Buellia Species with Pluriseptate Spores and the Physciaceae (Lecanorales, Ascomycotina)

Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 593

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Taxonomic, Phylogenetic and Ultrastructural Studies

ANDERS NORDIN

ACTA UNIVERSITATIS UPSALIENSIS UPPSALA 2001 Abstract Nordin, A. 2001. Buellia species with pluriseptate spores and the Physciaceae (Lecanorales, Ascomycotina). Taxonomic, phylogenetic, and ultrastructural studies. Acta Univ. Ups. Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 593. 15 pp. Uppsala. ISBN 91-554-4879-8.

This dissertation is primarily focussed on the Buellia species with pluriseptate spores, often referred to the section or genus Diplotomma, and secondarily on the Physciaceae. An ultrastructural study at family level gives new information on spore characters. Variations in the composition and structure of the spore wall layers and septa are described. A phylogenetic analysis of the Physciaceae, based on morphological and chemical characters, is presented. In the resulting trees the species with pluriseptate spores appear in a poorly resolved clade together with other crustose species with 1-septate spores. A subsequent analysis, excluding foliose and fruticose species, shows that the species with pluriseptate spores do not form a monophyletic group, and that only a few are grouped together with the type species of Diplotomma. Due to low resolution, poor branch support, and uncertainty concerning the typification of Buellia all species treated are retained in Buellia. Altogether 35 species with pluriseptate spores are treated, including ten new to science, viz. B. aeruginosa, B. morsina, B. muriformis, B. oidaliella, B. pallidomarginata, B. romoletia, B. rubroreagens, B. terricola, B. tombadorensis, and B. triseptata. Historical outlines are presented, and information on life strategy, morphology, chemistry, ecology, and distribution is given. All species are similar and macroscopically not easily distinguished, having crustose thalli and blackish apothecia, but there is a great variation in microscopic characters, secondary chemistry, substrate preferences, habitat ecology, and distribution range. Several names are typified and many synonyms are listed. References are given to all species referred to Diplotomma in literature.

Key words. Buellia, Diplotomma, phylogenetic analysis, pluriseptate spores, Physciaceae, taxonomy, ultrastructure.

Anders Nordin, Department of Systematic Botany, EBC, Norbyvägen 18D, SE-752 36 Uppsala, Sweden

ISSN 1104-232X ISBN 91-554-4879-8

Printed in Sweden by Uppsala University, Tryck & Medier, Uppsala 2000 To Lotte, Jonas and Edith the keel and the two sails of my frail vessel This thesis is based on the following papers, which will be referred to in the text by their respective Roman numerals:

I. Nordin, A. 1996. Buellia species (Physciaceae) with pluriseptate spores in Norden. – Symb. Bot. Ups. 31(3): 327–354.

II. Nordin, A. 1997. Ascospore characters in Physciaceae: an ultrastructural study. – Symb. Bot. Ups. 32(1): 195–208.

III. Nordin, A. 1999. Buellia species with pluriseptate spores: new and unrecorded species in North America. – Bryologist 102: 249–264.

IV. Nordin, A. Owe-Larsson, B. & Elix, J. 1999. Buellia aeruginosa, a new Australian species with pluriseptate spores. – Mycotaxon 71: 399–404.

V. Nordin, A. & Mattsson, J.-E. 2000. Phylogenetic reconstruction of character development in Physciaceae. – Lichenologist 32 (in press).

VI. Nordin, A. 2000. Taxonomy and phylogeny of Buellia species with pluriseptate spores (Lecanorales, Ascomycotina). – Symb. Bot. Ups. 33(1).

Published and accepted papers are reproduced with the publishers’ kind permission. CONTENTS

Introduction...... 6

Morphology and anatomy ...... 6

Habitat ecology and distribution...... 8

Chemistry...... 8

Phylogeny ...... 9

Taxonomy...... 11

Acknowledgements...... 14 Nordin

INTRODUCTION

The family Physciaceae (Lecanorales, Ascomycotina) is mainly characterized by the brown, septate spores and the lecanoralean asci with thickened tips. Morphologically and chemically there is a great diversity in the family. Most thallus types found among lichenized fungi are represented: crustose, placodioid, umbilicate, subfoliose, foliose, and fruticose; and a great number of secondary substances, representing different metabolic pathways occur. Most foliose genera have been thoroughly studied, viz. Heterodermia, Physcia, Phaeophyscia, and Physconia. Those containing crustose lichens, on the other hand, have not attracted the same interest. This particularly concerns Buellia, a heterogeneous genus rich in species, and in particular the group of species in the main focus here: the Buellia species with pluriseptate spores, often referred to as the section or genus Diplotomma (I, II, IV, VI). The project also includes studies at the family level: an ultrastructural study mainly focused on spore wall structures (III), and a phylogenetic analysis based on morphology and chemistry (V). The work has to a great extent been based on revisions of herbarium material, but field studies have been performed in different parts of the world (e. g. the Nordic countries, North America, and South Africa), where fresh material has been collected. In the co-authored papers (IV, V) I am responsible for the major parts.

