CRYPTOGAMIE

Mycologie

ANCIENNE REVUE DE MYCOLOGIE

Fondée par R. Heim en 1936

Directeur scientifique: Mme J. Nicot

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TARIFS DES ABONNEMENTS Tome 16, 1995

CRYPTOGAMIE comprend trois sections: Algologie, Bryologie-Lichénologie, Mycologie.

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CRYPTOGAMIE, Mycologie est indexé par Biological Abstracts, Current Contents, Geo Abstracts,

GEOBASE, Publications bibliographiques du CNRS (Pascal)

Source : MNHN, Paris

CRYPTOGAMIE

MYCOLOGIE

TOME 15 FASCICULE 4 1994

CONTENTS

S.M. BADALYAN, S. RAPIOR, L. DOKO, J. LE QUANG, M. JACOB, J.J

SERRANO and C. ANDARY - Investigation of primary and secondary

metabolites in a chemical study of Cortinarius armillatus (Cortinaria-

сезе, Тиетоша) 2.2222 put

I. KALYANASUNDARAM, L. MENON and P. LOGANATHAN - Occurrence

of melanin in bright-spored MyxOMYCEIES. coins

G. MORENO, F. ARENAL and V. GONZÁLEZ - Agaricales growing on shores

тот репіпѕшаг Ѕраіп. ............ uhi

G. MORENO, E. HORAK & M. LAGO. - Descolea maculata Bougher

(Agaricales), first report in Europe.

B. PAUL - Some species of Pyrhium isolated from cultivated soils in northern

France ..

S.A. OMAR and A.-L.E. MAHMOUD - Post-harvest rots of tomato in relation to

lyases and mycotoxin production in vitro and in vivo.

Bibliography

Index of volume 15 eene 550220222222 2-2

BIBL. DU

MUSEUM.

PARIS

Bibliothèque Centrale Muséum

Source : MNHN, Paris

3 3001 00226839 8

Source : MNHN. Paris

Cryptogamie. Mycol. 1994, 15 (4): 223-228 223

INVESTIGATION OF PRIMARY AND SECONDARY

METABOLITES IN A CHEMICAL STUDY

OF CORTINARIUS ARMILLATUS

(CORTINARIACEAE, TELAMONIA)

S.M. BADALYAN', S. RAPIOR *,L. DOKO', J. LE QUANG’, M. JACOB’, JJ. SERRANO”

and C. ANDARY*

* Department of Ecology and Nature Protection, Faculty of Biology,

Yerevan State University, 1 Alek Manukian Str., 375049, Yerevan, Armenia

Phone: (7) 88 52 57 21 19; Fax: (7) 88 52 15 10 87 Telex: 243388 HIMN SU

* Laboratoire de Botanique, Phytochimie et Mycologie, Faculté de Pharmacie,

15 Av. Ch. Flahault, 34060 Montpellier, France

Phone: (33) 67 52 30 89; Fax: (33) 67 41 19 40

* Laboratoire de Pharmacie Galénique, Pharmacotechnie et Biopharmacie,

Faculté de Pharmacie, 15 Av. Ch. Flahault, 34060 Montpellier, France

Phone: (33) 67 54 21 58; Fax: (33) 67 54 75 33

" Laboratoire de Pharmacodynamie, Faculté de Pharmacie,

15 Av. Ch. Flahault, 34060 Montpellier, France

Phone: (33) 67 63 55 25; Fax: (33) 67 54 75 33

ABSTRACT. - The fruit-bodies of Cortinarius armillatus were investigated for polyols, sugars,

phenolic acids, alkaloids and fungal toxins using thin-layer chromatography methods. Arabitol,

mannitol, trehalose, fructose, 4-hydroxybenzoic acid, choline and cortinarine A were detected from

aqueous and methanolic extracts of the mushroom. The fungal toxins, o-amanitin, orellanine,

muscarine, muscimol and bufotenine were not observed. These chemical investigations and acute

toxicity studies in mice supported the non-toxicity of C. armillatus.

RESUME. - Une étude chimique des métabolites primaires et secondaires a été réalisée sur

Cortinarius armillatus par chromatographie sur couche mince, L'arabitol, le mannitol, le tréhalose, le

fructose, l'acide parahydroxybenzoique, la choline et la cortinarine À ont été mis en évidence dans

les extraits méthanoliques et aqueux de C. armillatus. Les toxines fongiques, a-amanitine, orellanine,

muscarine, muscimol et bufotenine n'ont pas été observées dans ces mémes extraits. La non-toxicité

de C. armillatus est confirmée par des études chimiques et pharmacologiques sur la souris.

Key-words: Cortinarius armillatus; cortinarine A; arabitol; mannitol; 4-hydroxybenzoic acid;

choline

To whom correspondence should be addressed.

Source : MNHN, Paris

224 S.M. BADALYAN etal.

INTRODUCTION

Cortinarius armillatus (Fr. : Fr.) Fr. is a large, reddish-brown edible-like

mushroom with a clubshaped stalk and irregular reddish bands, single to several, in

deciduous woods (Cetto, 1978; Marchand, 1983). To the best of our knowledge, no data

exists as to the chemical constituents of C. armillatus except pigment composition

(Besl et aL, 1978). In this work, an examination of polyols, sugars, phenolic acids,

alkaloids and other fungal secondary metabolites was undertaken using mono- and two-

dimensional thin-layer chromatography (TLC) spraying with selective reagents. On the

other hand, acute toxicity studies were also carried out and had never been previously

reported.

MATERIAL AND METHODS

1. Material

C. armillatus was collected at Regensburg in Bavaria in october 1993 and

preserved by drying after morphological identification from fresh material.

2. Methods

Preparation of extracts

20 g of C. armillatus fruit-bodies finely powdered were extracted with methanol

(100 ml x 5) by sonication (Ultra Sonik 300 NEY) at room temperature for 1 h.

The methanolic combined extracts were filtered on Durieux filter. paper n°113

and evaporated to dryness in a rotary vacuum evaporator at 35-40*C. Aliquots of the

residue were added with methanol for thin-layer chromatography analyses (polyols,

sugars, alkaloids, fungal toxins) or resuspended in carboxymethyl cellulose for

pharmacological investigations. t

The powder of mushroom was then re-extracted with water (100 ml x 3) by

sonication in the same conditions as previously described. After filtration, the

combined aqueous solutions were freeze-dried. The residue was added with water for

TLC analyses (polyols, sugars, phenolic acids).

Qualitative determination of polyols and sugars

The methanolic and aqueous extracts were analyzed on silica 60 F254 plates

(Merck, ref. 5735) according to Andary et al. (1979) in the following solvent systems

(V/V): acetone-water (9:1) and methylene chlorure-acetic acid-methanol-water

(50:5:20:5) in the same dimension. The TLC profiles of the extracts were compared

with 0.015 % polyol and 0.1 % sugar standard solutions (W/V) in ethanol-water (98:

V/V). Rf values were: arabitol, Rf = 0.48; mannitol, Rf = 0.34; fructose, Rf = 0.4

glucose, Rf = 0.40 ; galactose, Rf = 0.36 and trehalose, Rf = 0.16.

Glucose, fructose and galactose were distinguished using TLC on cellulose

plates (Merck, ref. 5577) developed in n-butanol-ethanol-water (4:1:2.2, V/V) up to 4

cm solvent front, and sprayed with 0.2% naphthoresorcinol in ethanol (W/V) with 5%

sulfuric acid (Rapior er al., 1990). The three hexoses were detected as follows; fructose,

Source : MNHN, Paris

CHEMICAL STUDY OF CORTINARIUS ARMILLATUS 225

Rf = 0.36, dark fuchsia; glucose, Rf = 0.32, turquoise blue and galactose, Rf = 0.30,

pale blue. The polyols and the disaccharide were not revealed by this method.

Qualitative estimation of phenolic acids

A 100 mg aliquot of the water extract was added with water. The solution was

adjusted to pH 3 with 10% acetic acid and partitioned with diethyl ether. The organic

phase was evaporated to dryness and the residue added with 50% methanol (2 ml). The

hydromethanolic solution was analyzed by two-dimensional TLC on cellulose plates

(Merck, ref. 5577) developed in 2% acetic aqueous acid (AA) and toluene-acetic acid-

water (60:28: TAW, V/V) (Rapior et al., 1990). The chromatograms were sprayed

with 4-nitroaniline reagent (Stahl, 1969). The TLC profile of the extract was compared

with 0.1% phenolic acid standard solutions (W/V) in methanol.

Rf values were: 4-hydroxyphenylacetic acid (Rf = 0.81 in AA, Rf = 0.40 in

TAW, mauve); 4-hydroxybenzoic acid (Rf = 0.56 in AA, Rf = 0.46 in TAW, pink); 4-

hydroxycinnamic acid or p-coumaric acid (Rf = 0.41 in AA, Rf = 0.49 in TAW, blue-

grey); 3,4-dihydroxyphenylacetic acid (Rf = 0.75 in AA, Rf = 0.11 in TAW, purple);

3,4-dihydroxybenzoic acid or protocatechuic acid (Rf = 0.63 in AA, Rf = 0.16 in TAW,

mauve-grey) and 4-hydroxy 3-methoxybenzoic acid or vanillic acid (Rf = 0.50 in AA,

Rf = 0.67 in TAW, purple).

Qualitative detection of alkaloids, quaternary ammonium compounds and

o-amanitin

The methanolic extract was chromatographed on cellulose plates (Merck, ref.

5577) using methanol-water-acetic acid (90:5:5, V/V) as mobile phase. TLC plates

were sprayed with Dragendorffs reagent modified according to Bregoff-Delwiche

(Stahl, 1969) for alkaloids and quaternary nitrogen compounds and, with sulfanilic acid

diazotised for o-amanitin, bufotenine and muscimol (Andary er al., 1977).

Concentrations of standard solutions (W/V) and Rf values were: betaine (0.2%

in methanol, Rf — 0.56, orange); choline chloride (0.25 % in methanol-water (1:1), Rf =

0.70, brown); muscarine (0.01% in methanol, Rf = 0.82, orange pale); o-amanitin

(0.1% in methanol, Rf = 0.65, brownish-pink); bufotenine hydrogenoxalate (0.1% in

methanol-water (1:1), Rf = 0.71, pink) and muscimol (0.5% in methanol, Rf = 0.52,

pink orange).

Qualitative estimation of other fungal metabolites: cortinarine A and

orellanine

The methanolic extract of C. armillatus was analyzed for cortinarine A by TLC

on silica 60 F254 support (Merck, ref. 5735) in the following monodimensional solvent

systems (V/V) : n-butanol-acetic acid-water (4:1:1, BAW) and cyclohexane-ethyl

acetate (3:1, CHEA) (Caddy ег al., 1982) and compared with a methanolic extract of

Cortinarius orellanus Fr. Cortinarine A appeared as a blue fluorescent spot after

exposure to UV light at 254 nm.

Orellanine was detected from the methanolic extract of C. armillatus by TLC on

cellulose plates (Merck, ref. 5716) in n-butanol-hydrochloric acid-chloroform-water

(40:20:15:3.8, BCCE, V/V) as mobile phase and compared with a 0.002% orellanine

standard solution (W/V) in methanol-water (1:1, V/V) (Rapior er al., 1988). Orellanine

Source : MNHN, Paris

26 S.M. BADALYAN et al.

appeared in the form of a dark spot which, after exposure for 1-3 minutes to UV light at

366 nm, produced a bluish-white fluorescence characteristic of orelline, the

photodecomposition product of orellanine.

Rf values were: cortinarine A , Rf = 0.86 in BAW and Rf = 0.50 in CHEA;

orellanine, Rf = 0.60 in BCCE.

3. Acute toxicity studies

Adult, male and female SWISS mice from 4 to 5 weeks old weighing between

24-26 g and 15-20 g respectively, procured from the Department of Pharmacology

(Faculty of Pharmacy, Montpellier) were used for these experiments. The animals were

housed 3 male and 5 female per cage (25 x 45 x 15 cm). The room temperature was

22*C and humidity 60 + 10 %. Artificial light was the only source of light and the

animals were set on a 12 hour light/dark cycle. They had free access to commercial

pelleted diet (UAR A04) and tap water. The animals had been fasted 18 hours before

they were intraperitoneally (i.p.) injected with suspensions of the methanolic extract

residue from C. armillatus in 3% aqueous carboxymethyl cellulose. The following

single doses were given to 6 male (2000, 1000, 500 and 250 mg/kg) and 8 female

(2000 mg/kg). All suspensions were prepared in such a manner that 10 ml was given

per kg of body weight. The animals were weighed on day 1, 5, 10 and 15, and were

sacrified on day 15 for anatomical observations of thoracic and digestive organs.

RESULTS AND DISCUSSION

1. Polyol and sugar contents

Polyols were generally considered independent of other primary metabolites but

they frequently coexist in mushroom extracts with free sugars, which are related

carbohydrate compounds. Using the methods by Andary et al. (1979) and Rapior et al.

(1990), arabitol and mannitol were detected from the aqueous and methanolic extracts

of C. armillatus. Both extracts contained trehalose as also reported for Cortinarius,

subgenus Leprocybe, section Orellani (Rapior et al., 1990) and Boletus (Benedict and

Tyler, 1968). Fructose was present only in the methanolic extract of C. armillatus.

2. Phenolic acid content

Chromatographic examination of the water extract from C. armillatus

demonstrated only presence of 4-hydroxybenzoic acid. It was observed as the major

phenolic acid constituent from seven Cortinarius species of section Orellani (Rapior er

al., 1990).

3. Alkaloid and nitrogen compound contents

C. armillatus contained choline. As reported for Entoloma (Maki et al., 1985),

presence of choline proved to be of no significance chemotaxinomically. Betaine was

not detected by our method.

Source : MNHN, Paris

CHEMICAL STUDY OF CORTINARIUS ARMILLATUS

4. Other fungal metabolite content

Cortinarine A was detected from the methanolic extract of C. armillatus.

Similar result was obtained on analysis of 59 Cortinarius species from the seven

subgenera of the Cortinarius genus (Tebbett and Caddy, 1983).

On the other hand, the fungal toxins, o-amanitin, bufotenine, muscarine,

muscimol and orellanine were not identified from the fruit-bodies of C. armillatus. For

this reason it is suggested that C. armillatus fruit-bodies should now be considered to

be potentially non toxic.

5. Toxicity studies

All mice survived the observation period of 15 days. Male and female mice

given single i.p. doses of the methanolic extract from C. armillatus had no toxic

symptoms and no organic pathology up to 2000 mg/kg, two weeks after dosing. This

was confirmed by the evolution of the animal weight at different doses for the same

period of time. Autopsy of animals sacrified on day 15, revealed no digestive and

pulmonary changes.

CONCLUSION

TLC screening of the methanolic and aqueous extracts of C. armillatus

revealed the presence of polyols (arabitol, mannitol), sugars (trehalose, fructose),

phenolic acids (4-hydroxybenzoic acid), nitrogen compounds (choline) and other fungal

metabolites (cortinarine A). Mice given single intraperitoneal doses (from 2000 to 250

mg/kg) of a suspension from the methanolic extract of C. armillatus had no symptoms

and no organic pathology two weeks after dosing. These chemical studies and

pharmacological investigations supported the non-toxicity of C. armillatus.

ACKNOWLEGMENTS

This work was assisted by a convention between University Montpellier I and Yerevan State

University. We thank Dr. H. Besl (Institute of Botany, Regensburg) for the identification of

Cortinarius armillatus.

REFERENCES

ANDARY C, ENJALBERT F, PRIVAT G. and MANDROU B., 1977 - Dosage des amatoxines par

spectrophotométrie directe sur chromatogramme chez Amanita phalloides Fries

(Basidiomycètes). J. Chromatogr. 132: 525-532.

ANDARY C., PERSONNE D. and PRIVAT G., 1979 - Mise en évidence et dosage du mannitol et de

l'arabitol chez les Bolets granulés (Suillus granulatus, S. luteus et S. bellinii). Ann. Fals. Exp.

Chim. 72: 527-537

Source : MNHN, Paris

228 S.M. BADALYAN et al.

BENEDICT R. G. and TYLER V. E,, 1968 - Occurrence of sugars and sugar alcohols in the

Boletaceae. Herba Hungarica 7: 17-20.

BESL H., HALBAUER R. and STEGLICH W., 1978 - Neue Anthrachinonfarbstoffe aus Cortinarius

armillatus und Cortinarius miniatopus (Agaricales). Z. Naturf C33: 294-295.

CADDY B., KIDD C.B.M., ROBERSTON J., TEBBETT LR., TILSTONE W.J. and WATLING R.,

1982 - Cortinarius speciosissimus toxins - A preliminary report. Experientia 38: 1439-1440.

CETTO B., 1978 - / funghi dal vero. Arti Grafiche Saturnia, Trento, Italy, p 213.

MAKI T., TAKAHASHI K. and SHIBATA S., 1985 - Isolation of vomiting principles from ће

mushroom Rhodophyllus rhodopolium. J. Agric. Food Chem. 33: 1204-1205.

MARCHAND A., 1983 - Champignons du Nord et du Midi. Les Cortinaires. Société Mycologique

des Pyrénées méditerranéennes, Perpignan, France, 8: p 132.

RAPIOR S., ANDARY C. and PRIVAT G., 1988 - Chemotaxonomic study of orellanine in species

of Cortinarius and Dermocybe. Mycologia 80: 741-747.

RAPIOR S., VASSAS A., TARODO de la FUENTE A., COURTECUISSE R. and ANDARY C,

1990 - Investigation of polyols, amino acids and phenolic acids in a taxonomic study of

Cortinarius, subgenus Leprocybe, section Orellani. Mycologia 82: 243-248

STAHL E., 1969 - Thin-layer chromatography. A Laboratory Handbook. Springer Verlag, Berlin,

Germany, 1041 p.

