Fung. Sci. 20(1, 2): 1–9, 2005
New records of the genus Cunninghamella
(Mucorales) in Taiwan
Chung-Wen Liu1, Guey-Yuh Liou2, and Chiu-Yuan Chien3*
1. Kaohsiung Municipal Girls' Senior High School, Kaohsiung, 80148 Taiwan
2. Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu,
30062 Taiwan
3. Department of Life Science, National Taiwan Normal University, Taipei , 11677 Taiwan
(Accepted: March 31, 2005)
ABSTRACT
Four species of the genus Cunninghamella, namely, C. blakesleeana, C. bertholletiae, C. echinulata, and C.
elegans are described and illustrated. C. blakesleeana and C. bertholletiae are recorded as new to the fungal
flora of Taiwan.
Key words: Cunninghamella blakesleeana, C. bertholletiae, C. echinulata, C. elegans, Mucorales, Taiwan.
Introduction
Materials and Methods
The genus Cunninghamella Matruchot is one
of the most common members of the order Mucorales. They are frequently found in soils rich
in organic nutrients and have been reported predominating in warmer zones (Domsch and
Gams, 1980). During our study of Zygomycetes in Taiwan, four species of Cunninghamella, namely, C. blakesleeana, C. bertholletiae, C. echinulata, and C. elegans were
collected and identified. Characters of above
four species are described and illustrated. Cunninghamella blakesleeana and C. bertholletiae
are recorded as new to the fungal flora in Taiwan. Besides, C. echinulata and C. elegans
have been provided as additional records (Ho et
al., 1974; Yang and Liu, 1972).
By using Warcup’s method (Gams et al.,
1987), 34 strains of Cunninghamella were isolated from soils collected from 17 localities in
Taiwan. Soil samples were incubated on MEA
plates (malt extract agar: malt extract 2%, agar
1.6%) containing 50 ppm penicillin-G at room
temperature in dark. Pure cultures were established by isolating the mature sporangiola. A
piece of agar containing isolated sporangiola
was transferred to MEA plates under a binocular dissecting microscope.
For identification, cultures were inoculated
on SMA (synthetic mucor agar: dextrose 4%,
asparagine 0.2%, KH2PO4 0.05%, MgSO4
7H2O 0.025%, agar 1.6%, Thiamine chloride
0.5 ppm) and MEA at 25°C and 40°C in dark
*
Corresponding author
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Fung. Sci. 20(1, 2), 2005
for 6 days. Details of fungal characteristics
were examined, measured, described and photographed with dissecting binocular microscope
(WILD M7A), and light microscope (WILD
M20).
For scanning electron microscopic (SEM)
observations, sporangiola were fixed in 2.5%
glutaraldehyde for 2 h and washed with phosphate buffer (pH 7.2) three times. Then, specimens were gradually dehydrated with increasing concentrations of ethyl alcohol (25% to
100%). Finally, specimens were treated in a
critical point drying apparatus. Dried specimens were coated with evaporated gold (ion
coater. Eiko, Engineering) and examined under
scanning electron microscope (Akashi, DS130S).
The taxonomic systems of Baijal and Mehrotra (1980), Cutter (1946), Lunn and Shipton
(1983), Mil’ko and Beljakova, (1967), Samson
(1969), Weitzman (1984), and Weitzman and
Crist (1979; 1980) were used for identification.
Taxonomy
Cunninghamella bertholletiae Stadel, Über
neuen Pilz, Cunn. bertholletiae, (Diss., Kiel):
1–35. 1911. ( Fig. 1, A–G ).
