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Citation: Cano, M.J.; Jiménez, J.A.;
Martínez, M.; Hedenäs, L.; Gallego,
M.T.; Rodríguez, O.; Guerra, J.
Integrative Taxonomy Reveals
Hidden Diversity in the Aloina
catillum Complex (Pottiaceae,
Bryophyta). Plants 2024,13, 445.
https://doi.org/10.3390/
plants13030445
Academic Editor: Harald G.
Zechmeister
Received: 19 December 2023
Revised: 26 January 2024
Accepted: 26 January 2024
Published: 2 February 2024
Copyright: © 2024 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
plants
Article
Integrative Taxonomy Reveals Hidden Diversity in the Aloina
catillum Complex (Pottiaceae, Bryophyta)
María J. Cano 1, * , Juan A. Jiménez 1, Mónica Martínez 1, Lars Hedenäs 2, M. Teresa Gallego 1,
Omar Rodríguez 1and Juan Guerra 1
1Departamento de Biología Vegetal (Botánica), Facultad de Biología, Universidad de Murcia,
Campus de Espinardo, 30100 Murcia, Spain; jajimene@um.es (J.A.J.); monica.martinez11@um.es (M.M.);
mgallego@um.es (M.T.G.); omar.rodrigueza@um.es (O.R.); jguerra@um.es (J.G.)
2Department of Botany, Swedish Museum of Natural History, P.O. Box 50007, 104 05 Stockholm, Sweden;
lars.hedenas@nrm.se
*Correspondence: mcano@um.es
Abstract: Aloina catillum is a variable moss typical of xerophytic environments in the Neotropics,
characterized against other closely allied Aloina species with well-differentiated leaf border by its
setae twisted to the left throughout. In order to clarify its variability and its relationships with the
allied species with differentiated leaf border A. brevirostris,A. obliquifolia, and A. rigida, we performed
an integrative study including sequence data from four markers (nuclear ITS, plastid atpB-rbcL, trnG,
trnL-F), morphometry, and species assembling by automatic partitioning (ASAP) algorithm. Our data
suggest that A. catillum consists of at least three species: A. calceolifolia (an earlier name for A.catillum),
and two species described here as a new, A. bracteata sp. nov. and A. limbata sp. nov. This latter species
includes the specimens previously identified as A. obliquifolia from South America. Additionally,
some morphological and molecular variability was also detected in A. limbata, but was not consistent
enough to be recognized taxonomically. The study supports the presence of A. brevirostris in the
Neotropics and A. rigida is tentatively excluded from South America. Full descriptions of the A.
catillum s.l. species and a diagnostic key to this complex in South America are provided.
Keywords: ASAP; atpB-rbcL; bryophytes; cryptic diversity; ITS; NeighborNet split network;
phylogenetic
analysis; South America; taxonomy; trnG; trnL-F
1. Introduction
The incorporation of molecular phylogenetic studies into taxonomy has greatly im-
proved our understanding of relationships within bryophytes and has uncovered extensive
hidden diversity in groups with morphological characters that are difficult to interpret.
Cryptic species is a common and increasingly used term that refers to taxa that cannot
readily be distinguished morphologically, yet evidence indicates they are on different evo-
lutionary trajectories [
1
,
2
]. In addition, species with minor morphological differences found
after reinvestigations are usually named by terms such as semi-cryptic or pseudo-cryptic [
3
].
With the increasing use of molecular approaches in recent decades, many bryophyte
studies have been published on such variation (e.g., [
4
–
9
]), which represents a significant
portion of undiscovered biodiversity. However, many of these studies come from northern
temperate regions and very few treat cryptic or pseudo-cryptic species from the South-
ern Hemisphere [
10
,
11
]. Discoveries of cryptic or pseudo-cryptic diversity may alter the
assumed distribution of diversity and endemism, which in turn may change our under-
standing of evolutionary processes and could have profound implications for biogeography
and conservation planning [1].
Aloina Kindb. is a small genus of the moss family Pottiaceae adapted to xerophytic
environments [
12
]. The genus is well characterized morphologically by its leaves, usu-
ally differentiated into a sheathing base and a concave blade with the margins incurved
Plants 2024,13, 445. https://doi.org/10.3390/plants13030445 https://www.mdpi.com/journal/plants
Plants 2024,13, 445 2 of 29
and ventral chlorophyllous filaments that extend over the costa and the lamina surface.
However, the species level identification of some of their complexes such as A. ambigua
(Bruch & Schimp.) Limpr.-A. aloides (W.D.J. Koch ex Schultz) Kindb., or A. rigida (Hedw.)
Limpr.-A. obliquifolia (Müll. Hal.) Broth. is well known to be problematic. Thus, there still
exist disagreements regarding how many species to recognize in this genus, according to
different authors [
13
–
15
]. This genus is less frequent in the Southern Hemisphere than in
the Northern Hemisphere where, however, it appears to be strongly diversified in number
of species and endemism. In South America, seven species have been reported. One group
of species (A. bifrons (De Not.) Delgad., A. roseae (R.S. Williams) Delgad., and A. scindulosa
M.J. Cano, J.A. Jiménez & M.T. Gallego is characterized by unbordered leaves, with a
long, hyaline hair-point [
12
,
13
]. The other group, formed by A. brevirostris (Hook. & Grev.)
Kindb., A. catillum (Müll. Hal.) Broth., A. obliquifolia, and A. rigida, is characterized by leaves
bordered by elongate or oblate, thin-walled, hyaline cells disposed in several columns.
Aloina brevirostris is known from a single locality in the Magallanes Region of southern
Chile [
16
], and A. catillum,A. obliquifolia, and A. rigida have been reported from a few
localities in dry areas of the tropical Andes [
17
–
19
] (Figure 1). Two poorly known species,
due to the unavailability of type material [
18
], A. recurvipatula (Müll. Hal.) Broth. and
A. sedifolia (Müll. Hal) Broth., described from Argentina, seem to exhibit a differentiated
border, according to their protologues [
20
,
21
], and could be related with this group of
species [12].
Aloina catillum is a poorly known species described by Müller [
20
] from a single
Argentinian specimen. More recently, this species has been reported from a few localities
in the tropical Andes of Chile, Ecuador, and Peru [
13
,
19
]. Delgadillo [
13
], in his world
revision of Aloina, tentatively recognized this species, hoping that more collections were
available, because he only could study the type material and one additional specimen.
He characterized A. catillum by its suborbicular leaves, with marginal cells differentiated
and the long ovoid-cylindrical capsules. This author also indicated that the cladautoicous,
paroicous, or dioicous condition and the extremely broad costa may eventually be found to
be taxonomically useful characters. Regarding sporophytic differentiation, Delgadillo [
13
]
stated that in A. catillum, the seta turns to the left above, unlike the other species of Aloina
with leaves bordered, such as A. rigida (including A. obliquifolia) or A. brevirostris, where
it is dextrorse above. He also noted that most features of A. catillum show considerable
overlap with those of A. rigida. Thus, A. calceolifolia (Spruce ex Mitt.) Broth., treated as a
synonym of A. rigida by Delgadillo [
13
], had some leaves as broadly lingulate as some of
those in A. catillum, and the seta, at least in the type specimens of this taxon, twisted to
the left throughout. Unlike the Neotropical A. catillum,A. brevirostris exhibits a bipolar
distribution [
22
]. Aloina rigida is widespread in the Northern Hemisphere, but not reaching
the Arctic, with a few records in South America and tropical Africa [
13
,
15
]. Aloina obliquifolia
is easily distinguished by the leaf costa excurrent as a stiff awn or mucro. It was described
from China [
23
] as Barbula obliquifolia Müll. Hal., and later reported from Japan [
24
]. Aloina
rigida var. mucronulata (Bruch & Schimp.) Limpr. was considered to be conspecific with
A. obliquifolia, extending the distribution of this taxon to Europe [
15
]. Subsequently, it
was reported from Ecuador and Peru in the tropical Andes [
19
]. There is another taxon
with leaves bordered that is very close to A. obliquifolia, known only from two collections
from China, A. cornifolia Delgad., which is distinguished mainly by the presence of a
strongly cucullate leaf apex, with the mucro developing from the dorsal side of a hood-like
apex [
13
,
25
]. Despite the existence of a global monograph of the genus [
13
] and a taxonomic
revision in the Mediterranean area [
15
], there is no comprehensive molecular phylogeny of
this genus of the Pottiaceae and only a few Aloina sequences used as outgroups have been
published (e.g., [
26
,
27
]). Therefore, the current morphology-based species concept in this
genus has not been tested by molecular evidence.
Plants 2024,13, 445 3 of 29
Figure 1. Habit of plants from the Aloina catillum complex and allied species. (A)Aloina brevirostris
(Cano et al. 3403a, MUB 27728). (B)Aloina rigida (Cano 4843, MUB 28934). (C)Aloina obliquifolia
(A. limbata) (Cano et al. 7649, MUB 49149). (D)Aloina catillum s.l. (A. bracteata) (Cano et al. 4113,
MUB 29597). (E)Aloina catillum s.l. (A. limbata) (Cano 5065, MUB 32542). (F)Aloina catillum s.l.
(A. calceolifolia) (Cano and Alonso 8345, MUB 56422). Photos: A,C–FCarlos Aedo; BMaría J. Cano.
In our ongoing study of the Pottiaceae in South America, we have had the opportunity
to collect new material belonging to the genus Aloina in Andean dry areas. Most specimens
were dioicous without sporophytes, making the identification quite difficult. In addition,
we found a large morphological variation in the gametophytic and sporophytic characters
in the specimens identified as A. catillum, which may be due to overlooked diversity within
Plants 2024,13, 445 4 of 29
the genus. To clarify this, in the framework of our ongoing and integrative taxonomic
approach to the genus Aloina, a study of A. catillum and related species was performed.
