An integrative study of Pannaria hookeri (Ascomycota lichenized) from Argentina and the update of the taxon circumscription based on specimens from the Northern and Southern Hemispheres

Abstract

Pannaria hookeri (Borrer ex Sm.) Nyl. is reported as a bipolar cyanolichen that occurs in more than twenty countries in both Hemispheres. In the Northern Hemisphere, a minor variation is attributed to the species, whereas larger morphological and chemical variations have been reported in specimens from the Southern Hemisphere. Six P. hookeri DNA sequences are available in GenBank, and only one (from Australia) is associated with a published morphological description. In this study, we performed a preliminary phylogenetic analysis including seven nuITS and mtSSU sequences from eight specimens collected in south Argentina. In addition, we provide an update of the taxon circumscription based on 50 specimens from 13 countries. Here, the molecular analyses revealed a well-supported monophyletic group formed by the P. hookeri sequences from Argentina, Australia, Norway and USA (Alaska). Regarding the morphological and anatomical characteristics, the Southern Hemisphere specimens did not present the historical reported variations, resulting in the proposition of a concise species description that accomplishes most of P. hookeri known geographical distribution.
Keywords:
species circumscription; cyanolichen; lichenized fungi; nuITS; taxonomy; Tierra del Fuego

Full text

doi: 10.1590/0102-33062021abb0357
Acta Botanica Brasilica
, 2022, 36: e2021abb0357
Original article
1 Universidade Federal de Mato Grosso do Sul, 79200-000, Aquidauana, MS, Brazil
2 Programa de Pós-Graduação em Ecologia e Conservação, Universidade Federal de Mato Grosso do Sul, Instituto de Biociências, 79070-900,
Campo Grande, MS, Brazil
3 Programa de Pós-Graduação em Biologia Vegetal, Universidade Federal de Mato Grosso do Sul, Instituto de Biociências, 79070-900,
Campo Grande, MS, Brazil
* Corresponding author: junjimjk@gmail.com
.
An integrative study of Pannaria hookeri (Ascomycota lichenized)
from Argentina and the update of the taxon circumscription based on
specimens from the Northern and Southern Hemispheres
Marcos Junji Kitaura1*

, Mayara Camila Scur2

, Adriano Afonso Spielmann3

and Aline Pedroso Lorenz2, 3

Received: December 5, 2021
Accepted: August 29, 2022
ABSTRACT
Pannaria hookeri (Borrer ex Sm.) Nyl. is reported as a bipolar cyanolichen that occurs in more than twenty countries
in both Hemispheres. In the Northern Hemisphere, a minor variation is attributed to the species, whereas larger
morphological and chemical variations have been reported in specimens from the Southern Hemisphere. Six P.
hookeri DNA sequences are available in GenBank, and only one (from Australia) is associated with a published
morphological description. In this study, we performed a preliminary phylogenetic analysis including seven nuITS and
mtSSU sequences from eight specimens collected in south Argentina. In addition, we provide an update of the taxon
circumscription based on 50 specimens from 13 countries. Here, the molecular analyses revealed a well-supported
monophyletic group formed by the P. h o okeri sequences from Argentina, Australia, Norway and USA (Alaska).
Regarding the morphological and anatomical characteristics, the Southern Hemisphere specimens did not present
the historical reported variations, resulting in the proposition of a concise species description that accomplishes
most of P. ho o ke r i known geographical distribution.
Keywords: species circumscription, cyanolichen, lichenized fungi, nuITS, taxonomy, Tierra del Fuego
Introduction
Pannaria Delise ex Bory is one of the largest genera
within Pannariaceae (lichenized Ascomycota), in which
the lichen-forming fungi can be associated with dierent
photobiont partners (Ekman et al. 2014; Magain & Sérusiaux
2014). Usually, the thalli are bipartite and the mycobiont is
associated with Nostoc cyanobacteria. Still, tripartite thalli
are also found within the genus where the mycobiont is
associated with green algae, as the main photobiont, and the
cyanobacteria are conned in structures called cephalodia,