Colonies at 25°C spreading rapidly, grey on
maturity, growing well at 40°C; Sporangiophores erect, with solitary, opposite, verticillate or
irregular branches, the laterals sometimes
showing secondary branching; terminal vesicles subglobose to obovate, 14.8–41.2 µm in
width, average 30.6 µm, lateral vesicles smaller
than terminal ones, but no conspicuous difference in shape, 12–28.8 µm in width, average
19.1 µm; Sporangiola hyaline to pale brown,
spherical to ellipsoid, occasionally lacrymoid,
5.6–11.4 × 5.2–8.8 µm in diam., average 7.8 ×
7.0 µm, smooth to echinulate with very short
spines; Chlamydospores subglobose to broadly
ellipsoidal, average 21.86 × 18.83 µm in diam.;
Heterothallic.
Material examined. Taiwan, Taipei: Pitan,
soil, 31 Jul 1993, No. 12; Pingtung: Wan-Dan,
soil, 08 Aug 1993, No. 21.
Notes. Pispek (1929) has enumerated a new
scientific name of this species C. polymorpha
on the basis of variously branched sporangiophores and variously shaped sporangiola. However, Weitzman and Crist (1979, 1980) have
demonstrated C. polymorpha and C. bertholletiae as synonyms by their temperature tolerance and branching patterns. However, C.
bertholletiae can be distinguished from other
species by its diversely branched sporangiophores and lacrymoid sporangiola. The species is a
new record from Taiwan.
Cunninghamella blakesleeana Lendn., Bull.
Soc. bot. Genève, 2 sér. 19: 234. 1927. ( Fig.
2, A–G ).
Colonies at 25°C spreading rapidly, white to
light buff on maturity, not growing at 40°C;
Sporangiophores erect, with solitary, opposite
or verticillate branches; terminal vesicles
spherical to obovate, 14.8–42 µm in width, average 25.6 µm, lateral vesicles of similar shape
as terminal ones but smaller in size, 8–32 µm in
width, average 18.4 µm; Sporangiola hyaline,
spherical to ellipsoidal, 7.8–19.2 × 7.5–16.8
µm in diam., average 11.3 × 10.9 µm, smooth
to minutely echinulate, spines up to 1.6 µm in
length; Chlamydospores subglobose to ellipsoidal, average 22 × 19.1 µm in diam.; Zygosporangia spherical to subglobose, 46.4–54.4 ×
41.6–49.6 µm in diam., average 49.5 × 45.3
Cunninghamella found in Taiwan
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Fig. 1. Cunninghamella bertholletiae. A, sporangiophore under dissecting binocular microscope (bar = 150 µm); B, sporangiophores with two whorled or verticillate branches under light microscope (bar = 30 µm); C, sporangiola (bar = 15
µm); D, chlamydospore (bar= 15 µm); E–G, sporangiola smooth to echinulate under SEM, arrow shows a denticle (bar = 4
µm).
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Fung. Sci. 20(1, 2), 2005
Fig. 2. Cunninghamella blakesleeana. A, unbranched sporangiophore (bar = 30 µm); B, sporangiophore with lateral
branches (bar = 30 µm); C, sporangiola (bar = 15 µm); D, chlamydospore (bar = 15 µm); E, sporangiolum with spines under SEM (bar = 4 µm); F, zygosporangium with unequal suspensors (bar = 4 µm); G, zygosporangium under SEM (bar = 4
µm).
Cunninghamella found in Taiwan
µm, light reddish brown, warted; Suspensors
hyaline, smooth, equal shape, but sometimes
unequal, 30.8–54 × 14.8–34 µm in size, average 42.3 × 22.4 µm. Heterothallic.
Material examined. Taiwan, Kaohsiung:
Kaohsiung Park, soil, 09 Aug 1993, No. 19 (+);
Pingtung: She-Ting, soil, 16 May 1993, No. 04
(–).
Notes. The sporangiola up to 35 µm long
mentioned by Cutter (1964) and Baijal and
Mehrotra (1980) are not found in the two
strains studied. The two opposite suspensors of
C. blakesleeana are sometimes unequal, while
other species did not have such phenomenon.
So it can be distinguished from other species by
this characteristic. The species is newly reported from Taiwan.
Cunninghamella echinulata (Thaxt.) Thaxt.,
Rhodora 5: 98. 1903. ( Fig. 3, A–F ).
