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Trans. Br. mycol. Soc. 63 (2), 289-294 (1974)

Printed in Great Britain

CHLOROSPLENIUM IN INDIA

By HARNEK SINGH

Department of Microbiology, Punjab Agricultural University,Ludhiana, Punjab

(With Plate 49 and 1 Text-figure)

An illustrated account is given of three Chlorosplenium species, on the basisof so collections made from the North-Western Himalayas and Eastern Hima­layas during extensive forays to the hills from 1964 to 1969. Fruiting bodiesare described from fresh material supplemented by others preserved dry andin formalin-alcohol. For anatomical study both free-hand and microtomesections were prepared. C. indicum sp.nov. is described.

All collections have been deposited in various herbaria (PUI, BotanyDepartment, Punjab University, Chandigarh, India; BPI, NationalFungus Collections, Beltsville, Maryland, U.S.A.; K, Royal BotanicGardens, Kew, England) as mentioned under each species.

Fries (1849) proposed the generic name Chlorosplenium based on C.chlora (Schw.) Curtis (=C. sduoeinitzii Fr.), De Notaris (1864), withoutseeing the type (Seaver, 1936), took up this genus and added C. aerugino­sum (Fr.) de Not. and C. versiforme de Not. Nannfeldt (1932) thought thatC. aeruginosum and C. chlora were not congeneric and suggested that theformer be put in Ciboria. However, Seaver (1936) published Chlorociboriaas a new generic name for Chlorosplenium aeruginosum and other similarspecies but did not provide a Latin diagnosis. This was done by Rama­murthi, Korf & Batra (1957), thus validating the name. However, a fewmonths earlier in the same year Svercek (1957) had erected a monotypicnew genus Piceomphale based on Perirabulgarioides Rab., a specieswhich wasincluded by Ramamurthi et al. (1957) among the species ofChlorociboria.This name has therefore to be rejected under Art. 63 of the InternationalCode of Botanical Nomenclature.

Korf (1959a) published a nomenclatural note indicating that Chloro­ciboria must be rejected in favour of Piceomphale but deciding also to revertto Chlorosplenium as an acceptable genus for all the species referred sofar to Chlorosplenium, Chlorociboria and Piceomphale. Finally in the light ofhis studies of various Japanese collections of Chlorosplenium chlora, Korf(1959b) clearly established that this species differs from other species ofChlorociboria and Piceomphale only in having minute asci. Thus in hisopinion, this single character does not justify a distinction at genericrank. Korf's concept has been accepted by later workers, including Dennis(1968), and also in the present paper. Thus the generic synonymy is asfollows:

Transactions British Mycological Society

CHLOROSPLENIUM FR., Summa Veg. Scand. 356 (1849).

Piceomphale Svrcek, Ceska mykologie II, 237, 239 (1957).

Chlorociboria Seaver apud Ramamurthi et al., Mycologia 49, 857 (1958),nom. illegit.

Chlorociboria Seaver, Mycologia 28, 390, 391 (1936), nom.nud.

The genus is recognized by its yellowish-green, bluish-green to olivace­ous black, mostly Cupulate and stipitate (stipe mostly centric sometimesexcentric) apothecia, 8-spored, J+ asci, and fusoid to ellipsoid, hyaline,aseptate ascospores.

Type sp.: Chlorosplenium chlora (Schw.) Curtis.Chlorosplenium has a world-wide distribution (U.S.A., 6 species­

Ramamurthi et al. 1957; Australia, 3 species - Dennis, 1958; Japan, 4species - Korf, 1958; England, 4 species - Dennis, 1956, 1968). Threespecies are recorded from India in this paper, of which two have bluish­green apothecia which turn the infected wood verdigris green and onewith olivaceous-green apothecia which lack the ability to stain infectedwood. All the collections were made on dead angiosperm and gymno­sperm wood. Illustrated descriptions have been provided for the threespecies on the basis of Indian collections.

KEY

Apothecia olivaceous green, infected wood remains unstained; asci 95-140 pmlong C. rugipes

Apothecia bluish green, infected wood stains characteristic bluish green; ascismaller than in C. rugipes

Apothecia cupulate, always centrally stipitate; asci 4'5-6'5 pm wide; Asco-spores 8-12 x 2'2-3'2 pm, biguttulate C. aerugineum

Apothecia cupulate to spathulate, centrally stipitate to eccentrically stipitate;Asci 3-4 pm wide; ascospores 5-7 x I'4-I'6pm eguttulate ..... C. indicum

CHLOROSPLENIUM RUGIPES (Pk) Ramamurthi, Korf & Batra, Bull. Nat.Sci. Mus. 4, 391 (1959) (PI. 49, fig. C; Fig. 1C, D).