MORPHOLOGY AND ANATOMY (I–VI)

In the Physciaceae there is a great diversity in the organisation of the thallus. It is crustose (Amandinea, Buellia, Hafellia, Mobergia, Rinodinella, Rinodina), placodioid (Australiaena, Diploicia, Dimelaena), foliose (Anaptychia, Dirinaria, Heterodermia, Hyperphyscia, Physcia, Phaeophyscia, Physconia, Pyxine), subfoliose (Phaeorrhiza), umbilicate (Dermatiscum, Dermiscellum), or fruticose (Santessonia, Tornabea). In the Buellia species with pluriseptate spores it is usually inconspicuous but there is considerable intraspecific variation. The thallus is heteromerous and has a central medullary part. There are different types of upper and lower cortical layers (paraplectenchymatous, prosoplectenchymatous, scleroplectenchymatous). In the crustose species there is no lower cortex and the upper cortex can be characterized as a phenocortex, consisting of anticlinally arranged hyphal ends covered by residues of dead hyphae and collapsed algal cells. The photobionts are green-algal and trebouxoid. The ascomata are apothecioid. In some of the genera they are lecanorine, i. e. a thalline exciple surrounds the apothecia (e. g. Anaptychia, Physcia, Physconia, and Rinodina), in others they are lecideine, i. e. no thalline exciple surrounds them. The thalline rim surrounding the apothecia of some of the Buellia species with pluriseptate spores lacks algae and cannot be characterized as a thalline exciple. The proper exciple of the species with lecideine apothecia varies with respect to thickness and pigmentation. The paraphyses are simple or branched in the uppermost part and the

6 Buellia species with pluriseptate spores uppermost cells are usually widened. The asci are layered and nonfissitunicate and the innermost layer forms a thick amyloid tholus with a nonamyloid central part. In some genera this part has diverging flanks and reaches through the entire tholus (e. g. Anaptychia, Physcia, Physconia, and Rinodina). In others the flanks converge inside the tholus (e. g. Amandinea, Buellia, Diploicia, and Santessonia). In some species the flanks are more distinctly amyloid than the surrounding parts of the tholus. The subhymenial layers are brown to blackish brown in most species of some genera (e. g. Amandinea, Buellia, Diploicia, and Santessonia), consistently pale in other genera (e. g. Anaptychia, Physcia, Physconia, and Rinodina). The spores are brown and septate at maturity. In the great majority of the species they are 1-septate, but in Buellia, Rinodina, Hyperphyscia, and Pyxine some species have pluriseptate spores. More or less temporary, uneven thickenings of the walls are common, often only seen at premature stages. These are apical, subapical, and lateral and occur singly or combined. A great number of different spore types have been distinguished mainly due to the appearance of these thickenings. The spore walls are layered (Fig. 1). In all species there is a pigmented proper wall, inside which there is an unpigmented endospore, and a gelatinous sheath surrounds the pigmented parts. In most species the spores also have a perispore outside the proper wall, from which it is separated by a thin and often indistinct, unpigmented intermediary layer. The perispore is rugulate or more or less smooth. There is great interspecific variation in the thickness of the layers.

The pycnidia are globose to flask-shaped, often secondarily compartmentalized, and partly or almost entirely immersed in the thallus. The conidia are bacilliform and straight (the majority of the genera), filiform and curved (Amandinea, Australiaena, Hyperphyscia), or ovoid (Mobergia, Phaeophyscia).