Based on preliminary morphological identification of the specimens from South America,
we hypothesized that A. catillum s.l. could represent several distinct evolutionary entities.
Our aims were: (1) To test if the molecular topology is in accordance with the current
morphology-based species concept in A. catillum s. str. and related species; (2) To quantify
the morphological variation in A. catillum s.l. and to determine if this variation could be
correlated with discrete taxonomic entities.
2. Materials and Methods
2.1. Study Species and Material
A total of 37 specimens currently referable to Aloina catillum were selected for the
molecular and morphological evaluation in order to span its morphological variability and
geographical range. In addition, eleven specimens of A. obliquifolia, nine of A. brevirostris,
and six of A. rigida were included on the basis of their well-differentiated leaf border. With
the intention to assess the position of A. catillum s.l. and related species within the genus,
three additional species, A. aloides (one specimen), A. ambigua (one specimen), and A. bifrons
(two specimens), were included. Moreover, three representatives of other genera of the
subfamily Pottioideae (Aloinella Cardot, Crossidium Jur., and Erythrophyllopsis Broth.) were
selected to form the outgroup of the molecular analysis based on previous phylogenetic
studies [
27
,
28
]. Most specimens were collected during several fieldwork visits by the
authors in Argentina, Bolivia, Cape Verde, Cyprus, Ecuador, Greece (Crete), Peru, Spain,
and Venezuela. Full details on the fieldwork localities are available at the website of the
group (www.pottiaceae.com; accessed on 15 September 2023). In addition, herbarium
material from CAS, MUB, UBC, the private herbarium of T.L. Blockeel, and type material
of the taxa currently attributed to Aloina deposited at E, H, NY, PC, and S were examined.
Details of the plant material, including voucher information, accession provenance, and
GenBank accession numbers, are provided in Appendix A.
2.2. DNA Extraction, Amplification, and Sequencing
Plant tissue from the distal portions of gametophore shoots was isolated from herbar-
ium specimens or recent collections. Whole genome DNA was then extracted, either using
a modified version of the CTAB [
29
] or the protocol for extraction by Suzuki et al. [
30
], and
stored at
−
20
◦
C until the polymerase chain reaction (PCR) was carried out. We selected
four loci; three from the chloroplast genome, the atpB-rbcL intergenic spacer region (atpB-
rbcL), the trnG
UCC
G2 intron (trnG), and the trnL
UAA
exon trnF
GAA
region (trnL-F), as well
as the nuclear internal transcribed spacers 1 and 2 (ITS1-5.8S-ITS2). The ITS1 and ITS2 were
either amplified and sequenced separately or in a single amplification. All loci have been
shown to be useful for phylogenetic reconstruction in the Pottiaceae [
27
,
28
,
31
]. Although
paralogous ITS haplotypes have been reported in this family [
32
,
33
], no specimens with
different paralogues were found in Aloina. Therefore, the revealed limited ITS variation
was interpreted as being among homologous haplotypes.
The primer pairs used for each locus were atpB-1/rbcL-1[
34
], trnG-F/trnG-R [
35
],
trnC/trnF [
36
], ITS5-bryo/ITS4-bryo [
37
], ITS1-F/ITS1-R [
38
], and seqITS2 [
39
]. Ampli-
fication reactions were performed using an Eppendorf Mastercycler in a 25
µ
L volume
containing 10
µ
L Supreme NZYTaq II 2x Green Master Mix (Nzytech, Lisbon, Portugal),
2
µ
L of each primer (10
µ
m), and 1
µ
L of the DNA extract. Thermocycling conditions for
atpB–rbcL, trnG, and trnL–F were 95
◦
C for 4 min linked to 35 cycles at 94
◦
C for 30 s, 52
◦
C
for 30 s, and 72
◦
C for 7 min, with a final extension of 72
◦
C for 7 min. The amplification
cycle for nrITS was 95
◦
C for 4 min linked to 35 cycles at 94
◦
C for 1 min, 55
◦
C for 1 min,
and 72
◦
C for 1 min and 30 s, with a final extension of 72
◦
C for 7 min. PCR products were
visualized on a 1% agarose gel. Successful amplifications were purified using the GenElute
PCR Clean-Up kit (Sigma-Aldrich, St. Louis, MO, USA), and sequenced at Macrogen
Spain (Madrid, Spain). Nucleotide sequence contigs were edited and assembled for each
Plants 2024,13, 445 5 of 29
DNA region using Geneious 9.1.8 [
40
]. Consensus sequences were aligned using default
parameters of MUSCLE [
41
] implemented in Geneious with subsequent manual adjust-
ments. Regions of partially incomplete data at the beginning and end of the sequences were
identified and excluded from subsequent analyses. Insertions and deletions (indels) were
coded using SeqState v.1.4.1 [
42
] using a simple coding model as suggested by Simmons
and Ochoterena [
43
]. The indels provided additional phylogenetic evidence and we present
the analyses with these included. Each gene partition was tested for the best-fit substitution
model using jModelTest v.2.1.7 [
44
] under the Akaike information criterion (AIC). The
selected models were TPM3uf + I [
45
] for trnG, HKY + I [
46
] for trnL–F, TIM3ef + I + G [
47
]
for nrITS, and TPM1uf + I + G [45] for atpB-rbcL.
2.3. Phylogenetic Analysis
Phylogenetic relationships were estimated using both maximum likelihood (ML) and
Bayesian inference (BI). Analyses were performed separately on each dataset and the
plastids were combined afterwards. To check for incongruence among the plastid datasets
and between plastid versus ITS partitions, phylogeny reconstructions under ML and BI
were visually compared. Because some incongruence between the nuclear and plastid
markers for the studied taxa was found, we analyzed ITS and plastid markers separately.
Maximum likelihood analyses were performed with RAxML [
48
] through the graphical
front-end raxmlGUI v.2.0 [
49
]. A rapid bootstrap option with 1000 replicates and search for
the best-scoring ML tree were conducted under the GTRCAT model for the concatenated
and individual datasets. Nodes with bootstrap (BS) values of 70–89% were treated as
moderately, and of 90–100% as well supported.
Bayesian inference was performed using MrBayes v.3.2.6 [
50
], running a partitioned
analysis and specifying a substitution model for each block. The data were analyzed using
Markov chain Monte Carlo (MCMC), running two parallel analyses with four chains each
for 30 million generations, sampling trees and parameters every 1000 generations. Chain
convergence and stationarity were checked in Tracer v.1.7 [
51
], making sure the average
standard deviation of split frequencies remained below 0.01. Twenty-five percent of the trees
were discarded as burn-in, and a 50% majority-rule consensus tree was constructed. The
resulting trees for both ML and BI analyses were visualized and partially edited in FigTree
v.1.4.4 [52]. Posterior probability (PP) of 0.95–1.00 was considered to be strong support.
Since the topologies inferred from the nuclear and plastid markers may imply reticu-
late evolution or incomplete lineage sorting, we used the NeighborNet (NN) method as
implemented in SplitsTree v.4.16.2 [
53
] to visualize affinities between lineages within the
Aloina catillum complex and the outgroups included.
Assemble Species by Automatic Partitioning (ASAP) [
54
] was applied to delimit
molecular entities that may be considered a hypothesis for species within the Aloina catillum
complex. This method proposes species partitions using a hierarchical clustering algorithm
based on pairwise genetic distances. The analyses were performed on the ASAP web
server (https://bioinfo.mnhn.fr/abi/public/asap/; accessed on 12 June 2023), running
the concatenated chloroplast and ITS datasets separately, with default parameters. ASAP
delimitation was defined evaluating both the partitions with best and the second-best
asap-score [54].
2.4. Morphological-Anatomical Analysis
After the molecular relationships were established within Aloina catillum s.l., the mor-
phology of the specimens belonging to the distinguished molecular entities of this complex
was examined. In addition, the specimens of A. brevirostris,A. obliquifolia, and A. rigida, all
of them with a distinct leaf border, were also included in the analysis. Microscopic exami-
nations and measurements were taken with an Olympus-BX41 light microscope (Olympus,
Tokyo, Japan), while microphotographs were obtained with a Jenoptik ProgRes C7 camera
(Jenoptik, Jena, Germany) mounted on this microscope. Specimens were examined in 2%
potassium hydroxide.
Plants 2024,13, 445 6 of 29
We studied morphological-anatomical quantitative and qualitative characters, includ-
ing several ratios, which could be used for the identification of the species. Preference was
given to those characters which Delgadillo [
13
] and Gallego et al. [
15
] considered important
for taxa distinction, as well as those representative of the morphological variability among
the studied specimens. To avoid developmental deviations, descriptions and measures
were made from upper vegetative leaves (Figure 2). Cross-sections of the leaves were made
at mid-leaf. The number of cell columns at the leaf border was scored in the upper part of
the sheathing base. Leaf width was measured at the base. In addition, the width and length
of the apical cells of ventral surface costal filaments were measured in cross-section. Three
shoots were selected at random from each collection, and measurements were performed
on three randomly selected leaves. Measurements of sporophytic traits were performed
on 1–3 capsules depending on their availability for each specimen. To represent the vari-
ability of each character, boxplots containing medians and percentiles were performed.
The analyses were implemented using the STATISTICA v. 10 package (StatSoft Inc., Tulsa,
OK, USA).
Figure 2. A schematic vegetative leaf of Aloina, indicating the parts measured or assessed in the
morphological study: (A) Cross section at mid-leaf; (B) Number of columns at the leaf border; (C) Leaf
width; (D) Costa width; (E) Sheathing base with two shoulders highlighted as 1 and 2; (F) Limb;
(G) Leaf length measure; (H) Mucro/awn length; (I) Leaf margins showing 3, margins bent towards
the costa in both sides, and 4, margins parallel to the costa in both sides.