Colonies at 25°C spreading rapidly, white to
pale buff on maturity, powdery in appearance,
growing at 40°C; Sporangiophores erect, with
solitary, opposite, verticillate, pseudoverticillate or irregular branches, the laterals sometimes showing secondary branching; terminal
vesicles spherical to subglobose, 24.8–44 µm in
width, average 34.8 µm, lateral vesicles smaller
than terminal vesicles, but no conspicuous difference in shape, 18–29.6 µm in width, average
23.5 µm; Sporangiola hyaline, predominantly
spherical, sometimes subglobose to broadly ellipsoidal, 9.6–20.5 × 9.4–20.4 µm in diam., average 13.5 × 13.4 µm, usually thickly beset
with spines up to 4 µm in length, sometimes
smooth; Chlamydospores subglobose to ellipsoidal in shape, average 26.8 × 18.2 µm in
diam.; Zygosporangia spherical to ellipsoidal,
26.8–46 × 25.6–38 µm in diam., average 36.9 ×
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32.7 µm, reddish brown, warted; Suspensors
hyaline, smooth, 10.8–42.4 × 8.8–36.6 µm in
size, average 20.1 × 16.2 µm. Heterothallic.
Material examined. Taiwan, Taipei: Pali,
soil, 30 Jul 1993, No. 09 (+); Pingtung: SheTing, soil, 16 May 1993, No. 05 (–).
Notes. Cutter (1946) had indicated that there
was no verticillate branched sporangiophore in
this species. The present cultures studied here,
the verticillate form could be discerned. Samson (1969) and Baijal and Mehrotra (1980) also
have described this branching pattern. This
species can be distinguished from other species
of Cunninghamella by its pseudoverticillately
branched sporangiophores and powdery appearance.
Cunninghamella elegans Lendn., Bull. Herb.
Boissier, 2 sér. 7: 250. 1907. (Fig. 4, A–G ).
Colonies at 25°C spreading rapidly, grey on
maturity, not growing at 40°C; Sporangiophores erect, with solitary, opposite or verticillate
branches; terminal vesicles subglobose to
broadly clavate, 15.2–45.2 µm in width, average 27.6 µm, lateral vesicles of similar shape as
terminal ones but smaller in size, 11.2–31.2 µm
in width, average 19.5 µm; Sporangiola hyaline
to pale brown, spherical to ellipsoidal, 5–20 ×
5–15.4 µm in diam., average 10 × 9.4 µm,
smooth, verrucose or spinulose, spines up to
0.8 µm in length; Chlamydospores subglobose
to broadly ellipsoidal in shape, average 18.3 ×
13.9 µm in diam.; Zygosporangia subglobose to
broadly ellipsoidal, 45.6–53.6 × 36–48.8 µm in
diam., average 48.8 × 42.3 µm, dark reddish
brown, warted; Suspensors hyaline, smooth,
usually apposed, 33.4–52.8 × 13.2–16.8 µm in
size, average 40.8 × 15.3 µm. Heterothallic.
Material examined. Taiwan, Taipei: Shen-
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Fung. Sci. 20(1, 2), 2005
Fig. 3. Cunninghamella echinulata. A, sporangiophore with verticillate or irregular branches (bar = 30 µm); B, sporangiola (bar = 15 µm); C, chlamydospore (bar = 15 µm); D, sporangiolum with spines under SEM (bar = 15 µm); E, zygosporangium (bar = 25 µm); F, zygosporangium under SEM (bar = 4 µm).
Cunninghamella found in Taiwan
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Fig. 4. Cunninghamella elegans. A, sporangiophore with solitary branch (bar = 30 µm); B, sporangiophore with verticillate branches (bar = 30 µm); C, sporangiola (bar = 15 µm); D, chlamydospore (bar = 15 µm); E, sporangiolum with spines
under SEM (bar = 4 µm); F, zygosporangium (bar = 25 µm); G, zygosporangium under SEM (bar = 10 µm).