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Marcos Junji Kitaura, Mayara Camila Scur,
Adriano Afonso Spielmann and Aline Pedroso Lorenz
for example in Pannaria durietzii (P. James & Henssen)
Elvebakk & D.J. Galloway (Elvebakk et al. 2008). e Pannaria
species are characterized by squamulose or foliose thalli,
apothecia with thalline margins, an amyloid hymenium
(I+ blue), asci without internal amyloid apical structures,
secondary metabolism with pannarin and related substances,
and occurrence in tropical, subtropical, temperate and polar
regions from both Hemispheres (Ekman et al. 2014).
Pannaria hookeri (Borrer ex Sm.) Nyl. is a species with
bipolar distribution (Jørgensen 1978; Kantvilas & Gueidan
2018), which has already been reported from Austria,
Canada, Faeroes Island, Finland, France, Greenland, Iceland,
Ireland, Italy, Norway, Russia, Sweden, Switzerland, and
U.S.A. in the Northern Hemisphere (Jørgensen 1978; 2000b;
2003; 2007; omson 1984); as well as from the Antarctic
Peninsula, Argentina, Australia, Chile, New Zealand, and
for the subantarctic islands and archipelagos Bouvetøya,
Kerguelen, Marion, South Georgia, South Orkney, and South
Shetlands in the Southern Hemisphere (Lindsay 1974;
1976; Redón 1985; Jørgensen 1978; 1986; 2000a; 2007;
Olech 1989; 2001; Øvstedal & Lewis Smith 2001; Calvelo
& Fryday 2006; Kantvilas & Gueidan 2018; Passo et al.
2020). In addition, P. h oo k e r i was recorded at high altitudes
in Ecuador (5050 m. a.s.l., Z. Palice 4598; Jørgensen &
Palice 2010) and points from Kenya (4430 m a.s.l. and
at 4640 m a.s.l.; Frisch & Hertel 1998). e species was
originally described from Scotland (Smith & Sowerby 1811),
and its rst record in the Southern Hemisphere was made
from a South African specimen, initially identied as P.
leucolepis (Wahlenb.) Nyl. (Stizenberger 1890; Doidge 1950)
and posteriorly included in the synonym list of P. h o okeri
(Jørgensen 2006). Nevertheless, there is no consensus about
the identication of the South African material, and also
from others localities from Southern Hemisphere, which
can also be an extreme form of P. rubiginosa (unb.) Delise
(Jørgensen 2003).
Regarding the morphological descriptions, a minor
variation is reported to P. hooke r i from Europe (Jørgensen
1978), while a larger variation has been described to P.
hookeri from the Southern Hemisphere (Jørgensen 2000a).
In the same way, the chemical prole diers according to the
region studied. Traces of atranorin and ergosterol peroxide
was found in specimens from Robert Island (Quilhot et al.
1989), the absence of secondary substances to Bouvetøya
specimens (Jørgensen 1986), and pannarin or related
substances were reported to specimens from Argentina
(Passo et al. 2020), Australia (Kantvilas & Gueidan 2018),
and Europe (Jørgensen 1978; 2007), inferring cryptic
lineages and taxonomical problems in the morphological