3. Results
3.1. Molecular Analyses
In total, the analysis included 231 sequences, of which 212 (91.8%) were newly gen-
erated for this study; 19 additional sequences, that were sequenced previously in other
phylogenetic studies [
26
–
28
,
55
], were downloaded from GenBank. Summary characteristics
of each dataset are presented in Table 1.
Plants 2024,13, 445 7 of 29
Table 1. Statistics of the nuclear and chloroplast datasets analyzed in this study.
Locus Number of
Specimens
Newly Generated
Sequences Sequence Length
Parsimony
Informative
Characters
ITS 70 65 1012 204 (20%)
atpB-rbcL 52 48 530 37 (7%)
trnG 55 50 34 34 (5.4%)
trnL-F 54 49 435 29 (6.7%)
The ML and BI analyses of each individual marker resulted in largely congruent trees.
Therefore, only the Bayesian topologies are shown here (Figures 3and 4), with ML and
BI support values added where applicable. The plastid and nuclear datasets were not
combined due to some topology incongruences revealed when the plastid and nuclear
consensus tree were compared.
The topologies of trees inferred from the combined plastid, and nuclear data support
that the genus Aloina is monophyletic. On the other hand, plastid and ITS data, as well as
NeighborNet split networks (Figure 5), support the recognition of three lineages within
Aloina catillum s.l. These are labeled clade A, which includes accessions identified previously
as A. catillum or A. rigida; clade B, which includes two distinctive samples of Aloina; and
clade C, which includes the majority of the accessions previously determined as A. catillum
and the specimens identified as Aloina obliquifolia from South America (Figures 3–5).
In the phylogenetic tree derived from ITS sequences (Figure 3), the clade A
(PP = 1,
BS = 100)
is moderately supported by ML (BS = 86) as sister to a large clade strongly
supported (PP = 1; BS = 100), which includes A. aloides,A. ambigua,A. rigida, clade B,
A. brevirostris, and part of the specimens of A. obliquifolia.Aloina ambigua, and A. aloides
(PP = 1, BS = 98)
are resolved sister to a well-supported clade of A. rigida (PP = 1, BS = 100).
In turn, this latter clade is sister to a clade well supported (PP = 1; BS = 98), including
clade B
(PP = 1,
BS = 100) and a clade with the accessions of A. obliquifolia from Northern
Hemisphere nested with the accessions of A. brevirostris (PP = 1, BS = 100).
Clade C comprises most of the specimens identified as A. catillum and the South
American specimens identified as A. obliquifolia (PP = 1, BS = 100). This clade is moderately
supported as sister to A. bifrons by ML (BS = 83). In clade C, two well-supported subclades
can be distinguished: C1 (PP = 0.99, BS = 79), and C2 (PP = 1, BS = 100).
The phylogenetic tree derived from combined plastid sequences (Figure 4) shows a
moderately supported clade (BS = 73) with the accessions included in clade A sister to clade
B, both with maximal support. The remaining accessions of Aloina were included in a well-
supported clade (PP = 1, BS = 98). Within this latter clade, two well-supported subclades
can be identified: (1) A clade formed by all accessions of A. brevirostris plus A. obliquifolia
from the Northern Hemisphere, sister to the accessions of A. aloides,A. ambigua,A. bifrons,
and A. rigida (PP = 1, BS = 92); (2) The clade C with the accessions of A. catillum and the
South American specimens identified as A. obliquifolia (PP = 1, BS = 100). In this latter clade,
the subclade C1 (PP = 1, BS = 95), also revealed in ITS, is supported with two subclades:
C1a (BS = 82) and C1b (PP = 1, BS = 100), including two accessions (1218 and 1302) which
were not placed in clade C1 in the ITS topology. The remaining accessions included were
accommodated in two subclades, named C3 (PP = 1, BS = 97) and C4
(BS = 70).
The lineage
C2 (Figure 3), fully supported in the ITS tree, is resolved in the plastid tree in a ‘polytomy’
within the subclades C1b, C3, and C4.
The NN split networks (Figure 5) retrieved the same three clusters in the A. catillum
complex as the trees based on the ITS and plastid markers. The clade C represents a highly
divergent and complex structured dataset which splits into two supported subclades (C1
and C2) by ITS (Figure 5A) and three subclades (C1, C3, and C4) by plastid markers
(Figure 5B).
Plants 2024,13, 445 8 of 29
Figure 3. Bayesian tree of the Aloina catillum complex inferred from the ITS dataset. Bayesian posterior
probabilities (PP), followed by maximum likelihood bootstrap values (BS), are shown above the
branches. Support values of BS < 70 and PP < 0.95 are not shown. The clades that include species
considered in A. catillum s.l. in this study are labeled A to C and are referred to in the text. Letter and
numbers C1–C2 indicate nodes discussed in the text. Colored bars on the right show the putative
species inferred by ASAP.
Plants 2024,13, 445 9 of 29
Figure 4. Bayesian tree of the Aloina catillum complex inferred from the combined three-plastid
dataset. Bayesian posterior probabilities (PP), followed by maximum likelihood bootstrap values (BS),
are shown above the branches. Support values of BS < 70 and PP < 0.95 are not shown. The clades
that include species considered in A. catillum s.l. in this study are labeled A to C and are referred to in
the text. Letters and numbers C1, C1a, C1b, C3, and C4 indicate nodes discussed in the text. Yellow
triangles in the clades C1b, C3, and C4 indicate the specimens of A. catillum which were grouped in
clade C2 in the ITS analysis. Colored bars on the right show the putative species inferred by ASAP.
Plants 2024,13, 445 10 of 29
Figure 5. Neighbor-Net network built using SplitsTree, originating from the nr ITS (A) and plastid
(B) datasets. Colors indicate revealed lineages of the Aloina catillum complex.
The best ASAP for dividing the ITS dataset returned 10 partitions, which recognizes
the three lineages in A catillum s.l. (Clades A, B, C) as separate putative species (Figure 3),
in addition to the already existing species represented (with the particularity that A. aloides
and A. ambigua are grouped and A. brevirostris includes the A. obliquifolia samples from
the Northern Hemisphere). The second-best score suggested a division into seven species,
with Clade B, A. brevirostris (including A. obliquifolia NH), A. ambigua,A. aloides, and
A. rigida lumped.
Plants 2024,13, 445 11 of 29
The best scoring for the plastid dataset proposed 12 species, which split A. catillum s.l.
into six entities (clades A, B, and C, the latter split into four partitions). The second-best
ASAP included an 11-species scheme, which split A. catillum s.l. into five entities: Clades A,
B, and C, which split into three partitions (C1a, C1b, and C3 + C4) (Figure 4). In both cases,
inconsistencies between described and delimited species were identified because A. bifrons,
A. ambigua,A. rigida, and A. aloides sequences were merged.
3.2. Morphological Evaluation
Our morphological observations showed that the Aloina catillum complex can be
distinguished from A. rigida and A. brevirostris by the seta twist direction: twisted to the
left throughout in A. catillum s.l., and twisted to the left below and to the right above
in A. rigida and A. brevirostris. Without sporophytes, A. rigida could be separated from
these species that also have bordered leaves mainly by its higher number of dorsal stereid
rows in the costa (Figures 6A and 7B) and lower number of cell columns at the leaf border
(Figures 6B and 7C).
On the other hand, Aloina rigida has longer leaves (Figures 6C and 7A)
than A. catillum s.l.
We have not been able to study all previous records of A. rigida from the Neotropics,
but the studied specimens were identified as belonging either to the A. catillum clade A or
to A. brevirostris, representing the first reports of the latter from this geographic area.
Aloina brevirostris could be separated from A. catillum clade C and A. rigida by its usually
monoicous condition (synoicous or rhizoautoicous vs. dioicous) and its larger spores
(Figure 8A). However, it could be more difficult to distinguish from A. catillum s.l. without
sporophytes. In general, A. brevirostris usually has longer leaves
(Figures 6E and 7A),
a
higher ratio of basal laminal cell length/width (Figure 7D), and in some cases the central
strand of the stem is slightly differentiated (Figure 6G), which is not the case in A. catillum
complex (Figure 9E). Other characters suggested to be diagnostic in A. brevirostris, like
the short conic or conic-rostrate operculum, have been found to be variable as already
indicated by other authors [13,56].
In A. catillum s.l., sporophytic characters such as spore diameter (Figure 8A), ratio
urn length/width (Figure 8B), and height of the basal membrane (Figure 8C), have been
revealed as distinctive in the separation of the species. However, these data should be con-
sidered with caution, since the number of sporophytes measured in the specimens of clade
C was small. Among all studied features, the most significant gametophytic characters
in the A. catillum complex were leaf length (Figures 7A and 9A–C), ratio upper laminal
cells length/width (Figure 7E), costa width (Figure 7F), number of cell columns forming
the leaf border (Figure 7C), and number of differentiated perichaetial leaves (Figure 8D).
The qualitative variables that contributed most to the separation of species were the twist
direction of the upper seta, differentiation of perichaetial leaves
(Figures 9N and 10F,J),
shape of the urn (Figures 9J and 10C,H), costa ending (subpercurrent, percurrent, or excur-
rent) (Figures 9F–I and 10I), and ornamentation and curvature of the upper leaf margins
(Figures 9G,H and 10I).
It should also be noted that several of these morphotypes of A.
catillum s.l can grow together on the same turf and with other Aloina species such as A. bre-
virostris, which further complicates their discrimination when specimens lack sporophytes.
Several of the morphological characters examined were congruent with the clades
obtained in the molecular study and provide synapomorphies or combinations of character
states that support the recognition of the three lineages in A. catillum s.l. (named A, B, and
C in Figures 3–5).
