Tolypocladium japonicum (C. G. Lloyd) Quandt, Kepler & Spatafora, IMA Fungus 5: 126. 2014. Figures 1–8.
≡ Elaphocordyceps japonica (C. G. Lloyd) G. H. Sung, J. M. Sung & Spatafora, Stud. Mycol. 57: 37 2007.
Stromata one to five emerging directly from underground ascomata of Elaphomyces sp., clavate, unbranched, with a rounded apex, on a cylindrical, pale gray to pale brown stipe, 3.5–7.0 cm in length, 1.8–3.0 cm long by 4–6 mm broad at fertile part, 1.5–5.0 cm long by 0.2–0.4 cm broad at stipe; surface plane, roughened by ostioles, with the fertile part tan-colored when immature, blackened when mature; interior dull yellowish green, hollow at center. Perithecia ovoid to ellipsoid, 400–500 × 180–230 μm, with papillate ostioles. Asci clavate to cylindrical, 300–350 × 9–13 μm, with a conspicuously thickened cap. Ascospores hyaline, long cylindrical, disarticulated into part-spores within asci; part spores cylindrical, with flattened ends, 10–15 × 3.0–3.4 μm.
Cultures and anamorph. Colonies on PDA at 25 C attaining 3 cm in 3 wk, thick, white to grayish, cottony, with brownish pigments diffusing into the media. Anamorph simplicillium-like. Conidiogenous cells solitary, long, slender, tapering toward the apex, 25–50 × 1.7–2.2 μm, about 1 μm broad at the apex. Conidia produced enteroblastically, one-celled, hyaline, smooth, elliptical to obclavate, 6.5–10.0 × 3.5–4.5 μm.
Specimens examined. New Taipei City, Wu-lai, Nei-dong, on Elaphomyces sp., 26 Apr 2014, Ju, Y.-M., Hsieh, H.-M., Ke, Y.-H., Sung, A.-N., Fan, Y.-C., Hung, S.-C. & Lin, J.-R. 103042601 (HAST); New Taipei City, Wu-lai, Nei-dong, on Elaphomyces sp., 20 May 2014, Ju, Y.-M., Hsieh, H.-M., Ke, Y.-H. & Chang, Y.-Y. 103052001 (HAST; culture accession number: BCRC FU30561; GenBank accession number of ITS: KT873533).
Notes. Records of T. japonicam are fairly rare even in Japan (Kobayasi and Shimizu 1960) and appear sporadic in other countries, including Austria (Mains 1957; Kobayasi and Shimizu 1960) and China (Liang et al. 2003). Reflected in studies of recent years is the paucity of available material of this fungus: the DNA sequences deposited at GenBank resulting from two Japanese sources only, a specimen OSC 110991 and a culture IFO 9647. Among the three ITS sequences of T. japonicum cited below, AB027366 and EU039882 are from IFO 9647, while JN049824 is from OSC 110991.
The Taiwan collections fit well the descriptions of T. japonicum given by Kobayasi and Shimizu (1960). Besides T.
japonicum, four other Tolypocladium species also parasitize Elaphomyces fruiting bodies and produce clavate stromata, including T. ophioglossoides (Ehrhart) Quandt et al., T. jezoense (S. Imai) Quandt et al., T. tenuisporum (Mains) Quandt et al., and T. szemaoense (M. Zang) Quandt et al. (Mains 1957; Kobayasi and Shimizu 1960.; Zang 2001). Tolypocladium ophioglossoides and T. jezoense differ from T. japonicum in forming a rhizomorphous structure on the stromatal base, and T. tenuisporum and T. szemaoense differ in having smaller part-spores.
A MEGABLAST query in GenBank using the ITS sequence obtained from the specimen 103052001 resulted in the following five top matches: Elaphocordyceps japonica (AB027366, query coverage = 100 %, identities = 596/601 [99 %], gaps = 0/601 [0 %]); Elaphocordyceps japonica (EU039882, query coverage = 100 %, identities = 596/602 [99 %], gaps = 1/602 [0 %]); Elaphocordyceps japonica (JN049824, query coverage = 93 %, identities = 393/404 [97 %], gaps = 2/404 [0 %]); Cordyceps guangdongensis (EU039881, query coverage = 100 %, identities = 569/608 [94 %], gaps = 28/608 [4 %]); and Tolypocladium inflatum (JF796050, query coverage = 95 %, identities = 511/578 [88 %], gaps = 32/578 [5 %]). The query result reinforces the Taiwan material being T.
This is the first report describing cultures and anamorph of T. japonicum. The conidiogenous cells of T. japonicum are long, slender, lacking an inflated base. Tolypocladium, typified by T. inflatum W. Gams, was originally characterized by conidiogenous cells with an inflated base and a narrow neck. However, the genus is currently circumscribed on the basis of molecular phylogeny rather than morphology (Quandt et al. 2014).