and chemical studies.
Despite the observed inconsistencies, the wide
geographical distribution of P. hookeri was confirmed
when the sequences of the nuITS and mtSSU regions were
generated from an Australian specimen and clustered with P.
hookeri sequences from Norway (Kantvilas & Gueidan 2018).
Nowadays, only six specimens have available sequences
in the GenBank: one from Australia, three from Norway,
one from USA (Alaska) and one of unknown origin (Ekman
& Jørgensen 2002; Spribille & Muggia 2013; Kantvilas
& Gueidan 2018; Park et al. 2018; Marthinsen et al.
2019). erefore, better knowledge is still needed for the
circumscription characters that belong to P. h o o k er i , including
genetic analysis with Southern Hemisphere specimens.
e aim of this study was to a) perform a phylogenetic
analysis of P. h o okeri from Argentina and sequences
available on GenBank, generating the first molecular
sequences from the South America; and b) to update the
species circumscription based on specimens from the
Northern (including topotypes) and Southern Hemispheres,
establishing a unied morphological description.
Material and methods
Sampling
Fresh samples were collected in the National Park of
Tierra del Fuego, Ushuaia, extreme south of the Argentina,
during the summer of 2016-2017 (Proyecto Número 105-
CPA-2016, supported by CONICET and PROANTAR), and
specimens were found on rock and saxicolous mosses.
Phylogenetic analysis
Eight fresh-collected specimens were used in the
molecular analysis. DNA extraction, amplication, and
sequencing of the nuITS (nuclear internal transcribed
spacer) and mtSSU (mitochondrial small subunit) regions
were performed according to Kitaura et al. (2018). However,
the specimens requested as loans have more than 10 years,
and the DNA extraction was not performed. In addition,
four sequences of P. hooker i of each nuITS and mtSSU
regions were retrieved from GenBank. ese sequences
were generated from Australian, Norwegian, U.S.A., and
unknown origin specimens (Tab. 1).
Sequences of Staurolemma oculatum P.M. Jørg. &
Aptroot (GenBank accession numbers KC618738 and
GQ259045) and S. omphalarioides (Anzi) P.M. Jørg. &
Henssen (KJ533487 and KJ533439) were used as outgroups
according to Kantvilas & Gueidan (2018) (Tab. 1).
e alignments were performed in Geneious v9.1.2
(Kearse et al. 2012) with the MAFFT v7.308 algorithm
(Katoh et al. 2002) using default settings, and the
Gblocks web server (http://molevol.cmima.csic.es/
castresana/Gblocks_server.html) was used to exclude
unreliable aligned sites using the less stringent options.
The alignments were used to infer trees from nuITS,
mtSSU, and concatenated regions. Phylogenetic trees
were estimated using the Bayesian (BA) and Maximum
Likelihood (ML) approaches, and analyses for each region
separately and for a combined data matrix were performed.