The study of the type material of available published names allowed the identification
of the correct names for the specimens included in clade A. This study showed that the types
of A. catillum and A. calceolifolia belong to the same species and, applying the principle
of priority, the second name should be used. Aloina calceolifolia was described under
Tortula [
57
] from a specimen collected by Spruce in the Ecuadorian Andes and was later
transferred to Barbula Hedw. [
58
]. Finally, Brotherus [
59
] considered it as Aloina. Later,
Delgadillo [13] included it among the synonyms of A. rigida.
Plants 2024,13, 445 12 of 29
Figure 6. Distinctive morphological characters in Aloina brevirostris,A.obliquifolia, and A. rigida:
(A)A. rigida, cross-section of the costa at the mid-leaf (Cano 11198, MUB 62244); (B)A. rigida, leaf
border at the base (Cano 10198, MUB 62244); (C)A. rigida, vegetative leaf (Aedo 16324f, MUB 29388);
(D)A. obliquifolia, vegetative leaf (Sato s.n., MUB 52915); (E)A. brevirostris, vegetative leaf (Cano
et al. 4102, MUB 29586); (F)A. brevirostris, cross-section of the costa at the mid-leaf (Cano et al. 3403
(MUB 27728); (G)A. brevirostris, cross-section of a stem (Cano 7578, MUB 49074); (H)A. brevirostris,
leaf border at the base (Cano 7578, MUB 49074). Photos: María J. Cano.
Plants 2024,13, 445 13 of 29
Figure 7. Boxplots showing the variation ranges of the most discriminate morphological gametophyte
characters among the species studied: (A) Leaf length (mm); (B) Number of rows of dorsal stereids at
the costa; (C) Number the columns at the leaf border; (D) Ratio of basal laminal cells length/width;
(E) Ratio of upper laminal cell length/width; (F) Costa width (
µ
m). The boxes represent the data from
the 25th to 75th percentiles, with the median indicated within the box, and the lines above and below
boxes are the total range. The small circles outside the bar represent “outlier” and the black stars are
extremes. NH: Northern Hemisphere; SH: Southern Hemisphere. Note that
“catillum + obliq
uifolia
SH” is A. limbata.
Plants 2024,13, 445 14 of 29
Figure 8. Boxplot showing variation ranges of the most discriminate morphological sporophyte char-
acters among the species studied: (A) Spore diameter (
µ
m); (B) Ratio urn length/width; (C) Height
of the basal membrane of the peristome (
µ
m); (D) Number of differentiated perichaetial leaves. The
boxes represent the data from the 25th to 75th percentiles, with the median indicated within the box,
and the lines above and below boxes are the total range. The small circles outside the bar represent
“outlier” and the black stars are extremes. NH: Northern Hemisphere; SH: Southern Hemisphere.
Note that “catillum + obliquifolia SH” is A. limbata.
The type material of A. calceolifolia studied from NY and E exhibits all the morphologi-
cal characters of specimens grouped in clade A, although both plants and leaves (1.8 mm
length) were slightly longer and the costa was more robust, with three layers of dorsal
stereids. However, these characters are considered to be within the variation in the species.
Aloina calceolifolia can be distinguished from the other species of Aloina with a leaf marginal
border by its lingulate to suborbicular leaves, the leaf border developed only in the sheath-
ing base (Figure 10A,B,E), margins usually strongly sinuose-papillose above (Figure 10B),
and the leaf apex rounded to widely obtuse. Further, it has 1(2) inner perichaetial leaves
usually differentiated, seta strongly twisted to the left throughout, a long cylindrical capsule
(Figure 10C) [ratio urn length/width (3.3)3.9–6.6(7)] (Figure 8B)], spores 9.6–14.5
µ
m in
diameter, and the usually monoicous condition.
Distinctive characters for specimens of lineage C ascribed to A.catillum are the ovoid-
cylindrical urn (Figure 9J) [ratio urn length/width 2–2.8(4.2) (Figure 8B)], costa percurrent
or excurrent as a mucro or awn, leaf margins strongly differentiated throughout the limb in
4–12 columns of cells (Figure 9D), and undifferentiated perichaetial leaves or occasionally
with the innermost ones differentiated (Figure 9N). Consequently, it is here proposed as a
new species, Aloina limbata.
Plants 2024,13, 445 15 of 29
Figure 9. Distinctive morphological characters in Aloina limbata: (A–C) Vegetative leaves (A) Cano
et al. 7649, MUB 49149; (B) Cano and Gallego 2903a, MUB 22314; (C) Cano and Jiménez 5168, MUB
32545; (D) Leaf border at the base (Cano et al. 7150, MUB 48588); (E) Cross-section of a stem (Cano
and Gallego 2903a, MUB 22314); (F–I) Leaf apices (F) Cano et al. 2814b (MUB 59251); (G) Cano et al.
7075, MUB 48503; (H) Cano et al. 7150, MUB 48588; (I) Cano et al. 5076b (MUB 32548); (J) Urn and
peristome (Cano et al. 5065, MUB 32542); (K–M) Cross-sections of the costa at mid-leaf (K) Cano et al.
4082, MUB 29585; (L) Cano et al. 7075, MUB 48503; (M) Cano et al. 7150, MUB 48588; (N) Perichaetial
leaf (Cano and Gallego 2903a, MUB 22314). Photos: María J. Cano.
Plants 2024,13, 445 16 of 29
Figure 10. Distinctive morphological characters in Aloina calceolifolia and A. bracteata.Aloina calceolifo-
lia: (A) Vegetative leaf (Cano and Alonso 8144, MUB 56217); (B) Leaf border at the middle (Cano 2353,
MUB 20586); (C) Capsule (Cano 2353, MUB 20586); (D) Cross-section of the costa at mid-leaf (Cano
and Alonso 8144, MUB 56217); (E) Leaf border at the base (Cano 2353, MUB 20586); (F) Perichaetial
leaf (Cano et al. 3383, MUB 27722). Aloina bracteata: (G) Vegetative leaf (Cano and Alonso 8409a,
MUB 56491); (H) Urn and peristome (Cano et al. 4113, MUB 29597); (I) Leaf apex (Cano and Alonso
8409a, MUB 56491); (J) Perichaetial leaf (Cano and Alonso 8409a, MUB 56491); (K) Cross-section of
the costa at mid-leaf (Cano and Alonso 8409a, MUB 56491); (L) Leaf border at the base (Cano and
Alonso 8409a, MUB 56491). Photos: María J. Cano.
Plants 2024,13, 445 17 of 29
The subclades C1–C4, defined by the molecular data, can be weakly recognized mor-
phologically, with C1 being the best characterized. These C1 specimens have small, lingulate
leaves (Figure 9C), with a narrow costa (100–150
µ
m wide at base) and
0–1(2)
dorsal stereid
layers. Although in the plastid tree these specimens are divided into two subclades C1a and
C1b, no morphological differentiation has been found between them. The rest of the speci-
mens of clade C included specimens with a tendency to have ovate to oblong-ovate leaves.
The subclade C2, well supported in the ITS topology and split in the plastid tree, and clade
C3 could not be morphologically delimited. The subclade C4 in the plastid tree includes the
specimens identified as A. obliquifolia from the Southern Hemisphere
(Figure 9A)
. In gen-
eral, these specimens have the leaf border with many developed columns, the costa more
strongly differentiated, with 2–3 dorsal stereid rows (Figure 9K), and usually excurrent as a
mucro or stout awn (Figure 9F,H). The character of the development of an excurrent costa
as a mucro or awn, which mainly defined A. obliquifolia [
15
,
19
], has been found here to be
variable. Thus, it has been possible to find populations, even in the same plant, with a leaf
costa percurrent, excurrent on a short mucro or into a stout awn. These South American A.
obliquifolia can be easily differentiated from specimens of A. obliquifolia from the Northern
Hemisphere (Figure 6D) because the seta is twisted to the left throughout and usually has
developed papillae on the mucro or apex, a feature most readily seen in young leaves
(Figure 9H).
Specimens of clade B are unambiguously distinguished from A. calceolifolia and A.
limbata by 2–4 strongly differentiated perichaetial leaves (Figure 10J), ratio of upper laminal
cell length/width 0.6–1 (Figure 7E) and the ovoid to elliptical capsule (Figure 10H) [ratio
urn length/width 1.9–2.1 (Figure 8B)]. Other distinctive character states are the lingulate
leaves, with strongly inflexed margins, subpercurrent costa, border differentiated from the
sheathing base and part of the limb (Figure 10L), seta twisted to the left throughout, spores
13–16
µ
m in diameter, and operculum 0.6–0.8 mm long. These plants are here described as
a new species, Aloina bracteata.
In the search for a name among those published that would fit these three lineages,
we located the type material of Barbula recurvipatula Müll. Hal. (Aloina recurvipatula) at
S (B4615). The specimen exhibits leaves with marginal border differentiated, only one
shoot with undifferentiated perichaetial leaves, and a broken seta. Because it is difficult to
identify it with confidence, we prefer to treat this taxon as one with doubtful identity.
4. Discussion
This study represents the first integrative approach to a group of Aloina species charac-
terized by leaves bordered by hyaline cells. Based on our morphological and molecular
analyses, we congruently identify the existence of hidden diversity in A. catillum and
reveal the confusion that has existed among taxa identified as A. brevirostris,A. catillum,
A. obliquifolia, and A. rigida in South America.
The existence of three well supported lineages in the phylogenies, which can be char-
acterized morphologically within Aloina catillum s.l., indicates that this complex comprises
three entities, recognized in this study at the species rank as Aloina bracteata sp. nov., A. cal-
ceolifolia, and A. limbata sp. nov. The ASAP delimitations inferred largely coincided with
these clades based on the molecular phylogenetic results and morphological considerations.
