Fumarprotocetraric Acid Research Articles

Quantitative Distribution of Lichen Products in Australian Scyphose Cladonia Species

AbstractThe quantitative distribution of five lichen acids, including fumarprotocetraric acid, in seven specimens of Australian scyphose Cladonia species was determined by reverse phase high pressure liquid chromatography, using butylated hydroxy-toluene as internal standard. The highest concentrations of lichen acids were found in the scyphi and it is suggested that this localized distribution of lichen acids is due to repeated upward capillary flow, of saturated solutions of the acids, in the podetia followed by evaporation in the upper parts of the podetia.

Lichen Products of Cladonia Stellaris and C. Rangiferina Maintained Under Artificial Conditions

AbstractLichen thallus fragments were maintained in a growth cabinet under four differing conditions of light intensity. Levels of usnic acid and atranorin did not differ significantly from the control under any of the treatments. However, perlatolic acid and fumarprotocetraric acid per unit dry weight of thallus were dependent upon light availability.

A Standardized TLC Analysis of β-Orcinol Depsidones

Many p-orcinol depsidones with low Rf values in the three stan- dardized TLC solvents commonly used in lichenology are better resolved by a solvent system of toluenelethyl acetate/formic acid. Standardized chromatographic data and examples of specific problems resolved by one- and two-dimensional TLC with this solvent system are given. Chemical analyses are reported for 25 lichen species from 17 genera. A vexing problem in the identification of lichen products by thin-layer chromatography (TLC) has been the separation of some 35 low-Rf, p-orcinol compounds many of which are still unknowns. In the three standardized solvent systems (Culberson & Kristinsson 1970; Culberson 1972, 1974; Culberson & Ammann 1979), most of these compounds run below the control substance norstictic acid, which has Rf values between 28 and 40. Even if the spots for these substances were evenly spaced, the Rf values would differ by only about one millimeter. In addition, we now know that these compounds occur in complicated mixtures and consequently accurate identifications will require more sophisticated chro- matographic techniques than those routinely used. Reports in the recent literature of low- Rf compounds (even common ones like salazinic, constictic and connorstictic acids) may be misidentifications based upon insufficient TLC evidence. The purpose of the present paper is to give standardized TLC data for a fourth solvent system (Solvent G) to assist the resolution of difficult mixtures of low-Rf, p-orcinol depsidones by one- and two-dimen- sional TLC. The chromatographic procedure followed the standardized method (Culberson 1972) except that toluene (dried over CaCl2 and redistilled) was used in place of benzene. Solvent G, toluene/ethyl acetate/formic acid (139:83:8, v/v/v; 230 ml), is modified from previously suggested systems (Stahl & Schorn 1961; Hirayama et al. 1976). Formic acid (95-97%) and ethyl acetate were not purified or redistilled; it is not necessary to purify a good grade of toluene for satisfactory results with Solvent G. As with Solvent B, however, a grade of formic acid with low water content is essential. TLC plates were Merck Silica Gel 60 F254 (0.25-mm layer; No. 5765). Chromatographic tanks (Desaga- Heidelberg) were insulated from external temperature fluctuations but were not lined with filter paper. Pre-equilibration of the plates in 60% formic acid (Culberson 1974) did not improve the quality of chromatograms in Solvent G. Unless noted otherwise, the standardized data in Table 1 were from shortened plates (12.5 x 20 cm) developed to 10 cm. Initial spots were 2 cm from the bottom of the plate and 0.5 cm above the solvent. Controls for standardization were included at three positions on every plate. The Rf data in Table 1 are standardized with respect to fumarprotocetraric acid in addition to the usual control substances, atranorin and norstictic acid. Two-dimensional chromatograms were prepared by the standardized method (Culberson & John- son 1976) except that full plates (20 x 20 cm) were used with seven one-dimensional comparisons (including two atranorin-norstictic acid controls) in each direction. In addition, for the second direc- tion, an atranorin-norstictic acid control was added just to the left of the origin on the two-dimensional portion of the plate. This control helps to align spots and to monitor effects of residual first-direction solvent on R, values in the second direction. (In the present report, first- and second-direction solvents

On Bryoria Chalybeiformis and Some Related Species

AbstractBryoria chalybeiformis, as commonly interpreted, is an almost exclusively saxicolous or terricolous species with a largely arctic-alpine distribution; it produces small amounts of fumarprotocetraric acid in the soralia. The terricolous form of B. fuscescens var. positiva, distributed from sea level to the high mountains, is distinguished by the production of abundant fumarprotocetraric acid accompanied by protocetraric acid. B. intricans appears to be closely related to B. chalybeiformis and may possibly represent an environmentally induced morphotype of it. Attention is drawn to nomenclatural problems connected with B. chalybeiformis.

