Phylogenetic Relationships and Species Richness
of Coprophilous Ascomycetes
Åsa Nyberg Kruys
Department of Ecology and Environmental Science
Umeå University
Umeå 2005
AKADEMISK AVHANDLING
som med vederbörligt tillstånd av rektorsämbetet vid Umeå universitet för
avläggande av filosofie doktorsexamen
framläggs till offentligt försvar i Stora hörsalen, KBC,
fredagen den 25 november 2005, kl. 9.00.
Avhandlingen kommer att försvaras på engelska.
Examinator: Dr. Mats Wedin, Umeå Universitet
Opponent: Ass. Prof. Thomas Laessøe, Department of Microbiology,
University of Copenhagen, Denmark.
ISBN 91-7305-949-8
© Åsa Nyberg Kruys
Printed by Solfjädern Offset AB
Cover: Ascus of Sporormiella antarctica with eight 13-celled ascospores.
Design by Åsa and Nic Kruys.
Organization
UMEÅ UNIVERSITY
Department of Ecology and Environmental Science
SE-901 87 Umeå, Sweden
Document name
DOCTORAL DISSERTATION
Date of issue
November 2005
Author
Åsa Nyberg Kruys
Title
Phylogenetic relationships and species richness of coprophilous ascomycetes.
Abstract
Coprophilous ascomycetes are a diverse group of saprobes, of which many belong to three families,
Delitschiaceae, Phaeotrichaceae and Sporormiaceae, within the large order Pleosporales. The natural
relationships and circumscription of these families are unclear, especially within the family Sporormiaceae,
where the generic delimitation have been questioned. There is also a need to understand how different ecological
processes affect species richness and occurrence of coprophilous ascomycetes in general. The aim of this thesis
was therefore to test earlier classifications of coprophilous taxa within Pleosporales, using phylogenetic analyses
of DNA sequences; and to study how the habitat, dung type and herbivores´ food choice may affect the species
richness and species composition of coprophilous ascomycetes.
A phylogenetic study shows that coprophilous taxa have arisen several times within Pleosporales.
Sporormiaceae and Delitschiaceae are separate monophyletic groups and should continue to be recognized as
two distinct families within Pleosporales. Phaeotrichaceae forms a monophyletic group, and is, unexpectedly, a
strongly supported sister-group to Venturiaceae, but if they belong to Pleosporales or not, remains unresolved.
Testudinaceae and Zopfiaceae, which previously had an unclear position in Ascomycota, are shown to be
members of Pleosporales and should be treated as two separate families. The genus Eremodothis is, however, not
related to Testudinaceae, but is nested within Sporormiaceae and should be transferred to Westerdykella.
The natural relationships within Sporormiaceae are still not fully resolved and consequently, I suggest a
rather conservative generic classification, accepting Preussia, Sporormia, W e s t e r d y k e l l a , as well as
Sporormiella, despite that the latter is not conclusively well supported as monophyletic. Characters previously
used in the taxonomy and classification of Sporormiaceae, as choice of substrate, presence or absence of an
ostiole, presence or absence of germ slits, and spore ornamentation, were all homoplastic and not very useful for
circumscribing monophyletic groups.
Field-studies of moose (Alces alces), mountain hare (Lepus timidus) and roe deer (Capreolus capreolus)
dung resulted in several new species records, which suggests that coprophilous ascomycetes in boreal Sweden
are poorly known. Fungal species richness and occurrence on moose dung varied significantly between habitats.
Species diversity was negatively associated with amount of insect attack, and insects feeding either on the dung
and/or the fungi may be an important factor explaining the observed pattern. Species richness of coprophilous
fungi varied also significantly between different dung types. A study of moose, mountain hare, and roe deer
dung did not show any consistent patterns in respect to the animals´ digestive system. There was, however, a
general strong positive relationship between the total number of ascomycete species and the number of plant
species foraged by the three herbivores. Fungal species with large spores (≥ 50 µm) were over-represented on
roe deer dung, and under-represented on moose dung, while the reverse was found for species with small spores
(<10µm). This suggests that the foraging level of the herbivore, which in turn mirrors species-specific
differences in spore dispersal of the fungi, may be an important factor in explaining species richness and
diversity of the coprophilous community.
Key words: ß-tubulin, Bayesian analysis, dung, forage, fungi, moose, parsimony, phylogeny, Pleosporales, spore
dispersal, Sporormiaceae
Language: English
Signature:
ISBN: 91-7305-949-8
Number of pages: 28 + 4 papers
Date: 25 Oktober 2005
List of papers
This thesis is based on the following papers, which will be referred to in the text by
their Roman numerals:
I.
Kruys Å, Eriksson OE, Wedin M. Phylogenetic relationships of
coprophilous Pleosporales (Dothideomycetes, Ascomycota), and the
classification of some bitunicate taxa of unknown position. Manuscript.
II.
Kruys Å, Wedin M. A study of the natural relationships and
traditionally used taxonomic characters in Sporormiaceae (Pleosporales,
Ascomycota), utilizing multi-gene phylogenies. Manuscript.
III.
Nyberg Å, Persson I-L. 2002. Habitat differences of coprophilous
fungi on moose dung. Mycological Research 106: 1360-1366.
IV.
Kruys Å, Ericson L. Species richness of coprophilous ascomycetes
in relation to variable food intake by herbivores. Submitted manuscript.
Paper III is published with the kind permission of the publisher.
TABLE OF CONTENTS
1. INTRODUCTION .............................................................. 5
1.1 Natural relationships of the coprophilus Pleosporales.................. 6
1.2 Focus on Sporormiaceae................................................. 7
1.3 Species richness of coprophilous ascomycetes ......................... 8
2. OBJECTIVES.................................................................. 9
3. MATERIAL AND METHODS .................................................10
3.1 Molecular methods......................................................10
3.2 Field studies.............................................................10
4. MAJOR RESULTS AND DISCUSSION........................................11
4.1 Phylogenetic relationships of coprophilous families in Pleosporales..11
4.2 Phylogenetic relationships within Sporormiaceae.....................13
4.3 The effect of habitat on coprophilous fungi on moose dung ..........16
4.4 Species richness in relation to variable food intake by herbivores ..17
4.5 New records of coprophilous ascomycetes in Sweden ................19
5. FINAL CONCLUSIONS.......................................................19
6. ACKNOWLEDGEMENTS.....................................................21
7. REFERENCES ................................................................21
8. POSTSCRIPT .................................................................27
APPENDICES: PAPER I-IV
1. INTRODUCTION
A dung pile is a tremendously rich substrate sparkling with life! Fungi, bacteria,
arthropods, and mosses compete for space and nutrients, and interact in the
degradation of the heterogeneous substrate. Coprophilous (dung-loving!) fungi are a
diverse group of saprobes including taxa from most major fungal groups. This
thesis will, however, focus on Ascomycota, which is the largest group of fungi with
more than 32 000 species described (Kirk et al. 2001). The number of coprophilous
ascomycetes has not been estimated, though.
