Bryophytes of the cloud forest of Pico do Marumbi State Park , Paraná , Brazil

Bryophyte diversity in tropical forests is closely related to elevation. A survey was undertaken of the bryoflora of a poorly known cloud forest environment from 1,000 to 1,500 m.a.s. in Pico do Marumbi State Park, state of Paraná, Brazil. The aim was to analyze the geographical range of the bryoflora and significance of this elevational zone to bryophyte diversity. We found 364 bryophyte species, including 187 species of mosses, 175 liverworts and two hornworts. This diversity represents 40% of the bryoflora of Paraná and 24% of that of Brazil. Forty-one Brazilian endemic species were found, representing a highly relevant feature for the preservation of the area. These results emphasize the importance of Pico do Marumbi State Park to the conservation of bryophyte diversity and to understanding the species of the state of Paraná, as well as demonstrating the importance of bryophyte surveys in the Atlantic Forest of Southeast Brazil.


Introduction
Bryophytes are avascular terrestrial plants whose sexual reproduction is dependent on water because of their flagellated anterozoids.In their lifecycle, the gametophytic generation (haploid) is predominant over their sporophytic generation (diploid) (Gradstein et al. 2001).They are the second largest group of terrestrial plants (Frahm 2003), and currently, they comprise 3 divisions, the Anthocerothophyta (hornworts), the Marchantiophyta (liverworts), and the Bryophyta (mosses) (Shaw and Goffinet 2009).
There are 1548 species of bryophytes reported from Brazil, with 541 species occurring in the state of Paraná (Yano 2013, 2014, Ristow et al. 2015), 570 species in the state of Rio Grande do Sul, and 542 in the state of Santa Catarina (Costa and Peralta 2017).
The first studies of bryophytes of Paraná were those of Angely (1961Angely ( , 1965Angely ( , 1968)), who produced a list of taxa.Kummrow and Prevedello (1982) published a list of samples deposited in the herbarium MBM (Municipal Botanical Museum, Curitiba).Hirai et al. (1998) undertook a floristic survey in a small secondary Araucaria forest, and Yano and Colletes (2000) investigated the

LIST OF SPECIES
bryophytes of an area of Atlantic Forest.Recently Yano (2013Yano ( , 2014) ) published a compilation of the literature on bryophyte taxa of t Paraná and listed 706 taxa.Since then, Ristow et al. (2015) added 203 species records for the state.
Studies of bryophytes in Brazilian cloud forests remain scarce and are non-existent for the state of Paraná.The studies of Santos and Costa (2010) and Costa et al. (2015) represent the only bryophyte investigations conducted above 1000 m above sea level in Brazil.These authors indicated that bryophtes were the richest and most diverse above this elevation.The results of several other authors are consistent with this finding (Enroth 1990, Frahm 1990, Gradstein 1991, Kessler 2000, Santos and Costa 2010, Costa et al 2015).
Pico do Marumbi State Park (PMSP) is located in southern Brazil, and is 1 of the 10 centers of diversity and endemism in tropical America (Tan and Pócs 2000), and thus, it offers an excellent environment for the study of bryophyte diversity.PMSP is completely within the Atlantic Forest biome and comprises Dense High Mountain Ombrophylous Forest phytophysiognomy.These unique high-elevation montane forests possess high endemism and, in general, are subjected to minimal anthropogenic disturbance due their difficult access (Scheer et al. 2011).
The forest vegetation of PMSP decreases in size with increasing elevation and because soil is replaced by rocky outcrops near 1400-1500 m elevation.At elevations of 1000-1400 m the vegetation in the park is cloud (IBGE 2012).forest.The term "cloud forest" derives of the high condensation of evaporated water in the air, which forms a haze and maintains high precipitation.The features of cloud forest correspond to an arboreal mesophanerophytic formation (reaching >30 m), which is typically located on the summits of higher mountains (1000-1500 m elevation) (Roderjan et al. 2002, Scheer et al. 2011).
Upper Montane Forests and High-Altitude Meadows are fundamental to the conservation of bryophyte diversity in the Atlantic Forest, and Santos and Costa (2010) and Costa et al. (2015) emphasized the importance of their protection and conservation.The objective of our study is to record and analyze the floristic composition of bryophytes occurring in the cloud forest of Pico do Marumbi State Park, as well as to evaluate the geographic distributions of bryophyte species found.