TEBBETT LR. and CADDY B, 1983 - Analysis of Cortinarius mushrooms by high-performance

liquid chromatography. J. Chromatogr. 268: 535-538

TEBBETT LR., KIDD C.B. M., CADDY B., ROBERTSON J. and TILSTONE W.J., 1983 - Toxicity

of Cortinarius species. Trans. Br. mycol. Soc. 81: 636-638.

Source : MNHN, Paris

Cryptogamie, Mycol. 1994, 15 (4):229-237 229

OCCURRENCE OF MELANIN IN BRIGHT-SPORED

MYXOMYCETES

Indira KALY ANASUNDARAM, Lata MENON and P. LOGANATHAN

Centre for Advanced Study in Botany, University of Madras,

Guindy Campus, Madras 600 025, India

ABSTRACT - In a study of spore wall pigments of eleven species of the Orders Liceales and

Trichiales - taxa which are traditionally separated from the dark-spored Orders Physarales and

Stemonitales by their variously coloured spores - it was found that even in these bright-spored

Orders, melanin was present in the spore walls of all the species. In addition, there were several

organic-solvent-extractible pigments, usually in low amounts

RÉSUMÉ - Ce travail a eu pour but l'étude des pigments pariétaux de 4 espèces appartenant aux

ordres des Liceales et des Trichiales, taxons habituellement isolés des Physarales et Stemonitales,

ordres à spores pigmentées, en raison de leurs coloration variable. Il a été démontré que les parois

des spores de ces 4 espèces. à pigmentation claire, contenait de la mélanine. En plus, la présence en

faible quantité de pigments extractibles par les solvants organiques a pu être démontrée.

KEY-WORDS - Myxomycetes, spore pigments, melanin, Liceales, Trichiales.

INTRODUCTION

The occurrence of melanin as the only pigment in the spore walls of

Myxomycetes belonging to the dark-spored orders Physarales and Stemonitales, has

been reported (Loganathan el al., 1989). We studied the spore wall pigments of some

species belonging to the Liceales and Trichiales - traditionally the bright-spored group -

on which relatively new studies have been made (Liaane-Jenson, 1965; Blackwell &

Busard, 1978; Czeczuga, 1980; Steglich er al., 1980; Kopanski el al., 1982, 1987).

MATERIALS AND METHODS

Materials

For extraction of pigments, spores are required in large quantities. The Liceales

and Trichiales generally favour temperate climates. Specimens were collected by the

first author in the foothills of the Himalayan mountains in the state of Himachal

Pradesh (H.P.), at 31°15’ to 31° 45° N latitude, 77° 25’ to 77° 60° longitude, at

elevations of 2300 to 3000 m, in September 1989. They were identified after Lister

(1911), Martin & Axelopoulos (1969) and Emoto (1977). Twelve species were selected

for study, Spore colours were determined with reference to Rayner (1970). Details of

Source : MNHN, Paris

230 I. KALYANASUNDARAM

the material are listed in Table I. One species of the Stemonitales was included for

comparison.

Table 1 - Material used

Order and specific name Herbarium N° Source

LICEALES

*Lycogala epidendrum (L.) Fries MUBL/IK/FC/185 | Fresh collection from Н.Р. (а)

Reticularia lycoperdon Bull HPUB/12750 From the collection of Dr TNL (b)

Cribraria atrofusca Martin & Lovejoy | MUBL/IK/FC/194 | Fresh collection from H.P.

TRICHIALES

Arcyria ferruginea Fuckel HPUB/12604 From the collection of Dr TNL.

A. nigella Emoto MUBL/IK/FC/182 | Fresh collection from H.P.

*A. occidentalis (Macbr.) G.Liser — | MUBL/ZIK/FC/176 x

A. stipata (Schw.) A. Lister MUBL/IK/FC/178 2

*Trichia decipiens (Pers.) Macbr. MUBL/IK/FC/173 n

T. favoginea Schum MUBL/IK/FC/170 "

T. varia (Pers.) Pers. MUBL/IK/FC/172 "

*Hemitrichia serpula (Scop.) Ros, — | MUBL 2882 Earlier collection from Coorg, Karnataka

STEMONITALES

Stemonitis fusca Roth MUBL/IK/FC/196 | Fresh collection from H.P.

* Non-melanin pigments also studied

(a) Himachal Pradesh

(b) Prof. Dr T.N. Lakhanpal, Biosciences Department, H.P. University, Shimla.

Extraction of pigments

Whole spores were used for all extractions. These were separated by manually

shaking dry sporangia in small closed boxes until the sporangial structures separated

from spores, rolled up into a ball and could be removed en masse. The spores were

collected in small test tubes, sealed and stored at 4%C until needed. Non-melanin

pigments were extracted only from four species (marked with an asterisk in Table 1),

the amount of spores collected from the others being insufficient for two types of

extraction.

Melanin was extracted by standard methods (Thomas, 1955) from pre-weighed

spores with 1 M KOH, and purified as detailed earlier (Loganathan er al., 1989). After

drying, the weight of this purified pigment was determined. Non-melanin pigments

were extracted by boiling about 10 mg of spores in 5 ml of ethanol in a closed tube for

1 h. After cooling, the material was centrifuged at 2000 rpm and the supernatant

collected. The process was repeated until no more pigment could be extracted. The

extracts were pooled and concentrated over a water bath. As very little pigment could

be extracted by this method, the method described by Steglich et al., (1980) was tried

with Trichia decipiens, of which there was a plentiful supply. Following ethyl acetate,

Source : MNHN, Paris

MELANIN IN BRIGHT-SPORED MYXOMYCETES 231

several other solvents such as methanol, acetone, diethyl ether and dimethyl sulfoxide

were tried in succession, but only part of the pigment could be extracted and the spores

still remained coloured.

Analysis

Melanin was subjected to the various physical and chemical tests as detailed in

Loganathan et al. (1989).

Non-melanin pigments were separated by thin-layer chromatography on silica

gel-G (Merck). The plates were developed with Toluene:Formic acid-ethyl

acetate:Formic acid (10:10:3) (Kopanski et al., 1982). The spots thus separated, how-

ever, were too faint in most cases for further elution and testing. The chromatographs

of T. decipiens were developed in methanol, after initial testing with the above solvent

system and a few others. A single spot which developed from each extract, was eluted

and read in a Beckmann DU-40 spectrophotometer at the range of 200 to 500 nm.

RESULTS

Melanin

A brown pigment was extracted with 1 M KOH from all the 12 species, and a

range of tests (Loganathan er al., 1989) showed the pigment in all the species to be

melanin.

The ultra-violet spectra of the melanins from the 12 species showed a fairly

uniform pattern (Fig. 1). The Liceales showed the highest absorption at 220 to 223 nm,

as most clearly exemplified in Cribraria atrofusca. In the Trichiales the peaks were

also seen at 221-223 nm, and were sharply defined in all species. A hump at 280 to 310

nm, and were sharply defined in all species. A hump at 280 to 310 nm was seen in most

of the species in both the orders, but it was not very prominent except in the Trichias

and in Hemitrichia serpula. In Stemonitis fusca, the absorption peak was seen at 222

nm.

The infra-red spectra of the melanins of the 12 species showed variations (Fig.

2). The prominent characteristics were:

a. Sharp absorption peaks at 3.3 to 3.4 um (2860 & 2930 cm”)

b. A broad band at 4.3 to 5.0 pm (2000-2400 cm”)

c. An absorption peak at 5.7 to 6.0 um (1715 cm”).

In addition to these, absorption at 4.1-4.3 um (2300-2500 cm") was uniformly

seen in all the 12 species, but this was never prominent. Based on the presence or

absence/ prominent or diffuse nature of these features, the 12 species could be broadly

separated into four groups, but this grouping had no bearing on their taxonomic status

(Table II).

The percentage of melanin in relation to the fresh weight of the spores was

determined only in two species of the Liceales, amounting to 7-10%, and four species

of the Trichiales amounting to 4.5% in Trichia and 3% in Arcyria (Table Ш). Тһе dry

weight of the spores was determined only in 7. decipiens where a separate extracion

gave 6.8% melanin.

Source : MNHN, Paris

1 KALYANASUNDARAM

Arcyria stipata

Cribraria atrofusca

i Trichia decipiens

Fig. 1 - Ultraviolet spectra of melanin

та ers from three representative species of the

250 350 450 Trichiales and Liceales, showing

WAVELENGTH (nm) characteristic absorption patterns.

Source : MNHN, Paris

MELANIN IN BRIGHT-SPORED MYXOMYCETES 233

АУ Е NS А ШӘЙ)

30 40 5:0 60 70 80 10

Group IV. Arcyria occidentalis

100

20 Grouplil. Cribraria atrofusca

2 100

o 60]

z 4

= 0

100 Group ll.Arcyria nigella

= 60

100 P 2

a Group l.Trichia decipiens

A |

3500 2500 1800 1400 1000

WA w m NE E «Wu»

Fig. 2 - Infra-red spectra of melanin from the Trichiales and Liceales, representing the four groups

described in the text.

Source : MNHN, Paris

234 I. KALYANASUNDARAM

Table II - Infra red patterns of melanins of Liceales, Trichiales and Stemonitis.

Grouping species Features

a b c

Peaksat | Troughat | Peaksat Remarks

3.3-3.4 um | 4.3-5.0 um | 5.7-6.0 um

T Lycogala epidendrum + + +

Trichia decipiens + E +

П Arcyria stipata + +

A. nigella + = + Additional peak at

9um

Trichia favoginea F : *

T. varia * х +

Stemonitis fusca * : *

m Cribraria atrofusca + = Е

Reticularia lobata + Е E

Arcyria ferruginea + E :

IV Arcyria occidentalis = + Diffuse | Additional sharp peak

at7 um

Hemitrichia serpula = + Diffuse

Table III - Summary of spore wall pigments in the twelve species studied.

Spore colour* Percentage | Number of other

Order and Species (Nearest shade: Rayner) | Melanin | (W/W) pigments in

(Fresh wt) | ethanol extract

Term Notation.

LICEALES

Lycogala epidendrum | Ви? 19"f 8 100 6

Reticularia lobata Dark brick d) * Е =

Cribaria atrofusca Umber 13m + 70 Б

TRICHIALES +

Arcyria ferruginea Bay 5k * А :

A. nigella Sepia 13"k * 3.0 -

A. occidentalis Umber 13m * - 4

A. stipata Fawn 0 + 3.0

Trichia decipiens Umber 13m + 4.5 3

T. favoginea Amber 19 + 45 =

T. varia Ochreous 13'b * = -

Hemitrichia serpula Umber 13m + - 5

STEMONITALES

Stemonitis herbatica Chesnut 5'm * : None

+ : present; - : not done

* [n a few cases, spores were somewhat faded at the time of colour determination.

Source : MNHN, Paris

MELANIN IN BRIGHT-SPORED MYXOMYCETES 235

Non-melanin pigments

The solvents for pigment extraction and for developing the chromatographs

were selected after preliminary trials with several solvent combinations. Ethanolic

extracts were analysed from Lycogala epidendrum, Trichia decipiens, Arcyria

occidentalis and Hemitrichia serpula. For the other species, the spores were used up for

melanin extraction. Boiling with ethanol did not give complete extraction, as the spores

still remained coloured. The pigments of Т. decipiens were extracted with a series of

solvents but the spores still remained coloured. Chromatographic separation of the

extracts revealed six fractions in Lycogala, three in Trichia, four in Arcyria and five in

Hemitrichia, with Rf values ranging from 0.2 to 0.9. The UV absorption of the

pigments extracted from Trichia in different solvents is presented in Table IV. When

the dried pigments were pooled together, they formed 7.5% of the dry weight of spores.

Table IV - UV absorbance of pigments extracted from Trichia decipiens.

Absorbance at (wave length in nm)

Solvent 230 250 | 270 290 300

Ethyl acetate y + E = »

Dimethyl sulfoxide - z + > >

Diethyl ether + E : + 5

Ethanol - * = A +

Water + A Е , A

DISCUSSION

Melanin

Although melanin has previously been reported in a Liceaceous species

(Loganathan er al., 1989), its occurrence in all the other species including the bright-

spored Arcyrias and Trichias, was a surprise. Melanin forms 3 to 10 per cent of the

fresh weight, or up to 7 per cent of the dry weight of whole spores. In the dark-spored

orders where melanin is the only pigment, it constitutes 7 to 15 per cent of the dry

weight of separated spore walls (McCormick et aL, 1970; Chapman er al., 1983;

Paramasivan, 1990). From our results in Trichia decipiens, there is an indication that

the melanin and non melanin pigments may occur in equal amounts.

Apparently, in the bright-spored species, melanin is masked by other pigments.

Sporangia of Trichias, still in the process of development, are of a shiny black colour

and if crushed, would exude the still semi-liquid contents as a purplish-black fluid. The

characteristic brownish yellow colour of the mature sporangia develops much later. The

non-melanin pigments could be extracted from spores before, but not after the

extraction of melanin, apparently being destroyed by the drastic procedures of melanin

extraction. It seems reasonable to presume that the bright pigments appear after the

completion of melanin synthesis, and that they occur at the surface.

Source : MNHN, Paris

236 L KALYANASUNDARAM

The UV spectra of melanin from both the Liceales and Trichiales showed a fair

degree of uniformity, with peaks at 221-223 nm. In this respect they were similar to the

melanins of the Stemonitales (Loganathan ef al., 1989). Of the hump at 280-310 nm,

seen in several species, there had been only a faint suggestion in the Physarales and

Stemonitales.

The ir. spectra showed similarities to and differences from those of the

Physarales and the Stemonitales. The absorption around 3 um (2860 8: 2930 cm’),

which was seen in all but two species, had been seen in both the dark-spored orders,

although it was not equally prominent in all the species. The broad absorption at 4-5

um, seen in Lycogala epidendrum, Trichia decipiens, Arcyria occidentalis and

Hemitrichia serpula, and as low bands in all the species, had been seen as well-defined

peaks in all the Physarales and Stemonitales (Loganathan er al., 1989). The peak at

1715 cm’, clearly seen in Lycogala epidendrum and Trichia decipiens and in a diffuse

way in some Arcyrias and Trichias, had been seen in all the members of Physarales and

the Stemonitales.

The absorption at 9 pm (1000-1300 cm"), seen in Arcyria nigella, is similar to

that described by Rast er al. (1981) as being characteristic of the GDHB melanin of

Agaricus bisporus, and we have reported it earlier in Reticularia lycoperdon

(Loganathan et al., 1989).

Non melanin pigments

The separation into 3 to 6 fractions in our study is thus in accordance with

earlier findings. Blackwell & Busard (1978) reported 3 to 6 fractions in the ethanolic

extracts of some Trichiaceous species, with considerable intra- and interspecific

variation. Steglich and his associates obtained several fractions from Arcyria denudata

(Steglich et al., 1980), Trichia floriformis and Metatrichia vesparium (Kopanski et al.,

1982, 1987). Czeczuga (1980) reported several carotenoid pigments from each of eight

species of Myxomycetes, which represented all the four major orders. In the earlier

studies, however, whole sporangia were used for pigment extraction and not the spores

alone. The chemical nature of these pigments has been definitely established only in a

few cases, through the studies of Steglich and his associates (Steglich er al., 1980;

Kopanski et al., 1982, 1987).

Considering that the yellow and brown pigments of the Liceales and Trichiales

become non-extractible after melanin extraction, and are not completely extractible

even before melanin extraction, it is possible that they occur closely bound to the

melanin molecule. Verification of such a hypothesis, however, has to await further

study.

ACKNOWLEDGEMENTS

We are very thankful to Prof, T.N. Lakhanpal of the H.P. University, Shimla and his students

Meera Thakur, Lal Singh Chauhan and Anand Sagar and to Mr and Mrs K.S. Thakur of the H.P.

Forest Service, for the help extended to the first author in the collection of specimens; to our

colleagues P. Jayaraman and N. Mubarak Ali for helping us in different ways; and to the University

Grants Commission, New Delhi, for the award of a grant (F. 3-44/86) to the first author. The third

Source : MNHN, Paris

MELANIN IN BRIGHT-SPORED MYXOMYCETES 237

author thanks the Council of Scientific and Industrial Research, New Delhi for the award of a Senior

Research Fellowship.

LITERATURE CITED

BLACKWELL M. and BUSARD A. 1978 - The use of pigments as a taxonomic character to

distinguish species of the Trichiaceae (Myxomycetes). Mycotaxon 7: 61-67.

CHAPMAN C.P., NELSON R.K. and ORLOWSKI M., 1983 - Chemical composition of the spore

case of the acellular slime mold Fuligo septica. Exp. Mycol. 7: 57-65.

CZECZUGA B., 1980 - Investigations on carotenoids in fungi. VII. Representatives of myxomycetes

genus. Nova Hedwigia 32: 347-354,

EMOTO Y., 1977 - The Myxomycetes of Japan. Sangyo Yosho Publishing Company, Tokyo.

KOPANSKI L., KARBACH D., SELBITSCHKA G. und STEGLICH W., 1987 - Vesparion, ein

Naphthaol (2,3-b]pyrandion-Derivat aus dem Schleimpilz Metatrichia vesparium

(Mycomycetes). Liebigs Annalen der Chemie 1987: 783-796.

KOPANSKI L,, LI G.R., BESL H. und STEGLICH W., 1982 - Naphthaquinone-Farbstoffe aus dem

Schleimpilzen Trichia floriformis und Metatrichia vesparium (Myxomycetes). Liebigs

Annalen der Chemie 1982: 1722-1729.

LIAANE-JENSEN S., 1965 - On fungal carotenoids and the natural distribution of spirilloxanthin.

Phytochemistry 4: 925-931

LISTER A., 1911 - The Mycetozoa. Second edition, revised by G. Lister. British Museum, London.

LOGANATHAN P., PARAMASIVAN P. and KALYANASUNDARAM L, 1989 - Melanin as the

spore wall pigment of some Myxomycetes. Mycol. Res, 92: 286-292.