Coprophilous ascomycetes can be found on various dung types from all
over the world, but are more frequent on dung of herbivores than carnivores
(Lundqvist 1972, Richardson 2001). In addition, they have seldom been reported on
reptile or amphibian dung, indicating that coprophily of fungi developed among the
warm-blooded animals (Webster 1970). Some species are strictly coprophilous,
others occur on different ephemeral substrates. The strictly coprophilous
ascomycetes have a remarkable life-cycle, dung Æ plant Æ animal gut Æ dung
(Fig. 1). Thus, they do not disperse between habitat patches (Wicklow 1992).
Instead they have to pass the digestive system of an animal, and spore germination
may even be triggered by gastric juices (Sussman & Halvorson 1966, Furuya &
Naito 1980). The spores of coprophilous ascomycetes are often darkly pigmented,
with thick walls, and are probably well protected against both gastric juices and the
harmful UV-light of the sun (Ingold & Hudson 1993).
Fig. 1. The life-cycle of a strictly coprophilous pyrenomycete. A fruit-body that is
produced on a dung pile, will discharge its spores when mature. The spores are
often surrounded by mucilage or have gelatinous appendages, and attach easily to
the plant parts on which they land. When a plant is foraged by a herbivore, the
spores will follow and be transported through the digestive gut. Finally, when
ending up in a new dung pile, the spores will germinate and produce new fruitbodies. Drawing inspired by Bell (1983).
5
Coprophilous ascomycetes are advantageous organisms to study; a great
range of various morphologically and systematically different taxa can appear on a
fresh dung pile within few weeks, there is also no shortage of dung in nature, and a
majority of the species are possible to grow in vitro. Consequently, coprophilous
fungi have been subject to early and extensive studies (Massee & Salmon 1901,
Lundqvist 1972, Richardson 2001). The dark, thick-walled spores can be found in
ancient soil samples and sediments, and are like pollen grains, useful tools in
paleoecological studies (Hausmann et al. 2002, van Geel et al. 2002, Burney et al.
2003). A number of the coprophilous ascomycetes, such as Podospora anserina and
Sordaria macrospora are utilized as model systems for genetic, biochemical and
molecular studies (Esser 1974, Silliker et al. 1996, Nowrousian et al. 2005), and
have therefore been investigated very thoroughly. However, saprobic ascomycetes in
general are poorly known organisms, the circumscription of many groups are unclear
and they often lack good morphological features to distinguish them. Phylogenetic
information based on DNA sequences revolutionised the knowledge of the
classification of Ascomycota during the 1990´s (as continuously summarised in
Systema Ascomycetum between 1983-1998 by Eriksson or Eriksson &
Hawksworth, and later in Myconet 1997-2005 by Eriksson & Winka, Eriksson or
Eriksson et al.). However, several groups of coprophilous ascomycetes are still in
need of basic molecular systematic information. Unclear circumscriptions of taxa
have also complicated ecological studies of coprophilous ascomycetes, and there is a
need to reveal distribution patterns and how different ecological processes affect
species richness and occurence.
1.1 Natural relationships of the coprophilus Pleosporales
Coprophilous ascomycetes are found in numerous non-related taxa. However, a large
group belong to the bitunicate order Pleosporales in Dothideomycetes. Pleosporales
(sensu Eriksson 2005) includes 13 families, most of the members are saprobes or
parasites on vascular plants, but three of the families are solely or predominantly
coprophilous, viz. Delitschiaceae, Phaeotrichaceae and Sporormiaceae. These
families have often been assumed to be closely related, due to fimicolous lifestyle
and shared combination of morphological characters (Munk 1957, Luttrell 1973,
Hawksworth et al. 1983).
Sporormiaceae is characterized by dark brown, septate spores with germ slits
(Fig. 2). Several of the genera in Sporormiaceae are morphologically very similar
and the circumscriptions of these are unclear (cf. treatments in Arenal et al. 2004,
Doveri 2004). Barr (2000) has re-circumscribed Sporormiaceae and excluded
Delitschia and Semidelitschia.
Delitschiaceae resembles the Sporormiaceae in spore characters (dark brown,
septate, germ slits; Fig. 2), but differs in features of the ostiole, endotunica and the
ocular chamber (Barr 2000). Delitschiaceae is, however, still treated as a synonym of
Sporormiaceae in Kirk et al. (2001).
Phaeotrichaceae was described to accommodate the single genus Phaeotrichum,
characterized by dark brown, septate spores with terminal germ pores (Cain 1956).
The ascomata are setose cleistothecia, but Cain (1956) pointed out that it otherwise
had several similarities with the setose perithecial genus Trichodelitschia (Fig. 2).
This genus was placed in Sporormiaceae by Munk (1957), but Lundqvist (1964)
suggested it to be transferred to Phaeotrichaceae, a measure accepted by Luck-Allen
6
(1970), Eriksson (1981, in Eriksson & Hawksworth 1988), Parguey-Leduc (1974)
and Barr (2000). Trichodelitschia has very characteristic, bitunicate asci. Dehiscence
is fissitunicate and the ectotunica slides down to the base of the ascus and becomes
strongly wrinkled.
The taxonomical position and circumscription of Delitschiaceae,
Phaeotrichaceae, and Sporormiaceae have varied over time and previous molecular
studies included a very limited number of taxa, making detailed sister-group
relationships difficult to infer. Several authors have also suggested a close
relationship between one or more of these three families, and members of Zopfiaceae
and Testudinaceae (von Arx & Müller 1975, Malloch 1981, von Arx 1981).
Members of Zopfiaceae and Testudinaceae have mainly been isolated from soil. In
the case of Zopfiaceae, this has mostly been on or in association with roots. A
majority of the species in Zopfiaceae and all Testudinaceae have non-ostiolate
ascomata and brown, smooth to variously ornamented, usually septate spores
(Hawksworth 1979, Hawksworth in Eriksson & Hawksworth 1987). Both families
have at present an unclear position in the classification of Ascomycota (Kirk et al.
2001, Eriksson 2005).
Fig. 2. Spores of Trichodelitschia (Phaeotrichaceae), Delitschia (Delitschiaceae),
Sporormiella (Sporormiaceae), Zopfia (Zopfiaceae) and Neotestudina
(Testudinaceae). The drawings are not proportional in size.
1.2 Focus on Sporormiaceae
Sporormiaceae is the largest family among coprophilous fissitunicate ascomycetes.
