Low-Level Bioluminescence Detected in Mycena Haematopus Basidiocarps

LOW-LEVEL BIOLUMINESCENCE DETECTED IN MYCENA HAEMATOPUS BASIDIOCARPS

DAVID BERMUDES1

University of Wisconsin-Milwaukee, Center for Great Lakes Studies. 600 East Greenfield Avenue, Milwaukee, Wisconsin 53204

RONALD H. PETERSEN

Botany Department, University of Tennessee, Knoxville, Tennessee 37916

AND

KENNETH H. NEALSON

University of Wisconsin-Milwaukee, Center for Great Lakes Studies, 600 East Greenfield Avenue, Milwaukee, Wisconsin 53204

Bioluminescence has been reported to occur of low-level activity with which to make reliable in approximately 40 species of fungi (see Was correlations. Petersen and Bermudes (1992) have sink, 1948, 1978, 1979), nearly two thirds of which reported that some single spore isolates of Pa are members of Mycena. Luminescence, varying nel/us stypticus (Bull:Fr.) Karst. emit vary low among species, occurs in either the basidiocarp, levels of bioluminescence quantifiedusing a pho the mycelium, the spores, or in combinations tometer. We report here low levels of biolumi thereof (see O'Kane et al., 1990). Photometers nescence in the basidiocarps of Mycena hae have long been used to study bioluminescence matopus (Pers. :Fr.) Qm!let and compare its level (see Reynolds, 1972); however, perception of lu of mycelial luminescence with cultures of P. styp minescence in fungi has been largely by eye and/ ticus (monokaryotic and dikaryotic), Lamptero or documented photographically. Use of pho myces japonicus (Kawamura) Sing. (monokar tometric measurements has generally been re yotic and dikaryotic), Mycena citricolor (Berk. & stricted to physiological studies of common spe Curt.) Sacc. (dikaryotic), Omphalotus olearius cies and examinations of the possible occurrence DC:Fr. (dikaryotic) and Armillariella me/lea of bioluminescence in species suggested to be (Vahl:Fr.) Karst. (dikaryotic). luminous (e.g., Bermudes et al., 1990). Knowl Specimens of Mycena collected and examined edge of the distribution of fungal luminescence, photometrically were: Mycena haematopus, therefore, has been generally restricted by the Wisconsin, Milwaukee Co., Grant Park, 4-X- detection limits of the human eye. 90, coli. D. Bermudes, Milwaukee Public Mu Bioluminescence in fungi ranges from readily seum [MIL] catalog 146910; M. haematopus, detectable to dim and difficult to perceive, even Wisconsin, Green Co., Abraham Woods, 14-X- to the dark-adapted eye. Variation in human vi 90, coli. D. Bermudes; M. leaiana (Berk.) Sacc., sual acuity and the length of time allowed for Wisconsin, Milwaukee Co., Grant Park, 4-X- dark-adaptation make comparisons of total light 90, coiL D. Bermudes; and M. leaiana, Wiscon output based on visual observation difficult. Lu sin, Green Co., Abraham Woods, 14-X-90, coli. minescence is sometimes described as weak (see D. Bermudes. Mycena haematopus was found to Wassink, 1948, 1978, and references cited there possess low levels of luminescence in both young in) but there are few quantitative measurements and mature specimens. In M. leaiana, also col lected in varying stages of development, no lu 1 Current address: Yale University, School of Med icine, Infectious Diseases Section, 333 Cedar Street, minescence was detected. We used a Pacific Pho New Haven, Connecticut 06510. tometries photomultiplier coupled to an EMI type

799 800 MYCOLOGIA

FIGS. l, 2. Mycena haematopus. I. M. haematopus. A TCC 62052. grown on bread crumb agar (3% bread crumbs. 1.5% agar) under normal laboratory lighting conditions. 2. X-ray film exposed to the same culture as FIG. l showing maximum light emission occurs at the periphery of the culture. Cultures in petri plates with the lids on were inverted over the X-ray film.