Methods
Study area.PMSP (Fig. 1) is located between the cities of Morretes, Quatro Barras, and Piraquara, and covers an area of 8745 ha (IAP 2012).It is within the Atlantic Forest domain (IAP 1996) and includes areas of Dense Ombrophylous Submountain Forest, Dense Mountain Ombrophylous Forest, and Dense High Mountain Ombrophylous Forest in good states of preservation.The floristic composition and tree layer structure varies along the elevational gradient (Figs 2-7).
Floristic surveys.Sampling was permitted by the Instituto Ambiental do Paraná (IAP), and we followed the methodology of Frahm (2003).Random collects were performed using pre-existent trails (approximate coordinates plotted in Fig. 1) with an effort made to cover the range of habitats and microhabitats available in cloud forests between 1000 and 1400 m above sea level.Eleven expeditions were taken from March 2014 to June 2015.The samples were deposited in the Maria Eneyda P. K. Fidalgo (SP) herbarium with duplicates in Departamento de Botânica da Universidade Federal do Paraná (UPCB) herbarium.
Bryophyte identification.Bryophytes were identified from the preparation of slides and their observation with stereo-and optical microscopes using specialized literature according to each family (Frahm 1991, Sharp et al. 1994, Yano and Carvalho 1995, Buck 1998, Gradstein et al. 2001, Gradstein and Costa 2003, Yano and Peralta 2009, Yano and Peralta 2011) and comparisons with herbarium specimens.
The classification systems follow Söderström et al. (2016) for Anthocerotophyta and Marchantiophyta and Goffinet et al. (2009) for Bryophyta.Author abbreviations for the specific epithets followed Brummitt and Powell (1992).The species list provided here is in alphabetical order by division, family and species.The geographic distributions in Brazil follow the standardization of Valente and Porto (2006), who classify species as Endemic, Rare, Moderate or Wide, while the occurrences in states followed Costa and Peralta (2015) and Gradstein and Costa (2003).
Here we provide, in alphabetical order, brief characterizations of the 41 Brazilian endemic species found in PMSP.This list follows the characterization of cloud forest by Costa et al. (2015), with species tolerant to wide environmental variation, such as low temperatures, high humidity, and high light intensity.

Barbula riograndensis E.B. Bartram
Comments: distinguishable from any other known Brazilian species of Barbula by costa with dorsal and ventral bands of stereids in cross section, and short, yellow and strongly dentate apiculi.Habitat: soil, rocks and streams.Geographic distribution: Atlantic Forest (lowlands).Brazilian geographic distribution: PR, RS.

Bazzania heterostipa (Steph.) Fulford
Comments: forked, terminal branching scarce and plants without terminal branches may be mistaken with species of Calypogeia; the presence of a vitta in the leaves immediately separates this species from Calypogeia.Habitat: soil, rocks and corticicolous.Geographic distribution: Atlantic Forest.Brazilian geographic distribution: BA, ES, MG, PE, PR, RJ, RS, SC, SP.
Comments: conical protuberances restricted to the keel and sometimes a few apical cells of the lobe, never exceeding 10 μm in the height.Habitat: epiphyllous and decomposing tree trunks.Geographic distribution: Atlantic Forest.Brazilian geographic distribution: SP.

Lejeunea oligoclada Spruce
Comments: at first glance resembles a small species of Cheilolejeunea due to dull greenish color, long lobule tooth, and thick leaf cell walls; also resembles Lejeunea phyllobola Nees & Mont., but the latter is larger (0.6-1.1 mm wide), and with the leaf lobe apex widely rounded; plants pale and yellowish or light green.Habitat: corticicolous and decomposing tree trunks.Geographic distribution: Atlantic Forest.Brazilian geographic distribution: AL, BA, ES, MG, PE, PR, RJ, SC, SP.

Lepidopilidium brevisetum (Hampe) Broth.
Comments: distinguished by leaves with acute apexes, margin serrate at the apexl and entire toward the base; costa reaching approximately ½ of the leaf length.Habitat: corticolous.Geographic distribution: Atlantic Forest (lowlands to montane) Brazilian geographic distribution: AL, ES, MG, RJ, RS, SC, SP.