Most members of the family are strictly coprophilous, but some occur on other
substrates, e.g. wood, soil, and plant debris. In the recent treatment by Barr (2000),
the family comprises eight genera; Chaetopreussia, Pleophragmia, Preussia,
Pycnidiophora, Sporormia, Sporormiella, Spororminula, and Westerdykella.
Sporormia, Sporormiella, and Preussia are considered to be closely related
and are easily confused by their shared morphological features. The type genus
Sporormia is characterized by globose pseudothecia, which open with an ostiole.
The spores are typically joined together in a bundle within one common gelatinous
sheath, when released from the ascus (Ahmed & Cain 1972, Dissing 1992). Germ
7
slits, otherwise common and characteristic for the family, are absent in the type
species S. fimetaria. Sporormiella, the most species-rich genus in the family,
includes species with ampulliform ascomata and spores with distinct germ slits
(Ahmed & Cain 1972). Preussia differs morphologically from 4-celled
Sporormiella species only by the ascomata being non-ostiolate (Cain 1961, Barr
2000). The taxonomic importance of the ostiole has, however, been questioned (von
Arx 1973, Guarro et al. 1997). Von Arx & van der Aa (1987) instead used substrate
choice as a diagnostic character when separating Preussia and Sporormiella.
Preussia, in their sense, included species on plant debris, wood or soil, while
Sporormiella was restricted to comprise only strictly coprophilous species.
However, several Sporormiella species are not only generalists among coprophilous
substrates, but may occur on wood and soil, according to Guarro et al. (1997).
Westerdykella, Pycnidophora, and Chaetopreussia form, like Preussia,
non-ostiolate ascomata (Clum 1955, Stolk 1955, Locquin-Linard 1977). The spores
of Westerdykella, Pycnidiophora and Chaetopreussia separate at a very early stage
in the ascus and the lack of germ slits is also considered a taxonomically important
character (von Arx & van der Aa 1987, Barr 2000). Pycnidiophora has been
included in Westerdykella by several authors (Cejp & Milko 1964, von Arx &
Storm 1967, von Arx 1981). Both von Arx & van der Aa (1987) and Barr (2000),
however, suggested that species with smooth spore walls should be included in
Pycnidiophora, while Westerdykella should be restricted to the type W. ornata, the
only species with ornamented spores. Barr (2000) has also suggested a close
relationship between Pleophragmia, Sporormia, and the monotypic Spororminula,
because of their ostiolate ascomata, and the spores that lack germ slits, and should
be triangular in transverse view.
The classification of taxa within Sporormiaceae is apparently based on a
comparatively small number of easily observed ecological and morphological
characters. Several of these traditionally used characters have, however, been claimed
to be poor predictors of phylogenetic relationships, both in Sporormiaceae as well as
in other groups of Ascomycota (von Arx 1973, Guarro et al. 1997, Dettman et al.
2001, Hansen et al. 2002), and their applicability for circumscribing genera, need to
tested with molecular methods.
1.3 Species richness of coprophilous ascomycetes
Species richness and community composition of coprophilous ascomycetes vary
with abiotic and biotic factors. Temperature and moisture affect growth rate, fruiting
and species richness of coprophilous fungi (Wicklow & Moore 1974, Harrower &
Nagy 1979, Yocom & Wicklow 1980, Kuthubutheen & Webster 1986). Intra- and
interspecific interactions occur at the scale of the individual dung pile and have been
shown to affect fungal development, as well as species composition (Ikediugwu &
Webster 1970, Wicklow & Hirschfield 1979, Weber and Webster 1998).
Coprophagous arthropods can graze on fungal mycelia or indirectly affect fruiting by
hastening the decay process (Lussenhop et al. 1980, Wicklow & Yocom 1982,
Stevenson & Dindal 1987). These factors are expected to vary in nature, e.g.
depending on the habitat and the season, however, the majority of previous studies
have been conducted in vitro. Also, most studies of coprophilous ascomycetes have
focused on dung of domesticated animals (Dickinson & Underhay 1977, Wicklow
& Hirschfield 1979) and rabbits in grasslands (Yocom & Wicklow 1980, Angel &
8
Wicklow 1983), while dung from wild boreal animals, and especially forest-living
species, has been much less studied.
Many coprophilous ascomycetes are most common on only one or a few
dung types (Lundqvist 1972) and dung from closely related animals generally show
a similar species composition (Richardson 2001). Angel & Wicklow (1983) found
that the fungal community varied more between dung types (rabbit and cattle dung)
than between various grassland habitats. This suggests that the digestive system of
the herbivore may be another important factor for species richness, as differences in
digestion will affect both the passage of the spores through the digestive gut, and
the dung quality, e.g. consistency, moisture and nutrient content. Studies
addressing the importance of different digestive systems have been limited, though.
However, Wicklow et al. (1980) compared dung of a single rabbit and a single
sheep, fed with hay from the same bale, and found marked differences in species
compositions, differences that they attributed to differences in digestion.
Alternatively, differences in species richness and composition may mirror
differences in feeding habits and/or food choice between herbivores. Ebersohn and
Eicker (1992) in their study on coprophilous fungi on African herbivores concluded
that feeding habits and food choice were more important than the animals´ digestive
system for the species composition. Dung from generalist herbivores, feeding upon
a large number of plant species and within a large height interval, was generally
more species rich and also showed the highest species diversity. In contrast, dung
from specialist grazers foraging within a narrow height interval was characterised by
fewer species and lower species diversity. However, whether this reflects a wider
variation in food quality per se, or that generalist feeders while foraging both lower
and taller plants are inoculated with higher amounts of spores, or spores from more
species, remains unclear. That spore dispersal may be one important factor in
explaining species composition is supported by the fact that the the elephant, which
showed the broadest foraging level, feeding from grass root to tree top level, also
had the highest species diversity of fungi. In another study, Bell (1975) observed
that the coprophilous flora of the tree-feeding brush-tailed opossum showed a
predominance of small, hyaline spores in opposite to ground-dwelling herbivores.
Thus, there is some support that the coprophilous community mirrors the foraging
level of the herbivore.
2. OBJECTIVES
The main objectives with this thesis was to gain more knowledge in a difficult
group of coprophilous ascomycetes with unclear natural relationships, and to study a
number of factors that may have an effect on the species richness and species
composition of these organisms. To reveal these topics I, therefore, aimed to:
ß
ß
ß
ß
study phylogenetic relationships between coprophilous families in Pleosporales,
and test their earlier classifications (paper I).
clarify the circumscriptions of taxa within Sporormiaceae, and test the
homology of traditionally used characters (paper II).
study the effect of habitat on species richness of coprophilous fungi (paper III).
investigate how dung type and food choice may affect species richness and
species composition of coprophilous ascomycetes (paper IV).