MARTIN G.W. and ALEXOPOULOS C.J. 1969 - The Myxomycetes. University of Iowa Press,

Towa.

McGORMICK J.J., BLOMQUIST J.G. and RUSH H.P., 1970 - Isolation and characterization of a

galactosamine wall from spores and spherules of Physarum polycephalum. J. Bact. 104:

1119-1125.

PARAMASIVAN P., 1990 - Some physiological aspects of the life cycle of Stemonitis herbatica, Ph.

D. thesis, University of Madras, India.

RAST D.M., STUSSI H., HEGNAUER H. and NYHLEN L.E., 1981 - Melanins. /n: TURIAN G. and

HOHL H.R., The Fungal spore: morphogenetic controls. Academic Press, New York, pp.

507-531

RAYNER R.W., 1970 - A mycological colour chart. Commonwealth Mycological Institute, Kew,

Surrey

STEGLICH W., STEFFAN B.; KOPANSKI L. and ECKHARDT G., 1980 - Indole pigments from the

fruiting bodies of the slime mold Arcyria denudata. Angewandte Chemie, International

Edition in English, 19: 459-460.

THOMAS M., 1955 - Melanins. /n: PAECH K. and TRACEY M.V., Modern methods of plant

analysis, 4. Berlin, Springer-Verlag, pp. 661-675.

Source : MNHN, Paris

Source : MNHN. Paris

Cryptogamie, Mycol. 1994, 15 (4): 239-254 239

ALGUNOS AGARICALES DE LAS PLAYAS DE ESPAÑA

PENINSULAR

G. MORENO, F. ARENAL & V. GONZÁLEZ

Dpto. de Biología Vegetal, Universidad de Alcalá,

28871 Alcalá de Henares, Madrid, España.

RESUMEN - Se citan o describen catorce táxones de Agaricales s. l. que fructifican en arenas

marítimas. En general son especies poco conocidas y apenas citadas en la Península Ibérica. Desde el

punto de vista corológico son de interés para la distribución de la micoflora europea. Se aportan

microfotografias al microscopio óptico y electrónico de barrido (MEB) de sus características más

importantes. Se propone la nueva combinación Calocybe chrysenteron var. juncicola (R. Heim) G.

Moreno y la nueva variedad Melanoleuca polioleuca var. confusa G. Moreno.

ABSTRACT - Fourteen taxa of Agaricales s. 1. growing in maritime sands are recorded or described

upon, Allogether they are poorly known and most of them have only been recorded a few times in

the Iberian Peninsula. From a chorological point of view, they are interesting for the distribution of

the European mycoflora. Microphotographs, made under at the optical and scanning electronic

microscope (SEM) of their most striking features are added. One new combination, Calocybe

chrysenteron var. juncicola (R. Heim) G. Moreno and one new variety Melanoleuca polioleuca var

confusa G. Moreno are proposed.

RÉSUMÉ - Nous citons ou décrivons quatorze espèces d'Agaricales s. I. qui fructifient sur les sables

maritimes. Généralement ce sont des espèces peu connues et à peine citées dans la Péninsule

Ibérique. Du point de vue corologique elles sont intéressantes pour la distribution de la mycoflore

européenne. Nous adjoignons des microphotographies au microscope optique et électronique à

balayage de leurs caractéristiques les plus importantes. Nous proposons la nouvelle combinaison

Calocybe chrysenteron var. juncicola (R. Heim) G. Moreno et la nouvelle variété Melanoleuca

polioleuca var. confusa G. Moreno.

KEY WORDS: Taxonomy, Chorology, Psammophylous fungi, Agaricales s. l., Spain.

INTRODUCCION

Con motivo de la asistencia por parte de uno de nosotros (GM) a las

exposiciones micológicas de Santander en los años 1992 y 1993, tuvimos ocasión de

recorrer las excelentes playas de Liencres, y recoger abundante material de Agaricales

s. lato. Algunas de estas especies, junto con otras recogidas de otros lugares pero de

similar ecología, son descritas en este trabajo. Destacamos su hábitat característico

psamófilo o arenícola, y su rareza en la Península Ibérica.

Source : MNHN, Paris

240 G. MORENO, F. ARENAL y V. GONZÁLEZ

El material se conserva en el herbario AH (Dpto. Biología Vegetal, Universidad

de Alcalá de Henares).

Las microfotografías han sido obtenidas en un microscopio Nikon modelo

Optiphot, con sistema automático de fotografía incorporado y contraste de fases.

Hemos utilizado hidróxido amónico 1096, rojo congo amoniacal y reactivo de Melzer

principalmente.

Agaricus devoniensis P. D. Orton, Trans. British Mycol. Soc. 43: 177. 1960

Poco frecuente en dunas estabilizadas con Ammophila arenaria, Ribera de Cabanes

(Castellón), leg. J. Ayllón & G. Moreno, 6-ХП-1993, АН 16624; En dunas fijadas con Juniperus

phoenicia subsp. turbinata, Punta Entinas (Almería), leg. V. González, C. Illana & A. Altés, 3-XII-

1993, АН 16622; Ibidem, leg. G. Moreno, C. Illana & A. Altés, 15-11-1994, AH 16623.

Observaciones: Agaricus devoniensis se caracteriza por su sombrero

blanquecino con máculas amarillentas más o menos abundantes y persistentes en

herbario, por su porte pequeño a mediano (2-7 cm de diám.) y por el anillo, situado en

la zona media a basal del pie que rompe con facilidad, originando un segundo anillo

inferior característico.

Nuestras recolecciones coinciden con la descripción e iconografía de Cappelli

(1984).

Calocybe chrysenteron var. juncicola (R. Heim) G. Moreno, comb. nov. (Fig. 1)

richoloma chrysenteron var. juncicola R. Heim, Treb. Mus. Ci. Nat. Barcelona 15

(3): 101. 1934.

Gregario cerca de Juncus maritimus y Plantago crassifolia, Ribera de Cabanes (Castellón),

leg. A. Burguete, 17-XI-1991, AH 16514.

Sombrero de 1-1,5 cm. de diám., de color amarillo uniforme con tonos

amarillos crémeos en el centro, seco o débilmente viscoso. Margen decurvado a plano.

Láminas adnatas con lamélulas, poco apretadas, concoloras al sombrero inclusive la

arista. Pie cilíndrico, de 2,5-3,5 x 0,3-0,4 cm, concoloro al sombrero, a veces con una

pruina blanquecina en ejemplares jóvenes. Carne de sabor dulzaino y olor farinoso

típico. Color al corte no observado.

Pileipellis formada por hifas cilíndricas estrechas, 2-3 pm de diám., fibuladas,

entrelazadas. Basidios tetraspóricos, de 22-25 x 6-7,5 um, claviformes. Esporas

elipsoidales, de 5-6 x 3-3,5 (4) um, hialinas, lisas, no amiloides ni dextrinoides.

Cistidios faciales y marginales no observados. Al realizar la preparación en KOH 10%

se observa una tonalidad violácea en la solución

Observaciones: Calocybe chrysenteron var. juncicola, pertenece al complejo de

C. chrysenteron caracterizado por la pileipellis filamentosa y esporas pequeñas. C.

chrysenteron es una especie que suele fructificar en humus de coníferas y posee esporas

más pequeñas (2,5-3,5 x 2-3 um) y la carne es de color amarillento, (Kühner &

Romagnesi, 1974 ; Breitenbach & Kränzlin, 1991)

Heim (1934), describe una variedad nueva: Tricholoma chrysenteron var.

juncicola, para una recolección muy parecida por las coloraciones amarillentas del

Source : MNHN, Paris

ALGUNOS AGARICALES DE ESPAÑA 241

10pm

Fig 1. - Calocybe chrysenteron var. juncicola (AH 16514); a: fibula; b: basidio tetraspórico; c-d:

esporas.

cuerpo fructífero a Calocybe chrysenteron, y la diferencia después de consultar con

Maire, por el hábitat higrófilo, (en restos de Juncus acutus), carne al corte de color

blanquecino y esporas de 5,5 x 3-3,5 um.

Posteriormente esta variedad ha sido interpretada como especie diferente, por

Kühner & Romagnesi (1974) y Singer (1977).

El material recogido en Castellón tiene un hábitat semejante al indicado por

Heim (loc. cit). Nuestro colaborador (A. Burguete) nos envió el material (tres

carpóforos maduros) y una excelente diapositiva, pero no precisó el color de la carne, la

cual no hemos podido observar en el material desecado, por lo que no hemos podido

verificar este último carácter; sin embargo, el resto de las características, incluyendo las

dimensiones esporales, coinciden bien con la descripción original de Heim. Creemos, a

pesar de las limitaciones que conlleva tener una sola recolección, que esta especie es

muy próxima a Calocybe chrysenteron, y bien pudiera tratarse de una variedad que

crece en áreas sin vegetación arbórea, en humedales (albuferas) entre plantas higrófilas

como Juncaceae, con esporas ligeramente mayores que el tipo. El color de la carne al

corte convendría controlarlo en sucesivas recolecciones para precisar su constancia.

Estas razones nos hacen tratarlo en este trabajo de forma semejante a la de Heim (loc.

cit.) y realizar la combinación válida como variedad de C. chrysenteron.

Conocybe dunensis T. J. Wallace in P. D. Orton, Trans British Mycol. Soc. 43:192.

1960. (Fig. 2)

Abundante, pero localizado en dunas fijadas, Liencres (Santander), leg. G. Moreno, 19-X-

1993, AH 16515,

Píleo de 2-3 cm de diám., convexo-campanulado, mamelonado, liso, de color

pardo dátil a ocráceo. Margen entero a ligeramente decurvado. Láminas adnatas,

apretadas, de color ocráceo y con lamélulas. Pie de 2-9 x 0,1-0,2 cm, cilíndrico, recto,

Source : MNHN, Paris.

242 G. MORENO, F. ARENAL y V. GONZÁLEZ

Fig. 2. - Conocybe dunensis (AH 16515); a-b: cistidios; c-d: esporas.

radicante, enterrado en la arena hasta los 2/3 de su longitud, de color crema pálido, con

pruina blanquecina copiosa en su parte superior. Olor y sabor no apreciables.

Pileipellis himeniforme, formada por células claviformes de 40-65 x 17-20 um.

Esporas elipsoidales, de 12-14 x (5,5) 6-8 jun, ocráceas, con poro germinativo hialino,

apical y central. Basidios tetraspóricos. Queilocistidios lecitiformes de 23-25 x 10-12

um con cabeza de 2,5-5 um de diám. Caulocistidios semejantes a los queilocistidios

abundantes en el ápice del pie.

Observaciones: Conocybe dunensis se caracteriza por el tamaño de sus esporas,

presencia de cistidios lecitiformes en la arista y pie, y por su hábitat psamófilo

característico. C. ammophila M. Lange fructifica en semejantes áreas y se diferencia

por sus esporas de menores dimensiones (Watling, 1982).

Solamente conocemos la cita de Ortega & al. (1991), de la provincia de

Almería, aunque la medida de las esporas indicadas por dichos autores es diferente (11-

12 x 7-8 ym).

Hygrocybe conica var. chloroides (Malençon) Bon, Doc. Mycol. 15(59): 52. 1985.

Frecuente junto con Hygrocybe conicoides en dunas fijadas, Liencres (Santander), leg. G.

Moreno, 19-X-1993, AH 16574 y 16575.

Píleo de 3-6 cm de diám., cónico convexo, de color amarillo citrino uniforme o

amarillo sin tonos anaranjado rojizos. Láminas blanco amarillentas a amarillentas,

libres y con lamélulas. Pie de 4-7 x 0,5-1 cm de color amarillento, blanquecino en la

base, Olor y sabor no apreciables. Carne ennegreciendo en la madurez muy lentamente

en el sombrero, láminas y pie.

Pileipellis en cutis de hifas cilíndricas, fibuladas, de 2-5 um de diâm. Esporas

elipsoidales a faseoliformes,de 10,5-13 (15) x 5-7,5 pm, hialinas. Basidios de 35-45 x

9-11 um, claviformes y tetraspóricos.

Observaciones: Hygrocybe conica var. chloroides se caracteriza por el color

amarillento de sus cuerpos fructíferos, láminas amarillentas y hábitat psamófilo.

Hygrocybe conicoides P. D. Orton, es una especie que fructifica en los mismos

hábitats, incluso a veces juntos. Se diferencia por presentar las láminas de color rojizo a

asalmonado y el ennegrecimiento rápido de sus cuerpos fructíferos.

Source : MNHN. Paris

ALGUNOS AGARICALES DE ESPANA. 243

Hygrocybe konradii Haller, se diferencia por fructificar en praderas calizas y

poseer esporas más anchas (9-11 um.).

Hygrocybe persistens (Britzelm.) Singer, se diferencia por fructificar en humus

de bosque mediterráneo o en praderas y los basidios son bispóricos.

Hygrocybe persistens var. cuspidata (Peck) Arnolds (= H. aurantiolutescens P. D.

Orton), comparte el hábitat psamófilo y coloraciones amarillentas en sus cuerpos

fructiferos, pero estos no ennegrecen (Arnolds, 1990).

En la zona de estudio, al igual que Orton (1969), hemos observado otras

colecciones con colores variados, amarillo-anaranjados, láminas amarillas, que en un

principio podrían tratarse como H. persistens var. cuspidata. Sin embargo, algunas

colecciones (no todas) ennegrecen lentamente lo que las encuadra en H. conica s. lato.

Debemos precisar con nuevas recolecciones estos extremos, y observar la variación de

coloraciones del basidiocarpo con el grado de hidratación y con la insolación.

Hygrocybe conicoides P. D. Orton, Trans. Br. Mycol. Soc. 43: 262-263. 1960.

Muy frecuente y copioso junto con Hygrocybe conica var. chloroides en duna fijadas,

Liencres (Santander), leg. G. Moreno, 30-X-1992, AH 16625; Ibidem, 19-X-1993 y 20-X-1993, AH

16627 y 16626

Observaciones: Esta especie es muy parecida a Hygrocybe conica (Schaeff.:

Fr.) P. Kumm,, se diferencia por presentar un color rojizo a asalmonado en sus láminas

tanto en la juventud como en la madurez y un hábitat psamófilo característico.

Inocybe arenicola (R. Heim) Bon, Doc. Mycol. 12 (48): 44 (1982) 1983. (Fig. 3)

=I. fastigiata Ё. arenicola R. Heim, Genre Inocybe: 178. 1931.

Frecuente en dunas fijadas cerca de Pinus pinaster, Liencres (Santander), leg. G. Moreno,

19-X-1993, AH 16511.

Píleo de 3-7 cm de diám., convexo-campanulado a campanulado, con un amplio

mamelón central, de color crema pajizo a crema ocráceo, más claro a blanquecino en el

mamelón, con fibrillas longitudinales adpresas. Margen incurvado a plano no rimoso.

Láminas de color blanquecino, amarillo-crémeo a pardo-ferruginoso. Pie de 3-6 x 0,5-

1,5 em, cilíndrico a veces bulboso, de color blanquecino a crema pálido, enterrado en la

arena al menos 2/3 de su longitud.

Pileipellis en cutis formada por hifas fibuladas, cilíndricas, con pigmento

intracelular, Esporas elipsoidales a faseoliformes, de 12-15,5 x 6-8 um, amarillentas y

lisas. Basidios de 35-50 x 12-15 pm, tetraspéricos. Queilocistidios de 48-70 x 10-15

jm, cilíndricos, flexuosos. Caulocistidios semejantes a los queilocistidios.

Observaciones: /nocybe arenicola nos recuerda a [. fastigiata, pero las láminas

sin tonos oliváceos en la madurez, el margen pileico no rimoso y hábitat en dunas

costeras lo diferencian claramente.

Bon (1983) es el primer autor que considera la variedad de Heim como una

especie independiente. Kuyper (1986) en su monografía europea mantiene este

tratamiento.

Source : MNHN, Paris

244 G. MORENO, F. ARENAL y V. GONZÁLEZ

g^ ré =

bees e

Fig. 3. - Inocybe arenicola (AH 16511); a-b y d: queilocistidios; c: fíbulas; e-f: esporas.

Lepiota brunneolilacea Bon & Boiffard, Bull. Soc. Mycol. France 88: 18. 1972.

(Fig. 4)

Frecuente en dunas marítimas, directamente en la arena cerca de Ammophila arenaria,

Euphorbia paralias, Liencres (Santander), leg. G. Moreno, 19-X-1993,AH ; Ibidem 20-X-1993, AH

16510; Ibidem Playa de Bolonia, Tarifa (Cádiz), leg. I. Pereira, 22-XII-1993, AH 16577; Ibidem

Ribera de Cabanes (Castellón), leg. J. Ayllón & G. Moreno, 6-XII-1993, AH 16511.

Píleo de 3-5,5 cm de diám., convexo a plano convexo, mamelonado, escamoso

de color pardo rojizo a pardo lilacino, mamelón más oscuro. Margen plano, excedente.

Pie de 4-5,5 x 0,5-1,5 cm, cilíndrico, recto, ensanchandose hacia la base, de color

blanquecino con tonos liláceos o vino burdeos sobre todo hacia el ápice, el resto es

Source : MNHN, Paris

ALGUNOS AGARICALES DE ESPAÑA. 245

Fig. 4. - Lepiota brunneolilacea (AH 16510); a-d: queilocistidios; e-f: esporas.

concoloro con el sombrero. Anillo más o menos bien delimitado rojizo oscuro. Carne

blanquecina.

Pileipellis formada por una tricodermis de hifas de longitud variable, alargadas

con un sustrato basal himeniforme, pigmento intracelular presente. Esporas ovales a

elipsoidales de 9-11 (12) x 5-6 um, hialinas y dextrinoides. Queilocistidios claviformes.

Fíbulas presentes.

Observaciones: Lepiota brunneolilacea se caracteriza por su porte medio,

carnoso, tonalidades liláceas o vinosas en la pileipellis y ápice del pie, anillo sencillo y

hábitat en dunas litorales. Es una especie frecuente, segán hemos podido comprobar

personalmente en las costas españolas peninsulares.