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Acta Botanica Brasilica
, 2022, 36: e2021abb0357
An integrative study of Pannaria hookeri (Ascomycota lichenized) from Argentina and the update
of the taxon circumscription based on specimens from the Northern and Southern Hemispheres
Table 1. Dataset used in the present study for the phylogenetic analysis. Species, geographical origin, voucher information, nuITS
and mtSSU GenBank accession codes and references. Sequences generated in this study are marked in bold.
Species Origin Voucher
GenBank Accession numbers
Reference
nuITS mtSSU
Pannaria andina Chile Elvebakk 06-245 GQ927268 - (Elvebakk et al. 2010)
Pannaria athroophylla Argentina Passo 181 EU885295 EU885317 (Passo et al. 2008)
Pannaria athroophylla Argentina Passo 251 EU885303 EU885325 (Passo et al. 2008)
Pannaria calophylla Argentina Passo 101 EU885296 EU885318 (Passo et al. 2008)
Pannaria conoplea Norway Ekman 3188 AF429281 - (Ekman & Jørgensen 2002)
Pannaria contorta Argentina Passo 142 EU885297 EU885319 (Passo et al. 2008)
Pannaria farinosa Argentina Passo 119 EU885299 EU885321 (Passo et al. 2008)
Pannaria hookeri Australia GK102/16 MG786563 MG792317 (Kantvilas & Gueidan 2018)
Pannaria hookeri Norway Jørgensen s.n. AF429282 KC608083 (Ekman & Jørgensen 2002)
Pannaria hookeri Norway NK-325 KY350562 - (Park et al. 2018)
Pannaria hookeri Norway, NO, Nordland JT Klepsland MK812291 - (Marthinsen et al. 2019)
Pannaria hookeri USA, Alaska Spribille 29292 (KLGO) - JX464134 (Spribille & Muggia 2013)
Pannaria hookeri Unknown Jørgensen s.n. (BG) - KC608083 Unpublished
Pannaria hookeri Argentina MJK3314 MT755913 MN63424 is study
Pannaria hookeri Argentina MJK3319 MT755912 MN634242 is study
Pannaria hookeri Argentina MJK3323 MT755911 MN634243 is study
Pannaria hookeri Argentina MJK4022 - MN634244 is study
Pannaria hookeri Argentina MJK4028 MT755910 MN634245 is study
Pannaria hookeri Argentina MJK4036 MT755909 MN634246 is study
Pannaria hookeri Argentina MJK4041 MT755908 MN634247 is study
Pannaria hookeri Argentina MJK4045 MT755907 - is study
Pannaria immixta Unknown Elvebakk 02-352b - KC608084 (Kantvilas & Gueidan 2018)
Psoroma implexa Argentina Passo 84 - EU885333 (Passo et al. 2008)
Pannaria insularis Japan Kashiwadani 43760 KC618716 KC608085 (Kantvilas & Gueidan 2018)
Pannaria leucosticta Unknown Hur 041227 EU266107 - (Kantvilas & Gueidan 2018)
Pannaria lurida subsp. lurida Unknown Kashiwadani 43861 - KC608086 (Kantvilas & Gueidan 2018)
Pannaria lurida subsp. russellii Unknown Tønsberg 22565 - KC608087 (Kantvilas & Gueidan 2018)
Pannaria microphyllizans Argentina Passo 264 EU885300 EU885322 (Passo et al. 2008)
Pannaria multida Unknown Schumm & Frahm s.n. KC618717 KC608088 (Kantvilas & Gueidan 2018)
Pannaria pallida Argentina Passo 249 EU885301 EU885323 (Passo et al. 2008)
Pannaria rubiginella Canada or 10050 - GQ259037 (Wedin et al. 2009)
Pannaria rubiginella Unknown Tønsberg 32508 KC618718 KC608089 (Kantvilas & Gueidan 2018)
Pannaria rubiginosa Unknown Anonby 870/Purvis s.n. KC618717 AY340513 (Kantvilas & Gueidan 2018)
Pannaria sphinctrina Argentina Passo 221 EU885302 EU885324 (Passo et al. 2008)
Pannaria subfusca Unknown Tønsberg 33592 KC618719 - (Kantvilas & Gueidan 2018)
Pannaria tavaresii Argentina Passo 122 EU885294 EU885316 (Passo et al. 2008)
Pannaria tavaresii Unknown Schumm s.n. KC618720 - (Kantvilas & Gueidan 2018)
Staurolemma oculatum China Aptroot 55941 KC618738 GQ259045 (Kantvilas & Gueidan 2018)
Staurolemma omphalarioides Turkey Tibell s.n. KJ533487 KJ533439 (Bendiksby et al. 2014)