9
3. MATERIAL AND METHODS
3.1 Molecular methods
I utilized several independent genomic markers for the phylogenetic studies (paper I,
II). Nuclear ribosomal genes (e.g. nSSU and nLSU) are commonly used in fungal
phylogenetics, as the high number of copies and the availability of general primers
(White et al. 1990, Gargas & DePriest 1996) make them easy to amplify.
Ribosomal encoding genes can also be amplified from the mitochondrial genome
(e.g. mtSSU), although in fewer copies. Nuclear protein-coding genes (e.g. ßtubulin) are becoming increasingly used and have several advantages compared to
ribosomal genes. Since they code for proteins, the alignment is likely to contain
less ambiguity due to codon constraint, and both nucleotides and amino acids can
be analysed. However, only nucleotide sequences were analysed in these studies.
In order to reveal phylogenetic relationships on the family level within
Pleosporales, I amplified nSSU rDNA, nLSU rDNA, and mtSSU rDNA (paper I).
The more variable gene partitions, nITS-LSU, mtSSU and ß-tubulin, were used to
resolve relationships on the generic level within Sporormiaceae (paper II). For the
sequencing of ß-tubulin, I designed a new primer, BT1903R (paper II). The data sets
were analysed with parsimony (paper I, II) and Bayesian (paper II) methods
(Kitching et al. 1998, Ronquist 2004, Huelsenbeck et al. 2001). Alternative tree
topologies (paper I) were investigated with constrained parsimony analyses followed
by the Shimodaira-Hasegawa test (Shimodaira & Hasegawa 1999).
3.2 Field studies
The field work conducted for paper III and IV were done in boreal forest in
Västerbotten and Ångermanland in northern Sweden.
To study the effect of habitat on species richness and occurrence of
coprophilous fungi I used fresh moose (Alces alces) dung from a nearby moose
farm, where moose calves were kept in a natural pasture and all were fed the same
diet (paper III). Due to the homogenous origin, composition and age of the dung, it
was assumed that the dung piles had the same inoculum of species at the start of the
experiment. The dung piles were placed along line transects in three different
habitats. As previous studies have frequently shown that temperature and humidity
are key factors for fungal communities, the following habitat types were chosen: one
sunny and dry habitat (pine forest), one sunny and wet habitat (mire) and one shady
and mesic habitat (spruce forest). The water content of the dung was used as a
measurement of habitat moisture, and the number of holes visible on the surface of
the dung, which could be attributed to insect larvae, was used as an estimate of
insect attack. The percent cover of discomycetes was visually estimated in field,
while the species number of fungi was determined in laboratory.
Differences in species richness and composition of coprophilous
ascomycetes between dung types (paper IV) was studied by sampling dung of three
herbivores which differ with regard of their digestive system; two ruminants, moose
and roe deer (Capreolus capreolus), and one hindgut fermentor, mountain hare
(Lepus timidus). Winter dung was sampled from three areas in the coastal region
near Umeå. The three areas have a constant occurrence of the three herbivores, and
hence they were expected to have a similar species composition of coprophilous
10
fungi. Sampling of dung piles took place on one occasion in late winter after which
the dung piles were inoculated in vitro. This was done in order to minimize effects
on species composition depending on humidity and temperature (Yocom and
Wicklow 1980), occurrence of various arthropods (Stevenson & Dindal 1987), as
well as age of the dung and season (Angel and Wicklow 1983, Richardson 2001).
Moose, mountain hare, and roe deer forage within the same habitat, although within
different height intervals. During the winter season, all three animal species are
browsers and forage mainly on woody plants. In order to describe the food choice of
the three herbivores in each area, I followed their tracks and scored visible signs of
recent foraging.
4. MAJOR RESULTS AND DISCUSSION
4.1 Phylogenetic relationships of coprophilous families in Pleosporales
The coprophilous families Delitschiaceae (Fig. 3; clade H) and Sporormiaceae (Fig.
3; clade F) clustered on separate branches in a strongly supported clade,
corresponding to Pleosporales, and they were not closely related (paper I).
Coprophilous taxa have apparently arisen several times within Pleosporales, as in a
number of other fungal groups. Delitschiaceae forms a distinct monophyletic group,
well supported also by a constrained-topology analysis (p<0.001), and should be
accepted as a separate family. The bootstrap support for the monophyletic
Sporormiaceae is just below the level of significance (bs 68). This clade also
includes one species that has been accommodated in Testudinaceae, Eremodothis
angulata. It has one-celled spores with four blunt ends, projecting in caltrop spine
directions, but without germ pores (Udagawa & Ueda 1981), which makes it
morphologically distinct from other Testudinaceae. Eremodothis shares the lack of
ostiole and germ slits with Westerdykella, to which it is apparently most closely
related, but it has eight one-celled spores, contrary to the rest of Westerdykella.
Trichodelitschia grouped with Phaeotrichum with high support, and this
confirms the opinion of Lundqvist (1964), Eriksson (1981), and Barr (2000). An
unexpected result was the close relationship between Phaeotrichaceae (Fig. 3; clade
Q) and Venturiaceae (Fig. 3; clade P), which are strongly supported sister-groups on
a branch outside Pleosporales. This position of Venturiaceae is not in agreement
with some previous molecular studies (Silva-Hanlin & Hanlin 1999, Olivier et al.
2000, Eriksson & Hawksworth 2003). However, this result is also supported by
morphological traits, since Venturiaceae and Phaeotrichaceae both have setose
ascomata and coloured, one-septate spores (Eriksson 1981). Phaeotrichaceae and
Venturiaceae both belong to the class Dothideomycetes, but if they belong to the
order Pleosporales or not, is unclear. A constrained-topology analysis did not reject
such an alternative (p=0.10-0.57).
All taxa included in this study that have an unclear position in Eriksson
(2005), were nested within Dothideomycetes. Zopfiaceae and Testudinaceae belong
to Pleosporales, but they do not form a monophyletic group and should be treated
as two separate families. Zopfiaceae is polyphyletic, the type Zopfia is sister-group
to Delitschiaceae (with no support), while Didymocrea was nested within a mixed
clade (Fig. 3; clade B), consisting of one member of each of the families
Melanommataceae, Cucurbitariaceae, and Tubeufiaceeae. All members of the
11
Testudinaceae, except for Eremodothis angulata, form one monophyletic clade (Fig.
3; clade G), which also includes Verruculina enalia (now in Didymosphaeriaceae).
