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Home , Aloina, Andreaeaceae, Barbula unguiculata, Chenia (plant), Microbryum, Pottia

Plant Syst. Evol. 235: 197–207 (2002) DOI 10.1007/s00606-002-0230-0

Tortula and some related genera (, Musci): phylogenetic relationships based on chloroplast rps4 sequences

O. Werner, R. M. Ros, M. J. Cano, and J. Guerra

Departamento de Biologı´ a Vegetal (Bota´ nica), Facultad de Biologı´ a, Universidad de Murcia, Spain

Received May 23, 2002; accepted August 23, 2002 Published online: November 22, 2002 Ó Springer-Verlag 2002

Abstract. According to chloroplast rps4 sequence by a peristome of 16 mainly erect cleft teeth, data the genus forms a monophyletic stretching from near the base or from a low clade clearly separated from , while is basal membrane up to the apex, becoming shown to be polyphyletic and the Rhynchostegiae divided into two filaments. Pottia (Rchb.) species (with rostrate lid) are very similar to Ehrh. ex Fu¨ rn. has been separated by the Tortula. Crossidium is probably polyphyletic. The presence of a cleistocarpous or stegocarpous close affinity of Desmatodon, Stegonia, Pterygoneu- capsule (gymnostomous or with a peristome of rum and some of the species of Phascum with Tortula is confirmed by the molecular data. 16 imperfect or lanceolate bifid teeth) and Tortula Hedw. has been characterised by 32 Key words: Bryophyta, Pottiaceae, Tortula, Cros- filiform twisted teeth, united at the base into a sidium, Pottia, molecular phylogeny, rps4. tubular membrane of varying length. Also, some authors have considered the genus Syn- trichia Brid. for including the species of Tortula with a twisted peristome from a high spirally Introduction tessellated tube (Agnew and Vondra´ cˇ ek 1975). The generic boundaries in the Tribe Pottieae However, this separation has not been satis- (Limpr.) Dixon of the subfamily Pottioideae factory and there is controversy on the taxa (Limpr.) Broth., family Pottiaceae Bruch & that should be included in each genus. Schimp., notably in the series Pottia-Desmato- In his new treatment of the Pottiaceae don-Tortula-Syntrichia are particularly difficult genera, in which the gametophytic characters to delimit (Blockeel 1990), as the of were emphasized, Zander (1989, 1993) con- these genera is mainly based on their sporo- vincingly proposed some characters to segre- phytic characters, more specifically on the gate genera such as R. H. Zander and height of the basal membrane of the peristome Syntrichia from Tortula. According to this and on the patterns which the teeth follow author, Chenia differs from Tortula in its when they divide above this membrane dentate upper leaf margins, large epapillose (Limpricht 1890, Mo¨ nkemeyer 1927, Lawton laminal cells, thin costa, and red KOH 1971, Smith 1978, Nyholm 1989). Thus, the reaction of the upper laminal cells, while genus Desmatodon Brid. has been characterised Syntrichia differs in the crescent shape of the 198 O. Werner et al.: Phylogenetic relationships of Tortula and related genera stereid band in the leaf transverse section, Material and methods lack of differentiated dorsal epidermal cells, material. 36 sequences representing 35 taxa and red KOH reaction. On the other hand, were included in this study. A previous study this author included in Tortula sect. Pottia based on rps4 sequences representing a great (Rchb.) Kindb. taxa that had traditionally range of the Pottiaceae (unpublished data) sug- been placed in other genera, such as Pottia gests that the clade studied here is monophyletic. (the species with rostrate lid, named by Hedw. was chosen as out- Warnstorf (1916) as Rhynchostegiae)or group species because it was situated in the sister Phascum Hedw. (Phascum cuspidatum clade. Hedw.), but with gametophytic characters Table 1 lists all taxa used in this study, the very similar to those of Tortula. For the voucher specimen data or reference, the origin same reason, he included the genus Desmato- and GenBank accession numbers. Vouchers are don in Tortula sect. Tortula Broth. deposited at MUB. DNA isolation. Total DNA was extracted There is general agreement concerning from dry material using the DNeasy Plant Mini some taxonomic changes and phylogenetic Kit of Qiagen (Hilden, Germany). The DNA was considerations provided by Zander (1993) eluted in 100 ll of 10 mM Tris-buffer (pH 8.5) and (e.g. the separation of Syntrichia from Tortula) stored in the freezer until amplification. although other aspects such as the relation of Amplification. The chloroplast rps4 gene was Pottia, Phascum and Desmatodon with Tortula amplified in a 50 ll final volume with the primers are more debatable. Also, the relationships rps5 (Nadot et al. 1994) and trnas (Buck et al. between some genera of Pottieae, whose origin 2000). 