Gelatinous sheath Perispore Intermediary layer Proper wall Endospore

Proper Endospore septum septum

Figure 1. Spore wall layers and septa

7 Nordin

CHEMISTRY (I, III, IV, V, VI)

A great number of secondary metabolites have been recorded in the Physciaceae. The majority of the substances are products of the acetate-polymalonate pathway, but there are also representatives of the mevalonic acid and the shikimic acid pathways. The most common substance is atranorin, found in a great number of species in many different genera. Several substances not uncommon outside the Physciaceae are restricted to one of the genera, e. g. in Buellia barbatic acid, miriquidic acid, several xanthones, and calycin; in Dirinaria alectoronic acid, divaricatic acid, and sekikaic acid; and in Rinodina 5-O-metylhiascic acid, isosphaeric acid, lecanoric acid, pannarin, squamatic acid, thiomelin, and variolaric acid. The anthraquinones cinnamomeic acids are exclusively found in the Physciaceae (in the medulla of Rinodina mniarea var. cinnamomea and Buellia capitis-regum respectively). The substances are found in the medulla and/or the cortex, sometimes also in the apothecia. Spot test reactions with K, P, and C have been indicated in the papers dealing with Buellia species with pluriseptate spores (I, III, IV, VI). The K- and P- reactions are mainly due to the presence of atranorin and norstictic acid and the C-reactions to the presence of xanthones. In the apothecia of some species there are also accessory pigments with typical reactions with K and HNO3.

HABITAT ECOLOGY AND DISTRIBUTION (I, III, IV, VI)

This part only concerns the Buellia species with pluriseptate spores. Most of these species occur on bark and lignum of broad-leaved and coniferous trees and shrubs. One species, Buellia cedricola, grows exclusively on lignum of conifers. Seven species are obligately saxicolous: B. aeruginosa, B. scheideggeriana, B. subdispersa, and B. venusta are restricted to calciferous rocks, while B. capitis-regum, B. granulosa, and B. mexicana seem to be less specific. Two further species, B. alboatra and B. vernicoma, are saxicolous, but they also occur on bark and lignum of trees. There are also a few terricolous species growing on plant debris or over mosses and other lichens (B. geophila, B. graminicola, and B. terricola). Most species prefer semi-open to open habitats not exposed to direct sunlight. The majority of the species are mainly found in temperate areas of the northern hemisphere. Some of these species, such as Buellia alboatra, B. geophila, B. pulverulenta, B. subdispersa, B. terricola and B. venusta also extend farther north into the arctic tundra area. Seven species, Buellia alboatrior, B. lauricassiae, B. pallidomarginata, B. proximata, B. rubroreagens, B. sorediata and B. tombadorensis, mainly occur in the humid parts of the tropics, presumably in areas with a savannah climate (dryer in winter), but at least B. lauricassiae also extends into temperate areas. Only two species, B. oidaliella and B. romoletia, are restricted to the arid parts of the subtropics, but some additional species with a wider range, such as B. alboatra, B. morsina, B. oidalea, B. subdispersa and B. venusta, also

8 Buellia species with pluriseptate spores occur in arid areas. Only a few of the species are known from the southern hemisphere outside the tropics and subtropics. The Australian B. aeruginosa occurs in the warm temperate zone, and the South Georgian B. graminicola, together with the widespread B. pulverulenta, occurs in the cold temperate zone, while the ubiquitous B. alboatra is found in both. B. granulosa is an Antarctic tundra species. Some species are more or less restricted to highland areas, climatically variable but distinct from the surrounding lowlands. B. himalayensis, B. megaspora, and B. submuriformis are known from the Himalayas; B. cedricola from mountainous areas in SW North America, NW Africa and SW Europe; B. lauricassiaeoides and B. submuriformis from the highlands of Papua New Guinea; and B. penichra from the Rocky Mountains and mountain ranges farther west. These are all found in subalpine forests. B. mexicana, on the other hand, is an alpine species known from Arizona and Mexico.

PHYLOGENY (V, VI)