99
99
100
58
100
64
74
52
100
99
98
83
63
Pleosporales
85
100
62
100 99
100
71
68
60
100
98
99
63
100
100
100
100
82
69
100
100
93
100
100
100
Arthonia dispersa
Combea mollusca
Dendrographa leucophaea
Schismatomma decolorans
Arthopyrenia salicis
Bimuria novae-zelandiae
Didymocrea sadasivanii
Letendraea helminthicola
Curreya pityophila
Clathrospora diplospora
Lewia infectoria
Pleospora herbarum
Cochliobolus heterostrophus
Pyrenophora tritici-repentis
Leptosphaeria cf. macrospora
Leptosphaeria doliolum
Ophiobolus herpotrichus
Phaeosphaeria avenaria
Setomelanomma holmii
Shiraia bambusicola
Byssothecium circinans
Herpotrichia diffusa
Herpotrichia juniperi
Melanomma pulvis-pyrius
Lophiostoma macrostomum
Pleomassaria siparia
Eremodothis angulata
Westerdykella dispersa
Westerdykella cylindrica
Preussia terricola
Sporormia lignicola
Trematosphaeria heterospora
Lepidosphaeria nicotiae
Verruculina enalia
Ulospora bilgramii
Neotestudina rosatii
Delitschia didyma
Delitschia winteri
Zopfia rhizophila
Tubeufia helicoma
Botryosphaeria dothidea
Botryosphaeria ribis
Guignardia bidwelli
Delphinella strobiligena
Dothidea sambuci
Stylodothis puccinioides
Sydowia polyspora
Elsinoë veneta
Myriangium duriaei
Piedraia hortae
Microxyphium citri
Mycosphaerella populorum
Mycosphaerella punctiformis
Raciborskiomyces longisetosum
Hysteropatella clavispora
Farlowiella carmichaeliana
Metacoleroa dickiei
Spilocaea oleaginea
Venturia chlorospora
Venturia inaeqalis
Phaetrichum benjaminii
Trichodelitschia munkii
Capronia mansonii
Ceramothyrium carniolicum
Eurotium herbariorum
Spiromastix warcupii
Cladonia rangiferina
Xanthoria parietina
Hypocrea lutea
Xylaria hypoxylon
ARTHONIOMYCETES
A: Arthopyreniaceae
B: Cucurbitariaceae, Melanommataceae,
!!!!!Tubeufiaceeae and Zopfiaceae
C: Diademaceae and Pleosporaceae
D: Leptosphaeriaceae,
!!!!!Phaeosphaeriaceae,
!!!!!and genera of uncertain position
E: Lophiostomataceae,
!!!!Melanommataceae,
!!!!and Pleomassariaceae
F: Melanommataceae, Sporormiaceae,
!!!!!and Testudinaceae
G: Didymosphaeriaceae
!!!!!and Testudinaceae
H: Delitschiaceae and Zopfiaceae
I: Tubeufiaceeae
J: Botryosphaeriaceae
!!!!and Mycosphaerellaceae
K: Dothideales
L: Myriangiales and Piedraiaceae
M: Capnodiales, Mycosphaerellaceae,
!!!!!!and Pseudoperisporiaceae
N: Hysteriales
O: Hysteriales
P: Venturiacceae
Q: Phaeotrichaceae
EUROTIOMYCETES incl.
CHAETOTHYRIOMYCETES
LECANOROMYCETES
SORDARIOMYCETES
Fig. 3. A strict consensus of the 30 most parsimonious trees, based on nSSU,
nLSU and mtSSU rDNA sequences. Numbers represent bootstrap support. Bold
lines represent branches with 100% support. Arrows indicate the extent of the
clades and bold family names indicate clades including the type species of the
family. Clades discussed in paper I are marked with letters A-Q.
12
4.2 Phylogenetic relationships within Sporormiaceae
Within Sporormiaceae (paper II), Westerdykella (including Pycnidiophora),
Sporormia, Preussia (including Sporormiella alloiomera), and several groups
within Sporormiella are significantly supported as monophyletic (Figs. 4, 5).
However, the circumscription of the difficult Sporormiella–Preussia complex is not
revealed with convincing support. Several characters traditionally used as diagnostic
features within Sporormiaceae are here shown to be poorly correlated with
monophyletic groups. The absence of germ slits is not a homologous feature, and I
found no support for a close relationship between Sporormia and the monotypic
Spororminula, as suggested by Barr (2000). The results do not support a separation
between Pycnidiophora and Westerdykella based on spore ornamentation, as
suggested by von Arx & van der Aa (1987) and Barr (2000). The type of
Pycnidophora, P. dispersa (=Westerdykella dispersa), is nested within the
Westerdykella clade, close to the type of Westerdykella, W. ornata. Pycnidiopora
should, therefore, remain as a synonym of Westerdykella.
The members of Preussia and Sporormiella included in this study are
intermixed in the tree, and at present, I know few morphological features that are
useful for circumscribing the well supported clades presented here. My results
confirm the findings of Guarro et al. (1997), namely that the presence or absence of
the ostiole is not a useful character when delimiting Preussia and Sporormiella,
because non-ostiolate species are mixed with ostiolate species with a distinct neck.
Further, the results do not support a distinction between Preussia and Sporormiella
with regard to substrate choice, as suggested by von Arx & van der Aa (1987). For
practical reasons, one could therefore argue that Preussia and Sporormiella should
be treated as one genus. However, there is no support for one major monophyletic
group including both Preussia and Sporormiella, and a majority of the Preussia
species (including the type P. funiculata) still comprise a very well supported clade,
which could be treated as Preussia in a restricted sense. Therefore, I suggest a
conservative approach and continue to treat the two genera as separate, at least as an
ad interim solution until the knowledge of the relationships within the PreussiaSporormiella complex has been improved. One character that may be of importance
for the circumscription of some of the Sporormiella-Preussia groups is the shape of
the ascus, especially if used in combination with other features, such as the shape of
the spores.