Apothecia up to 1'5 cm across, highly gregarious to rarely scattered,solitary but closely appressed together, soft, fleshly, cupulate to irregularin shape, shortly stipitate; external surface brown vinaceous, squamulose,each squamule composed of heaps of globular cells, margin entire, hy­menium olivaceous green, concave, smooth; stipe small, concolorouswith the external surface, squamulose, squamules akin to those of theexternal surface. Asci 95-140 x 4'5-6 psu, 8-spored, clavate-cylindrical,tapering below into a long stem-like base, J+, apices obtuse. Ascospores7-9'5 x 2-3 tuu, mostly uniseriate, ellipsoid, hyaline, thin-walled, asep­tate, biguttulate, guttules sometimes disappearing at maturity. Paraphysesup to 2'2 flm wide, filiform, stout, hyaline, thin-walled, septate, projectingup to 15 flm beyond tips of asci. In the ascigerous region the Ectal excipu­lum is up to 90 flm thick, deep brown, textura globulosa, cells thick-walled,up to 12 X 10 usu ; medullary excipulum up to 450 flm thick, differenti-

Chlorosplenium. Harnek Singh 291

:\

F

o

E [)

Fig. I. (A, B) Chlorosplenium aeruginuem; (A) Ascospores; (B) asci and paraphyses.(0, D) C. rugipes, (0), ascospores, (D), asci and paraphyses. (E, F) C. indicum; (E), asciand paraphyses; (F) ascospores.

ated into distinct zones, outer textura porrecta, hyphae compact, up to3/tm wide, inner zone more prominent and bigger, textura intricata,hyphae loose, running in all directions, up to 3 /tm wide; hypotheciumprominent, up to 160 pm thick, deep brown, densely textura intricata,hyphae running mostly in longitudinal direction, up to 2 /tm wide. Stipeis differentiated into cortex and medulla; cortex deep brown, texturaglobulosa, cells thick-walled, up to 12/tm wide, covered externally bysquamules extending up on the ectal excipulum up to the margin ofapothecium, medulla divided into two zones, outer textura porrecta,hyphae compact, up to 3'2 /tm wide; inner zone textura intricata, hyphaerunning in the longitudinal direction only, loose, up to 3'2 /tm wide.

This species was first recorded by Batra & Batra (1963) from India.

292 Transactions British Mycological SocietyHowever, an adequate description was not provided for the species on thebasis of a Mussoorie collection. I have since then collected it from twowidely separated localities in the North-Western-Himalayas. Bothcollections are quite typical of C. rugipes. It is characterized by olivaceousgreen, short stipitate, squamulose apothecia, long, clavate-cylindricalasci, fusoid, hyaline ascospores and stout, septate paraphyses. It does notstain the infected wood.

Specimens examined. On dead angiosperm wood, Glen, Simla, H.P., 22 Sept. 1967,Harnek Singh (3455), PUI and BPI: Jabber Khet, Mussoorie, U.P. 7 Sept. 1968,Harnek Singh (3595)PUI and BPI.

CHLOROSPLENIUM AERUG1NEUM (Berk.) Sacc., Syll. Fung. 8, 318 (1889)'(PI. 49, fig. B; Fig. 1A, B).

Apothecia up to 6 mm across, up to 3'5 mm in total height, mostly gre­garious, rarely scattered, solitary, soft, fleshy, shallow cupulate to cupu­late, regularly stipitate; external surface bluish green, smooth to minutelyscurfy, margin entire; hymenium lighter-concolorous when fresh, dryingyellowish, concave; stipe up to 2'5 mm x 600 11m, cylindrical, concolorouswith external surface above, bluish black at the point of attachment,smooth to minutely roughened. Asci 70-95 x 4'5-6'5 11m, 8-spored, cla­vate, tapering below into stem-like base, J+, apices obtuse. Ascospores8-12 x 2'2-3'2 11m, mostly uniseriate, fusoid, hyaline, aseptate, thin­walled, biguttulate, guttulae invariably disappear at maturity. Paraphysesup to 1'2 11m wide, filiform, hyaline, thin-walled, aseptate, simple orbranched, projecting up to 25 11m beyond tips of asci. In the ascigerousregion the ectal excipulum is up to 120 11m thick, dark green, texturaglobulosa, cells perpendicular to the surface, thick-walled, up to 9 x 611m; medullary excipulum up to 400 11m thick in the middle decreasingtowards the margin, textura intricata, hyphae thick-walled, intertwined,loose, up to 2'2 11m wide; hypothecium narrow but distinct, up to 1511mthick, textura intricata, hyphae up to 1'2 11m wide. Stipe is differentiatedinto cortex and medulla; cortex up to 85 11m thick, textura globulosa,cells perpendicular to the surface, thick-walled, up to 9 x 6 11m, coveredby a thick opaque, dark green layer extending up to the margin of theapothecium; medulla up to 350l1m thick; textura intricata, hyphaeloose, thick-walled, intertwined, up to 2'2 11m wide.