Esta especie se conoce de Francia e Italia y ha sido descrita e iconografiada

recientemente en las obras de Candusso £ Lanzoni (1990) y Bon (1993).

Una excelente fotografía ha sido publicada por Lanzoni & Candusso (1983).

En España solo conocemos la cita de Cataluña, con una excelente fotografía de

J. Carbí, lám. 576 (Bolets de Catalunya, XII colección, 1993).

Marasmiellus mesosporus Singer, Mycologia 65:469. 1973.

=M. dunensis Robich, G. Moreno & Póder, Mycotaxon 42:181. 1991.

Copioso en rizomas y restos herbáceos de Sporobolus pungens (Poaceae), Ribera de

Cabanes (Castellón), leg. G. Moreno £: A. Burguete, 10-XI-1990, AH 12680.

Source : MNHN, Paris

246 G. MORENO, F. ARENAL y V. GONZÁLEZ

Observaciones: Marasmiellus mesosporus, es una especie muy poco citada en

la literatura micológica, y siempre sobre diversas Poaceae en dunas litorales.

En Europa se conoce de unas pocas localidades de España e Italia, (Robich &

al.,1991; 1994)

Marasmiellus trabutii (Maire) Singer, Lilloa 22: 300 (1949) 1951. (Fig. 5)

= M. caespitosus (Pat.) Singer, Pap. Michigan Acad. Sci. 32: 129 (1946) 1948

Muy abundante, disperso o en pequeños fasciculos (2-4 cuerpos fructíferos) sobre tallos de

Juncus maritimus, Ribera de Cabanes, Castellón, leg. G. Moreno £ A. Burguete, 9-XI-1990, АН

16199.

Observaciones: Marasmiellus trabutii, se caracteriza por el sombrero

blanquecino con fibrillas oliváceas a obscuras, las láminas distantes, anastomosadas de

color blanquecino, el pie cilíndrico con tonos oliváceos oscuros en toda su longitud y

hábitat característico.

Ha sido descrito ampliamente por Honrubia (1984) y Antonín & Noordeloos

(1993). Las esporas de nuestra recolección se corresponden con la morfología indicada

por Antonín & Noordeloos (loc. cit.), para Marasmiellus trabutii var. longisporus Bas

& Noordel., sin embargo las medidas se corresponden con M. trabutii var. trabutii, lo

que confirma la variabilidad de la especie.

En España es un taxon que pasa desapercibido por su pequeño tamaño y

ecología particular. Ha sido citado anteriormente por Honrubia (loc, cit.).

Fig.5. - Marasmiellus trabutii (AH 16199); a-b: pileipellis; c-d: esporas.

Marasmius epiphyllus (Pers.: Fr.) Fr., Epicr.:386. 1838. (Fig. 6)

=M. epiphyllus var. plantaginae R. Heim, Treb. Mus. Cien. nat. Barc. 15 (3): 89. 1964.

Muy abundante sobre hojas secas aún en la planta y vivas de Plantago crassifolia, Ribera de

Cabanes, Castellón, leg. J. Ayllón £: G. Moreno, 11-XI-1990, AH 16630; Ibidem, Plat de Llobregat

(Barcelona), leg. A. Mayoral, R. Póder, J. Boada, C. Illana & G. Moreno, 29-X-1993, AH 16629.

Observaciones: A nivel macro y microscópico coinciden nuestras

recolecciones con Marasmius epiphyllus. Es una especie muy abundante en hojas de

Source : MNHN, Paris

ALGUNOS AGARICALES DE ESPAÑA 247

1% Ys

SALA

1 т,

ж % уз

Fig. 6. - Marasmius epiphyllus (AH 16630); a-c: pileipellis; d: células de la pileipellis y pileocistidio;

ey g: pleurocistidios; f: basidio; h-k: esporas.

10pm

Plantago crassifolia, en áreas marítimas, sobre todo en el otoño cuando la humedad

ambiental es alta.

Coincidimos con el tratamiento taxonómico de Antonín & Noordeloos (1993) al

sinonimizar la variedad plantaginae con la variedad tipo de la especie.

Tal como indican Rivas-Martínez & Losa-Quintana (1969), en España es una

especie típica de la asociación Crucianelletum maritimae

Melanoleuca polioleuca (Fr.) Kiihner & Maire, Bull, Soc. Mycol. France 50: 18.1934.

(Fig. 7)

Aislada o dispersa en dunas marítimas con Ammophila arenaria, Liencres (Santander), leg.

G. Moreno, 19-X-1993, AH 16516; Ibidem, 20-X-1993, AH 16573; Ibidem, Oyambre (Santander),

leg. G. Moreno, 20-X-1993, AH 16571

Source : MNHN, Paris

G. MORENO, F. ARENAL y V. GONZÁLEZ

Source : MNHN, Paris

ALGUNOS AGARICALES DE ESPAÑA 249

Píleo de 4-6 cm de diám., convexo, plano convexo a ciatiforme de color gris

pardo más claro hacia el margen, más oscuro en el centro, no estriado, afieltrado a la

lupa. Láminas blanquecinas a color crema pálido, apretadas con lamélulas. Pie de 4-6 x

0,3-0,4 cm, concoloro, cilíndrico con la base engrosada. Carne al corte blanco parduzca

por encima de las láminas, parda en el centro del píleo a parda muy obscura o negruzca

en el pie y sobre todo en su base.

Epicutis formada por hifas entremezcladas de 10-15 um de diám., y sin fíbulas.

Esporas elipsoidales de 6,5-10 x 4-5,5 um, verrugosas, amiloides. Al MEB aparecen

constituidas por verrugas que se unen en cortas crestas, que a veces se unen en un

pequeño retículo. Basidios claviformes tetraspóricos. Pleurocistidios y queilocistidios

abundantes, de 50-70 x 8-12 um., de morfología variable, fusiformes a lageniformes,

generalmente no tabicados, con cristales en el ápice. Caulocistidios semejantes a los

cistidios himeniales.

Observaciones: Las recolecciones estudiadas se caracterizan por sus cuerpos

fructíferos de porte medio a grande, cistidios fusiformes a lageniformes que la

encuadran en el subgénero Macrocystis Boekhout, sect. Strictipedes Bon y esporas

alargadas (Q=1,6-1,8). Estos caracteres la encuadran en Melanoleuca polioleuca grex.

Aceptamos el sensu estricto para esta especie indicado por Bon (1991), y no

amplio y sumamente variable como Boekhout (1988), que incluye en ella M. vulgaris

Pat., taxon parecido microscópicamente pero muy diferente por el color blanquecino a

blanco crémeo de la carne del pie al corte, carácter que se manifiesta constante en las

recolecciones españolas. La coincidencia de características microscópicas de M.

vulgaris con M. polioleuca, nos hacen proponerla como una nueva variedad de M.

polioleuca especie prioritaria. Preferimos crear una variedad nueva para Melaleuca

vulgaris Pat, Hyménomycétes d' Europe: 96. 1887, especie cistidiada, que fue

confundida posteriormente por el propio Patouillard con una especie acistiada,

Melanoleuca vulgaris (Pat.) Pat., Catal. rais. pl. cellul. Tunésie :22.1897, taxón este

último que se corresponde con Melanoleuca melaleuca (Pers. : Fr.) Murrill, epíteto

prioritario.

La nueva variedad la denominamos: Melanoleuca polioleuca var. confusa G

Moreno, nov. var., differt a M. polioleuca colore carnis stipitis albido a cremeo.

Humus de Pinus sylvestris, Candelario (Salamanca), leg. M. Ladero, 25-X1-1989, AH

16706 Holotypus

Melanoleuca albifolia Boekhout, es una especie muy próxima

microscópicamente, diferenciándose por sus láminas más blancas y el sombrero muy

oscuro (rojizo pardo a sepia oliva pardo).

Melanoleuca cinereifolia (Bon) Bon y su var. maritima (Huijsman) ex Bon, son

táxones muy próximos en sus caracteres microscópicos con M. polioleuca, pero se

separan por no presentar la carne del pie obscura

Melanoleuca excissa (Fr.) Singer s. Kühner, descrita en Breitenbach & Kränzlin

(1991), posee la carne del pie con características similares a nuestra especie, pero

presenta el sombrero de color grisáceo uniforme y los cistidios son más estrechos.

Fig. 7. - Melanoleuca polioleuca (AH 16571); a-c: pleurocistidios; d-g: variación de la ornamen-

tación y placa suprahilar lisa (técnica del punto crítico)

Source : MNHN, Paris

250 G. MORENO, F. ARENAL y V. GONZÁLEZ

Omphalina galericolor (Romag.) Bon, Doc. Mycol. 19: 22. 1975. (Fig. 8)

= Omphalia galericolor Romagn., Rev. Mycol. 17: 42.1952.

En grupos dispersos a gregarios, entre briófitos, en dunas marítimas fijadas, Liencres

(Santander), leg. G. Moreno, 19-X-1993, AH 16513.

Píleo de 0,5-2 cm de diám., infundibuliforme, de color pardo a pardo miel,

estriado por transparencia, higrófanos. Margen incurvado. Láminas de color crema,

apretadas, decurrentes y con lamélulas. Pie de 1-3 x 0,1-0,3 cm, cilíndrico, central,

concoloro o más claro que el píleo.

Fig. 8. - Omphalina galericolor (AH 16513); a-c: pileipellis fibulada con pigmento en placas; d:

basidiolos; f-g: esporas.

Source -MNHN Paris

ALGUNOS AGARICALES DE ESPAÑA 251

Pileipellis formada por hifas cilíndricas, fibuladas, de .10-15 um de diám, con

pigmento parietal en placas que le dan un aspecto cebrado. Esporas elipsoidales, de 6-

10(11) x 4-5 (6) um, hialinas, no amiloides ni dextrinoides. Basidios tetraspóricos,

largamente claviformes.

Observaciones: Omphalina galericolor se caracteriza por los colores pardos,

sombrero higrófano y estriado, láminas decurrentes y por fructificar en briófitos de

dunas litorales fijadas.

Solamente conocemos la cita de Cataluña, con una excelente fotografía de J.

Carbó, lám. núm. 585 (Bolets de Catalunya, XII colección, 1993).

Psathyrella ammophila (Durieu & Lével.) P. D. Orton, Trans. Brit. Mycol. Soc.

43:180.1960.

En dunas fijadas con Cynodon dactylon (Poaceae), Playa de los Genoveses (Almería), leg,

G. Moreno, E. Gallego, C. Illana & A. Altés, 24-11-1994, AH 16621

Observaciones: Especie saprófita sobre restos de Poaceae, frecuentemente

Ammophila arenaria, en áreas litorales de la Península Ibérica.

Rhodocybe malençonii Pacioni & Lalli, Doc. Mycol. 14 (56): 56. 1984. (Fig. 9)

= Rhodocybe ammophila ( Malengon) Pacioni & Lalli, Micol, Ital. 13: 78. 1984, non R.

ammophila Horak, Sydowia 31 :61. 1978.

= Clitopilus ammmophilus Malengon, Champignons supérieurs de Maroc 2: 19-20

1975.

Un ejemplar aislado en dunas marítimas con Salsola kali, Juncus maritimus, Eryngium

maritimum y Juniperus oxycedrus subsp. macrocarpa, Playa de Bolonia, Tarifa (Cádiz), leg. 1

Pereira, 22-XII-93, AH 16512; Un ejemplar aislado junto a Ammophila arenaria, Punta Entinos

(Almería), leg. C. Illana, G. Moreno & A. Altés, 15-11-1994, AH 16628.

Sombrero de 3,5 cm de diám., plano convexo, de color blanquecino a gris pardo

claro, a gris pardo oscuro, según la insolación o que se encuentre más o menos

enterrado en la arena. Margen plano a recurvado. Láminas de color crema, de arista

sinuosa, decurrentes y con lamelulas. Pie de 3 x 1,1 cm, ligeramente excéntrico, de

color blanquecino grisáceo, radicante, ensanchándose hacia la base que se hace bulbosa.

Carne blanquecina. Olor no apreciable. Sabor amarescente.

Pileipellis en cutis formada por hifas alargadas sin fíbulas y con pigmento

parietal en placas. Esporas de (6,5) 7,5-9 x 6-7 (8) um de contorno irregular. AI MEB

aparecen anchas verrugas de distribución irregular. Basidios tetraspóricos de 30-37 x 8-

9 ym. Cistidios faciales y marginales no observados.

Observaciones: Rhodocybe malengonii se caracteriza por el sombrero

blanquecino a grisáceo, pie radicante ensanchado inferiormente, espora de contorno

irregular y hábitat psamófilo en dunas costeras.

Aparece citado de Cataluña (Tarragona), con una excelente fotografía de M.

Tabarés (Bolets de Catalunya, VII colección, 1988) y de Andalucía (Almería) por

Ortega & al. (1991).

Source : MNHN, Paris

252 G. MORENO, F. ARENAL y V. GONZÁLEZ

Source : MNHN. Paris

ALGUNOS AGARICALES DE ESPAÑA 253

Fig. 9. - Rhodocybe malengonii (AH 16512); a: hifa de la pileipellis; b: basidios: c-

variación de la ornamentación esporal (técnica del punto crítico).

: esporas; е-һ:

AGRADECIMIENTOS

Uno de nosotros (GM) agradece a la Sociedad Micológica Cántabra (Srs. A. Pérez, J. L.

Alonso y V. Castafera) la invitación para participar en sus Jornadas Micológicas. Trabajo en parte

financiado por el Proyecto de Investigación ref. 93/10, concedido por la Universidad de Alcalá de

Henares. Los autores agradecen a los Drs. M. Heykoop y F, Esteve-Raventós sus comentarios y

aportación bibliográfica.

BIBLIOGRAFIA

ANTONÍN V. and NOORDELOOS M.E., 1993 - A monograph of Marasmius, Collybia and related

genera in Europe. Part 1: Marasmius, Setulipes and Marasmiellus. IHW. Verlag.

ARNOLDS E., 1990 - Tribus Hygrocybeae (Kühner) Bas & Arnolds. In: C. Bas, Th.W. Kuyper, M.E.

Noordeloos & E.C. Vellinga, Flora Agaricina Neerlandica Vol. 3. Rotterdam, A.A. Balkema:

70-115

BOEKHOUT T., 1988 - Notulae ad Floram Agaricinam Neerlandicam-XVI. New taxa, new

combinations in Melanoleuca Pat. and notes on rare species in the Netherlands. Persoonia

13: 397-431.

BON M., (1982) 1983 - Validations de taxons. Doc. Mycol. 12(48): 44.

BON M. 1991 - Tricholomataceae (Fayod) Heim (lére partie) (Tricholomoideae et

Leucopaxilloideae). Doc. Mycol., Mém. Hors Sér. n* 2. Lille.

BON M, 1993 - Lepiotaceae Roze, Genres: Cystolepiota, Melanophyllum, Echinoderma, Lepiota,

Chamaemyces, Sericeomyces, Leucoagaricus, Leucocoprinus, Macrolepiota, Chloro-

phyllum. Doc. Mycol., Mém. Hors Sér. n° 3. Lille.

BREITENBACH J. et KRÄNZLIN F., 1991 - Champignons de Suisse. Tome 3 Bolets et champi-

gnons à lames ère partie, Edition Mykologia. Lucerne.

CANDUSSO M. e LANZONI G. 1990 - Fungi Europaei 4. Lepiota 5. 1. Saronno. Libreria editrice

Giovanna Biella

CAPELLI A., 1984 - Fungi Europaei 1. Agaricus L.:Fr. (Psalliota Fr.). Saronno. Libreria editrice

Biella Giovanna.

HEIM R., 1934 - Fungi Iberici. Observations sur la Flore Mycologique Catalagne. Treb. Mus. Ci. Nat.

Barcelona 15(3): 1-146.

HONRUBIA M.,1984 - Micromphale (Collybiopsis) trabutii (Maire) Honrubia, nov. comb., in Spain.

New Marasmiaceae Roze, family names. Crypytog. Mycol. 5: 51-58.

KÜHNER R. et ROMAGNESI H., 1974 - Flore analytique des champignons supérieurs (Agarics,

Bolets, Chantarelles). Masson et Cie., éditeurs. Paris.

KUYPER T. W. 1986 - A revision of the genus Inocybe in Europe. I. Subgenus /nosperma and

smooth-spored species of subgenus Inocybe. Persoonia suppl. vol. 3: 1-247.

LANZONI G. e CANDUSSO M,, 1983 - Alcune lepiote del litorale toscano. Boll. Gruppo Micol.

Bresadola, Trento 26: 100-118

ORTEGA, A., VIZOSO M. T. y CONTU, M., 1991 - Notas sobre la micoflora xero-termófila y

sabulícola de Andalucía (Primera pane). Doc. Mycol. 21(82): 19-42.

ORTON, P. D., 1969 - Notes on British agarics III. Notes Roy. Bor. Gard. Edinburgh 29: 5-127.

Source : MNHN, Paris

G. MORENO, F. ARENAL y V. GONZÁLEZ

RIVAS-MARTINEZ S. et LOSA-QUINTANA J. M*, 1969 - Comportement sociologique des

champignons des dunes littorales du fleuve Llobregat (Barcelone). Bull. Soc. Mycol. France

85: 235-244

ROBICH G, MORENO G. and PÓDER, R. 1991 - Marasmiellus dunensis (Marasmiaceae,

Agaricales), a new species from the European Mediterranean. Mycotaxon 42: 181-186.

ROBICH G.; MORENO G. e PÓDER; R: 1994 - Marasmiellus mesosporus Singer nome correcte di

M. dunensis Rübich, Moreno & Póder. Boll. Assoc. Micol. Bresadola, Trento (en prensa).

SINGER R., (1977) 1978 - Key for the identification of the species of Agaricales 1. Ann. Mycol. 30:

192-279

WATLING R.1982 - British Fungus Flora Agarics and Boleti. 3 Bolbitiaceae: Agrocybe, Bolbitius de

Conocybe. Edinburgh, Royal Botanic Garden.