Contact allergy to atranorin in lichens and perfumes.

Atranorin, one of the most common lichen substances, gave positive patch test reactions in eight subjects (1%) in a routine series. These subjects also reacted to fumarprotocetraric acid and some of them to evernic acid. Stictic acid and usnic acid gave negative reactions. The lichen oak moss Evernia prunastri and an oak moss perfume gave positive reactions. Thin-layer chromatography and a spot test indicated that atranorin is present in oak moss perfumes which are made from oak moss and tree moss. Contact with oak moss perfumes and lichens in nature may cause atranorin allergy. None of the eight subjects had a history of light sensitivity or atopy and none had chronic facial eczema.

Nomenclatural Notes on Cladonia Species

AbstractThe nomenclature and in part also the taxonomy of the following European lichens is discussed: Cladonia cervicornis (Ach.) Flotow subsp. cervicornis and subsp. verticillata (Hoffm.) Ahti comb. nov., C. bacilliformis (Nyl.) Glück, C. portentosa (Dufour) Coem., C. anomaea (Ach.) Ahti & P. James comb. nov. (syn. C. pityrea (Flörke) Fr.), C. conista Robbins ex Allen, C. conoidea Ahti sp. nov. (syn. C. conistea sensu Asah.: contains atranorin and fumarprotocetraric acid, type from England), C. bacillaris Nyl., C. coniocraea auct., and C. ochrochlora Flörke.The African lichen ‘Cladonia gorgonina (Bory) Eschw.’ is typified and its nomenclatural status reconsidered. Under the valid name Cladonia gorgonea Eschw. it is reduced to the synonymy of Cladia aggregata (Swartz) Nyl. The correct name for Cladonia gorgonina auct. is C. glaucopallida Vainio. A list of the new lichen taxa proposed by J. B. Bory, which were invalidly published by Flörke in 1809, is also presented.

A new dibenzofuran and diphenyl ether from the lichen Gymnoderma melacarpum

The dibenzofuran melacarpic acid [1-heptyl-3-hydroxy-7-methoxy-9- methyldibenzofuran-2-carboxylic acid], the diphenyl ether congrayanic acid [2-(3'-carboxy-2'-heptyl-4',6'-dihydroxyphenoxy)-4-methoxy-6- methylbenzoic acid] and the depsidones grayanic acid and fumarprotocetraric acid have been shown to be constituents of the Australian lichen Gymnoderma melacarpum (Wils.) Yoshim.

The effect of thallus age and drying procedure on extractable lichen substances

Cladonia rangiferina (L.)Wigg., Cladonia mitis Sandst., and Umbilicaria muhlenbergii (Ach.) Tuck, were extracted with acetone while damp and after three different drying procedures. The extracts were chromatographed and scanned spectrophotometrically. Some of the C. rangiferina compounds were not extractable from damp material. All of the lichen substances studied here decreased significantly in thalli which were dried in an oven at 80 °C, some to the point of imperceptibility. There was no qualitative difference between the complement of compounds found in lichen material dried at 22 °C and at 40 °C. We recommend that drying be carried out at a temperature not exceeding 40 °C. Chemical variability among thallus regions of different ages was studied in Cladonia rangiferina, C. mitis, Umbilicaria muhlenbergii, and Lasallia papulosa (Ach.) Llano. Fumarprotocetraric acid, atranorin, and usnic acid were present in significantly higher amounts in younger tissue than in older. Protocetraric acid in C. rangiferina was detectable only in the older portions and would have been overlooked if young branches had been analyzed exclusively. In neither of the two umbilicate lichens was there any significant difference in the concentration of gyrophoric acid between younger and older thallus parts. Chemical variability among different-aged thallus parts was thus greater in the fruticose species than in the umbilicates examined here.

