Fung. Sci. 19(3–4): 125–131, 2004 Notes on Zygomycetes of Taiwan (IV): Three Absidia species (Mucoraceae)
Hsiao-Man Ho1*, Shu-Cheng Chuang1, and Shiang-Jiuun Chen2
1. Department of Natural Science Education, National Taipei Teachers College, No. 134, Sect. 2, Ho-Ping E. Rd., Taipei 106, Taiwan, R.O.C. 2. Department of Life Science, National Taiwan University, No.1, Sec. 4, Roosevelt Road, Taipei 106, Taiwan, R.O.C.
(Acceped: December 13, 2004)
ABSTRACT
During a survey of Zygomycetes in Taiwan, three species of the genus Absidia: A. cylindrospora Hagem, A. glauca Hagem, and A. spinosa Lendner were isolated and reported. The former two are new records to Tai- wan.
Key words: Absidia, Mucoraceae, Taiwan.
Introduction are spherical and the sporangial walls usually break. The sporangiospores of Absidia are The genus Absidia (Mucoraceae, Mucorales) smaller and never show the striations of many was characterized by van Tieghem (1876) by species of Rhizopus. Absidia is also like Phy- stolons with rhizoids; sporangiophores bearing comyces in its manner of sexual reproduction. apophysate, pyriform sporangia arising, mostly Both genera have zygospores with typically in small groups, from the elevated parts of the fingerlike structures develop from the suspen- stolons; and by zygospores enveloped in ap- sors and envelop the zygospore. Phycomyces pendages from the suspensors. has similar fingerlike structures, but they are Absidia is closely related to Rhizopus be- branched, and the zygospores are formed be- cause of its rhizoids, stolons, apophyses, and tween tonglike suspensors. Another related ge- the indefinite growth of the fertile aerial myce- nus is Gongronella, which is Absidia-like fungi lium. It differs asexually from Rhizopus with its with an apophysis and columella reduced in sporangiophores always arise along the stolons size (Hesseltine and Ellis, 1964). but never opposite the rhizoids as in Rhizopus. Furthermore, Hesseltine and Ellis (1964) di- Although both genera possess an apophysis be- vided the genus Absidia into two subgenera. neath the sporangium, Absidia has pyriform, Those with suspensors lacking appendages deliquesce sporangia, whereas in Rhizopus they were separated from those with appendages and
* Corresponding author. Email: [email protected] 126 Fung. Sci. 19(3, 4), 2004 placed in Absidia subgenus Mycocladus. Since sporangium was touched with a sterilized nee- zygospores are not easy to find, additional dle and transferred it to a fresh corn meal agar characters such as growth at 37°C were used to plate. After 4–7 days, pure culture of Absidia indicate the subgenus Mycocladus (Schipper, were obtained. Then, the cultures were grown 1990). A monograph of the genus Absidia was on potato dextrose agar (PDA) plates and incu- published by Hesseltine and Ellis (1964, 1966) bated at 15°C, 25°C, 28°C, 31°C, 37°C, and and Ellis and Hesseltine (1965, 1966). Up to 40°C for species identification. date, twenty Absidia species were reported (Kirk et al., 2001). Most of them are soil in- SEM habitants. Pertinent material were selected under a dis- Previously three species and one variety of secting microscope and fixed for 1 hr with Absidia were reported from Taiwan. They were 2.5% glutaraldehyde in distilled water and post- A. corymbifera Lichth, A. hyalospora (Saito) fixed for 1 h with 1% osmium tetraoxide in dis- Lendner, A. spinosa Lendner, and A. spionosa tilled water. The materials were washed with var. azygospora Boedijn (Ho and Chien, 1979; distilled water and dehydrated in a graded ace- Ho, 1993; Tseng and Chen, 1981). These fungi tone series. Specimens were dried in a critical were all isolated from soil. Recently, from a point dryer, coated with gold, observed, and survey of Zygomycetes in Taiwan conducted by photographed with a Hitachi S-520 scanning the authors, three species of Absidia were iso- electron microscope (SEM) at 20 KV. lated and identified. They are A. cylindrospora Hagem, Absidia glauca Hagem, and A. spinosa LM Lendner, among them the former two are re- Pertinent material were selected under a dis- ported as new records to Taiwan. The purpose secting microscope and