Source : MNHN, Paris

Cryptogamie, Mycol. 1994, 15(4): 255-261 255

DESCOLEA MACULATA BOUGHER (AGARICALES),

NUEVA CITA PARA EUROPA

С. MORENO”, E. HORAK' £ M. LAGO”

* Dpto. Biología Vegetal, Univ. Alcalá de Henares,

28871 Alcalá de Henares, Madrid. Spain

* Geobotanisches Institut ETH, Herbarium, Zollikerstrasse 107.

CH-8008 Zürich. Schweiz.

Univ. de Vigo. Dpto. de Recursos Naturales,

Apdo. 874. 36200 Vigo. Pontevedra. Spain.

RESUMEN - Se describe e ilustra Descolea maculata Bougher, especie australiana que representa la.

primera cita del género y de la especie en Europa. Fructifica en Eucalyptus globulus en

repoblaciones alóctonas en Galicia, en el noroeste de España

RÉSUMÉ - Description et illustration de Descolea maculata Bougher, espéce australienne. Il s'agit

de la première citation de ce genre et de cette espèce en Europe. Descolea maculata fructifie sur

Eucalyptus globulus, introduits en Galice et dans le Nord Est de l'Espagne.

ABSTRACT - Description and illustration of Descolea maculata Bougher, Australian species. First

mention of the genus and the species in Europe. Descolea maculata was fruiting on Eucalyptus

globulus introduced in Galicia and in North West Spain.

KEY-WORDS - Descolea maculata, chorology, ecology, taxonomy.

INTRODUCCION

El género Descolea descrito por Singer (1951), para D. antarctica Singer, se

caracteriza principalmente por sus cuerpos fructíferos medios a pequeños, con un anillo

estriado, por su pileipellis himeniforme, por sus esporas ocráceas, citriformes,

ornamentadas y por fructificar en Nothofagus spp.

Posteriormente se ha recogido sobre diversos sustratos, principalmente en los

bosques subantarticos del hemisfério Sur, en la India (Himalaya) y en la región más

oriental de Eurasia (Siberia y Japón) en Pinaceae (Abies, Pinus y Taxus), Fagaceae

(Quercus, Castanopsis y Nothofagus) y Myrtaceae (Eucalyptus, Leptospermum y

Melaleuca) principalmente. En Nueva Zelanda fructifica en bosques de Nothofagus

(Fagaceae) y Leptospermum (Myrtaceae); en Japon en Pinus y Larix (Pinaceae) y en

Quercus y Castanopsis (Fagaceae) (Horak, 1971). En la India en bosques de coníferas

puras de montaña (Abies, Pinus, Picea y Taxus).

Source : MNHN, Paris

256 G. MORENO, E. HORAK y M. LAGO

Su comportamiento como un hongo ectomicorrizógeno facultativo o saprófito

habia suscitado controversias desde su creación, Bougher & Malajczuk (1985) en

recolecciones sobre Eucalyptus spp. confirman su comportamiento ectomicorrizógeno.

Descolea es un género que se puede confundir con especies de Pholiotina

Fayod y con Rozites P. Karst, y que según Hawksworth & al. (1983), es un

representante de la familia Cortinariaceae. Sin embargo, Singer (1986), lo considera

incluido en la familia Bolbitiaceae. Amplia información al respecto podemos

encontrarla en el trabajo de Horak (1971).

Actualmente se conocen descritas nueve especies de este género, que son:

Descolea antarctica Singer y D. pallida Horak (en Nothofagus en el sur de América),

D. majestica Horak (en Nothofagus en Nueva Zelanda), D. recedens (Cooke & Massee)

Singer (en Eucalyptus en Australia), D. gunnii (Berk.) Horak (en Nothofagus y

Leptospermum en Nueva Zelanda), D. pretiosa Horak (en Abies, Pinus y Taxus en el

Himalaya, India), D. flavo-annulata (Vasilieva) Horak (en Larix, Pinus, Quercus y

Castanopsis en Siberia , Japón y Corea), D. rheophylla (Bertault & Malençon)

Malençon (en repoblaciones de Eucalyptus en Marruecos) y D. maculata Bougher (en

Eucalyptus de Australia), y una especie innominada descubierta recientemente en las

islas Hawaii (Horak & Desjardin, Agaricales of the Hawaiian Islands. 3. Mycologia, en

prep.).

El género Descolea, es un género típico del hemisferio Sur, y su presencia en

el hemisferio Norte (España y Marruecos) debe interpretarse como esporádica, y

posiblemente se traten de especies alóctonas, importadas con las repoblaciones de

Eucalyptus, con las semillas y árboles vivos. Así parece confirmarlo la presencia de

Descolea maculata en España, descrita recientemente de Australia en Eucalyptus

diversicolor y E. marginata (Bougher & Malajczuk, 1985). Sin embargo D. rheophylla

(Bertault & Malengon) Malengon, aparece en la actualidad como una especie solo

conocida en Eucalytus.globulus en Marruecos (Malengon, 1979), pero creemos que es

muy probable su presencia en Australia.

DESCOLEA MACULATA BOUGHER, Aust. J. Bot. 33: 620. 1985.

(Figs. 1-3)

Material estudiado: ESPANA: Galicia, Pontevedra, zona costera ria de

Arosa, en restos de Eucalyptus globulus, 6.11.1993, leg. M. Lago, (AH 16707, ZT

5323). AUSTRALIA: Western Australia: Perth, Floreat Park, in glasshouse pots with

Eucaliyptus diversicolor and E. marginata, 17.V.1983, leg. N. Malajczuk, (Isotypus

Descolea maculata, ZT 1843 ); same region, on soil, under Melaleuca sp. 28.VI.1985,

leg. True (ZT 2762).

Sombrero de (1)1,5-3,5 (5) cm de diám., convexo a plano convexo, con un

amplio mamelón obtuso central, de color pardo dátil a pardo rojizo, con tintes o

irisaciones doradas, con escuamulas muy pequeñas a subliso, sin fibrillas, seco, estriado

a debilmente acanalado, más marcado en periodos húmedos. Láminas adnatas a

sublibres, con lamélulas de color ferruginoso a pardo rojizo, más intenso que el

sombrero. Pie de 2-4 x 0,2-0,4 cm, cilíndrico a muy ligeramente ensanchado en la base,

Source : MNHN, Paris

DESCOLEA MACULATA 257

Fig. 1.- Descolea maculata AH 16707: a. cuerpos fructíferos. Barra 1 cm; b. trama laminal regular

Barra 20 um; c. sección de la pileipellis. Barra 20 jm; d.-e. detalle pileipellis. Barra 10 pm; f.

queilocistidios. Barra 10 pm; g. basidios. Barra 10 um; e, esporas. Barra 10 um.

concoloro al sombrero, fibriloso en el ápice. Anillo membranoso, con tonos amarillo

dorados, estriado exteriormente, en la madurez se desprende facilmente del pie, pero no

llega a caer.

Source : MNHN, Paris.

G. MORENO, E. HORAK y M. LAGO

Fig. 2.- Descolea maculata AH 16707: a-d. detalle ornamentación esporal al MEB. Barra lum.

Esporas de 10,5-12,5 x 6-7 pm (basidios tetraspóricos), -13,5 x -8 um

(basidios bispóricos), amigdaliformes a citriformes con una papila apical más o menos

marcada, ocráceas, sublisas a ligeramente punteadas al microscopio óptico. Al MEB la

MNHN. Paris

DESCOLEA MACULATA 259

Fig. 3.- Descolea maculata ZT 2762: a-b. ornamentación esporal al MEB. Barra lum; ZT 1843

Isotypus. c-d. omamentación esporal al MEB. Barra lum.

ornamentación esporal aparece formada por cortas crestas y pequeñas verrugas que

forman un subreticulo muy poco marcado, carece de placa suprahilar. Basidios de 28-

42 x 8-10 um, claviformes generalmente tetraspóricos raramente bispóricos, a veces

con un pigmento amarillo ocráceo plasmatico. Cheilocistidios de 20-55 x 5-10 um, de

Source : MNHN, Paris

260 G. MORENO, E. HORAK y M. LAGO

morfología variable, la mayoria cilíndricos o subclavados. Pleurocistidios ausentes,

Pileipellis himeniforme a epitelial, formada por células claviformes o globosas en

cortas cadenas, de 12-30 um de diám., con pigmento incrustado o vacuolar amarillo

ocráceo. Escamas del velo compuestas de hifas cilíndricas y cortas, de paredes gruesas,

fuertemente incrustadas con pigmento pardo amarillo, de 5-10 um de diám. Trama

laminal formada por hifas de 5-10 um de diám., tipicamente regular. Fíbulas presentes.

Observaciones: Descolea maculata, fructifica en restos lenosos, hojarasca y

tocones de Eucalyptus globulus, arbol alóctono introducido en Galicia en diferentes

repoblaciones forestales. Esta especie ha sido observada muy abundante (Castro, com.

pers.) en toda la provincia de Pontevedra (A. Guarda, Camposancos; Vigo, A. Guia;

Redondela, Rande), durante el otoño y el invierno, especialmente en el año 1993.

En el terreno puede confundirse con representantes de Pholiotina (Conocybe)

grupo blattaria, pero se diferencia principalmente por la morfología y ornamentación

esporal.

Descolea maculata se caracteriza principalmente por su pileipellis formada

por células globosas a piriformes, que se reunen en cortas cadenas, y por sus esporas

citriformes, sin apenás ornamentación visible al microscopio óptico. Esporas rugulosas

hasta lisas han sido descritas en Descolea phlebophora Horak, recogida en Nueva

Zelanda y Tasmania. En el resto de especies conocidas, la ornamentación esporal

observada al microscopio óptico es más fuerte, y está formada por verrugas bien

marcadas conectadas por crestas (Bougher, 1987).

El material australiano de Descolea maculata, es más robusto que el español

(sombreros de 2-5 cm de diám. y pie de 3-6 x 0,3-1,5 cm) y los queilocistidios son

ligeramente más cortos y claviformes (15-30 x 5-12 um), pero el resto de

características, sobre todo el hábitat, tamafio esporal y ornamentación son similares. La

morfología esporal es variable en el material español donde abundan las esporas

citriformes, en el material australiano son más frecuentes las esporas elipsoidales a

amigdaliformes, sin embargo en todas las muestras estudiadas existen las tres

posibilidades (elipsoidales, citriformes y amigdaliformes), en mayor o menor grado.

Las pequeñas variaciones morfológicas en las muestras australianas y españolas, no nos

parecen suficientemente significativas, y preferimos considerarlas como la misma

especie, siendo el caracter más importante y constante en las muestras estudiadas la

ornamentación esporal.

AGRADECIMIENTOS

Nuestra agradecimiento a la DGICYT por la concesión del proyecto de investigación n*

PB91-0165, dentro del cual se enmarca dicho trabajo. A la Dra. M. Castro por los datos corológicos y

fotografía del material estudiado. Al Dr. N. Bougher por el envio de material Isorypus de Descolea

maculata. A J. A Pérez y A. Priego, del Servicio de Microscopía Electrónica de la Universidad de

Alcalá de Henares, su apreciable ayuda al microscopio electrónico de barrido. Por último, a A. Dreze

(Jardin Botanique National de Belgique) por su ayuda en los cortes histológicos.

Source : MNHN. Paris

DESCOLEA MACULATA 261

BIBLIOGRAFÍA

BOUGHER, N,, (1987). The systematic position and ectomycorrhizal status of the fungal genus

Descolea. Ph. D. Thesis, Univ. Western Australia, Perth. 233 pp.

BOUGHER, N. L. & N. MALAJCZUK (1985). A new species of Descolea (Agaricales) from

Western Australia, and aspects of its ectomycorrhizal status. Aust. J. Bor. 33: 619-627.

HAWKSWORTH, D. L, B. C. SUTTON & G. C. AINSWORTH (1983). Ainsworth & Bisby's.

Dictionary of the fungi. Commonwealth Mycological Institute. Kew. Surrey.

HORAK, E., (1971).- Studies on the genus Descolea Sing. Persoonia 6: 231-248.

MALENÇON, G., (1979).- Champignons du Maroc. Sydowia Beih. 8: 258-267.

SINGER, R., (1951).- Descolea antarctica, género y especie nuevos de Tierra del Fuego (New

genera of fungi). Lilloa 23: 255-258.

SINGER, R., (1986).- The Agaricales in the modern taxonomy. 4° ed. Vaduz, J. Cramer, 981 pp.

Source : MNHN. Paris

Cryptogamie, Mycol. 1994, 15 (4): 263-271 263

SOME SPECIES OF PYTHIUM ISOLATED FROM

CULTIVATED SOILS IN NORTHERN FRANCE

Bernard PAUL

Laboratoire de Mycologie, UFR Science, Université de Bourgogne,

B.P. 138, 21004 Dijon, France.

ABSTRACT - Nine species of Pyrhium including P. echinulatum, P. irregulare, P. mamillatum, P.

minor, P. ostracodes, P. torulosum, P. oligandrum, P. ultimum, and P. rostratum isolated from

cultivated soils in northern France are described. Some of these are new to this country. Taxonomic

and morphological details of the 9 species of fungi are discussed

RÉSUMÉ - Neuf espèces de Pythium notamment P. echinulatum, P. irregulare, P. mamillatum, P.

minor, P. ostracodes, P. torulosum, P. oligandrum, P. ultimum, et P. rostratum ont été isolées à partir

de sols cultivés dans le nord de la France. Quelques unes d'entre elles s'avèrent étre nouvelles pour

ce pays. Les détails morphologiques et taxonomiques de ces neuf espèces font l'objet de ce présent

article.

KEY WORD - Pyrhium, sporangia, oogonia, antheridia, oospore.

INTRODUCTION

A perusal of the literature on the genus Pythium shows that very little work has

been done on this important genus in France. Although works on the ecological and

pathological aspect of Pythium have been carried out (Roze & Cornu, 1869; Moreau &

Moreau, 1958; Bouhot, 1975; Montfort & Rouxel, 1988), have a description of Pythium

violae as the causal organism for the «cavity spot disease» of carrots, and that of Forbes

& Davet (1990) on the pathogenicity of Pythium ultimum, P. sylvaticum, and P.

irregulare on soyabean roots, but none of these have a taxonomic treatment of the said

species. An attempt is underway towards the taxonomy of the genus Pythium in France.

A project on the isolation, identification, and preservation of pythiaceous fungi has

been undertaken at the university of Bourgogne in Dijon. A number of soil samples

from the region of Lille and Compiégne were examined and in this report nine species

of Pythium are treated. All the cultures of the described fungi are being maintained at

the Laboratoire de Mycologie, Université de Bourgogne, Dijon, France.

MATERIALS AND METHODS

Soil samples were collected in sterilized capped bottles and brought to the

laboratory. Fungi were isolated by baiting with boiled hemp-seed halves introduced to a

Source : MNHN. Paris

264 B.PAUL

soil suspension in water (Paul, 1986, 1987). Temperature/growth relations were

observed on potato carrot agar (PCA) and corn meal agar (CMA). Benomyl (5 mg/l)

was used to suppress the growth of Fusarium like fungi (Paul, 1991), Identification was

done with the help of keys and descriptions of Middleton (1943), Waterhouse (1967),

Plaats-Niterink (1981) and Dick (1990).

RESULTS AND OBSERVATIONS

Pythium torulosum Coker & Patterson (Figs. 1-5, & Fig. 59).

Colonies on CMA and PCA submerged, on PCA showing a rosette pattern and

growing with an average daily growth rate of 15 mm at 25°C. Main hyphae upto 5-6 ym

wide. Sporangia consisting of filamentous inflated, toruloid elements, vesicles and

zoospores readily formed between 15-20°C, encysted zoospores measure about 7 um in

diam. Oogonia terminal or at times intercalary, globose or sub-globose, 12-25 (x x 19

+ 0.6) um diam. Antheridia mostly monoclinous, 1-2 per oogonium, antheridial cells

making apical contact with the oogonia. Oospores globose, plerotic, single, 10-24

(x .16 0.6) um diam. Wall 1-2,5 um in thickness.

Pythium torulosum was isolated only twice from the region of Lille. The above

description is that of F-80. Apart from slightly bigger oogonia and oospores, all the

other characters of this isolates resembles with the description of P. torulosum found

elsewhere (Plaats-Niterink, 1981). This is the first taxonomic treatment of the species

in France.

Pythium rostratum Butler (Figs 6-9)

Colonies on CMA and PCA submerged, showing a chrysanthemum patten on

PCA. It is a slow growing fungus with an average daily rate of 8.5 mm at 25°C on this

medium. Main hyphae upto 7-8 um wide. Sporangia globose, ovoid, limoniform to

cylindrical, terminal, intercalary, or catenulate, 18-32 um diam. (x 263 + 0.6).

Oogonia smooth walled, mostly intercalary or in chains, globose, ovoid, limoniform,

ellipsoidal, 17-32 (x 23.9 + 0.6) um diam. Antheridia mostly hypogynous,

monoclinous and sessile, 1-2 per oogonium. Oospores globose, rarely oval, plerotic

and aplerotic, usually single, rarely two per oogonium, 16-25 (x 19.6 + 0.6) um diam.

Wall 1.5-3 um thick.

Pythium rostratum was originally isolated from garden soil in France (Butler,

1907). It is a very common species inhabiting the soil and was isolated from soil

samples taken in the Compiègne as well as the Lille areas. Most of the morphological

characters of the above described isolate (F-83) resemble to those found in the

literature. The only difference worth mentioning is the presence of longer somewhat

rectangular sporangia which can attain a length of up to 40 um instead of 27 um

recorded by Plaats-Niterink (1981).