13
61
100
100
52
68
100
99
55
100
100
67
100
62
100
77
55
100
100
100
99
100
Sporormiella heptamera
Sporormiella octomera
The Sporormiella
Sporormiella affinis
vexans clade
Sporormiella vexans
Sporormiella leporina
Sporormiella dakotensis
Sporormiella longisporopsis The Spororminula clade
Spororminula tenerifae
Sporormiella irregularis
Sporormiella tetramera
The Sporormiella
Sporormiella dubia
irregularis clade
Sporormiella splendens
Sporormiella borealis
Sporormiella intermedia
Sporormiella bipartis
The Sporormiella
"Sporormia" lignicola
intermedia clade
Sporormiella minipascua
Sporormiella australis
Sporormiella minima
Preussia isomera
Sporormiella antarctica
Sporormiella septenaria
Preussia fleischhakii
Preussia funiculata
Preussia terricola
The Preussia clade
Preussia typharum
Preussia vulgaris
Sporormiella alloiomera
Pycnidiophora dispersa CBS 297.56
Pycnidiophora dispersa CBS 508.75
Westerdykella multispora
The Westerdykella
Westerdykella ornata
clade
Westerdykella cylindrica
Westerdykella nigra
Sporormia fimetaria 2302-c
The Sporormia clade
Sporormia fimetaria 81.194
Preussia terricola CBS 527.84
The Sporormiella
Sporormiella megalospora
megalospora clade
Curreya pityophila
Pleospora herbarum
Herpotrichia juniperi
Melanomma pulvis-pyrius
Trematosphaeria heterospora
Lepidosphaeria nicotiae
Verruculina enalia
Fig. 4. A strict consensus tree of the 49 most parsimonious trees from an analysis
based on ITS-nLSU rDNA, mtSSU rDNA, and ß-tubulin sequences. Bootstrap
values above 50% are shown above branches. Branches with ≥70% bootstrap
support (level of significance) are marked in bold.
14
Preussia terricola
Preussia.typharum
99
Preussia funiculata
The Preussia clade
100
Preussia vulgaris
Preussia fleischhakii
Sporormiella alloiomera
67
100
Pycnidiophora dispersa CBS 297.56
100
Pycnidiopora dispersa CBS 508.75
Westerdykella multispora
100
The Westerdykella clade
67
68
Westerdykella ornata
Westerdykella cylindrica
Westerdykella nigra
100
67
Sporormia fimetaria 2302-c
100
The Sporormia clade
Sporormia fimetaria 81.194
100
Preussia terricola CBS 527.84
The Sporormiella
Sporormiella megalospora
megalospora clade
100
Sporormiella borealis
88
Sporormiella intermedia
94
Sporormia "lignicola"
94
Sporormiella antarctica
91
Sporormiella bipartis
The Sporormiella
60
Sporormiella minipascua
intermedia clade
100
Preussia isomera
100
Sporormiella australis
100
Sporormiella minima
100
Sporormiella septenaria
Sporormiella heptamera
85
99
Sporormiella octomera
The Sporormiella
100
Sporormiella affinis
vexans clade
100
Sporormiella vexans
Sporormiella leporina
100
97
Sporormiella dakotensis
90
100
Sporormiella longisporopsis
The Spororminula clade
Spororminula tenerifae
98
100
Sporormiella irregularis
Sporormiella tetramera
100
The Sporormiella
100
Sporormiella dubia
irregularis clade
Sporormiella splendens
Trematosphaeria heterospora
100
Curreya pityophila
100
Pleospora herbarum
Herpotrichia juniperi
100
Melanomma pulvis-pyrius
Lepidosphaeria nicotiae
Verruculina enalia
50 changes
86
Fig. 5. A fifty percent majority-rule consensus tree of the 45 000 sampled trees
with best likelihood from a bayesian analysis, based on ITS-nLSU rDNA, mtSSU
rDNA, and ß-tubulin sequences. Branches with ≥95% posterior probability support
(level of significance) are marked in bold.
15
4.3 The effect of habitat on coprophilous fungi on moose dung
The results in paper III revealed significant differences in occurrence and species
richness of coprophilous fungi, between the three habitats (Fig. 6). As I used
uniform moose dung, assumed to have the same inoculum of species, the
importance of the habitat for the community composition is clearly demonstrated.
This is also in accordance with earlier studies of coprophilous fungi in grasslands
(Lussenhop et al. 1980, Yocom & Wicklow 1980).
In this experiment, dung from the most shady habitat, the spruce forest,
contributed most to the observed differences. It had the significantly highest average
water content, the significantly highest number of insect holes, and the significantly
lowest number of fungal species, as well as percent cover of discomycetes. The
species poor coprophilous flora was opposite to that I expected as fungi are mostly
associated with moist habitats, and experiments in vitro have found an increased
species richness of coprophilous fungi when provided with freely available water
(Kuthubutheen & Webster 1986).
b
40
35
7
Sampling 2
30
25
Sampling 1
a
e
No. of fungal species
Water content/total weight (%)
45
c
d
20
f
15
10
5
Pine forest
Spruce forest
3
b
2
1
Pine forest
a
12
a
8
6
b
4
Spruce forest
Mire
Habitat type
7
No. of insect holes
Discomycete cover (%)
4
b.
14
b
6
5
4
a
3
a
2
1
2
0
0
c.
5
Mire
Habitat type
10
c
a
0
0
a.
6
Pine forest
Spruce forest
Habitat type
Pine forest
Mire
d.
Spruce forest
Mire
Habitat type
Fig. 6. The parameters measured on moose dung in three habitat types in northern
Sweden: water content (a), the number of fungal species (b), percent cover of
discomycetes (c), and number of insect holes on the surface (d). The figure has been
altered compared to paper III.
16
There was a significant negative correlation between water content and number of
species of fungi, as well as cover of discomycetes, whereas there was a significant
positive correlation between water content and number of insect holes. Furthermore,
a negative correlation, although not significant, was found between the number of
insect holes and number of fungal species, and between number of insect holes and
the cover of discomycetes. This suggests a negative effect of insects on the fungal
community, but the lack of significance may be due to differences in the insect
assemblage between the three studied habitats, since different insect species may
have different impact on the fungi. Especially larval dung beetles can contribute to
crumbling and disruption of the dung (Lussenhop et al. 1980, Stevenson & Dindal
1987) and the dung in the spruce forest, was considerably more disintegrated than
the dung in the other two habitats. Previous studies have also shown that
coprophagous insects not only feed on remaining plant debris, but also graze on
fungal mycelium and fruit bodies (Wicklow 1979, Wicklow & Yocom 1982).
4.4 Species richness in relation to variable food intake by herbivores
I found that species richness differed significantly between various dung types, both
for the total number of species and the mean number of species per sample, as well
as between localities (paper IV). The highest species richness was found for roe deer,
while the other ruminant, moose, did not differ from mountain hare, a hindgut
fermentor. Thus, the data did not show any consistent patterns in respect to the
animals´ digestive system, which is in accordance with a study by Ebersohn and
Eicker (1992).
For roe deer I found a higher species richness and species diversity in the
two areas where graminoids, rich in cellulose, constituted 1/3 of the bites, compared
to the third area where browsing constituted 96% of the bites. For both moose and
mountain hare the lowest species richness was observed where the bites were
dominated by a few deciduous species rich in phenolic compounds (Salix spp.,
Betula pubescens). These observations, although based on correlative data, suggest
that differences in plant chemistry may be of importance for the species
composition. The high species richness on roe deer dung correlated also to a much
wider food choice by roe deer (Fig. 7). For moose and mountain hare the most
striking pattern was that the lowest number of coprophilous species was observed at
the same locality, where these two herbivores showed the lowest food diversity.
