Situation of type specimens of Cordyceps and allies described by Dr Kobayasi.

Information about authentic specimens of Cordyceps spp. that were described by Dr Kobayasi were summarised. Dr Kobayasi, National Museum of Nature and Science, has described/proposed about 28% of the scientific names for the major two genera of entomopathogenic fungi (Cordyceps s. l. and Torrubiella s. l.) with Mr Shimizu before 2007. In total 44 authentic specimens were discovered at present: 19 in Torrubiella, 6 in Cordyceps spp. on Elaphomyces, 19 in Cordyceps spp. on cicada. Although the colour iconography books published by Dr Kobayasi and Mr Shimizu did not explain the information about the illustrated specimens, 22 among the discovered specimens have been illustrated in colour showing their fresh condition in the books.

Introduction

Cordyceps Forum 2016, which was held in Pinghu, provided a platform for crossover discussion on studies of Cordyceps. For studies on both the basics and the applications, the most important knowledge is the exact identification of the specimen(s). Preferably, the study specimens are compared with the type specimen for the identification. However, it is sometimes difficult to gain access to the specimens.

Dr Kobayasi (1907–1993), the Director of the Department of Botany in the National Museum of Nature and Science in Tokyo, has described more than 150 species of Cordyceps and its allies with the help of Mr Shimizu (1915–1998) who has excellent skills in collecting Cordyceps specimens. They mentioned that the type specimens were deposited in the herbarium of the museum. However, because of unfortunate situations, both of specimens (without accession numbers) and of the managing system (fund, space, human resources, data base, etc.), most of those specimens became untraceable. Since 2004, a reordering of the specimens has been conducted. Unregistered specimens scattered in several different lockers were gathered in one room. Several specimens were recognised as type candidates. Some of the specimens were identical to the colour illustrations in the books on Cordyceps spp. (Kobayasi and Shimizu 1983; Shimizu 1994). Unfortunately, no information regarding the illustrated specimens (even if it is a holotype or not) was described in the two books.

Some holotype specimens of Cordyceps on cicada had been deposited in a private museum of cicada founded by Dr Masayo Kato (1898–1967). He was a pioneer of the cicada studies in Japan and studied not only cicada taxonomy but also the natural enemies of cicadas, including Cordyceps. He asked Dr Kobayasi to identify Cordyceps specimens that were sometimes new species. After his death, the museum was closed, and all the specimens have been kept by his family. In 2010, all the specimens, including several Cordyceps specimens, were deposited by his granddaughter at the museum in the University of Tokyo.

A special effort to identify the types in the National Museum of Nature and Science was initiated since 2005. In addition, I had a chance to check the Cordyceps specimens in the museum of the University of Tokyo in 2013. The situation of the type specimens of Torrubiella and Cordyceps on cicadas and on Elaphomyces has been reported (Sato et al. 2010a, 2010b, 2012, 2014). Though Cordyceps type specimens from other host groups remain to be rediscovered, listing up the type specimens that are found is useful for taxonomy research. In this manuscript, I have prepared a list of the type specimens as a halfway report.

Scientific name proposed by Dr Kobayasi

Dr Kobayasi has both described new species and proposed new taxonomic treatments. The scientific names of both the genera Cordyceps and Torrubiella were sorted in the database “Index Fungorum (http://www.indexfungorum.org/)” during the years from 1801 to 2006. The scientific names proposed after 2007 (including 2007) were omitted because Sung et al. (2007) proposed a new system of taxonomy for the genus Cordyceps. The number of species names was counted every 10 years, and the species that were described by Kobayasi were independently counted.

Dr Kobayasi studied Cordyceps for about 50 years and has described Cordyceps species from 1939 to 1983 (Kobayasi 1939, 1983). His most important publication was Kobayasi (1941), in which he proposed a taxonomic system for global Cordyceps spp. Since the 1960s, Dr Kobayasi started to publish manuscripts along with Mr Shimizu who is an excellent collector of Cordyceps. Especially, the years between late 1970s and early 1980s were the most active (Figures 1 and 2).

Figure 1.

The total species number of every 10 years in Cordyceps.

Figure 2.

The total species number of every 10 years in Torrubiella.

There were 517 scientific names of Cordyceps in the Index Fungorum described during 1818–2006 (Scientific names without year information in the database were omitted). In total, 135 Cordyceps species name described or proposed by Dr Kobayasi were recognised. About 26% of the scientific names of Cordyceps were proposed by Dr Kobayasi, although the total number in the database may have some synonyms or new treatment at present. For the genus Torrubiella, he proposed 31 scientific names (including three new combinations), and among the 82 scientific names of Torrubiella across the world, about 38% were proposed by Dr Kobayasi. In total, about 28% of the species names for the major two genera of entomopathogenic fungi were proposed by Dr Kobayasi and his collaborator, Mr Shimizu.