Lepidopilum caudicaule Müll. Hal.
Comments: distinguished by serrulate leaf margin at the apex and entire to sinuous toward the base.Habitat: moist rocks.Geographic distribution: Atlantic Forest (lowlands to high montane).Brazilian geographic distribution: PE, RJ, RS, SC.

Metzgeria brasiliensis Schiffn.
Comments: similar to Metzgeria aurantiaca Steph., Metzgeria convoluta Steph.and Metzgeria cratoneura Schiffn.by having cells of verrucous lamina and medulla with thickened walls.Metzgeria aurantiaca differs by possessing rhizoids on the ventral surface of the lamina and on the margin, non-differentiated margin of the cells and male stem smaller than the female; Metzgeria con voluta differs by costa formed by 4 (-6) rows of ventral epidermal cells, rhizoids on uniform margin, male stem smaller than the female stem and male branch without rhizoids.Habitat: corticicolous, epiphyllous and rocks.Geographic distribution: Atlantic Forest.Brazilian geographic distribution: AL, BA, PR, RJ, RS, SC, SP.

Paranapiacabaea paulista W.R. Buck & Vital.
Comments: can only be identified with sporophyte; sterile resembles Donnellia, when fertile resembles Pterogoniopsis cylindrical Müll.Hal., but this latter genus has a higher basal membrane, various super-alar cells and leaves with a flat edge.Habitat: corticicolous.Geographic distribution: Atlantic Forest (submontane to montane).Brazilian geographic distribution: PR, SP.

Radula brasilica Yamada
Comments: characterized by recurved free margin of lobules; leaf cells with large trigones.Habitat: decomposing tree trunks.Geographic distribution: Atlantic Forest.Brazilian geographic distribution: SP.

Rhacocarpus inermis (Mull. Hal.) Lindb.
Comments: distinguished by oblong-lanceolate leaves with short acute apexes that are cuspidate or when piliferous with the upper margin not bordered.Habitat: exposed, moist or soil covered rocks.Geographic distribution: Atlantic Forest (montane to high montane).Brazilian geographic distribution: ES, MG, RJ, RS, SC.

Sphagnum divisum H.A. Crum
Comments: with 1 or 2 fasciculate branches, large leaves of branches with annular pores commissurally arranged in continuous rows on both surfaces, and leucocyst stem leaves consistently divided with more numerous pores on internal than external surface.Habitat: soil and rocks.Geographic distribution: Atlantic Forest (montane and high montane) and Cerrado.Brazilian geographic distribution: BA, GO, MG, RJ, SC.

Sphagnum exquisitum H.A. Crum
Comments: similar to S. ramulinum, but possessing smaller leaves, oval branches, and fibrillous leucocysts only on upper portions of both stem leaves and branches.Habitat: soil and rocks.Geographic distribution: Atlantic Forest (submontane to high montane).Brazilian geographic distribution: MG, PR, RJ, SP.

Sphagnum globicephalum Müll. Hal. ex Warnst.
Comments: characterized by possessing a simple stem and sparse branches with terminal globular structures formed by groups of imbricate leaves; stem leaves and branches are similar with few or numerous pores on both surfaces.Habitat: dry soil.Geographic distribution: Atlantic Forest (montane to high montane).Brazilian geographic distribution: RJ, SC.

Sphagnum multiporosum H.A. Crum
Comments: characterized by branches in fascicles of three; leaves broad-elliptic, broad-rounded apex, apex smooth, sometimes fringy or bordered by a reabsorption furrow; leucocyst of leaves of branches on outer surface with group of 3 conspicuous pores at adjacent angles; chlorocyst elliptical.Habitat: soil and rocks.Geographic distribution: Atlantic Forest and Pampa.Brazilian geographic distribution: BA, MG, RJ, RS, SP.

Sphagnum pseudoramulinum H.A. Crum
Comments: similar to S. ramulinum in Southeast Brazil, however, the pores of the external surface of the stem leaves and branches are large and arranged in a commissural row, leucoscysts of stem leaves are divided and secondary branches grow near base.Habitat: soil and rocks.Geographic distribution: Atlantic Forest (high montane).Brazilian geographic distribution: RJ, RS, SP.