According to our results, A. calceolifolia could be one of the basal groups of Aloina. It
is very similar to some morphotypes of A. limbata because both species have similar leaf
shapes, a rounded leaf apex, and the seta is sinistrorse throughout. In fact, specimens
of A. limbata (Cano 223, 3083c, 3077b, 2125b, Cano and Gallego 3015, 3003b, Cano et al.
2814a, 2903a) and A. calceolifolia (Cano 2353, Cano 2176) were identified as A. catillum by
Cano et al. [
19
]. Without sporophytes, these species are mainly distinguished by mem-
branaceus leaf margins differentiated only in the sheathing base in A. calceolifolia. Since the
neglected name Aloina calceolifolia is here resurrected, its distribution extends to dry areas
of Argentina, Bolivia, and Peru.
Plants 2024,13, 445 18 of 29
Aloina bracteata, described here as a new species, forms a sister clade to A. brevirostris
in the ITS tree and sister to A. calceolifolia in plastid topology. Aloina brevirostris as well as
A. bracteata has a developed leaf border in the sheathing base and limb, lingulate leaves,
and strongly inflexed margins. However, A. brevirostris usually has cylindrical capsules,
and the seta turns to the right above. In addition, the perichaetial leaves are not strongly
differentiated or have only the innermost ones slightly differentiated. Aloina bracteata is
similar to A. limbata, but the former can be distinguished by smaller operculum and larger
spores, longer upper and middle laminal cells, narrow leaf border (2–3 rows of cells), and
2–4 strongly differentiated perichaetial leaves. At the moment, it appears to be an endemic
species to the Andes of northwestern Argentina (Salta and Tucumán).
Aloina limbata is a variable species in terms of leaf shape, width, and excurrence of
the costa. In general, it is characterized by the strongly developed leaf margin in both
the sheathing base and limb, seta twisted to the left throughout, and an ovoid-cylindrical
capsule. Many of the A. limbata specimens studied lack sporophytes and, consequently, all
sporophytic characters have not been properly analyzed. Molecular analysis and ASAP
recognized some well-supported subclades. However, our morphological and molecular
data suggest that boundaries are not yet fully formed. The only exception is the subclade
C1, where molecular and morphological studies indicate that sympatric cryptic genetic
variation may exist. It includes morphological specimens which are divided into two
molecularly well-supported entities (C1a and C1b) in the plastid tree and in ASAP, but
which do not differ phenotypically. The subclade C1 could possibly be recognized as
a distinct variety or even species. However, thorough examination of a wider range
of specimens, including samples with sporophytes, together with additional molecular
sampling, will be necessary to confirm this. At present, we recognize the subclades as a
potential example of ongoing speciation [60].
In the case of specimens identified as A. obliquifolia from the Southern Hemisphere,
these can be easily distinguished morphologically from other specimens of this complex,
when the mucro or awn is fully developed. However, this character has been found to be
variable and our molecular analysis does not support this taxon as a Southern Hemisphere
one. Aloina obliquifolia was reported from South America extending its distribution to the
Southern Hemisphere, on the basis of two specimens collected in Ecuador and a Peruvian
sample, all without sporophytes [
19
]. The American specimens shared with Northern
Hemisphere specimens the leaf costa excurrent as a long mucro and the leaves bordered by
thin-walled and hyaline cells. The study of new collections has revealed that the specimens
identified as A. obliquifolia from the Southern Hemisphere have smaller spores, a smaller
ratio between basal cells length/width, shorter upper laminal cells, narrower superficial
ventral cells, more rows of marginal cells, and shorter leaves. Cano et al. [
19
] already
pointed out some of these features, such as the smaller size of the leaves and more ovate
leaves than in material from the Northern Hemisphere. The variability observed in the
differentiation of the mucro in specimens identified as A. obliquifolia in South America
has led us to consider these specimens within A. limbata. Further studies will determine
whether these specimens merit any taxonomic rank. The results obtained with the two
specimens of A. obliquifolia from the Northern Hemisphere included in the A. brevirostris
clade in the molecular study, together with the high morphological variability observed in
both sporophytic and gametophytic characters within A. brevirostris, also open the way for
further studies.
Other results obtained in this work, such as the phylogenetic incongruence of nuclear
and chloroplast DNA sequences in the positions of the accessions attributed to A. rigida-
A. brevirostris-A. ambigua-A. aloides and, surprisingly, A. bifrons, may suggest incomplete
lineage sorting, past hybridization, and historic introgression among populations in this
group of species. This has already been indicated in other genera of the Pottiaceae [
33
,
61
,
62
]
and calls for further future studies.
The tropical Andes are known as biodiversity hotspots and may be particularly con-
ducive to cryptic speciation. The South American specimens we have studied, previously
Plants 2024,13, 445 19 of 29
identified as A. rigida, are accepted here as A. brevirostris and A. calceolifolia. Consequently,
A. rigida is tentatively excluded from this area and the distribution of A. brevirostris is
enlarged to the Neotropics. After this study, seven species of Aloina have been confirmed
in the area, five of them endemic. These results emphasize the need for integrative studies
of numerous Pottiaceae species complexes in this area. Future fieldwork will be essential
to complete the phylogeny of Aloina and to clarify the relationship in several cryptic or
semi-cryptic complexes of this genus.
5. Taxonomy
Based on the morphological and phylogenetic analyses performed in the present study,
we propose the following taxonomic treatment for the Aloina catillum complex. Complete
descriptions, distribution data, and nomenclatural notes are also provided.
Aloinacalceolifolia (Spruce ex Mitt.) Broth., Nat. Pflanzenfam. in Engl. &
Prantl I(3): 428.
1902. Tortula calceolifolia Spruce ex Mitt., J. Linn. Soc., Bot. 12: 157. 1869. Barbula calceolifolia
(Spruce ex Mitt.) A. Jaeger, Ber. Thätigk. St. Gallischen Naturwiss.
Ges. 1871–72: 409.
1873.
(Figures 1F, 10A–F and 11).
Figure 11. Aloina calceolifolia (Spruce) Broth. (A) Habit. (B–D) Vegetative leaves. (E) Leaf apex
ventrally. (F) Cross-section of a leaf in middle part. (G) Detail of the ventral chlorophyllous filaments
Plants 2024,13, 445 20 of 29
of the leaf. (H) Basal laminal cells. (I) Marginal laminal cells at base. (J) Upper laminal cells. (K) Cross-
section of a stem. (L) Axillary hair. (M) Perichaetial leaf. (N) Calyptra. (O) Peristome. (P) Exothecial
cells. Scale bars: A = 1 mm; B–D = 0.4 mm; E = 0.2 mm; F = 90
µ
m; G, H, I = 47
µ
m; J = 52
µ
m;
K = 30 µm;
L = 78
µ
m; M = 0.35 mm; N = 0.4 mm; O = 160
µ
m; P = 42
µ
m. (A,O,P) from Cano 2353
(MUB 20586); (B,M) from Cano et al. 3383 (MUB 27722); (C–E,G–J,L) from Cano and Alonso 8144
(MUB 56217); (F,K) from Cano et al. 4061c (MUB 61776); (N) from Cano et al. 7060 (MUB 48489).
Illustration: Inmaculada Guijarro.
Type citation: “Andes Quitenses, Carguairazo (11,000 ped.), ad terram nigram ag-
grerum, Spruce,n. 155”. Type: Ecuador, Tungurahua, Andes Quitenses, Carguairazo [Cari-
huairazo], Spruce (Musci Amazonici et Andini nº 155) (lectotype, designated by Delgadillo [
13
],
corrected here: NY!; isolectotypes: BM000720000–image!, E00165175!, G00265578–image!,
G00113834–image!, PC0657869!).
Aloina catillum (Müll. Hal.) Broth., Nat. Pflanzenfam. in Engl. & Prantl I(3): 428.
1902.
≡
Barbula catillum Müll. Hal., Linnaea 42: 329. 1879. Type citation: “Argentinia
subtropica, in montibus altioribus inter Siambon et Tafi. 1872”. Type: Argentina, Tucumán,
Siambon-Tafi, 1872, P.G. Lorentz s.n. (lectotype, here designated: S B B3591! (P. Dusén’s
moss herbarium); isolectotypes: H–BR 80004!, NY 01128073 (web)!, S B3592!, S B3593!,
syn. nov.