Source : MNHN, Paris

PYTHIUM FROM NORTHERN FRANCE 265

ТІП

Figure 1-5: Pythium torulosum. 1: sporangia, 2-3: oogonia with monoclinous antheridia, 4-5:

Oospores; Figs 6-9: Pythium rostratum, 6: intercalary oogonia, 7: oogonium with hypogynous

antheridia, 8-9: oospores; Figs 10-13: Pythium minor. 10: hyphal body, 11: oogonia with branched

antheridia, 12-13; oospores; Figs 14-19: Pythium ostracodes. 14-15: sporangia, 16-17: oogonia:, 14-

1S: sporangia, 16-17: oogonia with antheridia, 18-19: oospores; Figs 20-23: Pythium irregulare. 20:

smooth walled oogonia, 21: spiny oogonia with antheridia, 22-23: spiny oogonia; Figs 24-26:

Pythium ultimum. 24: hyphal body, 25-26: oogonia with antheridia

Source : MNHN, Paris

266 B. PAUL

30 um

Source : MNHN, Paris

PYTHIUM FROM NORTHERN FRANCE 267

Pythium minor Ali-Shatayeh & Dick (Figs 10-13, & Figs 52-53).

Colonies on CMA and PCA submerged, showing an indistinct rosette pattern on

PCA. Daily growth rate on PCA at 25°C is 11 mm. Main hyphae upto 6 um wide.

Sporangia and zoospores not formed. Hyphal bodies produced abundantly on solid as

well as in water on hemp seed halves. These are usually intercalary, catenulate, rarely

terminal, globose, ovoid to cylindrical, the elongated ones are at times hardly thicker

than the vegetative hyphae but filled with denser protoplasm, measuring 9-20 (x 12.4 +

0.6) um diam. Oogonia terminal, infrequently intercalary, globose, ovoid, smooth

walled, measuring 14-25 (x 15.9 + 0.6) um diam. Antheridia monoclinous, much

branched, giving a coralioid appearance and growing towards the oogonium, providing

1-2 antheridial cells to the latter. Oospores globose, 1-3 per oogonium, plerotic, rarely

aplerotic, 10-15 (x 12.7 € 0.6) um diam. with a very thin wall of 0.5-1 um. Description

isolate no. F-10.

Pythium minor is a very common species in the north of France. It was isolated

twice in Compiègne and 5 times from soil samples taken at different places in the

region of Lille. After its discovery in England (Ali-Shtayeh & Dick, 1985) it has not

been reported from elsewhere. Branched antheridia forming a corolloid structure

around the oogonia, small oogonia and oospores, and slow growth makes it easily

distinguishable from other species. However there are some differences between this

isolate and the one described by Ali-Shtayeh & Dick (1985): the hyphal bodies in this

case are much smaller (9-20 pm instead of 20-40 um). More than 3 oospores per

oogonium were not observed in the isolates from France as compared with 6 from that

described from England. This is the first report of its occurrence in France.

Pythium ostracodes Drechsler (Figs 14-19, & 54-55).

Colonies on CMA and PCA producing abundant aerial mycelium. Daily growth

rate on PCA at 25°C 8-9 mm. Main hyphae upto 7 um wide. Sporangia spherical,

ovoid, at times with an apical papilla 13-33 um diam. (x 23 + 0.6). Oogonia smooth

walled, globose, terminal or intercalary, 15-40 (x 26 + 0.6) um diam. Antheridia 1-2

per oogonium, monoclinous with long antheridial cells that apply to the oogonium to

most of its surface, antheridial cells upto 25 um long and 6 um wide. Oospores

globose, plerotic, one per oogonium, 14-39 (x 24 + 0.6) um diam., and provided with a

very thick wall of 4-7 um.

Pythium ostracodes isolated only once, from a soil sample taken from a wheat

field on Lille (No F-66). It was first described from wheat in Texas (Drechsler, 1943)

and later on it was isolated from rhizomes of latus in Japan (Takahashi et al., 1965).

This is the first report of its presence in France. This species can be separated from

Fig. 27-33: Pyrhium oligandrum. 27-31: irregular elements of the contiguous sporangia, 32: terminal

oogonia, 33: aplerotic oospores; Figs 34-43: Pythium mamillarum. 34-38: sporangia, 39-41

omamented oogonia with antheridia, 42-43: oospores. Figs 44-51: Pythium echinulatum. 44:

vegetative hyphae, 45-47: sporangia, 48: oogonia with hypogynous antheridia, 49: oogonia with

monoclinous antheridia, 50: oogonia containing two oospores, 51: oogonia with single oospore.

Source : MNHN. Paris

268 B.PAUL

other species by its slow growth, large plerotic oospores, and long, laterally applied

antheridia. This isolate has all these characters, but it does not sporulate. Zoospores

were not observed, moreover its average oogonial and oosporal sizes are smaller (26 &

24 um instead of 35 & 32.5 um, respectively), and oospore wall thicker (upto 7 um

instead of 5 um) than those described by Plaats-Niterink (1981).

Pythium irregulare Buisman (Figs 20-23).

Colonies on CMA & PCA producing profuse aerial mycelium. Daily growth

rate on PCA at 25°C is 24.5 mm. Main hyphae up to 5-6 um wide. Hyphal swellings

globose to ovoid, sometimes provided with single papilla, terminal or intercalary, 14-26

um diam. (x. 19 + 0.6). Sporangia and zoospores were not formed. Oogonia terminal or

intercalary, globose to somewhat elongated, 13-21 x 20 + 0.6) um diam., smooth

walled or provided with a varying number of blunt conical projections up to 12 um

long, 0-6 per oogonium. Antheridia mostly monoclinous, 1-3 per oogonium. Oospores

globose, plerotic or aplerotic, 12-20 (x 16 x 0.6) diam. Wall 1.5-2 um thick.

Pythium irregulare was isolated only on three locations, once in Compiègne and

twice in the region of Lille. Because of the presence of both spiny and smooth- walled

oogonia, it is an easily recognisable fungus. However, in water cultures, the number of

ornamented oogonia was larger than that of the smooth walled ones. The features of the

three isolates agree well with the description of this species reported in literature

(Plaats-Niterink, 1981). The characteristics given above are those of culture no. F-75

isolated from a soil sample taken in the region of Lille.

Pythium ultimum Trow (Figs 24-26).

Colonies on CMA forming profuse aerial mycelium and on PCA showing an

indistinct radiate pattern. Average daily growth rate on PCA at 25°C is 28 mm. Main

hyphae up to 10 um wide. Sporangia and zoospores not formed. Hyphal swellings

formed plentifully and are globose, intercalary, sometimes terminal 18-25 um diam.

(x 20 + 0.6). Oogonia terminal or intercalary, spherical, smooth walled, 19-31 (x

23.4 + 0.6) um diam. Antheridia mostly monoclinous originating immediately | E

the oogonium, sessile, saclike, infrequently hypogynous, rarely diclinous, 1 2 per

oogonium. Oospores spherical, aplerotic, at times plerotic, single, 17-24 (x 20.6 + 0.6)

um diam. Wall 1-3 pm thick.

Pythium ultimum is an aggressive plant parasite and was isolated many times in

Compiègne as well as Lille. The above description is that of isolate F-39.1 isolated

from Lille. Inspite of some variations of oogonium and oospore dimensions, and the

presence of some plerotic oospores together with the usual aplerotic ones, all the other

characters of this fungus fit closely the description of P. ultimum found in the literature

(Plaats-Niterink, 1981).

Pythium oligandrum Drechsler (Figs 27-33 & Fig. 58)

Colonies on CMA forming some aerial mycelium, submerged on PCA showing

an indistinct radiate pattern. Average daily growth rate on PCA at 25°C is 25 mm. Main

hyphae upto 8 um wide. Sporangia intercalary, composed of spherical to irregular,

continguous elements of up to 35 um wide and 60 um long. Zoospores not formed.

Oogonia mostly terminal, spherical, ornamented with conical, acutely tipped spines,

Source : MNHN. Paris

PYTHIUM FROM NORTHERN FRANCE 269

Figs 52-53: Pythium minor, 52: elongated hyphal body, 53: oogonia with branched antheridia; Figs

54-55: Pythium ostracodes. 54: oogonia with antheridia, 55: plerotic oospore; Figs 56-57: Pythium

echinulatum. 56: oogonia with hypogynous antheridia, 57: elongated intercalary sporangia; Fig. 58:

Pythium oligandrum. contiguous sporangia, Fig. 59: Pythium torulosum: inflated elements of

Sporangial complex. (Fig. 53, bar = 40um, all other figures, bar = 16 um).

Source : MNHN. Paris

270 B.PAUL

20-31 (x 2444 £ 0.6) um diam., with spines of upto 6 jm long and upto 3.5 um in basal

diameter. Antheridia absent. Oospores spherical, aplerotic, one per oogonium, 17-27

(x 22.6 0.6) um diam. with a wall approximately 2 um thick.

Pythium oligandrum is abundant in the North of France. It was isolated from

soil samples in the Compiègne as well as Lille regions. The above description is that of

isolate F-81 isolated from Lille. There are some minor differences in the oogonial and

oosporal dimensions of this isolate and those found in the literature, however all the

other morphological characters of this fungus are identical. This is the first taxonomic

treatment of Pythium oligandrum isolated in France.

Pythium mamillatum Meurs (Figs 34-43).

Colonies on CMA & PCA produces some aaerial mycelium and shows a rosette

pattern on the latter. Average daily growth on PCA at 25°C is 20 mm. Main hyphae are

up to 7 um wide. Sporangia or hyphal swellings are globose, ovoidal, ellipsoidal to

somewhat cylindrical, intercalary, at times terminal, measuring 13 to 28 um diam. (x

19.4 + 0.6), Oogonia intercalary or terminal, globose to slightly ovoidal, provided with

blunt to conical, and at times, curved spines 2-6 ym long and 1-3 ym broad. Oogonia

13-24 um in diam. (x 18.3 + 0.6). Antheridia mostly monoclinous rarely diclinous,

usually one, infrequently two per oogonium, antheridial cells clavate making apical

contact with the female gametangia. Oospores globose, plerotic, one per oogonium,

measuring 12-22 (x 16.4 + 0.6) um diam., provided with a moderately thin wall of up

to 2 um.

The above description is of isolate no F-60, isolated from the region of

Compiègne. The other two isolates of this species were collected in Lille. All the

features of this fungus resembles to those found in the literature. The only difference is

the absence of zoospores inspote of the presence of sporangia. The fungus failed to

sporulate despite of repeated flooding in cultures with distilled water, soil extract water

and maintenance at different temperatures. This is the first taxonomic description of

Pythium mamillatum isolated in France.

Pythium echinulatum Matthews (Figs 44-51 & Figs 56-57)

Colonies on CMA & PCA submerged showing an indistinct radiate pattern.

Daily growth rate on PCA at 25°C 10 mm. Main hyphae upto 7-8 ym wide. Sporangia

globose to cylindrical, terminal or intercalary, more often in chains, 11-25 pm diam.

(x 17 + 0.6). Oogonia terminal, intercalary, catenulate, globose to cylindrical,

provided with acute conical spines 3-9 um long. Oogonia 9-26 (x 18 + 0.6) um diam.

(excluding spines). Antheridia mostly hypogynous at times mono and diclinous, 1-2

per oogonium. Oospores globose, rarely oval, plerotic or aplerotic, usually single,

rarely two per oogonium, 6-24 (x 15 + 0.6)u diam. Wall 1.5-2 um thick.

Pythium echinulatum was frequently isolated from the north of France.

Nevertheless this species shows a great morphological variation. Acute conical spines

on the oogonia, hypogynous antheridia, aplerotic and plerotic oospores were common

in all the isolates, but the presence of catenulate sporangia and oogonia were not

constantly found in all the isolates.

Source : MNHN, Paris

PYTHIUM FROM NORTHERN FRANCE 271

ACKNOWLEDGEMENTS

The author wishes to thank Dr. A.J. Van Der Plaats Niterink who has been kind enough to

review this article.

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TAKAHASHI M., OHUCHI A. & ALICBUSAN R.V., 1965 - Ecologic and taxonomic studies on

Pythium as pathogenic soil fungi. 6. Some species of Pythium causing rhizome rot of hindu

lotus. Ann. Phytopathol. Soc. Japan 30: 186-191.

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Source : MNHN, Paris

Source : MNHN. Paris

Cryptogamie, Mycol. 1994, 15 (4): 273-281 273

POST-HARVEST ROTS OF TOMATO IN RELATION

TO LYASES AND MYCOTOXIN PRODUCTION

IN VITRO AND IN VIVO

S.A. OMAR and A-L.E. MAHMOUD

Department of Botany, Faculty of Science, Assiut University, Assiut, Egypt

ABSTRACT - The post-harvest tomato rotting fungi were isolated from 100 samples of tomato

showing rot symptoms. A total of 220 isolates were recovered on Czapek's dextrose agar medium

comprising 6 fungal species among which Alternaria alternata, Aspergillus flavus and A. niger were

predominant. Asp. tamarii, Cochliobolus spicifer and Penicillium citrinum were isolated at low

frequency. Testing enzymatic abilities of some isolates showed that most of these isolates produced

cellulase, pectin lyase and polygalacturonase on synthetic media as well as on inoculated tomatoes.

Moreover, some isolates which did not show enzymatic activities on agar media produced enzymes

on tomatoes. However, most isolates of Alt. alternata, Asp. flavus and Asp. niger tested were good

enzyme producers and showed the highest enzymatic activity either in vitro or in vivo suggesting that

tomato rot is mainly brought about by members of these fungi. It is worthmentioning that the ealthy

fruits had no delectable enzymatic activity

During screening for mycotoxigenicity, Alternaria alternata proved to be the most able

mould to produce different mycotoxins in liquid medium as well as in infected tomatoes. In addition

to Alternaria toxins, aflatoxins B,, B, and citrinin were produced by Asp. flavus and Р. citrinum,

respectively. Results of this study clearly showed that tomato fruits infected naturally with the

moulds previously mentioned may contain different mycotoxins which may represent a potential

health hazard.

RESUME - Les agents fongiques responsables du pourrissement aprés récolte des tomates ont été

isolés de 100 tomates présentant des symptómes de pourrissement. Deux cent vingt souches ont été

isolées sur Czapek-Dextrose agar. Ces souches se répartissent parmi six espèces. Alrernaria

alternata, Aspergillus flavus et Aspergillus niger sont les espèces les plus fréquentes. Aspergillus

tamarii, Cochliobolus spicifer et Penicillium citrinum ont été isolés à de faibles fréquences, Des

essais enzymatiques réalisés sur certains de ces isolats ont permis de montrer que la plupart

produisent des cellulases, des pectine-lyases et des polygalacturonases, aussi bien sur milieu

Synthétique que sur tomates. Certains isolats ne présentent d'activité enzymatique que sur tomates. La

plupart des isolats d'Alt. alternata, Asp. flavus et Asp. niger se sont révélés être de bons producteurs

d'enzymes et montrent la plus haute activité, tant in vitro qu'in vivo, suggérant que le pourrissement

des tomates puisse être incriminé à des membres de ces espèces. Les tomates saines ne présentent pas

d'activité enzymatique.

Les études mycotoxicologiques ont montré qu'Af. alternata était l'espèce isolée

produisant le plus de mycotoxines différentes en milieu liquide ou sur tomates. En plus des toxines

produites par Alt. alternata, des aflatoxines Bl et B2, et de la citrinine étaient produites

respectivement par Asp. flavus et Penicillium citrinum. ll en résulte que des tomates naturellement

zontaminées pourraient contenir différentes mycotoxines, représentant un risque pour la santé.

Source : MNHN. Paris

274 S.A OMAR and A.-L. E. MAHMOUD

INTRODUCTION

Tomato fruits represent one of the essential vegetables all over the world

throughout the year. After harvesting, these fruits may be invaded by several moulds

that probably cause fruit rotting and extensive damage to the crop (Ayres et al., 1964;

Barkai-Golan, 1974). Extensive deterioration results in economic loss to commercial

marketers of these fruits. Tomato rot is favoured by the high temperature and hence it is

pronounced in tropical and subtropical regions (Adisa, 1980) however, some fungi can

infect tomatoes stored at 10-12°C causing their spoilage (Ayres et al., 1964).

Infection of fruits by pathogenic fungi is initiated by production of cell wall-

degrading and macerating enzymes (Weste, 1970). The role of polysaccharide degrading

enzymes in microbial pathogenicity has been reviewed (Wood, 1976). The ability of

many pathogenic moulds to produce these enzymes in culture is not sufficient reason to

ascribe them a role in pathogenicity (Byrde, 1979).

In addition to biodeterioration of tomato fruits, different mycotoxins may be

produced in these fruits by toxigenic moulds and this may constitute a potential health

hazard (Harwig er al., 1979; Stinson er al., 1980 and 1981).

The present work was designed to throw some light on the moulds that cause

post-harvest spoilage of tomato. Enzymatic abilities and mycotoxin-producing potential

of these fungi, both in vitro and in vivo, were also studied.

MATERIALS AND METHODS

Source of samples. A total of 100 tomatoes showing lesions of different

appearance were collected from markets in different localities of Assiut Governorate,

Egypt.

Isolation and identification of moulds. By using sterile scalpal, tissue

fragments were excised from lesions of infected fruits and were plated on Czapek's

dextrose agar medium supplemented with rose bengal (65 ppm) as a bacteriostatic

agent, Inoculated plates were incubated for 7-10 days at 28°C. The resulting moulds

were isolated and identified.

Enzymatic activity of the isolated fungi. A total of 40 fungal isolates from

fungi recovered during this investigation were screened for their ability to produce

some enzymes on solid media as well as on tomato fruits. These fungi were Alternaria

alternata, Aspergillus flavus, A. niger, A. tamarii, Cochliobolus spicifer and

Penicillium citrinum.