Together, these patterns suggest that a more species rich food intake may be one
explanation for higher species richness of coprophilous ascomycetes (cf. Ebersohn
and Eickner 1992). However, an increase in the number of foraged species also
correlates both to an increased variation in the chemical composition of the forage,
and foraging of low-growing as well as taller plant species. Thus the data does not
allow an evaluation of the relative importance of food quality or foraging level for
species richness. Nor is it possible to exclude the possibility that plant chemistry
matters.
I found also that the coprophilous community on roe deer and moose dung
differed with regard to spore size. Species with large spores (> 50 µm) were overrepresented on roe deer dung and under-represented on moose dung, while species
with small spores (< 10 µm) showed the reverse pattern (Fig. 8). These patterns also
correspond with moose foraging on taller plants, while roe deer at all localities
foraged within a broader height interval. For mountain hare the deviations between
17
observed and expected frequencies of species in different spore size classes showed
no clear pattern. However, the species found on mountain hare dung were either in
association with roe deer or occurred on all three substrate types, while none of the
species was associated with only mountain hare and moose. All toghether, this
suggests that the species composition mirrors the foraging level of the herbivore and
thus differences in spore dispersal (cf. Bell 1975). If species with large spores are
characterized by an inferior dispersal ability, one would expect them to be more
frequent on dung of mountain hare. However, at the time of sampling, mountain
hare foraged mostly deciduous shrubs protruding above the snow package, which
may be the reason why this was not the case.
Fig. 7. The number of coprophilous ascomycetes plotted against the number of
foraged plant species. A linear regression analysis revealed a general strong
positive relationship between the two variables.
Fig. 8. The observed (black dots) and expected (dashed line) occurrence of
ascomycete species of different spore size classes, for mountain hare, moose, and
roe deer. Data is pooled for the three localities.
18
4.5 New records of coprophilous ascomycetes in Sweden
In total, 61 species of ascomycetes were encountered in the two field studies (paper
III and IV). Five to six species represent undescribed species, Coniochaeta sp.,
Sporormiella sp., Trichodelitschia sp., and possibly also Sporormiella cf. ovina in
paper IV, as well as two species of Sporormiella in paper III. Fifteen species were
new to the province of Västerbotten, and 28 species were new to the province of
Ångermanland. Five species were new records on mountain hare dung, 14 species
were new on roe deer dung, and 19 species were new on moose dung. Cercophora
gossypina (paper IV) has only been found twice in Sweden (Eriksson 1992), while
Thelebolus caninus (paper III, IV) has been found in the country but the frequency
and distribution is unknown (Hansen and Knudsen 2000). Apparently, the
coprophilous ascomycetes in boreal Sweden are poorly known, in particular when
considering the number of encountered species and the limited sampling effort in
paper III and IV.
The highest species richness was found on roe deer dung (paper IV). For
the most species rich group, the pyrenomycetes, 39 species were found on roe deer
dung, 21 on moose dung and 19 on mountain hare dung (paper IV). A comparison
between this data and the number of coprophilous pyrenomycetes previously
reported from Sweden by Eriksson (1992) shows the opposite pattern, namely 15,
29 and 52 species, respectively. These opposite patterns are likely to mirror earlier
sampling efforts and show the difficulties involved in comparisons between different
data sets.
5. FINAL CONCLUSIONS
This thesis clarifies the natural relationships of coprophilous families within
Pleosporales, in particular of Sporormiaceae. It contributes with important pieces to
the whole Pleosporales-puzzle, and sheds new light also on the relationships of
other taxa within this large order.
Sporormiaceae and Delitschiaceae are separate monophyletic groups within
Pleosporales and should continue to be recognized as two distinct families.
Phaeotrichaceae forms a monophyletic group, and is, unexpectedly, a strongly
supported sister-group to Venturiaceae, but if they belong to the order Pleosporales
still remains unclear. The two families Testudinaceae and Zopfiaceae, which
previously had an uncertain position in Ascomycota, are demonstrated to be
members of Pleosporales and should be treated as two separate families. Zopfiaceae
is polyphyletic, while a majority of the members of Testudinaceae form one clade.
The monotypic genus Eremodothis is, however, not related to Testudinaceae, but is
nested within Sporormiaceae and should be transferred to the genus Westerdykella.
Several questions still remain to be solved though; the circumscription of
Pleosporales is not clear, a number of clades within Pleosporales do not have a
natural classification, and several taxa placed with uncertain position in
Dothideomycetes/Chaetothyriomycetes in Eriksson (2005), still need to be
investigated.
The relationships between genera within Sporormiaceae are still not fully
resolved and I therefore, suggest a rather conservative generic classification of the
family, including the well supported and monophyletic Sporormia (excluding S.
19
lignicola), Preussia (excluding P. isomera, but including Sporomiella alloiomera),
and Westerdykella (including Pycnidiophora and Eremodothis). I also suggest
accepting Sporormiella, despite not being conclusively well supported as
monophyletic, at least as a preliminary solution. Sporormiella should then include
the monotypic Spororminula, but exclude Sporormiella alloiomera. Characters
previously used in the taxonomy and classification of Sporormiaceae, as the choice
of substrate, the presence or absence of an ostiole, the presence or absence of germ
slits, and spore ornamentation, were all homoplastic and not very useful for
circumscribing monophyletic groups. I believe, however, that characters of the ascus
may be important for the circumscription of some of the Sporormiella–Preussia
groups. A number of features may be useful for circumscribing smaller clades if
used in combination with other characters, as for instance the shape of the spore
combined with the shape of the ascus. I believe that an extended search for useful
morphological characters is needed though, preferably in combination with
physiological and molecular studies. Future molecular studies of Sporormiaceae
should also include more taxa. The two genera Chaetopreussia and Pleophragmia
remain to be investigated, and considering that I found several undescribed
Sporormiella species in the field studies, an extended search may result in even
more taxa, which could help to resolve the phylogeny of Sporormiaceae in the
future.
Several new records of coprophilous ascomycetes were observed in the two
field studies, in spite of a limited sampling effort. This suggests that the
coprophilous ascomycetes in boreal Sweden are poorly known, and it is likely to
assume that more species can be revealed in the future. The species richness is,
however, affected by a number of ecological factors. When comparing moose dung
of uniform age and composition and thus assumed to have the same inoculum of
species, the fungal species richness and occurrence varied significantly between
habitats. There was a highly significant negative correlation between water content
of the dung and number of species of fungi, as well as percent cover of fungi, which
is opposite to what I expected. However, species diversity was also negatively
associated with amount of insect attack, which suggests that insects have a large
impact on the fungal community, and insects feeding either on the dung and/or the
fungi may be an important factor for species richness.