Situation of type specimens

Genus Torrubiella

Dr Kobayasi described 28 species of Torrubiella. A total of 19 holotype specimens were discovered. A new name, Torrubiella plana, has been given for the species T. minutissima Kobayasi, because of homonym problem (Sato et al. 2010a). Information of the type specimens is described in Table 1.

Table 1.

The list of rediscovered type specimens of Cordyceps and Torrubiella species described by Dr Kobayasi.

No.	Name by Dr Kobayasi		ID	Status	KSa	Sb	Current name	Literature
	Torrubiella
1	T.	alboglobosa	TNS-F-12,061	Holo	64–2	352
2		aurantia	TNS-F-12,069	Holo	59–1	330
3		corniformis	TNS-F-12,064	Holo
4		elIipsoidea	TNS-F-12,055	Holo	54–2	345
5		formosana	TNS-F-12,059	Holo
6		fusiformis	TNS-F-234,548	Holo	53–3	360
7		globosostipitata	TNS-F-12,057	Holo	53–2	340
8		longissima	TNS-F-12,071	Holo	64–1	339
9		mammiIlata	TNS-F-12,060	Holo	55–2	-
10		minutissima	TNS-F-12,061	Holo	56–1	347	Torrubiella plana	Sato et al. 2010a
11		miyagiana	TNS-F-12,062	Holo	56–3	326
12		neofusiformis	TNS-F-12,058	Holo	54–3	323
13		oblonga	TNS-F-12,070	Holo	60–2	322
14		ooaniensis	TNS-F-12,063	Holo	57–1	353
15		pallida	TNS-F-12,789	Holo	58–2	328
16		rosea	TNS-F-12,065	Holo	57–2	317
17		ryogamimontana	TNS-F-12,058	Holo	58–3	341
18		ryukyuensis	TNS-F-11,932	Holo	55–3	336
19		superficialis	TNS-F-12,072	Holo
	Cordyceps on Elaphomyces
20	C.	minazukiensis	TNS-F-197,989	Holo			Tolypocladiumminazukiense	Quandt et al. 2014
21		ophioglossides var. cuboides	TNS-F-230,312	Holo			T. ophioglossoides	Quandt et al. 2014
22		valvatistipitata	TNS-F-230,284	Holo	46–3	369	T. valvatistipitatum	Quandt et al. 2014
23		×jezoensoide	TNS-F-230,286	Holo
24		delicatistipitata	TNS-F-230,293	Lecto	44–2	375	T. delicatistipitatum	Quandt et al. 2014
25		ophioglossides f. alba	TNS-F-18,223	Lecto			T. ophioglossoides	Quandt et al. 2014
	Cordyceps on Cicada
26	C.	inegoensis	TNS-F-230,289	Holo			T. inegoense	Quandt et al. 2014
27		paradoxa	TNS-F-230,313	Holo			T. paradoxum	Quandt et al. 2014
28		toriharamontana	TNS-F-230,288	Holo			T. toriharamontanum	Quandt et al. 2014
29		heteropoda	TNS-F-230,294	Holo			Ophiocordycepsheteropoda	Sung et al. 2007
30		longissima	TNS-F-230,285	Holo			O. longissima	Sung et al. 2007
31		prolifica var. terminalis	TNS-F-230,295	Holo	13–2	43	Perennicordycepsprolifica	Matočec et al. 2014
32		pseudolongissima	TNS-F-197,983	Holo			O. pseudolongissima	Sung et al. 2007
33		takaoensis	TNS-F-3026	Holo			O. sobolifera	Sung et al. 2007
34		kanzasiana	TNS-F-198,015	Holo	11–1	37	Polycephalomyceskanzashianus	Kepler et al. 2013
35		sinclairii	TNS-F-212,384	Holo			Cordyceps kobayasii	Koval’ 1984
36		minuta	TNS-F-11,933	Holo
37		ramosipulvinata	TNS-F-197,979	Holo	13–3	36	Po. ramosopulvinatus	Kepler et al. 2013
38		ryogamimontana	TNS-F-230,292	Holo			Purpureocilliumtakamizusanense	Ban et al. 2015
39		prorifica	TNS-F-230,300	Lecto			Perennicordycepsprolifica	Matočec et al. 2014
40		yakusimensis	TNS-F-230,287	Lecto	24–2	16	O. yakusimensis	Sung et al. 2007
41		pleuricapitata	TNS-F-197,965	Para
42		imagamiana	TNS-F-197,966	Authentic
43		owariensis	KATM-Fun01	Holo
44		nipponica	KATM-Fun06, Fun 13	Authentic
	Other cicada pathogenic fungi
45	Isaria	nipponica	KATM-Fun02	Holo
46	I	oncotympanae	KATM-Fun16-3	Holo
47	Synnematium	graptopsaltriae	KATM-Fun03	Holo
48	Massospora	sp.	KATM-Fun17	Authentic