Sphagnum sucrei H.A. Crum
Comments: stem leaves large, oblong, with no divided hyaline cells and numerous pores in commissural rows on external surface, but absent on internal surface, contrasting in size and shape with leaves of the branches, which possess a similar pore distribution; the chlorocyst of the leaves of the branches are narrowly trapezoidal in cross-section, more widely exposed on the internal surface.Habitat: rocks.Geographic distribution: Atlantic Forest.Brazilian geographic distribution: MG, RJ.

Trichocolea argentea Herzog.
Comments: characterized by lamina of the leaf with scattered cilia on the surface.Habitat: soil, rocks, and decomposing tree trunks.Geographic distribution: Amazonia, Caatinga, Cerrado, Atlantic Forest, Pampa and Pantanal.Brazilian geographic distribution: PR, RJ, SC, SP.

Discussion
Bryophyte species richness.The bryophytes found in our study of PMSP correspond to 40% of all the species recorded from the state of Paraná, although we found fewer species than indicated for this elevational range by Santos and Costa (2010).However, we found 41 Brazilian endemic species, including mosses and liverworts, which is near twice the total of endemic species (21) reported by Santos and Costa (2010).A possible explanation for this difference in species richness and endemism at PMSP is the high incidence of mist, as also observed by Montfoort and Ek (1990) in lowland tropical forests.
Moss species richness at PMSP was greater than that of liverworts, which is in accordance with findings by Santos (2011) in Rio de Janeiro, where liverworts had a greater species richness at lower elevations (between 50 m and 950 m) and mosses had a greater species richness at higher elevations (above 950 m).This pattern is well described in literature, and Gradstein et al. (2001) related the morphology of mosses to the colonization of open sites, like rock cliffs and exposed soils.
In the PMSP, the family Sphagnaceae had the greatest richness of endemic moss species found.The genus Sphagnum was found to be dominant in the cloud forests that we surveyed, and this dominance was earlier noted by IAP (2006).Sphagnaceae was among the families with the greatest contribution to the high species richness in High Mountain Forest habitats and elevational meadows of Itatiaia National Park, state of Rio de Janeiro (Costa et al. 2015).Species of the genus Sphag num were found growing in dry habitats, such as on such rocks and tree trunks, as well as moist habitats, such as springs and bogs.
With 66 species, the family Lejeuneaceae represented 18% of our list.Citing 83 species from sea level to 1200 m above sea level, this family was identified by Santos (2011) as the most representative of the Atlantic Forest; Sonnleitner et al. (2001) recorded 59 species in a cloud forest in Monte Verde, Costa Rica but at a lower altitude, below 285 m.Lejeuneaceae is the richest family of the recent lineage of Porellales, with about 1000 species (Bechteler et al. 2016) and includes specialists at colonizing bark, branches, twigs, and leaves of trees.Thus, this family is frequently abundant in humid tropical forests (Lücking 1995, Pócs 1996, Gradstein 1997, Cornelissen and Ter Steege 1989, Sonnleitner et al. 2009).
The family Leucobryaceae was represented by 19 species (2 Brazilian endemics) in our study, and Costa et al. (2015) also observed this in Rio de Janeiro state.
Conservation.Our findings reinforce the importance of PMSP for the preservation of the bryophyte flora of Paraná and the recognition of diversity hot spots for prioritizing conservation efforts in areas with high species diversity and endemism (Kier et al. 2009, Costa et al. 2015).Furthermore, our findings expand the known distributions of several Brazilian endemic bryophytes.The bryophyte flora of PMSP has great importance to the conservation of these species with narrow to moderate distributions in Brazil.Of the species that we list, 95 are rare (50 liverworts and 45 mosses), 151 moderate (75 liverworts, 74 mosses, and 2 hornworts), and 118 widespread (60 mosses and 50 liverworts) (Table 1).
According to Costa and Peralta (2015), Brazil has 298 endemic species of bryophytes, with 73 species occurring in Paraná (24% of the total).In PMSP, 41 endemic species were found (25 mosses and 16 liverworts; see Results).Table 2 compares bryophytes richness in various regions of the world, and shows the significance of the PMSP for bryoflora.
Our study, based on new surveys, is a pioneering contribution to the knowledge of the bryoflora of the state of Paraná and Brazil.

Figure 1 .
Figure 1.Location map of Pico do Marumbi State Park in Paraná state.

Table 2 .
Bryophyte species number comparison among the Pico do Marumbi State Park with different geographic regions.