Species richness of coprophilous fungi varies also significantly between
different dung types. A study of the coprophilous ascomycetes on moose, mountain
hare, and roe deer dung did not show any consistent patterns in respect to the
animals´ digestive system, since the highest species richness was found for roe deer,
while the other ruminant, moose, did not differ from mountain hare, a hindgut
fermentor. However, there was a general strong positive relationship between the
total number of ascomycete species and the number of plant species foraged by the
three herbivores. Whether this mirrors the number of forage species per se, or
changed chemical composition of forage, or foraging of both low- and tall-growing
plants remains unclear as all these parameters were confounded. However, species of
different spore sizes differed in their occurrence. Species with large spores (≥ 50 µm)
were over-represented on roe deer dung, and under-represented on moose dung, while
the reverse was found for species with small spores (<10µm). This suggests that the
foraging level of the herbivore, which in turn mirrors species-specific differences in
spore dispersal of the fungi, may be the most important factor in explaining species
richness and diversity of the coprophilous community.
20
6. ACKNOWLEDGEMENTS
Financial support for the studies in this thesis was kindly provided by the Swedish
Research Council (NFR B5101-20005187, VR 629-2001-5756, VR 621-2002-349,
VR 621-2003-3038), the Kempe Foundation (JCK2026), Magn. Bergvalls
foundation, the Royal Swedish Academy of Sciences (Stiftelsen Th Kroks
donation), and the Gunnar and Ruth Björkman Foundation. I am grateful to Nils
Lundqvist for numerous specimens contributed to this study. The Directors and
Curators of cited herbaria and museums are acknowledged for the loan of specimens.
Many thanks also to Mary Berbee who kindly allowed us to use her extractions of
five taxa, and to Katarina Winka who contributed with the extraction of one taxon.
The staff at Umeå Plant Science Centre assisted with sequencing, and Carin
Olofsson is thanked for invaluable laboratory assistance. Mats Wedin, Lars Ericson
and Nic Kruys are acknowledged for valuable comments on earlier versions of this
thesis.
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8. POSTSCRIPT
Att snickra ihop denna avhandling, har varit ett långt snubblande, där jag ofta
tänkt efter, just efter, men med hjälp av följande personer har jag ändå mirakulöst
lyckats hålla mig på benen.
Mats, du har varit min vardags-handledare som med gott mod löst många problem
längs vägen. Du har fått mig att inse storheten med systematiken, och att du
dessutom tappert stått ut med mitt bristande lav-intresse och min förmåga att tycka
tvärtemot, är jag otroligt glad för! Ove, du är en skicklig lärare som fångade upp
mig som student och sedan dess har du varit min källa till mycket information och
inspiration. Allt jag har lärt mig om de förunderliga ascomyceterna kommer ju i
grunden från dig, och när du inte varit på plats så har jag hittat de mest knepiga
referenserna i din bokhylla. Lasse, utan din entusiasm och uppmuntran hade den här
avhandlingen varken påbörjats eller avslutats, du har också haft ett imponerande
tålamod i fält, det var inte bara en gång mina markeringar regnade bort…
Ett speciell tack vill jag ge till Katarina W, för att du har lärt mig labba, hjälpt mig
med undervisning, dragit med mig på iksu-träning och förklarat det här med
fylogenetiska analyser ett oändligt antal gånger. Du har helt enkelt varit en skön
kombination av extra-handledare och god vän. Tack också till min medförfattare
Inga-Lill P, för ett fint samarbete. Jag hoppas att vi får chansen att klappa söta
älgkalvar igen! Nils Lundqvist, du har hjälpt mig med många kollekter och varit en
ovärderlig guide när jag gått vilse i bestämningsnycklarna. Hans G, utan din hjälp
med statistiken hade jag faktiskt inte blivit klar.
Med Anna C & Elisabeth W har jag delat roliga kurser, kämpiga boktentor, lavexq i
snöyra… tillsammans har vi stått starka när ekorrsamlandet fångat vår handledare i
ett maniskt grepp! Anna du har blivit en jättebra kompis under dessa år och den som
otroligt nog orkat läsa alla mina manus! Carin O, du har hjälpt mig med labbandet,
men lika viktigt är att du alltid haft tid för prat och godis, tack för alla pauser. Våra
postdocs, Per I och Heidi D har bidragit med kul sällskap och många nya ideer. Jag
vill också tacka Karin D för snygga sekvenser, och Ulla C-G som peppat mig det
sista året.
En av de mest lärorika erfarenheterna från doktorandtiden är den undervisning jag
varit inblandad i, därför vill jag passa på att tacka systematikkollegorna som snöat
in på djur, Anders N & Johannes B, samt Åsa H, du har med ett otroligt tålamod
bankat in lite storsvampar i mitt huvud. Fast jag är fortfarande inte övertygad om
varför man ska äta dom.
Institutionen är ju en stor, och dynamisk arbetsplats där man hela tiden lär känna
nya personer (det liknar successionen av svampar på en stor spillningshög, faktiskt).
Alla bidrar på sitt lilla sätt till att det är en trivsam stämning, men jag vill särskilt
tacka alla doktorander, ni är för många att nämnas vid namn, men jag giller er alla
skarpt!
Som doktorand flyter ju fritid och jobb lätt ihop och några av dom som genom
jobbet, även lyst upp min tillvaro utanför, är: Lena D , ditt sällskap har varit
ovärderligt, en enkel fika eller en kväll i din kala, kalla källare, alltid lika kul! Tina
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N, tack för alla glada stunder, som på blåsiga klippor i Bodö eller en horisontell
flygelsång. Tuss Z, tack för avslappnande middagar och hetsiga spelkvällar i
vintermörkret. Mattias E & Anna-Maria E, Henrik & Kristina H, Jocke S & Hanna
O, Johan & Eva O, och Pernilla & Bent C, tack för alla trevliga tillställningar.
Marie, Gunilla och Annette, ni fick mig genom grundutbildningen och har varit
fenomenalt sällskap under roliga hajker och tradiga föreläsningar. Tack för att ni då
och då påminner mig om vilket sjukt kul projekt jag haft.
Mamma och pappa, er outtröttliga hjälp med läx-traglandet under första halvan av
skoltiden har gjort att jag på ren tjurighet orkat lika länge till. Tack för att ni alltid
ställer upp och på ett övertygande sätt uppmuntrar de flesta av mina ideér. Mikke,
ständig storebror som hetsar mig till de flesta stordåd och mindre katastrofer, tack
för år av sparring.
Nic, du förgyller allt, hela mitt hjärta till dig!
Nu ska jag ut i solen.
/ åsa
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