Physiography and Fumarprotocetraric Acid Production in the Cladonia chlorophaea Group in North Carolina

Physiography and Fumarprotocetraric Acid Production in the Cladonia chlorophaea Group in North Carolina

Paleogeography, Chemistry and Taxonomy of the Lichenized Ascomycetes Dimelaena and Thamnolia

World distribution of the species of Dimelaena and Thamnolia, and chemical variants, are considered in relation to current concepts of conti- nental drift. It is concluded that both genera date from the Permian-Triassic and paleogeographies of species and chemotypes are presented which are consistent with the established sequence of continental movements. In addition, long- distance dispersal has probably occurred in both species of Thamnolia. Seven chemotypes of Dimelaena oreina are now known and are interpreted as belonging to three races with differing paleogeographies. The races correspond to the fumarprotocetraric acid, gyrophoric/acid deficient chemotypes and the stictic/ norstictic chemotypes. They have not been given taxonomic recognition, how- ever, because the acid deficient chemotype may not be genetically homogenous, possibly incorporating acid deficient phases of the two other races in addition to the acid deficient phase of the gyrophoric acid chemotype. The fumarprotocetra- ric and gyrophoric acid chemotype incorporates substances of two races and does not occur outside the area of sympatry. This chemotype is best regarded as either a cross or a mycobiont chimera of its constituent races. Little attention has been paid to phytogeography of lichens relative to other groups of macroscopic organisms. There are two reasons for this. Firstly, few lichen genera are sufficiently well known taxonomically for worldwide distribution to be adequately documented and, secondly, the apparent widespread distribution of some taxa has lead to a belief that their phytogeography is inherently uninteresting. The cosmopolitan dis- tribution of such lichens may be a function of either their abilities for long-distance dis- persal or of their great age and passive dispersal by movements of the earth's crust. While plate tectonics provides an explanation for the present distribution of many groups of organisms, to the knowledge of this author, only W. L. Culberson (1972) considered the phytogeography of the lichens in this light. The two small genera Dimelaena Norm. and Thamnolia Ach. ex Schaer. are well known and have widespread distributions. These provide an opportunity for further examining the role of continental drift in lichen phytogeography. The species of Dime-

The Genus Dimelaena in North America North of Mexico

The Genus Dimelaena in North America North of Mexico

Morphological and Chemical Correlations in the Cladonia chlorophaea Complex

Measurements of granules from within the podetial cups of different chemical variants of the Cladonia chlorophaea complex in North Carolina show clear correlation between granule size and certain chemical characters. The results support the view that the replacement of cryptochlorophaeic acid by grayanic acid in this group is taxonomically more significant than the presence or absence of fumarprotocetraric acid. In the species tested, the soredia of C. conista are the smallest, those of C. perlomera and C. grayi are intermediate, and those of C. cryptochlorophaea are the largest. The Cladonia chlorophaea group was treated by Sandstede (1931) as including two species-C. chlorophaea (Flirke) Spreng., containing fumarprotocetraric acid and consequently distinguished by a bitter taste, and C. grayi Merr. ex Sandst., lacking fumarprotocetraric acid and consequently with a mild taste. Asahina (1940) discovered grayanic acid, cryptochlorophaeic acid, and merochlorophaeic acid in this group and divided the complex into four species: Cladonia grayi, C. cryptochlorophaea Asah., and C. merochlo?rophaea Asah. are characterized by the presence of grayanic acid, cryptochlorophaeic acid, and merochlorophaeic acid, respectively, while C. chlorophaea is considered to have fumarprotocetraric acid only. Within each of the first three species, the presence or absence of fumarprotocetraric acid is generally ignored from taxonomic considerations, but Asahina treated such fumarprotocetraric acid variants as forms. Although these four species are the most common ones of the chlorophaea group, many other variants have been recorded (Ahti, 1966; Culberson & Kristinsson, 1969). The morphologic variability has generally been considered to be the same within all the chemically different races in the complex. Ahti (1966), however, indicated some morphological differences between some of the plants called C. merochlorophaea and those of the other chemical species, and Krog (1968) claimed that the rangiformic acid variant in Alaska has a characteristic morphology. Wetherbee (1969) has demonstrated substrate specificity in some of the chemical species in Michigan. The present paper deals with the morphology and distribution of the chemical variants of the C. chlorophaea group in North Carolina. Since a preliminary survey indicated that there was a significant difference in the size of the soredia in the podetial cups between some of the chemical variants, a large-scale survey was made. Other 1 This study was supported by grant GB-6041X from the National Science Foundation. I thank Dr. William Louis Culberson for his interest in this work and for linguistic corrections and Dr. Chicita F. Culberson for her advice on the identification of the lichen substances. 2 Department of Botany, Duke University, Durham, North Carolina 27706. Present address: NMttlirugripasafnit Q Akureyri, P.O. Box 580, Akureyri, Iceland. This content downloaded from 157.55.39.153 on Mon, 19 Sep 2016 04:46:55 UTC All use subject to http://about.jstor.org/terms 14 THE BRYOLOGIST [Volume 74 100 Cladonia grayi PD+ s ---C. grayi PD?. C. cryptochlorophaea PD+ . .... .. ........ C. cryptochlorophaea PDo/ .... C)' I 0 100 200 300p LLI N A