This species was first recorded from India in 1854 by Berkeley (asPeriza aeruginea Berk.). Later Thind & Singh (1961) and Batra & Batra(1963) recorded it on the basis of few collections made from the Mus­soorie hills, India, under the name Chlorociboria aeruginosa (Oed. exGray) Seaver. I have collected from most localities in the Himalayas,where it grows luxuriantly on dead wood of both angiosperms andgymnosperms. It colours the infected wood a characteristic blue green.All my collections are quite typical of C aerugineum (Berk.) Sacco It canbe easily identified by its bluish green, stipitate apothecia, fusoid, hyaline,aseptate ascopores and filiform paraphyses.

Specimens examined. On dead wood of angiosperms and gymnosperms, Rangaroon,Darjeeling, W. Bengal, 19 Aug. 1964, Harnek Singh (305), PUI, BPI and K; Glen.

Chlorosplenium. Harnek Singh 293Simla, H.P., 23 July 1965, Harnek Singh (3008), PUI, BPI; Narkanda, H.P. 16 Aug.1965, Harnek Singh (3019), PUI, BPI; Banikhet, Dalhousie, H.P., 23 July 1966,Harnek Singh (3184), PUI, BPI; Lover's Walk, Dalhousie, H.P., 27 July 1966, HarnekSingh (3200), PUI, BPI; Banikhet, Dalhousie, H.P., 9 Aug. 1966, Harnek Singh (3235),PUI, BPI; Baloon, Dalhousie, H.P., 18 Aug. 1966, Harnek Singh (3259, 3260), PUI,BPI; Khajjiar, Dalhousie, H.P., 20 Aug. 1966, Harnek Singh (3282), PUI, BPI; Glen,Simla, H.P., Harnek Singh (3458), PUI, BPI; JabberKhet, Mussoorie, U.P., 7 Sept1968, Harnek Singh (3586), PUI, BPI.

Chlorospienium indicum Dumont, Korf & Singh sp.nov, (PI. 49,fig. A; Fig. I E, F)

Apothecia gregaria vel sparsa, stipitata, cupula vel spathula at a, careuleo-viridia.Stipes aequalis, altit 8 mm, refescens. Asci cylindrici, octospori, obturaculo judo caeru­lescente, 55-70 x 3-4 pm. Ascoporae distichae vel oblique monostichae, elongatae,eguttulatae, hyalinae, simplices, 5-7 x 1'4-1.6 ps»: Paraphyses filiformes, crassit 1'4 pm,interdum furcatae. Excipuli textura subporrectas vel textura intricata.

Ad truncis emortuis, Khadrala, Narkanda, H.P., II Octo. 1967, Harnek Singh 3490(holotypus) .

Apothecia up to 8 mm across, up to I em in total height, mostly gregari­ous, sometimes scattered, solitary, sometimes caespitose, more or lesscupulate, sometimes becoming one-sided and spathulate, soft, fleshy,bluish green, stipitate; external surface bluish green, pruinose, marginentire; hymenium concolorous, concave; stipe up to 8 mm in length,cylindrical, concolorous above, almost black at the point of attachment,minutely roughened on the outer surface. Asci 55-70 x 3-4 ps«, 8-spored,cylindrical, J+, apices narrow but obtuse. Ascospores 5-7 x 1'4-1.6 /hm,irregularly biseriate above, uniserate below, elongated, hyaline, thin­walled, aseptate, eguttulate. Paraphyses up to 1'4/hm wide, filiform, palebluish green in mass, lighter-coloured lower down, septate, thin-walled,branched, branching at lower levels, projecting up to 15 /hm beyond tipsof asci.

In the ascigerous region ectal excipulum is massive, textura sub­porrecta, hyphae impregnated with greenish amorphous matter especiallyin outer few layers, not easily discernible, up to 2·6/tm wide; medullaryexcipulum textura intricata, hyphae hyaline, glistening, loosely woven,up to 3 /hm wide; hypothecium narrow but distinct, hyphae compactlyinterwoven.