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Marcos Junji Kitaura, Mayara Camila Scur,
Adriano Afonso Spielmann and Aline Pedroso Lorenz
e nucleotide substitution and site heterogeneity models
were inferred following the Bayesian Inference Criterion
in jModelTest2 on XSEDE, CIPRES Science Gateway V.3.3
(Guindon & Gascuel 2003; Miller et al. 2010; Darriba et al.
2012). e BA was performed in MrBayes v.3.2.7a on XSEDE,
available in the CIPRES Science Gateway V.3.3 (Huelsenbeck
& Ronquist 2001; Ronquist & Huelsenbeck 2003; Miller et
al. 2010) using TrNef+I+G as substitution model for nuITS,
TPM2uf+I+G for mtSSU regions and TIM1+I+G for the
concatenated dataset, and their respective base frequencies
were settled. e settings were: two independent runs with
four chains (length of 10,000,000 generations), with trees
sampled every 1,000 steps. e rst 25 % of the generated
trees were discarded as burn-in, and a 50 %-majority-rule
tree was built. e ML trees were built with the RaxML-
HPC2 v.8.2.12 on XSEDE available in the CIPRES Science
Gateway V.3.3 (Miller et al. 2010; Stamatakis 2014), using
GTR+G+I as nucleotide substitution model and 1,000
bootstrap replicates. FigTree v1.4.2 (http://tree.bio.ed.ac.
uk/software/gtree/) was used to edit the trees. Branches
with posterior probability values above 0.95 (BA) and
bootstrap above 75 (ML) were considered signicantly
supported.
Morphological and anatomical studies
e AAS, C, COLO, E, H, M, O, and S herbaria sent 35
specimens from the Northern Hemisphere and seven from
the Southern Hemisphere. e specimens received as loan
were morphologically examined, and the determinations
were conrmed (Text S1), when compared with eight fresh
P. ho o ke r i specimens.
The P. h o o k e r i description was built according to
Jørgensen (1978). en, our description was compared
with other available in the literature (Lindsay 1974; Redón
1985; Jørgensen 1986, 2007; Øvstedal & Lewis Smith 2001;
Kantvilas & Gueidan 2018; Passo et al. 2020), and the
variable characters were highlighted in Table 2 and discussed
here. e specimens were tested chemically by applying
PD spot tests, and orange reactions were interpreted as
indicating the presence of pannarin. erefore, TLC and
HPLC analyses are necessary to conrm the substances.
Results
Molecular analysis
e dataset of the present study was composed of
sequences of the nuITS and mtSSU regions generated from
eight specimens of Pannaria hookeri collected in southern
Argentina. Furthermore, six P. h o oke r i sequences retrieved
from GenBank composed the dataset together with other
Pannaria species sequences and the selected outgroups
(Tab. 1). e nal dataset comprised 31 sequences and an
alignment 1105 base long, with 423 of the nuITS and 682
of the mtSSU regions.
Trees of both BA and ML approaches, among genes
and concatenated datasets, showed congruent topology,
so only the concatenated tree is presented, with the
values of the posterior probabilities and bootstrap values
shown (Fig. 1). e sequences of the specimens identied
as P. hookeri, newly produced and those obtained from
GenBank, presented 0.89/95 of posterior probability and
bootstrap values, respectively, grouping the specimens
from Argentina, Australia, Norway, and the USA in a well-
supported monophyletic group. e divergence in the nuITS
region, considered the universal barcode DNA of fungi
(Schoch et al. 2012), revealed a low variation, with the
mean percentage of identity of 99.34
% (98.25
% to 100
%)
among the sequences, without deeply divergent populations
or cryptic lineages.
e morphological descriptions of Pannaria hookeri from
the literature were gathered and used for the circumscription
of the species and for the observation of inconsistencies
in the species descriptions and identications, with the
dierences found highlighted in the sections “Description
of Species” and “Discussion” below (Tab. 2).
Description of species
Pannaria hookeri (Borr. ex Sm.) Nyl., Mém. Soc. Sci. Nat.
Math. Cherbourg 5: 109 (1857). (Fig. 2 A-F)
Description. Thallus placodioid. Lobes at the
circumference narrow, 0.5-1.0(-2.0) mm broad and 1.0-2.5
mm long, convex, enlarged in the apices, centrally verrucose
and fragmented or often partially detached. Upper surface
whitish beige to brownish-grey under uorescent light,
whitish beige to whitish brown under the stereomicroscope.
Hypothallus inconspicuous at marginal branches. Apothecia
frequent, 0.4-1.5(-2.0) mm diam., with black discs. alline
margin conspicuous, usually crenulate, concolorous with the
thallus. allus 200-400 µm thick. Upper cortex 10-50 µm (2-
12 layers of cells) thick, proso- to colloparaplectenchymatous
tissue; cells thick-walled cells, 2.5-5.0 µm diam. Photobiont
layer dense and massive, probably Nostoc clusters, spherical
cells ca. 5 µm diam.; numerous vertically orientated fascicles
of hyphae derived from the medulla penetrating into the
photobiont layer. Medulla is hyaline with some photobionts.
Lower cortex 12.5-25.0 µm (ca. 3 cells) thick when on the
moss, blackish. Apothecia with a thick thalline margin to
250 µm, containing a large number of algal cells; cortex of
thalline exciple colloplectenchymatous or with amorphous
cells, 5.0-10.0 µm (2 cells) thick at the apices, 7.5-35.0 µm
(2-8 cells) thick at the middle, indistinct at the base. Proper
exciple ca. 50 µm thick, colloplectenchymatous to elongated
cells. Subhymenial layers yellowish to hyaline, 25-65 µm
thick, usually prosoplectenchymatous. Hymenium 100-
200 µm thick, partially I+ persistently deep blue, mainly
in the vicinity of the asci. Simple paraphyses, up to 2.5 µm
thick, slightly clavate at the apices. Asci 50-65 × ca. 20 µm.