TNS-F: National Museum of Nature and Science; KATM-Fun: The University Museum, University of Tokyo.

Data from Sato et al. (2010a, 2010b, 2012 and 2014).

aThe colour illustration plate number in Kobayasi and Shimizu (1983).

bThe colour illustration plate number in Shimizu (1994).

Cordyceps species on Elaphomyces

Dr Kobayasi described nine Cordyceps taxa (species and forma) on Elaphomyces, including one invalid situation that was emended by Yao et al. (1995). Four holotype specimens were discovered, and two specimens were selected as lectotypes (Sato et al. 2010b) (Table 1). One of the two was the species emended by Yao et al. (1995). Sato et al. (2010b) used the name of the genus Elaphocordyceps after Sung et al. (2007). The latest genus name is Tolypocladium (Quandt et al. 2014). Table 1 also shows the current names of each species with references.

Cordyceps species on Cicada

Dr Kobayasi described 20 Cordyceps species on Cicada, of which 19 authentic specimens, including the holotype, were discovered (Sato et al. 2012, 2014) (Table 1). From the National Museum of Nature and Science, 13 holotype specimens were discovered. Two specimens were selected as lectotypes. A paratype and an authentic specimen were also discovered (Sato et al. 2012). From the museum of the University of Tokyo, one type specimen (C. owariensis) and several authentic specimens of C. nipponica were discovered (Sato et al. 2014) (Table 1). Dr Kato’s specimens can be viewed on the web-museum of the university (http://umdb.um.u-tokyo.ac.jp/DDoubutu/katomasayo/fungi_en/index.html). Any authentic specimens of C. polycephala, the last one species, should be found.

At the same time, in the university museum, holotypes of Isaria nipponica, I. oncotympanae, Synnematium graptopsaltriae and the voucher specimen for the first record of Massospora in Japan (Kobayasi 1951) were also discovered (Sato et al. 2014).

One-to-one correspondence between the type and the illustration

When the type specimens were discovered, we compared the outline of the specimens with colour illustrations in the books (Kobayasi and Shimizu 1983; Shimizu 1994). Fortunately, we recognised 22 type specimens that were illustrated in colour in the books, at present, comprising 16 cases in Torrubiella spp., 2 in Cordyceps spp. on Elaphomyces and 4 in Cordyceps spp. on cicadas (Table 1). Colour illustrations are very useful in recognising the fresh colour of the species, when the colour(s) of the types have been lost due to prolonged preservation period.

In a special case of a dried holotype specimen (C. ramosipruvinata), at first, we could not compare the specimen with the illustration in the books because of its shrunk stromata. Fortunately, the host morphology in the illustration, especially the traits of broken legs, was identical to those parts of the holotype specimen. The very precise illustration enabled us to associate the specimen with the illustration.

Handling of formalin-preserved specimens

Almost all specimens were stored in formalin liquid. As cork plugs become deteriorated due to a long storage period, we must transfer each specimen into new vials. Formalin is generally a popular chemical used for preserving specimens for a long period. Its vapour has a strong poisonous odour. Fortunately, to prevent vaporisation and to maintain the level of the liquid volume, the surface of the formalin liquid was covered with an over-layer of liquid paraffin.

On the other hand, while transferring the specimens from the formalin liquid with the over-layer to new bottles, it was difficult to take out the specimen without contacting the liquid paraffin. An oil-absorbent sheet for machine maintenance has shown good results (T-151J, 3M Company). We cut the oil-absorbent sheet into small-size pieces of 1 cm × 1 cm, and put the pieces into the specimen bottles, repeating this process until the oil was completely removed. After transferring the specimen, the top of the formalin liquid in the new vials was re-covered with liquid paraffin (Sato et al. 2011).

Ongoing survey

Specimens from Lepidoptera and Coleoptera, the major host insects of Cordyceps, are now being ordered. Next, the specimens of Cordyceps from other host groups will follow, and then, other entomopathogenic genera, including anamorphic taxa, will be the last.