9781 A phototube as previously described (Ber from above the cap and 3.7 x 1 07 quanta sec-1 mudes et al., 1990). In one collection of M. hae from below. Luminescence from the stipe mea matopus. luminescence from a single cap 17 mm sured only 6. 5 x I 05 quanta sec-1• In the second in diameter measured 1.6 x I 07 quanta sec 1 collection, luminescence from a single cap II mm in diameter measured I. 7 x I07 quanta sec-1 from above the cap and 1.5 x 1 07 quanta sec 1 from below while the stipe did not exhibit de tectable luminescence. In contrast, a basidiocarp of Panel/us stypticus approximately 10 mm av erage diameter that was collected at the Green Co. location measured 2 x I 09 quanta sec 1 from above the cap and 4.6 x 109 quanta sec 1 from below. Basidiocarps of P. st_vpticus were visible to the dark-adapted eye while those of M. hae matopus were not. However, light-emission from M. haematopus was detectable on X-ray film af 20 2 3 4 5 6 7 8 9 10 ter h exposure. CULTURE The mycelium of M. haematopus from Ger many has recently been reported to be dimly FIG. 3. Relative maximum levels of total biolu 1990), minescence observed in cultures grown on bread crumb bioluminescent (Treu and Agerer, al agar. l) Mycena haemaropus. ATCC 62052; 2) Panel though no quantitative measurements were made. Ius stypticus. ATCC 66462 (dikaryotic); 3) P. stypticus. Attempts to culture M. haematopus from the R. H. Petersen 2416-3 (monokaryotic); 4) P. stypticus. Wisconsin specimens either by spores or by tis R. H. Petersen 2416-19 (monokaryotic); 5) Lamptero sue culture (Molina and Palmer, 1982) using YM myces japonicus. R. H. Petersen 2305, polyspore-2; 6) L. japonicus. R. H. Petersen 2305-4 (monokaryotic); (0.4% yeast extract, I% malt extract, 0.4% sterile 7) L. japonicus. R. H. Petersen 2305-13 (monokar filteredglucose and 1.5% w/v agar) or MsY (2.5% yotic); 8) Mycena citricolor. ATCC 34884; 9) Om w/v molasses, 0.5% w/v yeast extract, and 1.5% phalotus olearius. University of Wisconsin Center for w/v agar) media were unsuccessful. Two cultures Forest Mycology Research HHB 2268-s; and 10) Ar 62052 mil/ariel/a mel/ea. University of Wisconsin Center for of M. haematopus were obtained, strain Forest Mycology Research GB 795-s. (ATCC) and one cultured from the tissue of a BRIEF ARTICLES 801 specimen from North Carolina, Macon Co., Blue acuity and attraction of invertebrates by the ap Valley, Forest Service rd. 79, 23-V-89, coil. S. propriate wavelengths and intensity have not been A. Gordon, field no. 1958. Both cultures were established. Further studies along the lines of also found to emit low levels of light, but only those performttd by Sivinski (1981) as well as the brighter of the two, ATCC 62052, was ca determinations of visual detection limits of the pable of exposing X-ray film after 48 h (Fros. 1, organisms hypothesized to be affected wouldbe 2). Maximum light emission occurred at the pe required in order to establish an effect. Another riphery of the culture when grown under normal hypothesis suggests that light emission is a by laboratory light conditions. Similar results were product of some other biochemical function reported by Bermudes et al. (1990) for Panellus (Buller, 1924). The presence of luminescence stypticus. Comparison of the light emission levels solely in the mycelium and base of the stipe of of some bioluminescent cultures is presented in A. mellea (Buller, 1924) might support this view. Fro. 3. The level of luminescence emitted by M. Fungal luminescence is oxygen-dependent (Bull haematopus is similar to some single-spore iso er, 1924; Hastings, 1952) and in cell-free systems lates of P. stypticus (Petersen and Bermudes, 1991) (Airthand Foerster, 1962, 1964) is stimulated by and Lampteromyces japonicus and significantly hydrogen peroxide (D. Bermudes, unpubl. re lower than dikaryotic cultures of L. japonicus, P. sults). After chemical activation, the substance stypticus, Omphalotus olearius, Mycena citrico panel isolated from P. stypticus is chemilumi lor or Armillariella me/lea. Culture origins and nescent in the presence of hydrogen peroxide the media used are given in the legend to Fros. (Nakamura et al., 1988; Shimomura, 1989), while 1, 3. the presence of other chemicals from crude ex Luminescence was not reported for the basid tracts of P. stypticus stimulated by superoxide iocarps of M. haematopus in the compilations anion have also been indicated (Shimomura, by Wassink (1948, 1978, 1979) or O'Kane et al., 1991). A relationship of the bioluminescence re (1990), nor is M. haematopus synonymous with action to lignin degradation has been suggested any of the species names listed as luminescent (Lingle, 1989) and may act as a detoxification based upon the taxonomy of Singer (1986). Lu method for peroxides, such as those formed in minescence has also been reported in the my the process. Many of the luminous fungi engage celium of morphologically similar species M. in wood and leaf litter decay, including M. hae sanguinolenta (Alb. & Schw.: Fr.) Quelet by Bo matopus. A third hypothesis for fungal lumines the (1931), but not in the basidiocarp. Photo cence, accounting for its presence in the myce metric measurements of these basidiocarps may lium and/or the basidiocarps, is the repulsion of also reveal low-level bioluminescence. heterotrophs which might otherwise ingest either The occurrence of low-level luminescence rais the fungus or its substratum (Sivinski, 1981). es questions both new and old. There are a num However, low-level bioluminescence poses the ber of nonexclusive hypotheses for the occur same difficulties to hypotheses involving an apo rence of bioluminescence of fungi discussed by sematic function as it does to ones involving Sivinski (1981). One hypothesis suggests that lu attraction. minescence occurs for the attraction of dispersal This study suggests that fungal luminescence agents (McAlpine, 1900; Ewart, 1906; Johnson, may occur more commonly than human visual 1919; Lloyd, 1974), although this has been perception has detected. Further attention to the doubted by some (e.g., Murrill, 1915; Buller, occurrence of luminescence may help reveal its 1924). However, the presence of luminescence taxonomic distribution and importance to those most strongly in the gills (e.g., P. stypticus; Buller, fungi which possess it. 1924, Watling, 1981; O'Kane et al., 1990) or spores (e.g., Mycena rorida var. lamprospora This work was supported by the National Institute Corner or M. pruinosoviscida var. rabaulensis of Environmental Health Science National Service Re Corner; see Wassink, 1978, 1979) and some at search Award Grant ES 07043 (for D.B.), the Hesler traction of invertebrates to luminous basidio Endowment Fund, University ofTennessee (for R.H.P.) carps (Sivinski, 1981) would seem to support this and by the Office of Naval Research Grant NOOO14- 88-0244 (for K.H.N.). We thank Dr. Martyn J. Dibben, view. Some specimens of M. haematopus we ex Milwaukee Public Museum for confirming the iden amined also emitted light most strongly from the tification of M. haematopus and Dr. Reiner Picard for gill-bearing side of the cap. However, the visual translation of Bothe's description of M. sanguinolenta. 802 MYCOLOGIA

Key Words: basidiocarps, bioluminescence, Mycena Murrill, W. A. 1915. Luminescence in the fungi. My haematopus cologia7: 131-133. Nakumura, H., Y. Kishi, and 0. Shimomura. 1988. Panal: a possible percursor of fungal luciferin. Tet LITERATURE CITED rahedron 44: 1597-1602.

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