mounted in water or of this paper is to describe the morphological lactophenol cotton blue. Photographs were characters of above three species and the dif- taken with a Leica MPS32 light microscope ferences between them and closely related taxa. (LM). All examined specimens, slides and pure cul- Materials and Methods tures are deposited in the mycology laboratory, Department of Natural Science Education, Na- Soil samples were collected from country tional Taipei Teachers College, Taipei, Taiwan. roadside or forest and brought to laboratory in Identification of the fungi was based on keys of polyethylene bags. Two to three milligrams of Ellis and Hesseltine (1965; 1966), Hesseltine soil samples were placed on corn meal agar and Ellis (1964; 1966) and Schipper (1990). (CMA) plates (Warcup, 1950). The plates were left on a bench at room temperature, incubated Taxonomy for ca. 2 weeks, and then observed by using a dissecting microscope. When the special whirl- Absidia cylindrospora Hagem, Skriftr u.a. arrranged sporangiophores and the pyriform- Vidensk.-Selsk. Crist. Math. Nat. K1., No.7. shaped sporangia of Absidia were found, the 45. 1908. (Fig. 1) Notes on Zygomycetes of Taiwan (IV) 127
Colonies growing on PDA rapidly at 25 and stolons, typically simple, occasionally branch- 28°C, reaching 9 cm in diam in 1 week, 1.5 cm ing, hyaline to light brown, always with a sep- high, growth slowing down at 15, and 31°C, no tum under the sporangium. Sporangia 15.1– significant growth at 37°C. Colony color at 22.5 × 17.5–25.0 µm, pyriform, hyaline to deep 25°C initially white, in 4 days becoming light gray, wall smooth, deliquescing at maturity. brown in the center portion, later turning to Columellae 12.2–21.2 × 12.3–15.1 µm, smooth light brown in one week, and pale grayish with a projection at the apex. Sporangiospores brown in 2 weeks. Odor none. Sporangiophores cylindrical, 3.5–4.1 × 1.1–2.0 µm, smooth, with 130–230 µm long, 2.5–4.5 µm wide, erect, rounded ends. Zygospores not found. Hetero- smooth, arising singly up to 7 in a whorl but thallic. generally 1–4 from the same place of the
Fig. 1. Absidia cylindrospora Hagem, A, B, LM, C, D, SEM. A. Mature sporangium, note the septum under the apophysis (arrow head). Bar = 20 µm; B. Columella with pointed spine and collar. Bar = 20 µm; C. Columella with pointed spine. Bar = 3.8 µm; D. Cylindrical sporangiospores. Bar =1.5 µm. 128 Fung. Sci. 19(3, 4), 2004
Specimen examined. Sym 0302, isolated Absidia glauca Hagem, Skrifter u. a. Vidensk.- from soil, Yangmingshan National Park, Taipei, Selsk. Christ. I. Math.-Nat. K1. No. 7, p. 43, Taiwan, March, 2002. 1908. (Fig. 2) Notes. Absidia cylindrospora can be distin- Colonies growing on PDA rapidly at 15, 25, guished from the closely related species A. and 28°C, reaching 9 cm in diam in 1 week, 1.5 pseudocylindrospora by the longer sporangio- cm high, no significant growth at 31°C; colony phores (longer than 170 µm in A. pseudocylin- color at 25°C initially white, later becoming drospora ) and the less number of sporangio- bluish green in center portion, turning to dark phores in a whorl (usually 2–4, vs 7–11 in A. green in 1 week and brownish olive in 2 weeks. pseudocylindrospora) (Hesseltine and Ellis, Odor none. Sporangiophores 60–210 µm long, 1964). 2.5–5.0 µm wide, erect, single or 2–3 arising
Fig. 2. Absidia glauca Hagem, A. LM, B-D, SEM. A. Two mature sporangia, note the septa under the apophyses (arrow heads). Bar =20 µm; B. Smooth-walled sporangium, note the septum under the apophysis (arrow heads). Bar = 7.5 µm; C. Columella with pointed spine and sporangiospores. Bar = 3.8 µm; D. Globose sporangiospores. Bar = 1.5 µm. Notes on Zygomycetes of Taiwan (IV) 129 from the same place on the aerial stolons, re- arising from the same place on the aerial peatedly branching, always with one septum stolons, usually 3–5; typically with a septum always present 10–15 µm below apophysis. below the apophysis, generally simple. Sporan- Hyphae at first hyaline through greenish be- gia 22.3–25.1 × 16.2–18.4 µm, pyriform, oval, coming brownish olive later, wall smooth. Spo- deliquescing at maturity. Columellae 10.1–20.2 rangia 