5
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Acta Botanica Brasilica
, 2022, 36: e2021abb0357
An integrative study of Pannaria hookeri (Ascomycota lichenized) from Argentina and the update
of the taxon circumscription based on specimens from the Northern and Southern Hemispheres
Figure 1.
Bayesian Maximum Clade Credibility tree based on nuITS and mtSSU datasets showing the phylogenetic relationships within the Pannaria and P. h o ok e r i sequences from Argentina,
Australia, Norway and USA (Alaska). Posterior probabilities and Bootstrap values are above branches, respectively. Bold branches have posterior probabilities (PP) >0.95 % or bootstrap
values >75. Sequences obtained in this study are marked in bold. Text after species names corresponds to voucher information (Tab. 1).

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Acta Botanica Brasilica
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Marcos Junji Kitaura, Mayara Camila Scur,
Adriano Afonso Spielmann and Aline Pedroso Lorenz
Table 2. Morphological descriptions of Pannaria hookeri reported in the literature and in the present study.
Lindsay (1974) Redón (1985) Jørgensen (1986) Jørgensen (2000a) Øvstedal & Lewis Smith
(2001) Jørgensen (2007) Kantvilas & Gueidan (2018) Passo et al.
(2020) This study
Upper surface
color
Greenish grey,
streaked with
white towards tips
of squamules
Greenish grey
Brownish, white-grey
striated and with distinctly
white-grey margins which
are sometimes blackened
Brownish grey, but
dominated by the whitish
margins and the laminal
white striae
Grey to brownish, also to
Pannaria caespitosa P.M.
Jørg
Grey-brown
though whitish
striate
Pale brownish grey to
smoky bluish grey
Bluish grey to
brownish
Whitish beige to
brownish-grey
Branches Not mentioned Up to 3 mm broad Up to 4 mm broad To 2 mm 3–4 mm, also to Pannaria
caespitosa P.M. Jørg Up to 3 mm 0.3–1.0 mm wide 0.5–1.0 mm diam. 0.5–1.0(–2.0) mm
broad
Marginal lobes Enlarged Enlarged Deeply divided, finger like
peripheral lobes Fan-shaped Radiated ± egurate and spathulate Discrete Enlarged in the
apices
Central part Areolate to
granulose Not mentioned Disintegrating in the
subcrustose Not mentioned
Centrally often
verrucosely
fragmented and
partially detached
Irregularly rhomboidal and
delimited by deep cracks Grouped
Verrucose and
fragmented or
often partially
detached
Apothecia Up to 5 mm diam. 0.5–2.0 (–5.0) mm
diam. Up to 3 mm broad To at most 2 mm diam
Up to 3.5 mm, also to
Pannaria caespitosa P.M.
Jørg
Up to 2 mm 0.4–1.3 mm diam. 0.4–1.3 mm diam. 0.4–1.5(–2.0) mm
diam.
Discs Some shade of
brown Brown to black Black or dark brown Usually with black Pale brown Black to rarely
dark brown Dark brown to black Black to dark
brown Black discs
Ascospores Ellipsoid, 14–16 ×
7–8 µm
Ellipsoid,
(11.5–)14–16(–17) ×
7–8(–11) µm
Ellipsoid to globular, 12–14 ×
10–12 µm
Nearly globular, 12–15 ×
8–11 µm
Subglobose, 12–15 ×
10–13 µm
Subglobose, 12–15
× 8–11 µm
Ellipsoid to ovate,
(10–)12–14.0–16(–18) ×
6–7.6–9(–10) µm
Ellipsoid to ovate,
12–16 × 6–9 µm
Ellipsoid to
spherical, (7.5–
)12.5–15.0(–17.5) ×
5.0–12.5 µm
Examined
specimens South Georgia South Shetland
Islands Bouvetøya Island
Canada, Greenland,
Argentina, Chile, Kerguelen,
Marion I., New Zealand, and
South Shetland Isl.
Antarctica and South
Georgia Island Nordic Region Tasmania Argentina
14 localities from
the northern
and southern
Hemispheres

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Acta Botanica Brasilica
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An integrative study of Pannaria hookeri (Ascomycota lichenized) from Argentina and the update
of the taxon circumscription based on specimens from the Northern and Southern Hemispheres
Figure 2. Pannaria hookeri from Argentina. (A): Specimen on the habitat. (B): Detail of laciniate branches on moss (MJK3314).
(C): Saxicolous specimen (MJK4041). (D): Transversal section of the thallus. (E): Section of apothecium on the squamule. (F): Diametral
section of apothecium. Symbols. apo = apothecium; black arrowhead = parahymenial tissue; black arrow = moss; hym = hymenium;
hyp = hypothallum; lc = lower cortex; mh = medullar hyphae; ph = photobiont layer; squ = squamule; sub = subhymenium; uc = upper
cortex; white arrow = P+ orange (pannarin).