Cellulase activity was studied by using the method described by Eggins &

Pugh (1962). The tested fungi were grown on medium contained (g/L), ammonium

sulphate, 0.5; L-asparagine, 0.5; potassium dihydrogen phosphate, 1.0; potassium

chloride, 0.5; magnesium sulphate, 0.2; calcium chloride, 0.1; yeast extract, 0.5;

cellulose, 10 and agar 20. After 7 days incubation at 28°C, plates were flooded with

chloro-iodide of zinc. The uncoloured zone gave a measure of the cellulolytic power of

the moulds

Source : MNHN, Paris

BIODETERIORATION OF TOMATO 275

The test isolates were screened on MP-7 and MP-5 media of Hankin et al.

(1971) for pectin lyase (PL) and polygalacturonase (PG), respectively. After growth of

organisms for 7 days at 28°C, pectolytic activities on both media were determined by

flooding plates with 7 mol/L HCI solution. This precipitate intact pectin and pectolytic

moulds were thus surrounded by clear zones against an opaque medium. The extent of

zone of clearing around moulds was used as a measure of the degree of pectolytic

activity.

Amylolytic activity of the test fungi were screened according to the methods

described in the Society of American Bacteriologists (1957). The experimental medium

consisted of 28 g of nutrient agar to which 2 g of soluble starch (Merck) were added per

litre. After incubation of the inoculated plates for 7 days at 28°C in darkness, they were

flooded with an iodine solution (KI, 15 g and I, , 3 g litre). A zone void of blue

indicated the production of amylase.

Proteolytic activity of moulds was determined using a casein substrate. Each

mould culture was inoculated onto the surface of mycological agar (peptone, 10 g; agar,

20 g per litre) to which sterile skim milk (10% solution of powder of defatted milk in

water) was added at the rate of 5 ml per 100 ml of medium. After incubation for 7 days

at 28°C, complete degradation of milk protein was seen as clearing zone in the

somewhat opaque agar around colonies. The extent of the clear zone represented the

degree of proteolytic activity.

Enzymatic activities of the tested fungi on tomatoes. Each tested mould

was inoculated on the surface of tomatoes (individual weight, w = 70-100 g) disinfected

with ethanol (90%). The inoculation was done by placing a square block of Czapek's

dextrose agar with the fungal spores in two windowshaped wounds per fruit (Vinas et

al., 1992). Inoculated fruits were placed in sterile plastic bags and incubated at 28°C for

10 days. Two tomatoes were utilized for each strain,

After incubation period, the decayed tomatoes were taken, blended with 0.9

(v/w) distilled water for 3 min. Fruit extracts were clarified by centrifugation at 15000 x

£ for 15 min at 4°C. The supernatants were employed as crude enzyme solutions.

Plates containing different solid media specific for detection of amylase,

protease, cellulase and pectinases were prepared as previously showed. Under sterilized

conditions, 0.5 ml of the enzyme solution was pipetted in a cup made in the center of

each plate. After incubation at 28°C for 24 h, the presence of these enzymes was

examined.

Screening for mycotoxigenicity. Ten isolates of both Alternaria alternata

and Aspergillus flavus and 5 isolates of each of Asp. niger, Asp. tamarii and Penicillium

citrinum isolated during this study were screened for mycotoxigenicity on liquid

medium as well as on tomatoes.

Inoculation and incubation procedures. The tested fungi were grown on

yeast extract sucrose medium (YES). Spore suspension of a 7-days old culture of each

mould was made and 0.5 ml (approx. 10° spores/ml) was used as an inoculum for each

50 ml quantity of YES medium in 250 ml Erlenmyer flasks. The flasks were incubated

Source : MNHN. Paris

276 S.A OMAR and A.-L. E. MAHMOUD

at 28°C for 10 days as stationary cultures under darkness. Two replicates of each strain

were analysed.

Like wise, each tested mould was inoculated on the surface of tomatoes.

Inoculation and incubation procedures were previously mentioned. Tomatoes were

frozen after they had reached the desired stage of rot, as estimated by the extent of

external discoloration and kept untill extraction.

Extraction procedures. In case of liquid medium, the contents of each flask

were homogenized with 50 ml chloroform for 5 min in a high speed blender (16000

rpm). Extraction was repeated three times. The combined chloroform extract was

washed with distilled water, dried over anhydrous sodium sulphate, filtered and dried to

near dryness on a rotary evaporator. The residue was diluted with chloroform to one ml.

The chloroform solution was analysed for the presence of aflatoxins [B, ,B, , б, and

G,), patulin and citrinin using thin-layer chromatography.

In case of Alternaria, liquid cultures were extracted twice with ethyl acetate

(30 ml) by overnight shaking under darkness and filtration. The two extracts were

combined, dried over anhydrous sodium sulphate and evaporated to near dryness. The

residue was dissolved in | ml of methylene chloride and analysed for the presence of

alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TeA) on

TEC:

The decayed frozen tomatoes were extracted according to Harwig e al.

(1979). The fruits were thawed, blended with 0.9 (v/w) methanol and 50 ml of n-hexane

for 3 min, and then centrifuged at about 2000 rpm for 5 min. for citrinin analysis (P.

citrinum) , 3 ml of 10 N sulfuric acid was added to the extraction solvent. An aqueous

methanol extract was removed by pipette and shaken with two 25-ml portions of

chloroform. In case of Alternaria alternata analysed for AOH, AME and TeA, 3 ml of

10 N sulfuric acid was added to the aqueous methanol portion. Chloroform extracts

were combined and evaporated to near dryness. The residue was dissolved in | ml of

chloroform and was analysed for the presence of toxins previously mentioned.

TLC analysis. This was performed on precoated silica gel plates of kieselegel

G type 60 (MERCK) of about 0.3 mm thickness using chloroform-acetone (9-1) as a

solvent system. The developed plates were examined under UV light at wave lengths of

254 and 366 nm. Mycotoxins were identified by comparison with appropriate reference

standards before and after treatment with p-anisaldehyde and ferric chloride solution as

described by Durackova et al. (1976). Alternaria toxins were analysed according to

Harwig er al. (1979). Authentic samples of aflatoxins and Alternaria toxins were

purchased from Sigma Chemical CO., U.S.A. Patulin and Citrinin were obtained from

U.S. Department of Agriculture, Northem Research Laboratories, Peoria, Minois,

USA.,

RESULTS AND DISCUSSION

During this study, Alternaria alternata, Aspergillus flavus, Asp. niger, Asp.

tamarii, Cochliobolus spicifer and Penicillium citrinum were isolated from deteriorated

Source : MNHN, Paris

BIODETERIORATION OF TOMATO 277

tomato fruits. As indicated in Table I, Alt. alternata was isolated from 70% of samples

followed by Asp. flavus (62%) and Asp. niger (54%) suggesting the responsibility of

these three moulds for biodeterioration of tomatoes. These results confirmed with those

of Adebanjo & Shopeju (1993) who isolated Asp. flavus and Asp. niger at high

frequencies from some sundried vegetables over a period of 8 weeks. Also, Asp. flavus

was reported to be associated with the spoilage of tomato fruits (Fajola, 1979). In a

similar study, Alternaria spp. were isolated from as many as 51.6% of decaying

tomatoes stored at 10-12°C (Ayres et al., 1964). The involvement of Alt. alternata in

spoilage of stored tomatoes was reported by other workers (Pearson & Hall, 1975). ,

Results presented in Table II revealed that most strains of isolated fungi

exhibited cellulolytic, pectolytic, amylolytic and proteolytic abi

ities when grown on

Table I: Fungi isolated from tomato fruit infected by rotting fungi

Number of cases of isolation

Fungi isolated (out of 100 samples)

Alternaria alternata 70

Aspergillus flavus 62

Asp. niger 50

Asp. tamarii 15

Cochliobolus spicifer 8

Penicillium citrinum 15

Table II: Enzymatic abilities of some moulds, isolated from deteriorated tomatoes, in agar media.

| tested isolates lyase turonase

| Alt, alternata 10 Set 10++ бек S++ 6++

| 3++ 2++ 2+ 2+

2-ve 2-уе 3-уе 2-уе

| Asp. flavus 10 [me Bene S+++ 3+++ S++

2-ve 2-ve 3 34+ 2+

2+ 2+ 3-уе

| 2-уе

Asp. niger 5 S++ S++ 3+++ pe 3++

| 2-ve 1-уе 2-уе

| Asp. tamarii 5 3er 4% 3er i mS

2+ l-ve 1+ 3+

lave 1-уе 2-ve.

C. spicifer 5 5++ 3+ 4+ 3+ 2er

2-ve lve 2-ve 2+

l-ve

| P. citrinum 5 2++ 5% 3+ ++ 2+

| 3+ 2-ve 2+ 3+

l-ve

+, low activity; +++, high activity; ++, fair activity; -ve, no activity.

Source : MNHN. Paris

278 S.A OMAR and A.-]

„E. MAHMOUD

synthetic media. On inoculated fruits (Table HI), the production of cellulase and

pectinases was more pronounced. On the other hand, protease and amylase production

was varied since some tested isolates which produced these enzymes on synthetic

media did not exihibt any enzymatic activity on tomatoes. In this respect, Adisa (1985)

found that cellulase, polymethylgalacturonase and pectinmethyltrans-eliminase were

identified in vivo and in culture filtrates of two tomato spoilage moulds, (Asp. flavus

and Asp. fumigatus. It was realised that the softening of tissues in ripening peaches was

correlated with increase in the pectinic acid content (Shewflet et al., 1971) and increased

pectic enzyme activity (Pressey & Avants, 1971). These enzymes bring about the

breakdown of polysaccharide components resulting in maceration of tissue and death of

host cells. Among cell wall-degrading enzymes, pectinolytic and cellulolytic enzymes

Table III: Enzymatic abilities of some moulds, isolated from deteriorated tomatoes, in inoculated

fruits.

tested isolates lyase turonase

Alr. alternata 10 10+++ 6++ Tert 4+ 5+

4+ 3+ 6-ve 5ле

Asp. flavus 10 9e Bee [m 3+ 6+

Іле 2+ 4+ 7-уе 4-уе

Asp. niger 5 5+ 5+ 4er 2+ 3+

1-уе 3-уе 2-ve.

Asp. tamarii 5 Em 4+ 4+ 3+ 3+

1+ 1-уе lve 2-ve 2-ve

1-ve

C. spicifer 5 5% 44 3+ 2+ 2+

lve 2-ve 3-ve 3-ve

P. citrinum 5 5+ 5% 5+ 4+ 4+

lve lve

+, low activity; +++, high activity: ++, fair activity; -ve, no activity

pose a unique position and several references referred the pathogenicity of plant

pathogens to the ability of secretion of these enzymes (Weste, 1970; Kachhawaha &

Ali, 1982), in spite of the involvement of other enzymes in cell wall degradation. This is

because cellulose and pectin represent the main and most complex components of plant

cell wall.

During screening for mycotoxigenicity of the tested fungi, 14 out of 35

isolates were toxigenic (Table IV). The toxigenic isolates belonged to Alternaria

alternata (8 isolates), A. flavus and P. citrinum (3 isolates for each).

Alternaria alternata proved to be the most able species in mycotoxin

production where 80% of its tested isolates produced toxins both in vitro and in vivo.

Four isolates produced tenuazonic acid (TeA), 2 isolates produced tenuazonic acid

(TeA) in addition to alternariol monomethyl ether (AME) and 2 isolates produced

alternariol monomethyl ether (AME) as well as alternariol (AOH). These results

suggest that tomato fruits infected naturally with Alternaria alternata may contain TeA,

Source : MNHN. Paris

BIODETERIORATION OF TOMATO 279

AME and AOH. These compounds are known metabolites of Alternaria alternata (Pero

et al., 1973).

These results agree with those of Stinson er al. (1980) who found that most

isolates of Alternaria produced TeA, AOH and AME on tomato fruits. They also stated

that the known toxigenic strains of Alternaria produced more of TeA on tomatoes than

the dibenzo-7o-pyrone toxins (AOH and AME). In another study, Stinson er al. (1981)

reported that TeA was the main mycotoxin produced in Alternaria-infected tomatoes

from commercial sources while AOH and AME were present in small amounts.

Similarly, Harwig et al. (1979) found that Alternaria alternata, isolated from decayed

tomatoes, produced TeA and AME in culture medium as well as in infected tomatoes

however, AOH was not detected.

Table IV: Ability of some moulds, isolated from deteriorated tomatoes, to produce mycotoxins in

liquid medium and in infected fruits.

On liquid medium On tomatoes

Numberof | Numberof | Mycotoxins | Number of | Mycotoxins

Organism tested isolates | toxigenic detected toxigenic detected

| isolates isolates

Alrernaria 10 8 Alternarial 8 Altemarial

alternata (AOH), (AOH),

Alternariol, Alternariol,

Monomethyl Monomethyl

| ether (AME) ether (AME)

| & Tenuazonic & Tenuazonic

acid (TeA) acid (TeA)

Aspergillus flavus 10 3 Aflatoxins BI 3 Aflatoxins BI

| & B2 & B2

Asp. niger 5 - - - -

Asp. tamarii 5 - - - -

Penicillium 5 3 Citrinin 3 Citrinin

| citrinum

From 10 isolates of Asp. flavus, one isolate produced aflatoxin B and 2 isolates

produced aflatoxins B, and B,. Published literatures dealing with the production of

aflatoxins in infected tomatoes are not available. However, the natural occurrence of

aflatoxins in tomato paste samples has been recorded (Saber et al., 1992). In a similar

study, Neelakantan er al. 1983) found that aflatoxin B was naturally present in apples.

Experimental production of aflatoxins in various fruits has been reported (Detroy et al.,

1971).

Citrinin was produced by 3 isolates (60%) of P.citrinum in YES medium and

in tomato fruits. These results agree, to some extent, with those of Harwig er al. (1979)

who recorded this toxin in culture filtrate as well as in tomatoes inoculated with P.

expansum.

Results of the present study clearly showed that most of the moulds that cause

post-harvest spoilage of tomatoes are enzymatically active. These moulds excrete an

array of enzymes which bring about the breakdown of organic matter resulting in

Source : MNHN, Paris

280 S.A OMAR and A.-L. E. MAHMOUD

maceration of fruit tissues. In addition to biodeterioration, different mycotoxins may be

produced in fruits by toxigenic moulds which represents a potential health hazard.

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Cryptogamie, Mycol. 1994, 5 (4): 283-287 283

ANALYSES BIBLIOGRAPHIQUES

REED C.F. & D.F. FARR : Index to Saccardo's Sylloge Fungorum Volumes I-XXVI

in XXIX, 1882-1972. - Contribution N? XXXI of the Reed Library and Herbarium,

Darlington, Maryland; Contribution N° 6 from the U. S. National Fungus Collection,

Beltsville, Maryland; Rose Printing Company, Tallahassee, Florida. Hardback $ 75,

Softback $ 60, plus $ 4 for shipping. Obtainable from Clyde Reed, 1222 Main St.,

Darlington, MD 21034, USA.

Le Sylloge Fungorum est un répertoire des noms et des descriptions des espèces

fongiques et bactériennes proposées entre la fin du XIXème et le début du XXème

siècle. Les vingt-cinq volumes de cette oeuvre, presque encyclopédique, du célébre

mycologue italien P.A. SACCARDO et de ses collaborateurs, furent publiés entre 1882

de base en taxonomie de champignons. On y trouve également les auteurs des taxa

proposés et les références bibliographiques majeures les concernant. Seul point négatif

de cette compilation critique magistrale, l'absence d'un mode cohérent de présentation

de l'énorme masse d'informations contenues dans ses divers volumes. Ce mode particu-

lier de présentation est à l'origine des difficultés constantes, rencontrées lors d'une

recherche des caractéristiques morphologiques d'un taxon donné; en particulier, si le

nom recherché n'est pas répértorié dans le premier volume, puisqu'un index cumulatif

n'était pas disponible, d'où l'impérieuse nécessité de préparer un index général couvrant

la totalité du Sylloge.

L'Index du Sylloge Fungorum volume I - XXVI dans XXIX, 1881-1972, de

Reed et Farr constitue ce document cumulatif, tant désiré, des nombreux volumes de

cette oeuvre magistrale. Seul le titre n? 13 afférent aux noms des plantes-hótes, n'a pas

été inclus dans l'index réalisé par ces deux auteurs. Dans une courte introduction de

trois pages, Reed et Farr passent rapidement en revue les problèmes rencontrés lors de

la préparation de cet index: problèmes de pagination, certaines pages du Sylloge n'ayant

jamais été numérotées; conformité de la pagination orignale avec celle de la première

réédition du Sylloge réalisée par Edwards en 1944, etc... D'ailleurs, une copie des qua-

tre pages manquantes dans l'édition d'Edwards a été ajoutée à la fin de cet Index; ceci

permet aux détenteurs de copie de cette dernière de combler cette lacune.

La réalisation de cet Index ne pouvait être envisagée sans l'existence des

moyens électroniques modernes de traitement de l'information. L'utilisation d'une ban-

que de données informatique rend possible le traitement de la totalité des 121 627 noms

figurant dans le Sylloge. L'informatisation des éléments taxonomiques de chaque bi-

nóme a permis, par ailleurs, la mise en évidence, de certaines corrélations intéressantes:

répartition des noms recensés par volumes de citation, définition de la masse des genres

traités et établissement des listes respectives des espèces afférentes, fréquences

d'utilisation des épithètes spécifiques, mode de répartition de ces épithètes d'après la

fréquence de la première lettre alphabétique employée, etc … On y découvre ainsi que

dix des genres traités comportent chacun plus de mille espèces, que l'épithète spécifique

Source : MNHN. Paris

284 ANALYSES BIBLIOGRAPHIQUES

elegans se distingue par une fréquence optimale d'utilisation (151), que la lettre c de

l'alphabet est la première lettre de 12,5 % des noms d'espèces repertoriés alors que la

lettre y ne concerne que 0,1% de ces noms, etc...