This fungus appears to be common in the North-Western Himalayasand has been collected from many different districts during extensiveforays in 1965-9. It belongs to the group of species which stain theinfected wood verdigris green. It comes near C. aeruginascens (Nyl.)Kanouse, which has cup-shaped, bluish green (sometimes assumingyellow tints) smaller apothecia (up to 5 mm across) broader asci (up to5 pm) and longer ascospores (6-10 /hm) containing oil drops. However,it does not resemble any species of the genus described by Korf (1958,1959b) or Dennis (1968). It can be easily diagnosed by the long stipitate,bluish green, pruinose, cupulate to spathulate apothecia, cylindrical,J+ asci, elongated, eguttulate, aseptate ascospores, and narrow filiform,branched paraphyses.

Additional specimens examined. On dead angiospermic wood, Batote, J. & K., 15 Oct.1966, Harnek Singh (3368), PUI, BPI; Khadrala, Narkanda, H.P., II Oct. 1967,

294 Transactions British Mycological SocietyHarnek Singh (3492), PUI, BPI; Khadrala to Rampur Road, Narkanda, H.P., I I Oct.1967 Harnek Singh (3494), PUI, BPI; Baghi, Narkanda, H.P., 13 Oct. 1967, HarnekSingh (3497), PUI, BPI; Hatoo Peak, Narkanda, H.P., 15 Oct. 1967, Harnek Singh(3502), PUI, BPI.

Thanks are due to the United States Department of Agriculture,Agricultural Research Service (Grant PL.48o), for financial help. Theauthor is deeply indebted to Dr R. P. Korf, Department of Plant Pathol­ogy, Cornell University, Ithaca, New York, U.S.A., for identification ofnew species, and to Prof. K. S. Thind, Botany Department, PunjabUniversity, Chandigarh, India, for helpful guidance and valuablesuggestion during the course of this work.

REFERENCES

BATRA, L. R. & BATRA, S. W. T. (1963). Indian Discomycetes. University of KansasScience Bulletin 44, 109-256.

BERKELEY, M. J. (1954). Decades offungi. 474. Hooker's London;Journal ofBotany6,210.DENNIS, R. W. G. (1956). A revision of the British Helotiaceae in the Herbarium of

Royal Botanic Gardens, Kew, with notes on related European species. MycologicalPapers 62, 1-216.

DENNIS, R. W. G. (1958). Critical notes on some Australian Helotiales and Ostropales.Kew Bulletin 2, 321-358.

DENNIS, R. W. G. (1968). British ascomycetes. Lehre: J. Cramer.DE NOTARIS, G. (1863)' Proposte di alcune rettificazionial Profilo dei discomiceti.

Commentario Societe Crittogamologica Italiana I (5), 357-388.FRIES, E. M. (1849). Summa vegetabilium scandinaviae. Sectio Posterior. Upsaliae.KORF, R. P. (1958). japanese Discomycetes. Notes 1-8. Science Reports Yokohama National

University 727-35.KORF, R. P. (1959a). Nomenclatural notes: III. Chlorociboria vs. Piceomphale Mycologia

51, 298-299.KORF, R. P. (1959b). japanese Discomycetes. Notes 9-16. Bulletin of the National Science

Museum, Tokyo 4, 38g-400.NANNFELDT, j. A. (1932). Studien tiber die Morphologie und Systematik der nicht­

lichenisierten inoperculaten Discomyceten. Nova Acta Regiae Societas ScientiarumUpsaliensis, ser, 4, 8, 1-369.

RAMAMURTHI, C. S., KORF, R. P. & BATRA, L. R. (1957). A revision of the NorthAmerican species of Chlorociboria (Sc1erotiniaceae). Mycologia 49, 854-863.

SEAVER, F. ], (1936). Photographs and descriptions of cup fungi. 24. Chlorociboria.Mycologia 28, 390-394.

SVRCEK, M. (1957). Piceomphale bulgarioides (Rabenh. in Kalchbr.) Svrcek Comb. n aPoznamky k Problematice diskomycetu Ombrophila strobilina v pojeti Rehmove.Ceska mykologie II, 235-24°.

THIND, K. S. & SINGH, P. (1961). The Helotiales of Mussoorie Hills: 1. Journal IndinnBotanical Society 40, 295-308.

EXPLANATION OF PLATE 49Fig. A. Chlorosplenium indicum, x 2.

Fig. B. Chlorosplenium aerugineum, x 0'75.Fig. C. Chlorosplenium rugipes, x I.

(Acceptedfor publication 28 January 1974)

Trans. Br. mycol. Soc.

A

Vol. 63. Plate 49

(Facing p. 294)