumurae Muroi (Nikko-narihira), S. okuboi Makino(Bizen-narihira) , S. tatebeana Muroi (Kenashi-yashadake), S. kagamiana
Makino (Rikuchu-dake), Pleioblastus gramineus (Bean) Nakai (Taimin-chiku),
Pleioblastus Sect. Nezasa sp. (Ne-zasa), Sasa Sect. Sasa sp. (Chimaki-zasa), andSasa Sect. Lasioderma sp. (Nambu-suzu). We also confirmed the natural infection on Sibataea kumasaca (Okame-zasa), which had been reported as a hostplant of the fungus by artificial inoculation (Chen, 1970; Shinohara, 1970) . From
the results of this study and the list of Hara (1936), it is clear that somePhyllostachys species and their horticultural cultivars, and the members of thegenus Semiarudinaria were the most frequently attacked by the fungus . However,Moso-chiku (Phyllostachys pubescens Mazel ex Houz.) and its horticultural
cultivars, which are widely cultivated in Japanese archipelago, especially in thesuburbs of Kyoto-shi, was free of the fungus in our survey. This plant has beenreported as a host of the fungus in Taiwan (Chen, 1970). This might be due toa difference in susceptibility of the plant depending on the geographical location
or a problem in identification of Moso bamboo. In the genus Semiarudinaria, we
could not find the fungus onS. yashadake (Makino) Makino (Yasha-dake). This

32

Mitsuya Tsuda et al.

Fig. 6. Naturally produced conidia of Aciculosporium take. A. Conidia. B. Conidiumwith middle septum (arrow head).
Fig. 7. Cultural type conidia produced by germination of natural conidia of Aciculosporium take. A. Cultural type conidial mass with end-to-end germination on the agar
medium (arrow heads) . B. Germination from basal part (arrow head).
Fig. 8. Mycelium colony of Aciculosporium take on the agar medium.
Fig. 9. Mycelium with cultural type conidia of Aciculosporium take on the agar medium.
(Bars indicate lO,um.)

may also reflect a problem in host taxonomy. These results were interesting andmay open the way to phylogenetical consideration of these species on the basis ofaffinity with the fungus.
Difference in conidial morphology in nature and in artificial culture. The

Host Range of Aciculosporium take33

conidia from field collections had the same morphological characteristics as
already reported (Fig. 6). When we tried to isolate the fungus monoconidially on
agar medium, it was difficult to establish cultural isolates. Then we tried to isolate
the fungus by mass inoculation of stromatic tissues on agar medium. Slowgrowing pustules appeared on the agar medium and almost all colonies stopped
growing at this stages (Fig. 8). Mycelial growth was very rare and occurred bychance. The reasons for this phenomenon were revealed by microscopic observations. The naturally formed conidia of the fungus germinated from both ends andformed two dichotomously branched appendages at their apex (Fig. 7), whichhave been referred to as germinating hyphae (Shinohara, 1965, 1966). The
appendages, however, seldom produced hyphae during our observations. Consequently, the germinated conidia stopped growing at this end-to-end stage (Fig. 7) .
When they separated by breaking at the middle septum, new germination occurred from the basal end and formed the same structures with apical appendages as
Fig. 7. This kind of structure was also produced from the hyphae as a lateralbranch as shown in Fig. 9. We conclude that this structure is one kind of
conidium. Next we tried to culture the fungus in liquid medium by shaking.
Growth proceeded by repeated separation at the bases of these conidia, and many
single conidia or unseparated few-celled conidia with appendages were observedin the culture. Thus the fungus has two types of conidia: naturally produced
filiform conidia with swollen ends and one septum at the middle (Fig. 6), andcultural type conidia with apical appendages (Fig. 7) . These cultural type conidia
with appendages might function for dispersal in the field by water droplets on thebamboos, like those of amphibious fungi in streams (Kendrick, 1992) .
Acknowledgements

We are grateful to the curators and authorities of the following botanic
gardens, who allowed us to survey their bamboo stands: The Kyoto Botanical
Garden, Rakusai Bamboo Park, Kamigamo Experimental Forest, Kyoto University, Kansai Research Center, Forestry and Forest Products Research Institute,
Yoro Bamboo Garden in Yoro-Kohen. We thank Prof. Dr. Shozo Shibata,
Kamigamo Experimental Forest, Kyoto University for identification of several
specimens of bamboo plants and valuable suggestions on bamboo taxonomy. We
also thank the members of the Pesticide Research Institute, Kyoto University andthe Department of Botany, National Science Museum for their encouragement
during this study. This work was partly supported by a research grant from theSumitomo Foundation.

34

Mitsuya Tsuda et al.

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