Description: Plants small, forming open turfs or gregarious, olive green to reddish
or reddish brown. Stems 0.3–2(–2.3) cm high, simple or branched; in cross-section cir-
cular, hyalodermis, sclerodermis, and central strand absent; axillary hairs of 4–10 cells,
all hyaline. Rhizoids densely covering stem, whitish to pale brown, smooth. Leaves in-
curved when dry, erect-patent to spreading when wet, broadly lingulate to suborbicular,
(1.1–)1.4–1.8 ×0.6–1.1(–1–3) mm
[ratio length/width (1.1–)1.5–2.1(–2.9)], not constricted,
concave; lamina unistratose, not fragile, reddish in KOH; apex rounded to widely ob-
tuse, not apiculate, cucullate; margins broadly incurved, often partially covering the
leaf blade, parallel to the costa in both sides above, strongly sinuose-papillose in up-
per mid-leaf, bordered by (2)3–4 columns of hyaline, thin-walled, membranaceous cells
differentiated only in the sheathing base, base oblong, with shoulders slightly marked,
not decurrent; costa
(250–)325–388(–450) µm
wide at base, ending 2–5 cells below the
apex, surface cells dorsally usually long-rectangular from apex to base, smooth, surface
cells ventrally forming chlorophyllous, uniseriate, usually branched filaments, which ex-
tend to the lamina; in cross-section at mid-leaf broadly flattened, 35–53 guide cells in
(1–)2–3 layers, ventral stereids absent,
(1–)2(–3) rows
of dorsal stereids, hydroids absent,
surface cells dorsally slightly differentiated, ventral surface cells filaments disposed in
4–5(–6) layers of barrel-shaped, thin-walled cells, apical cell conical, rarely subglobose,
(11–)21–28 ×11–13(–17.6) µm,
usually smooth; upper laminal cells oblate-rectangular to
oblate-fusiform,
(5–)6.5–9.5(–13) ×(16–)19–27(–40) µm
[ratio length/width 0.2–0.5(–0.7)],
thick-walled, not collenchymatous, smooth; middle laminal cells oblate-rectangular to
oblate-fusiform, 5–9.5(–15)
×
(14–)16–29(–32)
µ
m [ratio length/width (0.2–)0.3–0.8(–0.99],
thick-walled, not collenchymatous, smooth; basal paracostal cells rectangular,
(24–)32–61(–75)
×
16–24
µ
m [ratio basal paracostal cells
1.4–3.8(–4.7)],
thin-walled, collenchy-
matous, smooth; basal marginal cells rectangular,
(24–)32–61(–75) ×(6.5–)9.6–11(–13) µm
[ratio length/width 1.5–5(–7.2)], thin-walled, smooth, hyaline. Structures of asexual re-
production absent. Monoicous (cladautoicous or synoicous), dioicous (?, when perique-
tia not seen). Perigonial leaves ovate to suborbicular, concave, 0.6–0.8
×
0.5–0.6 mm;
perichaetial leaves usually differentiated from vegetative, mainly the inner, in number of
1(2), oblong-lingulate to spatulate, with plane margins, the most internal usually without
dorsal costal filaments, 1.1–1.3(–1.6)
×
0.4–0.7 mm [ratio length/width 1.9–2.7(–3.1)]. Setae
1 per perichaetium, 8–13.2 mm long, twisted to the left throughout, orange to brownish.
Urns cylindrical, 2.1–3.8(–4.2)
×
0.5–0.7 mm [ratio urn length/width (3.3–)3.9–6.6(–7)],
reddish brown to yellowish brown; exothecial cells rectangular, rarely oblong-hexagonal,
(31–)40–58
×
8–13
µ
m [ratio length/width 3.3–7], thin-walled; stomata phaneropores; an-
Plants 2024,13, 445 21 of 29
nulus in 3–4 rows of vesiculose cells; peristome 16 split nearly to the base into two filiform
teeth, 600–900 mm long, papillose-spiculose, straight or slightly twisted, orange, basal
membrane 60–83
µ
m long; operculum conic, 0.8–1 mm long. Calyptrae cucullate, 2.7–3 mm
long, yellowish brown, smooth. Spores spherical, (9.6–)11–13(–14.5)
µ
m in diameter, pale
brown, finely granulose.
Habitat: Banks, crevices of rocks and soils in puna, grassland, and interandean valleys;
2730–4215 m altitude.
Distribution: Argentina, Bolivia, Ecuador, and Peru. Endemic to tropical Andes.
Nomenclatural note: Delgadillo [
13
] considered material from NY as the holotype of
Tortula calceolifolia. As this publication predates 1 January 2001, it should be considered an
inadvertent lectotypification (Art. 7.11 and Art. 9.23, [63]) and is here corrected.
Aloina bracteata M.J. Cano, J.A. Jiménez & M.T. Gallego, sp. nov. (Figures 1D, 10G–L
and 12).
Figure 12. Aloina bracteata M.J. Cano et al. (A) Habit. (B–D) Vegetative leaves. (E) Perichaetial leaf.
(F) Leaf apex ventrally. (G) Leaf apex dorsally. (H) Middle laminal cells. (I) Basal laminal cells. (J) Cross-
Plants 2024,13, 445 22 of 29
section of a leaf in middle part. (K) Detail of the ventral chlorophyllous filaments of the leaf.
(L) Peristome. (M) Calyptra. (N) Axillary hair. (O) Detail of exothecial cells. (P) Cross-section of a
stem. Scale bars: A = 0.8 mm; B–D = 0.3 mm; E = 0.2 mm; F, G = 60
µ
m; H, I = 38
µ
m; J = 90
µ
m;
K = 25 µm;
L = 30
µ
m; M = 0.5 mm; N = 55
µ
m; O = 43
µ
m; P = 100
µ
m. (A–P) from Cano and Alonso
8409a (MUB 56491). Illustration: Inmaculada Guijarro.
Diagnosis: Differing from all other species of the genus Aloina by the following
unique combination of character states: lingulate leaves, with strongly inflexed margins,
border differentiated in the sheathing base and part of the limb, costa with two layers of
dorsal stereids, ending below the apex, ovoid capsule, seta twisted to the left throughout,
spores of 13–16
µ
m in diameter, operculum 0.6–0.8 mm long, and 2–4 perichaetial leaves
strongly differentiated.
Type: Argentina, Salta, Iruya, 22
◦
47
′
27
′′
N 65
◦
13
′
26
′′
W, 2800 m, 27–III–2014, M.J.
Cano and M. Alonso 8409a (holotype: MUB 56491!; isotype: CORD C0014154!).
Etymology: The specific epithet refers to the characteristic differentiated perichaetial leaves.
Description: Plants small, forming open turfs or gregarious, olive green to reddish
or reddish brown. Stems 0.9–3 cm high, simple or branched; in cross-section circular,
hyalodermis, sclerodermis, and central strand absent; axillary hairs of 7–10 cells, all hya-
line. Rhizoids densely covering the stem, pale brown, smooth. Leaves erect, slightly
incurved when dry, patent when wet, lingulate to obovate, occasionally oblong-lingulate
or oblong-ovate,
1.2–1.5 ×0.7–1 mm
(ratio length/width 1.6–1.8), not constricted, strongly
concave; lamina unistratose, not fragile, reddish in KOH; apex rounded to obtuse, not
apiculate, strongly cucullate; margins broadly inflexed, often partially covering the leaf
blade, usually bent towards the costa in both sides in upper part, sinuose to dentate, bor-
dered by
2–3 columns
of hyaline, thin-walled, membranaceous cells from base to upper
third, base oblong, with shoulders slightly marked, not decurrent; costa 250–413
µ
m wide
at base, ending
2–11 cells
below the apex, surface cells dorsally long-rectangular from
apex to base, smooth, surface cells ventrally forming chlorophyllous, uniseriate, usually
branched filaments, which extend to the lamina; in cross-section at mid-leaf broadly flat-
tened,
28–40 guide
cells in
1–2 layers,
ventral stereids absent, 1–2 rows of dorsal stereids,
hydroids absent, surface cells dorsally differentiated, ventral surface cells filaments dis-
posed in (4–)5–6 barrel-shaped, thin-walled cells, apical cell conical, occasionally subglobose
16–23
×
8–13
µ
m, usually unipapillose; upper laminal cells rounded to quadrate, or trans-
versely rectangular,
9.6–11 ×11–14(–19) µm
(ratio length/width 0.6–1), thick-walled, not
collenchymatous, smooth; middle laminal cells oblate-rectangular, occasionally quadrate,
6.5–13 ×9.5–13(–18)
(ratio length/width 0.6–1), thick-walled, not collenchymatous; basal
paracostal cells rectangular, 24–48
×
11–19
µ
m (ratio length/width 1.5–3.4), thin-walled,
collenchymatous, smooth; basal marginal cells rectangular, 18–24
×
5–6.5(–10)
µ
m, (ratio
length/width 2–3.8), thin-walled, smooth, hyaline. Structures of asexual reproduction ab-
sent. Probably dioicous. Perigonial leaves not seen; perichaetial leaves differentiated from
vegetative, in number of 2–4, oblong-lingulate, with plane margins and the most internal
without dorsal costa filaments, 1.5–1.8
×
0.5–0.8 mm (ratio length/width
2.4–3.3)
. Setae
1 per perichaetium,
6.3–7.9 mm
long, twisted to the left throughout, orange. Urns ovoid,
occasionally elliptical,
1.5–2.1 ×0.8–1.1 mm
(ratio urn length/width 1.9–2.1), pale brown;
exothecial cells rectangular to quadrate-hexagonal, 24–67
×
17–30
µ
m (ratio length/width
1.6–2.1), thin-walled; stomata phaneropores; annulus in 3–4 rows of vesiculose cells; peri-
stome of 16 split nearly to the base into two filiform teeth, 430–550 mm long, papillose-
spiculose, straight, orange, basal membrane 20–32
µ
m long; operculum conic, 0.6–0.8 mm
long. Calyptrae cucullate, 2.6–2.9 mm long, yellowish, smooth. Spores spherical, 13–16
µ
m
in diameter, light brown, finely papillose.
Habitat: Banks in dry interandean valleys, growing with A. brevirostris;
2110–2800 m altitude.
Distribution: Argentina (Salta, Tucumán). Endemic to tropical Andes.
Plants 2024,13, 445 23 of 29
Aloina limbata M.J. Cano, J.A. Jiménez & M.T. Gallego, sp. nov.
(Figures 1C,E, 9and 13).
Figure 13. Aloina limbata M.J. Cano et al. (A) Habit. (B–C) Vegetative leaves. (D) Leaf apex ventrally.
(E) Cross-section of a leaf in middle part. (F) Detail of the ventral chlorophyllous filaments of the leaf.
(G) Middle laminal cells. (H) Upper laminal cells. (I) Basal laminal cells. (J) Cross-section of a stem.
(K) Axillary hair. (L) Perichaetial leaf. (M) Calyptra. (N) Detail of exothecial cells. (O) Peristome.