Ophiocordyceps odonatae (Kobayasi) G. H. Sung, J. M. Sung, Hywel-Jones & Spatafora, Stud. Mycol. 57: 45. 2007. Figures 9–14.
≡ Cordyceps odonatae Kobayasi, Bull. Nat. Sci. Mus. Tokyo, Ser. B, 7: 6. 1981.
= Hymenostilbe odonatae Kobayasi, Sci. Rep. Tokyo Bunrika Daig., Sect. B 5: 223.1941.
Teleomorph not produced. Anamorph synnematous. Synnemata gregarious, protruding from abdominal and thoracic joints of dragonflies, pale yellow to pale orange, clavate, abruptly rounded on top, stipitate, curved towards the front of dragonflies, 3–6 mm long, 1–2 mm diam; interior white, consisting of densely interwoven hyphae of 3.5–4.5 μm in width. Sporulating region distributed mainly on convex side of synnemata, forming a more light-colored and slightly fluffy region. Conidiogenous cells hyaline, clavate, apiculate on tip, 13–21 × 2.5–3.2 μm, warted. Conidia produced holoblastically in sympodial sequence, one-celled, hyaline, cylindrical to fusiform, 10–12 × 2.5–3.5 μm.
Cultures and anamorph. Colonies on PDA at 25 C attaining 1 cm in 3 wk, white, overlain with short dense aerial hyphae, diffuse at margins. Sporulation absent.
Specimens examined. I-lan County, Da-jiao-si Experimental Forest, on Planaeschna sp. (Odonata: Aeshnidae), 27 Apr 2013, Ju, Y.-M. & Hsieh, H.-M. 102042701 (HAST; culture accession number: BCRC FU30560; GenBank accession number of ITS: KT873534). New Taipei City, Wu-lai, on Planaeschna sp. (Odonata: Aeshnidae), 14 Jul 2013, Ke, Y.-H. 102071404 (HAST).
Ophiocordyceps odonatae is the only species in the genus known to parasitize dragonflies. The two studied Taiwan collections were made from the dragonfly genus Planaeschna, from which the type specimen of Hymenostilbe odonatae Kobayasi was also collected (Kobayasi 1941, 1981). Only the anamorph was present in the Taiwan collections. It should be noted that the basionym of O. odonatae is the teleomorph-typified binomial Cordyceps
odonatae Kobayasi, which is predated by the anamorph-typified binomial Hymenostilbe
odonatae. Recombining the epithet of H. odonatae with Ophiocordyceps would result in an illegitimate later homonym of O. odonatae (Kobayasi) G. H. Sung et al.
Conidiogenous cells and conidia in Taiwan collections are slightly larger than those documented in the protologue of H.
odonatae (Kobayasi 1941). Also, unlike the more or less curved conidia described in Kobayasi (1941), conidia in Taiwan collections are not curved or only slightly curved. Modes of conidiogenesis were considered an important character in separating Hymenostilbe from Akanthomyces Lebert by Samson and Evans (1975), with the former producing conidia holoblastically in sympodial sequence and the latter enteroblastically. Samson and Evans (1975) related H.
odonatae to Akanthomyces with reference to the description in Kobayasi (1941). Our study clearly shows that the anamorph of O. odonatae produces conidia holoblastically in sympodial sequence and can be accommodated in Hymenostilbe.
A MEGABLAST query in GenBank using the ITS sequence obtained from the specimen 102042701 resulted in the following five top matches: Hymenostilbe odonatae (AB104725, query coverage = 100 %, identities = 576/581 [99 %], gaps = 1/581 [0 %]); Ophiocordyceps forquignonii (HQ662164, coverage = 33 %, identity = 187/198 [94 %], gaps = 3/198 [1 %]); Cordyceps forquignonii (AJ786562, coverage = 33 %, identity = 187/198 [94 %], gaps = 3/198 [1 %]); uncultured Volutella (HM136667, coverage = 42 %, identity = 175/187 [94 %], gaps = 3/187 [1 %]); and Phialophora sp. (AJ010039, coverage = 35 %, identity = 187/205 [91 %], gaps = 4/205 [1 %]). The query result shows a good match between our materials and O.
In addition to our report in Taiwan, O. odonatae had previously been recorded in Japan (Kobayasi 1941), northeastern and southern China (Song et al. 2006; Yang et al. 2004), and New Guinea (Kobayasi 1981), being widely distributed across climate zones from temperate regions to the tropics. Future collections may reveal that O. odonatae is distributed mainly along the Western Pacific Region.