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Marcos Junji Kitaura, Mayara Camila Scur,
Adriano Afonso Spielmann and Aline Pedroso Lorenz
Spores 8 per ascus, (7.5-)12.5-15.0(-17.5) × 5.0–12.5 µm,
colorless, ellipsoid to spherical, smooth-walled. Pycnidia
were not observed.
Spot tests: K-, C-, KC-, P+ orange (probably pannarin).
Notes: Pannaria hookeri is characterized by presence of
thallus placodioid with marginal lobes enlarged (less than 2
mm), central part often verrucose, fragmented and partially
detached, and by presence of pannarin (P+ orange). e
lecanorine apothecia have 0.4-1.5(-2.0) mm diam., with the
blackish discs, subhymenium of prosoplectenchymatous
tissue, and hypothecium and parahymenium of
colloplectenchymatous tissue and elongated cells. e
ascospores have always smooth epispores.
We examined specimens are from Austria (4 specimens),
Canada (2), Denmark (Faroe Islands, 1), Greenland (1),
Iceland (1), Italy (1), Norway (16), Russia (1), Scotland
(4), and Sweden (4) from the Northern Hemisphere;
and Argentina (11), Chile (1), Kerguelen Island (2), and
New Zealand (1) from the Southern Hemisphere. e
determination of the specimens received on loan from
herbaria was confirmed only through morphological
approach. Unfortunately, most of the received material
was collected more than 10 years ago (Text S1), and we
were not able to generate genetic sequences. When the P.
hookeri characters were not observed in the material, the
specimen was excluded of the study.
Discussion
In this study, we re-conrmed the wide geographical
distribution of Pannaria hookeri through phylogenetic and
morphological evidence, now including the rst P. hook e ri
sequences from the southern South America, Argentina.
e phylogenetic analysis resulted in a well-supported
clade with P. ho o ker i sequences from Argentina, Australia,
Norway, and the USA, whereas the morphological analysis
included specimens from Argentina, Austria, Canada,
Chile, Denmark (Faroe Islands), Greenland, Iceland, Italy,
Kerguelen Island, New Zealand, Norway, Russia, Scotland
and Sweden, generating a concise morphological description.
e materials from Scotland are topotypes (Text S1) and
were compared with fresh specimens.
e bipolar distribution has been concepted as species
that are disjunctly distributed in both hemispheres, mostly
in polar and subpolar regions, and are largely absent in the
tropics. However, many lichens reported as bipolar are also
distributed in temperate and tropical latitudes - usually in
high mountains, where their ecological requirements are met
(Garrido-Benavent & Pérez-Ortega 2017), which is the case
of P. ho okeri, and should be carefully revised and possibly
considered as (sub)cosmopolitan. Nowadays, the species is
mentioned in high altitudes of Ecuador and Kenya (material
not revised in the present study) (Frisch & Hertel 1998;
Jørgensen & Palice 2010), but eorts are still needed to
conrm or not the species distribution through the tropical
latitudes. Studies including DNA analysis of species reported
as bipolar has conrmed this remarkable distribution, for
example in Austroplaca, Cetraria and Cladonia (Myllys et al.
2003; Søchting & Castello 2012; Fernández-Mendoza &
Printzen 2013), while others has resulted in the discovery
of cryptic lineages or restricted distribution, such as in
Parmelia, Sphaerophorus and Rusavskia (Crespo et al. 2002;
Högnabba & Wedin 2003; Scur et al. 2022), indicating that
phylogeographical studies should be performed to revise
the distribution stated to P. ho o ke r i.
Historically, Pannaria hookeri specimens from the
Southern Hemisphere were reported with a larger
morphological plasticity than specimens from Northern
Hemisphere (Jørgensen 2000a), and the taxonomic
problems has invalited other results, e.g. the chemical
study (Quilhot et al. 1989). e variations in the color of
the thallus, branching, diameter of apothecia, and color of
the apothecium discs, which were reported in the literature
from Antarctic specimens (Lindsay 1974; Redón 1985;
Jørgensen 1986; 2000a; Øvstedal & Lewis Smith 2001), were
not observed in the material conrmed by us and can be
characters of other cryptic species. For example, apothecia
up to 5.0 mm diam. were reported to the specimens from
South Georgia and South Shetland Islands (Lindsay 1974;
Redón 1985), apothecia up to 3.0 mm diam. in specimens
from Bouvetøya Island (Jørgensen 1986), but only apothecia
up to 2 mm diam. were observed during the present study.
In the same way, Øvstedal & Lewis Smith (2001)
reported the species to South Georgia, Bouvetøya, South
Orkney, South Shetland Islands, and Antarctic Peninsula,
but posteriorly transferred the most of examined specimens
within P. caespitosa P.M. Jørg. (Jorgensen 2000a). Only
two specimens, SJA 168 and RILS 8347, were kept within
P. h ookeri, but they have the presence of pale brown discs
in the apothecia (Øvstedal & Lewis Smith 2001) which
do not match with the dark brown to blackish discs of P.
hookeri (Tab. 2). erefore, a revision is still required to the
both specimens.
e material received as loan that were not conrmed
as P. ho oker i are from Antarctic and subantarctic Islands
(Antarctica Peninsula and Kerguelen, Marion, South
Georgia, South Orkney and King George islands - Text
S1), highlighting the taxonomic problem in the region. In
addition, fresh specimens collected during four Brazilian
Antarctic Expeditions in the Antarctic Peninsula (near to
Esperanza and Primavera stations), James Ross Island,
Marambio Island, and South Shetland Islands (Deception,
Greenwich, King George, Livingston, Nelson, Robert, and
Snow Islands) did not cluster with the P. h ook e r i sequences
and they were also excluded from our analysis. An integrative
revision of the Psoroma-complex is also required to
Antarctica as well as a complete identication key of the
group; P. ho o keri in Antarctica can be more restricted than
previously thought.