Scale bars: A = 0.9 mm; B–C, L = 0.35 mm; D–E, N = 53
µ
m; F–H = 35
µ
m; I, J = 40
µ
m;
K = 50 µm;
L = 30 µm;
M = 0.5 mm; N = 0.5 mm; O = 100
µ
m. (A–O) from Cano et al. 5065 (MUB 32542).
Illustration: Inmaculada Guijarro.
Diagnosis: Differing from all other species of the genus Aloina by the following unique
combination of character states: leaf border strongly differentiated in the sheathing base and
Plants 2024,13, 445 24 of 29
most of the limb, costa with two layers of dorsal stereids, percurrent on excurrent as a mucro
or awn, seta twisted to the left throughout, ovoid-cylindrical urn, and undifferentiated
perichaetial leaves or occasionally with the most internal ones differentiated.
Type: Peru, Cajamarca, camino de Celendín, pr. Chinchín, 07
◦
09
′
15
′′
S 78
◦
24
′
54
′′
W,
3050 m, 16–VI–2009, M.J. Cano, J. Guerra, and J.A. Jiménez 5065 (holotype: MUB 32542!;
isotypes: USM, S B332546!).
Etymology: The specific epithet refers to the strongly differentiated leaf border.
Description: Plants small, forming open turfs, olive green to reddish or reddish
brown. Stems 0.3–1.9(–3) cm high, simple or branched; in cross-section circular, hyalo-
dermis, sclerodermis, and central strand absent; axillary hairs of 5–8(–11) cells, all hya-
line. Rhizoids covering the stem, pale brown, smooth. Leaves erect-incurved to in-
curved when dry, erect-patent to patent when wet, oblong-ovate to ovate or lingulate,
1–1.6(–2.5) ×0.5–1 mm
[ratio length/width 1.3–2.4)], not constricted, concave; lamina unis-
tratose, not fragile, reddish in KOH; apex rounded to obtuse, sometimes mucronate to
long apiculate, cucullate; margins broadly incurved, often partially covering the leaf blade,
parallel to the costa in both sides above, sinuose-crenulate to slightly crenulate, bordered
by 4–9(–12) columns of hyaline, thin-walled, membranaceous cells from base to upper
third or near apex, base with shoulders slightly or strongly marked, not decurrent; costa
(113–)148–325(–432) µm
wide at base, percurrent or excurrent as a mucro or awn, sur-
face cells dorsally long-rectangular to linear, rectangular at base, smooth, surface cells
ventrally forming chlorophyllous, uniseriate, usually branched filaments, which extend
to the lamina; in cross-section at mid-leaf broadly flattened, 15–36(–40) guide cells in
1–2 layers
, ventral stereids absent, 1–2(–3) rows of dorsal stereids, hydroids absent, surface
cells dorsally slightly differentiated or undifferentiated, ventral surface cells filaments
disposed in
(4–)5–6 layers
of barrel-shaped, thin-walled cells, apical cell conical to globose,
(11–)14–24(–27) ×(8–)9.5–13(–16) µm,
smooth or unipapillose; upper laminal cells trans-
versely rectangular to oblate or quadrate-rounded,
(3–)5–9.5(–14) ×(6–)9.5–21(–27) µm
[ratio length/width 0.2–1.8(–2)], thick-walled, not collenchymatous, smooth; middle
laminal cells transversely rectangular to oblate,
(3–)5–8(–9.5) ×(8–)9.5–21(–29) µm
[ratio
length/width 0.2–0.7(–1.2)], thin-walled, collenchymatous, smooth; basal paracostal cells
rectangular, rarely quadrate, (11–)13–50(–75)
×
(5–)8–14.5(–19)
µ
m [ratio length/width
0.8–3.7(–4.7)],
13–41(–75)
×
(5–)7–18
µ
m, thin-walled, collenchymatous, smooth; basal
marginal cells rectangular, (11–)13–35(–40)
×
(5–)6.5–9.5(–13)
µ
m [ratio length/width
(0.8–)1.2–4(–6.7)],
thin-walled, smooth, hyaline. Structures of asexual reproduction absent.
Dioicous. Perigonial leaves ovate, strongly concave, ca. 1.4
×
1.8 mm; perichaetial leaves
not or only inner scarcely differentiated from vegetatives, in number 0–1, oblong-ovate
or lingulate, with flat margins and costa less developed, 0.9–1.8
×
0.4–0.7(–0.8) mm (ratio
length/width 1.4–2.7). Setae 1 per perichaetium, 7.6–13(–15) mm long, twisted to the left
throughout, reddish brown to pale brown. Urns ovoid-cylindrical, occasionally elliptical
(1.1–)1.7–3.3 ×0.6–0.9 mm
[ratio length/width 2–2.8(–4.2)], reddish brown to pale brown;
exothecial cells rectangular, 23–53(–80)
×
8–24
µ
m [ratio length/width 2–5.6(–8)], thin-
walled; stomata phaneropores; annulus usually in three rows of vesiculose cells; peristome
of 16 split nearly to the base into two filiform teeth, 420–900(–1250) mm long, papillose-
spiculose, usually twisted one turn, yellowish brown, basal membrane 55–125
µ
m long;
operculum conic, 0.8–1 mm long. Calyptrae cucullate, 2.2–2.7 mm long, yellowish, smooth.
Spores spherical, 8–9.5(–11) µm in diameter, light brown, finely papillose.
Habitat: Banks, crevices of rocks and soils in puna, grassland, interandean valley, dry
forests, and Chaco serrano formations; 2730–4215 m altitude.
Distribution: Argentina, Bolivia, Chile, Ecuador, Peru, and Venezuela. Endemic to
South America.
Plants 2024,13, 445 25 of 29
6. Key to the South American Species of Aloina with Differentiated Leaf
Marginal Border
1. Seta twisted to the right in the upper part, occasionally to the left below; monoicous
(synoicous, or rhizoautoicous), rarely dioicous; leaves usually lingulate, more rarely subor-
bicular; spores 15–22
µ
m in diameter; central strand of the stem usually weakly differenti-
ated ........................................................................................................................... A. brevirostris
1. Seta twisted to the left throughout; dioicous or monoicous (cladautoicous, syn-
oicous); leaves lingulate to elliptical, oblong-ovate to suborbicular; spores 8–16
µ
m in
diameter; central strand of the stem absent ................................................................................ 2
2. Leaf border differentiated only in sheathing base; leaf margins strongly sinuose-
papillose distally; urns usually cylindrical; usually monoicous ......................A. calceolifolia
2. Leaf border differentiated from base to the upper third or near apex; leaf mar-
gins lightly crenulated to dentate; urns ovoid, elliptical to ovoid-cylindrical; usually
dioicous............................................................................................................................................. 3
3. Costa ending some cells below apex; 2–4 perichaetial leaves strongly differentiated
from vegetative leaves; urns usually ovoid ............................................................... A. bracteata
3. Costa usually percurrent or excurrent as a mucro or awn; perichaetial leaves not or
only the 0–1(–2) innermost ones scarcely differentiated; urns usually ovoid-
cylindrical ..................................................................................................................... A. limbata
Author Contributions: Conceptualization, M.J.C. and J.A.J.; methodology, M.J.C., J.A.J. and M.M.;
software, M.J.C., J.A.J., L.H., M.M. and O.R.; formal analyses, M.J.C., J.A.J., L.H., M.T.G., M.M. and
O.R.; investigation, M.J.C., J.A.J., M.T.G., L.H. and J.G.; data curation: M.J.C. and J.G.; sampling and
fieldwork, M.J.C., J.G., J.A.J. and M.T.G.; writing—original draft preparation, M.J.C.; writing—review
and editing, M.J.C., J.A.J., M.T.G., L.H., O.R. and J.G.; visualization, M.J.C., J.A.J., L.H., M.T.G. and
J.G.; supervision, M.J.C., M.T.G., J.A.J., L.H. and J.G.; funding acquisition, M.J.C., J.A.J., M.T.G. and
J.G. All authors have contributed substantially to the general discussion of results and taxonomic
treatment. All authors have read and agreed to the published version of the manuscript.
Funding: This research was funded by “Agencia Estatal de Investigación”, Spanish Ministry of Science
and Innovation (MCIN/AEI/10.13039/501100011033), under grant number PID2019-103993GB-I00.
Data Availability Statement: DNA sequences are available on GenBank and all authors agree with
MDPI Research Data Policies.
Acknowledgments: We are grateful to T.L Blockeel, J. Shevock, and T. McIntosh for the loan of
specimens for DNA studies and the curators and staff of the herbaria cited in the text (CAS, E, H, NY,
PC, S, UBC) for assistance during our visit to the herbaria or the loan of specimens. We also thank the
Botanical Museum of Córdoba (Argentina), the National Herbarium of La Paz (Bolivia), the National
University of Loja (Ecuador), the San Marcos University of Lima (Peru), the National Herbarium of
Venezuela in Caracas, the Natural History Museum of Crete (Greece), Cyprus Forestry Department of
Nicosia (Cyprus), and the Department of Botany, University of Concepción (Chile) for the assistance
during fieldwork and help with export of material. We are also grateful to Inmaculada Guijarro for
the illustrations.
Conflicts of Interest: The authors declare no conflicts of interest.
Appendix A
Vouchers and GenBank accession numbers for taxa used in the molecular phylogenetic
analysis. Taxon name, country and next division, voucher (collector, number, and where
the specimen is housed), and GenBank accession number for DNA sequences ITS, trnG,
atpB-rbcL and trnL-F; a dash (
−
) indicates missing data. An asterisk (*) is provided for
sequences retrieved directly from GenBank. Specimens of A. limbata, previously identified
as A. obliquifolia, are indicated as A. obliquifolia SH (SH: Southern Hemisphere).
Aloina aloides (W.D.J. Koch ex Schultz) Kindb.: 1185, Spain, Albacete, Gallego s.n.