Aprés cette courte introduction, le Conspectus du Sylloge Fungurom reproduit,

sur dix pages, les références bibliographiques de ses divers volumes et les sommaires

respectifs; ces derniers comportent la liste des ordres, familles et sections traités dans

chaque volume avec indication des numéros des pages afférentes. Cette synthèse per-

met une vue générale des groupes taxonomiques traités, rangés d'après le système parti-

culier mis au point par Saccardo lui-même

Le coeur de l'ouvrage se compose de plus de huit cents pages de format A4;

c'est l'Index des noms figurant dans le Sylloge Fungorum, repértoriés sur deux colonnes

par page, un mode de présentation favorisant un balayage visuel rapide du contenu de

chaque page. 4827 genres sont ainsi répérioriés par ordre alphabétique. Pour chaque

genre cité est précisé le nom(s) d'auteur(s), les éléments de la. référence bibliographique

originale et des informations d'ordre supragénérique: ordre et famille d'appartenance. A

l'intérieur des genres, les noms des espèces qui en relèvent sont également présentées

en ordre alphabétique, accompagnés chacun des numéros de volumes et des pages de

citations dans le Sylloge. Point important, les synonymes ont été également répértoriés

avec indication entre parenthèses du nom correct admis dans le Sylloge.

Tl ne fait aucun doute que cet index tant attendu devient un outil inestimable et

indispensable pour les mycologues poursuivant des recherches en taxonomie des

champignons; également pour tout chercheur amené à résoudre un problème d'ordre

nomenclatural concemant ces microorganismes. Sa réalisation a exigé un effort colos-

sal répartis sur plusieurs années et cela malgré le recours massif aux moyens modernes

de traitement de texte: d'ailleurs sans l'avènement de l'informatique, cet index n'aurait

sans conteste pas vu le jour. D'autres mycologues ont également contribué, de manière

bénévole, à la solution des problèmes soulevés par cet essai d'intégration en une masse

cohérente du contenu des volumes du Sylloge. Les descriptions originales des espèces

répertoriées ainsi que les références afférentes peuvent maintenant être rapidement

localisées en un minimum de temps, sans recherches bibliographiques vaines à travers

le dédale des volumes du Sylloge. Plus important encore, ce nouvel index permet aux

personnes ne possédant pas une copie originale ou une reproduction du Sylloge, de

démarrer une recherche bibliographique efficace et rapide pour un genre déterminé.

Il reste enfin à signaler qu'une liste des noms omis dans le Sylloge a été anté-

rieurement préparée par P. M. KIRK; cette liste fut publiée en 1985 par le Common-

wealth Mycological Institute.

Jean MOUCHACCA

ARORA D.K., ELANDER R.P. & MUKERJI K.G. (Eds) : Handbook of Applied

Mycology. Vol. 4 - Fungal Biotechnology, Marcel Dekker Inc., 270 Madison Ave.,

New York, NY 10016, 1992. 1114 pp. Price £ 150.00; by subscription $ 127.00.

La mycologie appliquée est actuellement devenue une discipline biologique

comparativement trés stimulante et un champ d'action en évolution rapide. Cette acti-

Source : MNHN, Paris

ANALYSES BIBLIOGRAPHIQUES 285

vité scientifique progesse grâce à une intégration harmonieuse d'un ensemble de spé-

cialités relevant de disciplines variées: agricoles, industrielles, pharmocologiques,

médicales et alimentaires. Cette définition de la mycologie appliquée est proposée par

D. K. ARORA, principal éditeur de la série des cing manuels des "Handbook of

Applied Mycology”. L'ouvrage "Fungal Biotechnology" en est le quatrième volume. Ce

livre de dimensions marquées, comporte quatre sections de volume inégal et plus de

mille pages de texte. Les trois volumes antérieurs ont porté sur la nature des liens entre

les Sols et les Plantes, entre l'Homme, les Animaux et les Insectes et, enfin, sur ceux

impliquant les Champignons dans l'Alimentation Humaine et Animale.

Selon les trois éditeurs du quatrième volume, la biotechnologie des champi-

gnons est l'utilisation des organismes fongiques ou de leurs composants subcellulaires,

dans des processus technologiques appliqués dans des domaines de productions indus-

trielles ou de gestion de l'environnement. Cette discipline requiert des connaissances

approfondies en génétique, biologie moléculaire et biochime des champignons et éga-

lement en chimie analytique ou autre. L'ouvrage traite successivement des thémes

majeurs suivants : Technologies moléculaires, Applications commerciales, Décomposi-

tion des résidus biologiques et chimiques et Collections de culture, aspects légaux et

sécurité biologique.

Cet ouvrage bénéficie d'un chapitre introductif traitant de la. biotechnologie des

champignons, une synthése liminaire qui constitut la premiére contribution de la sec-

tion Technologies moléculaires. Ses deux auteurs Elander & Lowe passent en revue les

processus fongiques commercialiés à ce jour et qui, globalement, font appel à des espè-

ces fongiques, toutes capables d'un bon développement sur des milieux de culture à

coûts de production excessivement faibles. Ce chapitre est suivi par une analyse judi-

cieuse des approches molécularistes en taxonomie des champignons: description des

stratégies développées et avantages et inconvenients des techniques analytiques mises

àu point; ses deux auteurs, Klich et Mullaney, insistent sur le fait que la biologie molé-

culaire n'offre pas de solutions miracles pour les controverses ou disputes d'école exis-

tant en taxonomie des champignons. Cette premiére section comporte dix chapitres

avec des contributions marquées sur la technologie des protoplastes, sur les plasmides

fongiques et la transformation et manipulations génétiques chez les champignons fila-

menteux.

La deuxiéme section rassemble une série de contributions sans lien apparent.

On y trouve deux articles sur les champignons thermophiles: rôle en agriculture et

potentiel biotechnologique et un autre sur les mycorhizes; ce dernier sujet a été large-

ment débattu dans le premier volume de cette série. La troisiéme section (quatre chapi-

ires), comporte des contributions bien structurées à l'échelle individuelle mais sans

aucune trame collective convaincante; le thème global traité aurait pu d'ailleurs faire

l'objet d'un volume distinct. Enfin, la dernière section relative aux collections de cultu-

res, propose trois contributions analysant les problèmes liés à l'établissement des bre-

vets et la conclusion des accords commerciaux secrets, seule forme légale de protection

dans le cas des inventions biotechnologiques.

Source : MNHN. Paris

286 ANALYSES BIBLIOGRAPHIQUES

Globalement et malgré une certaine dose de répétition, le quatrième volume de

cette série propose une masse importante d'informations sur les sujets abordés, surtout,

dans certains cas, au niveau des références bibliographiques afférentes. Les trente-

quatre contributions proposées, quoique de poids relatif inégal, constituent néanmoins

d'excellents articles de synthèse. Ceux-ci seront largement appréciés par les chercheurs

débutants ou confirmés désireux de parfaire leur connaissance sur un sujet approprié ou

d'acquérir des notions introductives sur un thème inédit. Il reste à souligner que le

faible prix demandé pour cet ouvrage est de nature à favoriser son achat par les biblio-

thèques institutionnelles.

Jean MOUCHACCA

BHATNAGAR D., LILLEHOY E. B. & ARORA D. K. (Eds) : Handbook of Applied

Mycology. Vol. 5 - Mycotoxins in Ecological Systems, Marcel Dekker Inc., 270

Madison Ave., New York, NY 10016, 1992. 443 pp. Price £ 150.00; by subscription $

127.00. prix à voir

Ce dernier volume de la série des manuels à multi-auteurs de mycologie appli-

quée a pour titre: les mycotoxicoses dans les systèmes écologiques. Dans la préface, les

éditeurs soulignent qu'aprés l'épopée héroïque des antibiotiques d'origine fongique, les

recherches en mycotoxines se sont rapidement développées pour devenir des thèmes

majeurs en mycologie appliquée. En efffet, la mycotoxicologie a définitivement acquis

ses lettres de noblesse; elle n'en demeure pas moins une activité scientifique à caractère

multidisciplinaire. Ce champ d'action soulève cependant une question fondamentale qui

reste encore un sujet de controverses : Quel est le fondement mystérieux de la logique

biologique conduisant à la production de métabolites secondaires par les champignons ?

Ces composés chimiques sont à l'origine des symptômes toxicologiques enregistrés.

Cet ouvrage de quatre cents pages comporte seize contributions, non rassem-

blées en sections autour de thèmes particuliers moins généraux. En parcourant les titres

proposés, il devient évident que le contenu de certains articles s'éloigne quelque peu du

titre général de l'ouvrage. Ces chapitres revêtent un caractère purement descriptif du

sujet traité sans qu'un système écologique donné y soit incriminé. Ceci dit la qualité de

l'ensemble des textes retenus reste de haut niveau: ce sont des synthèses actualisées de

grande importance.

Le fil conducteur de quelques chapitres semble être les effets intracellulaires

des mycotoxines, actives dans certains systèmes animaux. En particulier, ces chapitres

traitent des mécanismes de cytotoxicité et de genotoxicité attribués aux aflatoxines.

D'autre part, les mécanismes subcellulaires de toxicité, attribués à l'acide cyclopiazonic

et à l'ochratoxine, font également l'objet de deux chapitres intéressants. La biosynthèse

et la régulation de la production des aflatoxines et des trichothécenes sont également

traitées de manière exhaustive dans diverses contributions; celles-ci n'intègrent pas

Source : MNHN. Paris

ANALYSES BIBLIOGRAPHIQUES 287

malheureusement et de manière détaillée, des corrélations écologiques afférentes aux

sujets abordés.

On peut regretter que le panel des rédacteurs sollicités rassemble presque ex-

clusivement des spécialistes travaillant dans des institutions nord-américaines et euro-

péennes, où se réalise en réalité la quasi-totalité des recherches consacrées aux problè-

mes de toxicologie d'origine fongique. La parution de cet ouvrage sera, néanmoins,

largement acceuillie à l'échelle internationale, par tous les chercheurs spécialisés ou

non spécialisés, intéressés par les déréglements métaboliques résultant du développe-

ment végétatif des champignons filamenteux sur des productions industrielles animales

et agricoles. En effet, les mises à jour proposées permettent de très rapidement acquérir

une vue d'ensemble approfondie d'un sujet déterminé, et surtout, de dégager des futurs

points de recherches dans des thèmes en cours d'investigation.

La bibliograhie afférente à chaque article est abondante et comporte une bonne

proportion de titres récents. Le dernier volume de cette série devrait rapidement trouver

sa place dans les bibliothèques des laboratoires intéressés. Il sera consulté par un large

éventail de spécialistes travaillant dans des domaines diversifiés et touchant, de prés ou

de loin, à la production, la distribution, la commercialisation et la consommation de

productions agricoles et animales industrielles.

Jean MOUCHACCA

Source : MNHN. Paris

Cryptogamie, Mycol. 1994, 15 (4): 289-290

TABLE DU TOME 15

BADALYAN S.M., RAPIOR S., DOKO L., LE QUANG J., JACOB M., SERRANO J.J. and

ANDARY C. - Investigation of primary and secondary metabolites in a chemical

study of Cortinarius armillatus (Cortinariaceae, Telemonia).

BAÑARES A., BELTRAN E. y RODRIGUEZ J.L. - Estudio micologica de la reserva de la

biosfera «El Canal de los Tiles» (La Palma, islas Canarias). III. Agaricomycetidae

(2a parte). . іа : 2

BEKHOUCHE F, BRETON A. et GAILLARD-MARTINIE B. - Champignons cellulo-

lytiques du sol des zones arides du sahara algérien. Mise en évidence de l'activité

cellulasique.

BOIDIN J. et GILLES G. - us mu. E de l'ile de la Réunion. XVIII les

SO Tele ct AM

CHECA J., BLANCO MN. y BARRASA ІМ. - Estudio sobre ne sensu lato de

a AA nini

EL KADY LA., ABDEL-MALLEK A.Y., EL MARAGHY S.S.M., and HASSAN H.AH. -

Toxigenic moulds in pesticide treated liquid medium...

HENNEBERT G.L. PASCAL S. et COSYNS M. - Interactions d'incompatibilité entre

homocaryons du Basidiomycète bifactoriel Lenzites betulinus. Une phéromone de

répulsion sexuelle? OU

HEYKOOP M., ESTEVE-RAVENTOS F. y MORENO G. - Algunos sss interesantes

de la provincia de Guadalajara (Espana peninsular). I.. es

KALYANASUNDARAM I., MENON L. and LOGANATHAN P. - Occurrence of melanin in

bright-spored Myxomycetes.

KHABAR L., NAJIM L., JANEX-FAVRE M.C. et PARGUEY-LEDUC A. - L'ascocarpe de

Terfezia leonis Tul. (Discomycàtes, Tubérales)... к

LÉGER Ј.С. et LANQUETIN P. - Hymenochaete coffeana nov. sp. (Basidiomycetes,

Aphyllophorales): description et caractères culturaux. а. S

MAHMOUD A-LE. and OMAR S.A. - Enzymatic activity and mycotoxin-producing

potential of fungi isolated from rotted lemons ………

MICHAUT A., JACQUIER MJJ., BOUCHET F., DIJOUX M.G.,. LE MEN L., BOUCHET P.

- Etude de l'action antifongique de différents composés bromés extraits d'un annélide

polychète des îles Kerguelen : Thelepus setosus. ...... 1. ыы

MORENO G., ESTEVE-RAVENTOS F. y ORTEGA A. - Estudios micologicos en el Parque

Natural de los Alcomocales (Andalucía, España). I. Agaricales. M

141

133

125

229

187

117

Source : MNHN. Paris

290 TABLE DU TOME 15

MORENO G., ARENAL F. y GONZÁLEZ V. - Ages dun de las irae de España

peninsular. 239

MORENO G., HORAK E. y LAGO M.. - Descolea maculata Bougher o nueva

cita para Europa. .. аа 3 ; 255

MOUBASHER M.H. and EMAN MOSTAFA M. - Nutritional and environmental factors

affecting glycerol production by Aspergillus wentii IMI 023010. cuca 175

MOUCHACCA J. and ZUCCONI LE. - Fungi of New Caledonia III. Some interesting

dematiaceous hyphomycetes from leaf ltter.……. 27

OLUFOLAJI D.B. - Cultural condition for growth and sporulation of Colletorrichum

falcatum (sugarcane red-rot fungus). . = 207

OMAR S.A. - Utilization of phenols and biosynthesis of humic acid by Aspergilllus terreus. 57

OMAR S.A. and MAHMOUD A.-LE. - Post-harvest rots of tomato in relation to lyases and

mycotoxin production in vitro and IN VIVO. «ni 273

PAUL B. - Some species of Pythium isolated from cultivated soils in northern France... 263

Analyses bibliographiques. 63, 149, 219, 283

Instructions aux auteurs.. 65

Source : MNHN, Paris

Commission paritaire 16-1-1986 - N° 58611 - Dépôt légal 4° trimestre 1994- Imprimerie F. Paillart

Sortie des presses le 30 décembre 1994 - Imprimé en France

Editeur : A.D.A.C. (Association des Amis des Cryptogames)

Président : D. Lamy: Secrétaire : B. Dennetiére

Trésorier : E. Bury: Directeur de la publication : H. Causse

Source : MNHN. Paris

CRYPTOGAMIE

Diffusion de CRYPTOGAMIE

LE PÉRIODIQUE FRANCAIS

CONSACRÉ A LA CRYPTOGAMIE

CRYPTOGAMIE est un périodique édité

par l'A.D.A.C. (Association des Amis des

Cryptogames), dont le siége est au Labo-

ratoire de Cryptogamie du Muséum Na-

tional d'Histoire Naturelle. Les cher-

cheurs de tous pays y publient leurs

travaux en français, allemand, anglais, es-

pagnol et italien, après accord des

Comités de Lecture constituės de

spécialistes de réputation internationale.

CRYPTOGAMIE propose trois sections:

Cryptogamie, Algologie

Cryptogamie, Mycologie

Cryptogamie, Bryologie-Lichénologie

Chaque section publie 4 numéros par an

(rage: 450 exemplaires).

THE FRENCH JOURNAL

DEVOTED TO CRYPTOGAMY

CRYPTOGAMIE is a periodical

published by A.D.A.C. (Association des

Amis des Cryptogames), settled at Labo-

ratoire de Cryptogamie, Muséum National

d'Histoire Naturelle. Research workers

from the whole world publish their papers

in French, German, English, Spanish and

Jtalian, after acceptation by à selection

commettee that comprises experts of inter-

national renown.

CRYPTOGAMIE offers to its subscribers

three sections:

Cryptogamie, Algologie

Cryptogamie, Mycologie

Cryptogamie, Bryologie-Lichénologie

Each section publishes 4 numbers a year

(printing: 450 ex.).

[ E europe marine

| Bamérique E asie

O Australie

Origine des 453 articles publiés de 1986 à 1991

E Europe

mérique

EJ Francais Espagnol Ml italien

PA angieis [Allemand

Source : MNHN, Paris

SOMMAIRE

S.M. BADALYAN, S. RAPIOR, L. DOKO, J. LE QUANG, M. JACOB, J.J

SERRANO and C, ANDARY - Investigation of primary and secondary

metabolites in a chemical study of Cortinarius armillatus (Cortinaria-

ceae, Telemonia). .

1. KALYANASUNDARAM, L. MENON and P. LOGANATHAN - Occurrence

of melanin in bright-spored Myxomycetes.

G. MORENO, F. ARENAL y V. GONZÁLEZ - Algunos agaricales de las playas

de España peninsular.

G. MORENO, E. HORAK y M. LAGO. - Descolea maculata Bougher

(Agaricales), nueva cita para Europ:

B. PAUL - Some species of Pyrhium isolated from cultivated soils in northern

S.A. OMAR and A.-L.E. MAHMOUD - Post-harvest rots of tomato in relation to

lyases and mycotoxin production in vitro and in vivo. ...

Analyses bibliographiques

Table du tome 15 ....

Cryptogamie, Mycol. 1994, 15 (4): -223-290.

223

229)

255

263

213

283

289

MINH,

АР,