In order to get an understanding of the phylogenetic relationships in the Physciaceae (V) and between the Buellia species with pluriseptate spores (VI) respectively, cladistic parsimony analyses, based on morphological and chemical characters, were performed. In both cases, however, the resulting phylogenetic reconstructions were rather poorly resolved and poorly supported. In the strict consensus tree resulting from the family analysis, the unresolved ingroup only contained one group with more than two members (Anaptychia ciliaris, Tornabea scutellifera, A. runcinata, Physconia distorta, P. enteroxantha and Phaeorrhiza nimbosa), but in the great majority of the 3119 trees resulting from the analysis, there were two large major clades, mainly containing crustose lichens and foliose lichens respectively. The crustose clade was dominated by an unresolved branch, containing Amandinea, Buellia, Dermiscellum, Diploicia, Hafellia, and Santessonia, and the foliose clade was divided into two branches, one containing the group of the consensus tree, the other containing representatives of the remaining foliose genera. Rinodina species were found at basal positions. In the second analysis (VI) all the Buellia species with pluriseptate spores were included, together with a selection of other members of the crustose clade of the previous analysis, and with two Rinodina species as outgroup. As clearly indicated by the strict consensus tree resulting from the analysis (Fig. 2), the species with pluriseptate spores do not form a monophyletic group. Thus neither Diplotomma sensu Sing & Awasthi, Szatala, and others, nor section Diplotomma sensu Zahlbruckner are natural units. However, the type species of Diplotomma, Buellia alboatra, forms a monophyletic group together with B. pharcidia, B. venusta, B. scheideggeriana, B. subdispersa, and B. pulverulenta, a group that might merit taxonomic recognition, but as long as the relations to other groups remain unresolved and as long as the typification of Buellia has not been finally settled, all species have been retained in Buellia. A number of other validly described genera containing species with pluriseptate spore are also briefly discussed (Fig. 2).

9 Nordin

Strict R. bischoffii R. confragosa A.coniops Amandinea B. alboatrior B. submuriformis B. capitis-regum B.disciformis Buellia H. leptoclinoides H. parastata Hafellia B. oidalea B. oidaliella B. muriformis B. penichra B. cedricola B. erubescens B. griseovirens Aplotomma B. himalayensis B. lauricassiae Cratiria B. lauricassiaeoides B. rubroreagens B. pallidomarginata B. proximata D.canescens Diploicia B. sorediata B. megaspora B. morsina B. aeruginosa B. vernicoma Gassicurtia B. tombadorensis B. triphragmioides Mattickiolichen B.aethalea B. mexicana B. stellulata B. jugorum B. alboatra Diplotomma B. pharcidia B. venusta B. scheideggeriana B. subdispersa B. pulverulenta B. geophila Tetramelas B. terricola B. graminicola B. insignis B. granulosa B. romoletia B. triseptata

Figure 2. Strict consensus tree of 168 most parsimonious cladograms. Generic names added to the right of the terminal taxa indicate the positions of the type species or other representatives of validly described genera. Species with 1-septate spores are bold-faced.

10 Buellia species with pluriseptate spores

TAXONOMY (I, III, IV, VI)

The Buellia species with pluriseptate spores recognized, altogether 35, are briefly presented in alphabetical order. For new species and new combinations of paper VI the latin diagnoses and the basionyms are omitted and appear only in the original publication.

Buellia aeruginosa A. Nordin, Owe-Larsson & Elix (IV, VI) is a saxicolous, Australian species. It has weakly submuriform spores and the thallus (and apothecia) contains xanthones. An aeruginose pigment is found in the apothecia.

Buellia alboatra (Hoffm.) Th. Fr. (I, VI) is a widespread species occurring on different substrata. It is facultatively parasitic on other lichens (mainly Caloplaca and Xanthoria species). The spores are submuriform. It lacks secondary substances or contains norstictic acid.

Buellia alboatrior (Nyl.) Zahlbr. (VI) is a corticolous and lignicolous, tropical species, known from India and Brazil. The spores are submuriform and the thallus contains atranorin and diploicin.

Buellia capitis-regum W. A. Weber (VI) is a saxicolous, coastal, Californian species. The spores are submuriform and the secondary chemistry is complex, with diploicin as the major substance. The medulla contains anthraquinones, making it yellow in the lower part.

Buellia cedricola Werner (III, VI) is a lignicolous species occurring at high altitudes in south-western Europe, north-western Africa, and south-western North America. The spores are submuriform and the thallus contains norstictic acid and usnic acid, the latter giving the thallus a yellowish tinge.

Buellia geophila (Flörke ex Sommerf.) Lynge (I, VI) is a terricolous, arctic-subalpine, European species. The spores are 3-septate and the thallus contains 6-O- methylarthothelin.

Buellia graminicola Øvst. (VI) is a terricolous, South Georgian species. The spores are 3-septate and the thallus has irregularly dispersed, coarse soredia and contains 6- O-methylarthothelin and traces of two anthraquinones.

Buellia granulosa (Darb.) C. W. Dodge (VI) is a saxicolous, Antarctic species. The spores are 3-septate to weakly submuriform and the thallus contains 6-O- methylarthothelin.