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Fung. Sci. 20(1, 2), 2005
Gi-Zen, soil, 21 Feb 1993, No. 01 (+); Taipei:
Pitan, soil, 31 Jul 1993, No. 11 (–).
Notes. Because there are no significant differences in the morphological characteristics
between C. elegnas and C. bertholletiae, Zycha
(1935), Mil’ko and Beljakova (1967), Samson
(1969), and Baijal and Mehrotra (1980) have
considered them as the same species. However,
Cutter (1946), Weitzman and Crist (1979,
1980) and Weitzman (1984) have distinguished
them as two valid species by their conspicuously different temperature tolerance and
branching patterns, as well as the inability to
form zygospore when paired. The species can
be distinguished from the others by its maximal
temperature tolerance below 38°C.
Acknowledgements
We are grateful for the support grant
(NSC79-0211-B003(01)) from the National
Science Council in Taipei, Taiwan, R.O.C.
References
Baijal, U. and B.S. Mehrotra. 1980. The genus
Cunninghamella—a reassessment. Sydowia
33: 1–13.
Cutter, V.M., Jr. 1946. The genus Cunninghamella (Mucorales). Farlowia 2: 321–343.
Domsch, K.H. and W. Gams. 1980. Compendium of Soil Fungi. Vol. I. Academic Press,
London.
Gams, W., H.A. van der Aa, A.J. van der PlaatsNiterink, R.A. Samson, and J.A. Stalpers.
1987. CBS Course of Mycology, 3rd ed.
Centraalbureau voor Schimmelcultures,
Baarn, The Netherlands. 136 pp.
Ho, S.M., B.Y. Yang, and C.E. deVol. 1974.
Studies on the Mucorales isolated from
Yang-Ming-Shan humus. Taiwania 19: 75–
87.
Lunn, J.A. and W.A. Shipton. 1983. Reevaluation of taxonomic criteria in Cunninghamella. Trans. Brit. mycol. Soc. 81: 303–
312.
Mil’ko, A.A. and L.A. Beljakova. 1967. The
genus Cunninghamella Matruchot and taxonomy of the Cunninghamellaceae. Mikrobiologiya 36: 684–690.
Pispek, P.A. 1929. Edafske mukorineje Jugoslavije. Acta Bot. Inst. Bot. Univ. Zagreb 4:
77–112.
Samson, R.A. 1969. Revision of the genus
Cunninghamella (Fungi, Mucorales). Pro.
Kon. Ned. Akad. Wetensch. Amsterdam, Ser.
C, 72: 322–335.
Weitzman, I. 1984. The case for Cunninghamella elegans, C. bertholletiae and C. echinulata as separate species. Trans. Brit. mycol Soc. 83: 527–529.
Weitzman, I. and M.Y. Crist. 1980. Studies with
clinical isolates of Cunninghamella. II.
Physiological and morphological studies.
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Cunninghamella found in Taiwan
臺灣產小克銀漢黴屬 (毛黴目) 的新紀錄種
劉崇文1 劉桂郁2 簡秋源3
1. 高雄市立女子高級中學，高雄市 80148，臺灣
2. 食品工業發展研究所 生物資源保存及研究中心，新竹市 30062，臺灣
3. 國立臺灣師範大學 生命科學系，臺北市 11677，臺灣
摘
要
本文描述臺灣產小克銀漢黴屬 (Cunninghamella) 真菌，分別為短刺小克銀漢黴 (C. blakesleeana)、灰色
小克銀漢黴 (C. bertholletiae)、刺孢小克銀漢黴 (C. echinulata) 以及雅緻小克銀漢黴 (C. elegans)，其中
C. blakesleeana 及 C. bertholletiae 為臺灣新紀錄種。
關鍵詞：毛黴菌目、短刺小克銀漢黴、灰色小克銀漢黴、刺孢小克銀漢黴、雅緻小克銀漢黴、臺灣。
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