Template: Editora Letra1 | www.editoraletra1.com.br
9
Acta Botanica Brasilica
, 2022, 36: e2021abb0357
An integrative study of Pannaria hookeri (Ascomycota lichenized) from Argentina and the update
of the taxon circumscription based on specimens from the Northern and Southern Hemispheres
Previously, P. ho o k e r i was considered as strictly saxicolous
(Jørgensen 2000a). However, we observed in Argentina that the
species can grow on the rock (Fig. 1A), and also on saxicolous
moss (Fig. 1B), expanding the species’ known ecology.
Furthermore, the species were usually found on the mountains
in Argentina, Australia and Chile (Jørgensen 2000a; Kantvilas
& Gueidan 2018). Calvelo & Fryday (2006) reported P. h o o k e r i
on rock outcrops along a stream the sea-level in Argentina, but
the material has not been reviewed by us.
Our description was based on 50 specimens from 13
countries, corroborating with the descriptions provided by
Jørgensen (1978; 2000a; 2007), Kantvilas & Gueidan (2018)
and Passo et al. (2020). erefore, the large morphological
variations reported for specimens from the Southern
Hemisphere are potentially characters of dierent Psoroma
species. Studies including more genetic sequences, chemical
and morphological variation analysis should be performed
to dept the evolutionary history and concept of Pannaria
hookeri.
Acknowledgments
e authors thanks Dr. A.E., Dr. A.P. and others anonymous
reviewers that substantially corroborated to better the
manuscript. is work was nanced in part by the Conselho
Nacional de Desenvolvimento Cientíco e Tecnológico
(CNPq), under the “Ação Transversal nº 64/2013 – Chamada
MCTI/CNPq/FNDCT”; and by the Fundação Universidade
Federal de Mato Grosso do Sul – UFMS/MEC – Brazil. Marcos
J. Kitaura was supported in part by the Fundação de Apoio ao
Desenvolvimento do Ensino, Ciência e Tecnologia do Estado
de Mato Grosso do Sul (FUNDECT – Chamada 04/2019),
by the CNPq, and by the Coordenação de Aperfeiçoamento
de Pessoal de Nível Superior - Brasil (CAPES) - Finance
Code 001”. Mayara C. Scur was supported by the CAPES -
Finance Code 001”.
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