Buellia griseovirens (Sm.) Almb. (I, VI) is a corticolous and lignicolous, sorediate species, widespread in Europe and North America. It often lacks apothecia. The spores are submuriform and the thallus contains atranorin and norstictic acid.

Buellia himalayensis (S. R. Singh & D. D. Awasthi) A. Nordin (VI) is a corticolous

11 Nordin

Indian species. The spores are large and muriform and the thallus contains atranorin and norstictic acid.

Buellia lauricassiae (Fée) Müll. Arg. (VI) is a corticolous and lignicolous, widespread, tropical species. The spores are 3-septate and the thallus contains atranorin and norstictic acid, the latter also found in the excipulum of the apothecia.

Buellia lauricassiaeoides Aptroot (VI) is a corticolous species, known from Papua New Guinea. The spores are 3-septate and the thallus contains atranorin.

Buellia megaspora (S. R. Singh & D. D. Awasthi) A. Nordin (VI) is a corticolous Indian species. The spores are large and muriform and the thallus contains atranorin.

Buellia mexicana J. Steiner (III, treated under the name B. tolucae, VI) is a saxicolous, North American species, found at high altitudes in Mexico and Arizona. The spores are mainly submuriform and the thallus contains atranorin and norstictic acid. An aeruginose pigment is found in the apothecia.

Buellia morsina A. Nordin (VI) is a corticolous Mexican species. The spores are submuriform and the thallus contains atranorin and norstictic acid.

Buellia muriformis A. Nordin & Tønsberg (III, VI) is a corticolous and lignicolous, North American species, occurring on the Pacific coast between British Columbia and mid-California. The spores are muriform and the thallus contains atranorin, placodiolic acid, and some additional minor substances.

Buellia oidalea (Tuck.) Tuck. (VI) is a corticolous and lignicolous, North American species, occurring on the Pacific coast between Oregon and Baja California Sur. The spores are large and muriform and the thallus contains diploicin and some additional minor substances.

Buellia oidaliella A. Nordin (III, VI) is a corticolous and lignicolous, Mexican species, restricted to southernmost Baja California and Baja California Sur. The spores are submuriform and the thallus contains diploicin and some additional minor substances.

Buellia pallidomarginata A. Nordin (VI) is a corticolous Indian species. The spores are 3-septate and the thallus contains atranorin and diploicin. The apothecia usually have a pale rim.

Buellia penichra (Tuck.) Hasse (VI) is a corticolous and lignicolous, North American species, occurring in the Rocky Mountains and the mountain ranges farther west. The spores are submuriform and the thallus contains atranorin and sometimes placodiolic acid.

Buellia pharcidia (Ach.) Malme (I, VI) is a corticolous and lignicolous species, widespread in Europe. The spores are 3-septate to weakly submuriform. No secondary substances are produced.

12 Buellia species with pluriseptate spores

Buellia proximata H. Magn. (VI) is a corticolous and lignicolous, widespread but seemingly rare, tropical species. The spores are 3-septate and the thallus contains atranorin and diploicin.

Buellia pulverulenta (Anzi) Jatta (I, VI) is a widespread, arctic-subalpine species, obligately parasitic mainly in the thalli of Physconia and Physcia species. The spores are 3-septate. No secondary substances are produced.

Buellia romoletia A. Nordin (VI) is a corticolous species so far only known from westernmost South Africa. The spores are 3-septate and the thallus contains 6-O- methylarthothelin.

Buellia rubroreagens A. Nordin (VI) is a corticolous and lignicolous, tropical species, occurring in Brazil and the West Indies. The spores are 3-septate and the thallus contains atranorin and sometimes norstictic acid. The excipulum contains a yellow KOH+ red pigment.

Buellia scheideggeriana Bricaud & Cl. Roux (VI) is obligately parasitic on the saxicolous species Caloplaca chrysodeta and C. xantholyta. It occurs in southern Europe. The spores are 3-septate to weakly submuriform. No secondary substances are produced.

Buellia sorediata (Tuck.) H. Magn. (VI) is a corticolous, sorediate species, only known from the type locality in Nicaragua. The spores are 3-septate and the thallus contains atranorin.

Buellia subdispersa Mig. (III, VI) is a saxicolous species, mainly occurring in Europe but also found in America, and not seldom establishing on other lichens. The spores are 3-septate. No secondary substances are produced.