Chelating ability of fumarprotocetraric acid andParmelia conspersa

Fumarprotocetraric acid andParmelia conspersa interact with granite to produce soluble colored complexes which shows that chemical weathering of the rock material takes place. The chelating ability ofP. conspersa may be due to the presence of fumarprotocetraric acid. Contrary to popular belief, many lichen acids are not very insoluble in water.

The Chemistry and Systematics of Some Species of the Cladonia cariosa Group in North America

Cladonia polycarpia, an endemic of the coastal plain of the southeastern United States, consistently produces atranorin and norstictic and stictic acids. Records of this species from western North America are referable to C. cariosa and C. symphycarpa. Cladonia polycarpoides is abundant and widely distributed in the East but absent from most of the range of C. polycarpia. It produces norstictic acid and lacks stictic acid and atranorin. Cladonia subcariosa, a rare species producing atranorin and norstictic acid, is known from Cuba and a few localities in the Atlantic coastal plain. Morphologically, chemically, and taxonomically Cladonia is the best known of the large genera of lichens. In spite of the intensity with which the genus has been studied, numerous species groups within it remain poorly understood-the result of inadequate collecting in critical regions, analytic procedures too insensitive to assess the chemical variation, or insufficient study of morphological variation, or a combination of all of these factors. The Cladonia cariosa complex is one of these problem groups. The present study grew out of my interest in the relationship between Cladonia polycarpoides and C. polycarpia, two common, almost allopatric species of the eastern United States. Most of the effort of the present research went to the systematics of these two species, and data on other members of the group are included for comparative purposes only. Some of the species are well differentiated morphologically, such as C. apodocarpa and C. capitata. The rest of the species resemble each other greatly although subtle morphological differences, some of which might better be called mere tendencies, seem to accompany most of the major chemical types. Chemistry, however, shows an impressive variation. Chemically the C. cariosa group is quite distinct (Table 1). With the exception of C. cariosa itself, every species contains at least one depsidone of the p-orcinol series. While the p-orcinol depsidone fumarprotocetraric acid is widely distributed in many Cladoniae, other p-orcinol depsidones are rare outside of the C. cariosa group (Culberson, 1969). Psoromic acid is known in only a few species in addition to C. brevis. Norstictic acid, a constituent of four members of the C. cariosa group, is known additionally in Cladonia only in the single species C. acuminata. In the entire genus the only stictic acid-producing species is C. polycarpia. There are, moreover, no /orcinol depsidones in Cladonia which are not represented in the C. cariosa group. 1 Assistance from grant GB-8359 from the National Science Foundation is acknowledged with thanks. I also thank Dr. Teuvo Ahti and Dr. John W. Thomson for criticism of the manuscript and Dr. Ahti and Dr. Mason E. Hale, Jr., for the loan of specimens. 2 Department of Botany, Duke University, Durham, North Carolina 27706. This content downloaded from 207.46.13.124 on Wed, 22 Jun 2016 05:19:57 UTC All use subject to http://about.jstor.org/terms 378 THE BRYOLOGIST [Volume 72 TABLE 1. The major secondary natural products in the North American species of the Cladonia cariosa group. The substances are: atr = atranorin; nor = norstictic acid; sti stictic acid; fum = fumarprotocetraric acid; pso = psoromic acid.