22.8–37.5 × 17.8–30.0 µm, pyriform, × 9.3–17.4 µm, hemispherical with collars; hyaline to deep gray, wall smooth, deliquescing usually with a projection on upper surface of at maturity. Columellae 11.0–33.0 × 9.0–25.1 columella. Sporangiospores 2.9–3.9 × 1.1–2.0 µm, hyaline to gray, hemispherical above the µm, smooth, cylindrical with rounded ends. apophysis, smooth typically with a pointed Zygospores globose, usually 50.3–125.5 µm in spine on the top. Sporangiospores globose, 2.0– diam, brown, wall coarsely verrucose; suspen- 3.2 µm in diam. smooth, hyaline. Zygospores sors typically unequal and with fingerlike ap- not found. Heterothallic. pendages from the larger suspensors. Homo- Specimen examined. CHT0106, isolated thallic. from soil, Tainan County, Taiwan, Feb. 2003. Specimen examined. S42701, isolated from Notes. Absidia glauca can be distinguished soil, Pintung County, Taiwan, Aug, 2002. from other species of Absidia by its greenish Notes. This species is characterized by the colony color and its perfectly globose sporan- cylindrical sporangiospores and numerous zy- giospore. A. coerulea Bainier and A. californica gospores with finger-like projections from the Ellis and Hesseltine are the other two Absidia larger suspensors. Also it’s easily to be distin- species which have uniformly perfectly globose guished from the closely related species A. sporangiospores, but A. coerulea possesses anomala by the grayish brown colony color on purple-colored colonies and A. californica a PDA plate. The colony color of A. anomala on gray colony color (Ellis and Hesseltine, 1965). PDA plate is violet or purple. We identified this isolate as A. glauca mainly based on its green colony color and the globose Acknowledgements sporangiospores. This study was supported by a grant (NSC- Absidia spinosa Lendner, Bull. Herb. Boiss., 91-2621-B-152-001) from the National Science Ser. II, 7:250. 1907. (Fig. 3) Council of R.O.C. Colonies growing on PDA at 25 and 28°C rapidly, reaching 9 cm in diam in one week, References growth slowing down at 15 and 31°C, reaching 9 cm in two weeks and nearly no growth at Ellis, J.J. and C.W. Hesseltine. 1965. The genus 37°C. Colony color at 25°C initially white, in 4 Absidia: globose-spored species. Mycologia days becoming light grayish brown in the cen- 57: 222–235. ter portion, becoming grayish brown in 2 Ellis, J.J. and C.W. Hesseltine. 1966. Species of weeks. Ordor none. Sporangiophores 61–109 Absidia with ovoid sporangiospores. II. µm long, 2.0–2.8 µm wide, erect, smooth, 1–6 Sabouraudia 5: 59–77. 130 Fung. Sci. 19(3, 4), 2004
Fig. 3. Absidai spinosa Lendner, A-D, LM. A. Mature sporangium. Bar = 10 µm; B. Columella with spine. Bar = 10 µm; C. Cylindrical sporangiospores. Bar = 10 µm; D. Globose zygospore with finger-like projections. Bar = 30 µm.
Hesseltine, C.W. and J.J. Ellis. 1964. The genus Ho, H.-M. and C.-Y. Chien. 1979. Isolation, Absidia: Gongronella and cylindrical-spored culture and observation on Absidia spinosa species of Absidia. Mycologia 56: 568–601. (Mucorales). Bio. Bull. N.T.N.U. 14: 11–14. Hesseltine, C.W. and J.J. Ellis. 1966. Species of (In Chinese) Absidia with ovoid sporangiospores. I. My- Kirk, P.M., P.F. Cannon, J.C. David, and J.A. cologia 58: 761–785. Stalpers, edi. 2001. Dictionary of the fungi. Ho, H.-M. 1993. Morphological study of Ab- 9th. Edi. CABI Bioscience, U.K. sidia spinosa var. azygospora isolated in Schipper, M.A.A. 1990. Notes on Mucorales-I Taiwan. Bull. N.T.T.C. 6: 421–434. (In Chi- Observations on Absidia. Persoonia. 14(2): nese) 133–148. Notes on Zygomycetes of Taiwan (IV) 131
Tieghem, P. van. 1876. Troisime memoire sur study of fungal flora on stored feeds. Tai- les Mucorinees. Ann. Sci. Nat. Ser. VI, wania 26(3): 68–89. Botan. 4: 312–398. Warcup, J.H. 1950. The soil plate method for Tseng, H.-Y. and Z.-C. Chen. 1981. Preliminary isolation of fungi from soil. Nature 166: 117.
lĥRȎƣǏHȯά (IV), 'ɱฃͬ (ǔฃ͌) ƈƣ
̵ʎ1 Ŵ̯ߟ1 ॕଝț2
1. ϤlƔ ˓ॊbƛ͌˓ž˓Ň lƔyɧÁ˚Ѡ_Ƹ"'Ϡ; lĥ 9ƿǹ 2. ϤlĥŎ˓Eɜ͌˓Ň lƔyʵƷ̝ѠϠƸ"; lĥ 9ƿǹ Êǜ
ĭǂɱ ʗ'ɱฃͬƈƣ,̸˅ɱฃ (Absidia cylindrospora) 9ͬחਫ਼lĥRȎƣǏȯά"! ฃ (A. glauca) ǫ ė̒ɱฃ (A. spinosa) Ņøথ7ࢰ˩;Ģ³̛ŕlĥħƸŊߎ;Ɲ9ȶ֙ºɪƑ ǫŰֆઙࣥHtȒ;
खࠛɡ,ǔฃ͌:ɱฃͬ:lĥ;