Buellia submuriformis Aptroot & Diederich (VI) is a corticolous, tropical species, occurring in Papua New Guinea and India. The spores are large and mainly submuriform and the thallus contains atranorin and diploicin.

Buellia terricola A. Nordin (III, VI) is a terricolous, arctic-subalpine, American and Asian species. The spores are 3-septate and the thallus contains arthothelin and usually atranorin and norstictic acid (the latter mainly in the excipulum of the apothecia).

Buellia tombadorensis A. Nordin (VI) is a corticolous species, so far only known from the type locality in Brazil. The spores are 3-septate and the thallus (and apothecia) contains xanthones. The excipulum contains two accessory pigments. One is yellow and KOH+ red and the other is blackish and gives a yellow solution in HNO3. Buellia triphragmioides Anzi (I, VI) is a corticolous and lignicolous, subarctic- subalpine species, mainly occurring in Europe and North America. The spores are 3- septate and the yellowish thallus contains xanthones.

13 Nordin

Buellia triseptata A. Nordin (III, VI) is a corticolous and lignicolous species, known from western North America and Spain. The spores are 3-septate and the thallus rarely contains atranorin.

Buellia venusta (Körb.) Lettau (I, VI) is a saxicolous species, mainly occurring on calciferous rocks in Europe and North America and not seldom establishing on other lichens, such as Lecanora muralis. The spores are 3-septate and the thallus sometimes contains norstictic acid.

Buellia vernicoma (Tuck.) Tuck. (VI) is a saxicolous, corticolous and lignicolous species, occurring in eastern North America and Japan. The spores are small and 3- septate and the yellowish, granular thallus contains xanthones.

ACKNOWLEDGEMENTS

Leif Tibell and Roland Moberg are gratefully acknowledged for their continuous supervision and tuition during the different phases of my work. Under their leadership the lichen group of Uppsala has discussed not only the work in progress among the members of the group but also lichenological and mycological matters in general, which has been most instructive and valuable. The fact that Rolf Santesson has remained a member of the group has greatly contributed to the width and depth of the discussions, and I owe him much, particularly for good advice in nomenclatural matters. Other constant or temporary members who have contributed to the discussions and assisted in different ways are Starri Heiðmarsson, Jan-Erik Mattsson, Göran Thor, Mats Wedin, Sven-Gunnar Ryman, Kristina Articus, Liselott Skarp, and Lena Dahlman. The present and former heads of the department, Birgitta and Kåre Bremer, and the members of the Board and the teaching staff, are acknowledged for keeping high standards and for creating an intellectually stimulating atmosphere. I am also most grateful for the substantial financial support granted by a PhD-student position at the department. Special thanks go to Starri Heiðmarsson, my room-mate for six years, now back in Iceland. Constantly in good spirits and with an optimistic approach to life, he was always willing to help in various matters, and he never complained about my using his computer for time-consuming phylogenetic analyses. Also the other fellow PhD- students of the department, past and present, contributed to making the years at the department a rewarding time. I am also indebted to the directors and curators of the herbaria for sending material on loan; to Roland Moberg and the staff of the Botany Section, Museum of Evolution, Uppsala University for the administration of these loans; and to Pieter van den Boom, Ana Burgaz, Philipe Clerc, Javier Etayo, Tassilo Feuerer, Mireia Giralt, Trevor Goward, Klaus Kalb, Bruce McCune, Dag Olov Øvstedal, Björn Owe-Larsson, Mark Seaward and Tor Tønsberg for providing additional interesting material.

14 Buellia species with pluriseptate spores

I also wish to express my gratitude to Jack Elix for letting me include unpublished results from HPLC analyses; to Hannes Hertel, Rosemary Romaker, Ulrich Søchting, Walter Till, Mauro Tretiach, and Orvo Vitikainen for important information on literature, collections and type localities; to Tom Nash, Charis Bratt, Mónica Alegre Gonzáles, and Beth Kantrud for organisation and assistance in connection with the field work; and to Anette Axén, Stefan Gunnarsson, Karin Ryman, Ulla-Britt Sahlström and Gary Wife for technical assistance. Financial support was received from The Royal Swedish Academy of Science ("Karin och Axel Binnings fond ”, J. A. Wahlbergs minnesfond”, and ”P. F. Wahlbergs minnesfond”), and from the funds of "Göransson-Sandviken", "Regnells Botaniska", "Elias Fries", "Uddenberg-Nordinska Stiftelsen", and "Sernaders stiftelse".