Chemical and Morphological Variation in the Cetraria islandica Complex in Iceland

A study of collective lichen species Cetraria (L.) Ach. based upon some 3,000 individuals from 100 localities in Iceland shows that extreme morphological variation is not correlated with chemical variation (presence or absence of fumarprotocetraric acid). This is in striking contrast to high correlation of chemical and morphological characters in this species complex on European mainland, which serves there as basis for separation of C. sens. str. and C. ericetorum Opiz. Differences in geographical distribution of chemical characters, fertility, and range of morphological variability indicate that there are two ecological races in Iceland: 1) fumarprotocetraric acid-producing, nonfertile race with rather narrow morphological variability, mainly distributed along coasts and in more oceanic southern part of country, and 2) morphologically and chemically variable race (with or without fumarprotocetraric acid), frequently fertile, and distributed throughout Central Highlands and northern Iceland. In latter race, frequent occurrence of both chemical types within same morpho-phenotype at individual localities indicates that gene exchange may occur between fumarprotocetraric acid-producing and fumarprotocetraric acid-lacking plants. The related species Cetraria (L.) Ach. and C. ericetorum Opiz have long been considered distinct in Europe. Originally they were distinguished by morphological characters only, C. having broad, flat to involute lobes with numerous white pseudocyphellae all over lower side, and C. ericetorum having narrow, almost tubular lobes and lacking pseudocyphellae on lower side except along revolute edge. After thallus spot test with p-phenylendiamine (PD) was introduced, specific distinction of these taxa was strengthened by fact that all specimens of C. showed PD+ medullary reaction and all specimens of C. ericetorum PDreaction. But as later became apparent, this situation prevails only in Europe. Lynge (1921) considered C. ericetorum (then called C. crispa) a proper species, well separated from C. islandica after he studied lichen flora of Norway. His opinion changed, however, after experience with these taxa in Arctic. In his work on Greenland (1932) he wrote the specific difference is much less distinct in Arctic than in southern regions. After testing all of his specimens of C. 1 This study was supported by Grant No. GB-6041X from National Science Foundation. I thank Dr. William Louis Culberson for valuable suggestions and aid in preparing this paper and Dr. Chicita F. Culberson for help with chemistry. 2 Department of Botany, Duke University, Durham, North Carolina 27706. This content downloaded from 157.55.39.45 on Fri, 02 Sep 2016 04:46:26 UTC All use subject to http://about.jstor.org/terms 1969] KRISTINSSON: CETRARIA ISLANDICA IN ICELAND 345 complex from Spitsbergen and North-East Land with PD, he said (Lynge, 1938): ... well known morphological characters are so confluent, that they do not warrant reliable identification alone. We might, perhaps, quite as well unite . . . [C. and C. ericetorum] into one species with two types which are distinct chemically. Imshaug (1957) commented upon distinctness of variation in C. complex in Europe and confusing variation in it outside Europe and discussed possible history of group. Ahti (1964) pointed out that delimitation based upon chemical characters alone leads to very unnatural species grouping in this complex and suggested that separation based on morphological and ecological grounds may be possible-even though this would lead to inclusion of different chemical races within each species. Krog (1968) recorded different chemical constituents in C. specimens from Alaska, variation not only with regard to presence or absence of fumarprotocetraric acid but also with regard to rangiformic acid. The purpose of present paper is to analyze morphological and chemical variation in Cetraria group in Iceland. It is part of broad study of total lichen flora of that country.

Some Microchemical Tests for the Lichen Depsidone Hypoprotocetraric Acid

Hypoprotocetraric acid, a depsidone of the p-orcinol type recently found in a race of Ramalina siliquosa, can be identified by microcrystal tests and by chromatography. This substance replaces protocetraric acid in individuals in some populations of Ramalina farinacea in France. Hypoprotocetraric acid (Fig. 1) is a depsidone of the p-orcinol type closely related to protocetraric acid. It was first obtained by Asahina and Tanase (1933) by synthesis from fumarprotocetraric acid. Hypoprotocetraric acid was unknown in nature until recently when it was identified (C. F. Culberson 1965) in a race of Ramalina siliquosa (Huds.) A. L. Sm. from western Europe. It seems to occur in at least

A Summary of the Lichen Genus haematomma in North America

A Summary of the Lichen Genus haematomma in North America