(MUB 56946), MZ239210, MZ290025, MZ339865, MZ290077. Aloina ambigua (Bruch &
Plants 2024,13, 445 26 of 29
Schimp.) Limpr.: 1458, Cape Verde, Fogo, Cano 8114 (MUB 44972), PP002526, PP036083,
PP036131, PP036033. Aloina bifrons (De Not.) Delgad.: 939, Spain, Madrid, Cano 6086
(MUB 34263), PP002523, PP036077, PP036124, PP036027; 985, Tunisia, Mahdia Governorate,
Pocs et al. 03040/D (MUB 28451), PP002522, PP036078, PP036125, PP036028. Aloina
bracteata M.J. Cano et al.: 937, Argentina, Salta, Cano and Alonso 8409a (MUB 56491),
PP002524, PP036075, PP036122, PP036025; 1208, Argentina, Tucumán, Cano et al. 4113
(MUB 29597), PP002525, PP036076, PP036123, PP036026. Aloina brevirostris (Hook. &
Grev.) Kindb.: 1300, Argentina, Catamarca, Cano et al. 4125 (MUB 29594), PP002535,
PP036089, PP036137, PP036039; 1216, Bolivia, Oruro, Cano et al. 3403 (MUB 27728),
PP002537, PP036088, PP036136, PP036038; 1465, Canada, British Columbia, Mclntosh
NAD83 (UBC B239220), PP002536, PP036087, PP036135, PP036037; 1464, Canada, British
Columbia, Bjork 22763 (UBC B227649), PP002532, PP036086, PP036134, PP036036; 1219, Rus-
sia, Krasnoyarsk Krai, Fedosov 17–2–15–08 (CAS 1288359), PP002534, PP036084, PP036132,
PP036034; 1317, Russia, Krasnoyarsk Krai, Fedosov 16–0140 (CAS 1267137), PP002540,
PP036092, PP036140, PP036042; 1196, United Kindom, England, Blockeel 47/384 (Herb.
Blockeel), PP002533, PP036085, PP036133, PP036035; 1387, USA, New Mexico, Sargent
2591 (CAS 1290032), PP002539, PP036093, PP036141, PP036043; 1389, USA, New Mex-
ico, Dillingham 2646 (CAS 1259598), PP002538, PP036091, PP036139, PP036041. Aloina
calceolifolia (Müll. Hal.) Broth.: 1182, Argentina, Jujuy, Cano and Alonso 8345 (MUB
56422), PP002516, PP036071, PP036118, PP036021; 1836, Argentina, Salta, Cano et al. 8144
(MUB 56217), PP002513, PP036068, PP036115, PP036018; 1293, Argentina, Salta, Cano and
Alonso 8168b (MUB 56242), PP002515, PP036072, PP036119, PP036022; 1838, Argentina,
Tucumán, Cano et al. 4061c (MUB 61776), PP002521, –, –, –; 1478, Bolivia, Oruro, Cano
et al. 3383 (MUB 27722), PP002519, PP036069, PP036116, PP036019; 1190, Peru, Ancash,
Cano et al. 6879 (MUB 48303), PP002514, PP036073, PP036120, PP036023; 1451, Peru,
Ancash, Cano et al. 7060 (MUB 48489), PP002518, PP036074, PP036121, PP036024; 1484,
Peru, Arequipa, Cano 2176 (MUB 20394), PP002517, –, –, –; 1176, Peru, Puno, Cano 2353
(MUB 20586), PP002520, PP036070, PP036117, PP036020. Aloina limbata M.J. Cano et al.:
1304, Argentina, Catamarca, Cano et al. 4158 (MUB 28598), PP002490, PP036056, PP036106,
PP036007; 1302, Argentina, Catamarca, Cano et al. 4267 (MUB 29589), PP002495, PP036050,
PP036100, PP036001; 976, Argentina, La Rioja, Cano et al. 4324 (MUB 29599), PP002488,
–, –, –; 1218, Argentina, Mendoza, Cano et al. 4374 (MUB 29595), PP002487, PP036052,
PP036101, PP036002; 977, Argentina, Salta, Cano et al. 4082 (MUB 29585), PP002481,
PP036046, PP036096, PP035997; 1294, Argentina, Salta, Cano and Alonso 8152 (MUB 56225),
PP002491, PP036057, PP036107, PP036008; 1172, Argentina, Salta, Cano and Alonso 8205
(MUB 56282), PP002494, PP036054, PP036104, PP036005; 1180, Argentina, San Juan, Cano
et al. 4350 (MUB 29587), PP002498, –, –, –; 1303, Argentina, Tucumán, Cano et al. 4085 (MUB
29592), PP002482, PP036047, PP036097, PP035998; 1306, Argentina, Tucumán, Cano et al.
4061a (MUB 29593), PP002500, PP036059, PP036108, PP036010; 1213, Bolivia, Chuquisaca,
Churchill et al. 24638 (MUB 38330), PP002486, PP036051, PP036102, PP036003; 1207, Bolivia,
Cochabamba, Cano et al. 3561 (MUB 27720), PP002483, PP036048, PP036098, PP035999;
1206, Bolivia, Cochabamba, Cano et al. 3565 (MUB 27725), PP002484, PP036049, PP036099,
PP036000; 1476, Bolivia, Potosí, Cano and Jiménez 3699a (MUB 27723), PP002505, –, –, –;
1479, Ecuador, Azuay, Cano et al. 2814a (MUB 22313), PP002493, –, –, –; 1178, Ecuador,
Azuay, Cano and Gallego 2903a (MUB 22314), PP002499, PP036060, PP036109, –; 1477,
Ecuador, Loja, Cano and Gallego 3015 (MUB 22317), PP002479, –, –, –; 1191, Peru, An-
cash, Cano et al. 7075 (MUB 48503), PP002489, PP036055, PP036105, PP036006; 1449,
Peru, Cajamarca, Cano et al. 5157 (MUB 32544), PP002512, –, –, –; 1482, Peru, Cajamarca,
Cano and Jiménez 5168 (MUB 32545), PP002480, PP036045, PP036095, PP035996; 1189,
Peru, Huánuco, Cano et al. 7364a (MUB 48842), PP002506, PP036063,
−
, PP036013; 1485,
Peru, Junín, Cano 2073a (MUB 20357), PP002503, –, –, –; 1480, Peru, Junín, Cano 2069b,
(MUB 20581) PP002497, –, –, –; 1483, Peru, La Libertad, Cano and Jiménez 5357 (MUB
32546), PP002496, PP036058, –, PP036009; 1481, Peru, La Libertad, Cano and Jiménez
5373 (MUB 32547), PP002511, PP036067, PP036114, PP036017; 1214, Peru, Puno, Cano
Plants 2024,13, 445 27 of 29
2288b (MUB 23220), PP002478, –, –, –; 1205, Venezuela, Merida, Grande et al. 5938 (MUB
36828), PP002485, PP036053, PP036103, PP036004; 1452, Peru, Ancash, Cano et al. 7113
(MUB 48546), PP002492, –, –, –; 1209, Ecuador, Azuay, Cano et al. 2814b (MUB 59251),
PP002504, –, –, –; 1210 (A. obliquifolia SH), Ecuador, Imbabura, Cano 3200b (MUB 22585),
PP002510, PP036066, PP036113, PP036016; 1193, Peru, Ancash, Cano et al. 7150 (MUB
48588), PP002507, PP036065, PP036112, PP036015; 1212 (A. obliquifolia SH), Peru, Caja-
marca, Cano et al. 5076b (MUB 32548), PP002502, PP036061, PP036110, PP036011; 556
(A. obliquifolia SH), Peru, Cajamarca, Cano and Jiménez 5196 (MUB 32549), PP002501,
PP036062, PP036111, PP036012; 1292 (A. obliquifolia SH), Peru, Junín, Cano 2069a (MUB
20352), PP002508, PP036064, –, PP036014; 978 (A. obliquifolia SH), Peru, Lima, Cano et al.
7649 (MUB 49149), PP002509, –, –, –. Aloina obliquifolia (Müll. Hal.) Broth.: 1240,
China, Yunnan, Wen Zhang and Shevock 16–7978 (CAS 1283278), PP002542, PP036094,
PP036142, PP036044; 961, Japan, Nagano, Sato s.n. (MUB 52995), PP002541 PP036090,
PP036138, PP036040. Aloina rigida (Hedw.) Limpr.: 1462, Cyprus, Limassol, Cano 10652
(MUB 59090), PP002531, –, –, –; 1275, Greece, Creta, Cano et al. 10367 (MUB 58849),
PP002530, PP036082, PP036129, PP036032; 1459, Spain, Almería, Cano 9374 (MUB 51040),
PP002529, PP036081, PP036128, PP036031; 1276, Spain, Córdoba, Cano 10285 (MUB 56743),
PP002527, PP036080, PP036127, PP036030; 1453, Spain, Islas Canarias, Tenerife, Cano 8813
(MUB 49260), PP002528, PP036079, PP036126, PP036029; 125, Spain, Murcia, Cano 4839
(MUB 28930), MW398549, GU953707, PP036130, GU953732. Aloinella galeata var. and-
ina (Delgad.) Delgad. & Schiavone: 274, Argentina, Catamarca, Cano et al. 4124 (MUB
29607), MW398550, JN968401, MW433018, JN968437. Crossidium squamiferum (Viv.) Jur.:
272, Spain, Canary Islands, Cano 4801 (MUB 28896), MW398558. JN968402, MW433022,
JN968438. Erythrophyllopsis andina (Sull.) R.H. Zander: 86, Bolivia, La Paz, Cano and
Jiménez 3825 (MUB 27961), MW398546, GU953698, MW433015, GU953723.
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