1 ll of stock DNA was added as template. has been supposed to be close to a Tortula-like The amplification conditions were as follows: ancestor (Delgadillo 1975, Frey et al. 1990, 3min at 94 °C, 35 cycles with 15 sec 94 °C, Cano et al. 1993, Zander 1993) remain unre- 30 sec 50 °C and 1 min 72 °C, and a final 7- solved. This is the case with the genera min extension step at 72 °C. Amplification pro- ducts were controlled on 8% PAA gels, and Crossidium Jur. and Jur., successful reactions were cleaned with the help of which differ from Tortula in the presence of the High Pure PCR Product Purification Kit supranervial filaments or lamellae, respective- of Roche Molecular Biochemicals (Mannheim, ly. Kindb. also presents this kind of Germany). chlorophyllic structures on the ventral side of Sequencing and data analysis. Cycle sequenc- the lamina but is excluded from our study ing was performed with the Big Dyes Sequencing because previous DNA assays made by the Kit (Perkin Elmer) using a standard protocol and authors (Werner et al. unpublished) demon- the amplification primers. The aligning tempera- strated that this genus does not belong to the tures were set to 60 °C in the case of rps5 and 55 °C group of genera closely related to Tortula.In in the case of trnas. The reaction products were addition, Stegonia Venturi has been supposed separated on a ABI Prism 3700 automatic to be related to Pottia by some authors sequencer (Perkin Elmer). The sequences were edited using Bioedit 5.0.9 (Hall 1999) and aligned (Limpricht 1890, Mo¨ nkemeyer 1927, Chen manually. The data are available from the corre- 1941, Wareham 1972). However, it has been sponding author on request. The aligned sequences treated as an independent genus by other were analysed using Neighbor-Joining (NJ; Saitou recent authors, such as Smith (1978), Nyholm and Nei 1987), Maximum Parsimony (MP; Fitch (1989) and Zander (1993). 1971) and Bayesian inference (Bayes 1763). In the The purpose of the present paper is to case of NJ we used PAUP* (Swofford 1998), with provide a new and independent dataset (chlo- distance ¼ hky85, objective ¼ LSFIT, power ¼ 2. roplast rps4 sequence) to clarify the circum- A bootstrap analysis with 1000 replicates was run. scription of the genus Tortula and the The MP analysis, run with PAUP*, used the phylogenetic relationships between Tortula following settings: RANDOM additions, TBR and some related genera. branch-swapping, MULTREES ¼ yes, steepest Table 1. Taxa included in the rps4 sequence analysis. All sequences are submitted to GenBank of relationships Phylogenetic al.: et Werner O. Species Voucher Origin GenBank # or reference Barbula unguiculata Hedw. MUB 10325 Germany, Baden-Wu¨ rttemberg AF480952 Crossidium aberrans Holz. & E.B. Bartram MUB 6089 Spain, Albacete AF481006 Crossidium crassinerve (De Not.) Jur. MUB 11622 Morocco, Anti-Atlas AF481045 Crossidium davidai Catches. MUB 5349 Spain, Canary Islands, Lanzarote AF481005 Crossidium laevipilum The´ r. & Trab. MUB 11665 Morocco, Anti-Atlas AF481046 Crossidium seriatum H.A. Crum & Steere UNLV, Arizona U.S.A., Arizona AF481007 Crossidium seriatum H.A. Crum & Steere MUB 4354 Spain, Albacete AF481010 Crossidium squamiferum (Viv.) Jur. MUB 7982 Spain, Huelva AF481004 Desmatodon latifolius (Hedw.) Brid. MUB 11476 Spain, Granada AF480954 Leptophascum leptophyllum (Mu¨ ll. Hal.) MUB 10427 Spain, Murcia AF480960 J. Guerra & M.J. Cano curvicolle (Hedw.) R.H. Zander MUB 8269 Spain, Almerı´ a AF480971 (Sm.) R.H. Zander MUB 12537 Spain, Alicante AF480983 Microbryum rectum (With.) R.H. Zander MUB 11199 Spain, Valencia AF480970 Tortula Phascum cuspidatum Hedw. MUB 11344 Spain, Sevilla AF480972 Pottia intermedia (Turner) Fu¨ rnr. MUB 5430 Germany, Nordrhein Westfalen AF481013 n eae eea199 genera related and Pottia lanceolata (Hedw.) Mu¨ ll. Hal. MUB 10334 Spain, Murcia AF480988 Pottia pallida Lindb. MUB 8768 Spain, Almerı´ a AF480987 Pottia truncata (Hedw.) Bruch & Schimp. MUB 10336 Spain, Cadiz AF481044 Pottia wilsonii (Hook.) Bruch & Schimp. MUB 8740 Spain, Almerı´ a AF481011 Protobryum bryoides (Dicks.) MUB 4936 Spain, Alicante AF480986 J. Guerra & M.J. Cano Pterygoneurum lamellatum (Lindb.) Jur. MUB 11484 Spain, Murcia AF480959 Stegonia latifolia (Schwa¨ gr.) Venturi ex Broth. La Farge et al. (2000) - AF222901 Syntrichia ammonsiana (H.A. Crum MUB 12225 South Africa, AF480957 & L.E. Anderson) Ochyra Western Cape Prov. Syntrichia caninervis Mitt. MUB 10977 Spain, Guadalajara AF480958 Syntrichia leucostega (Mu¨ ll. Hal.) R.H. Zander MUB 12226 South Africa, AF480982 Western Cape Prov. Syntrichia montana Nees MUB 10881 Morocco, Rif AF481031 Syntrichia papillosa (Wils.) Jur. MUB 12227 South Africa, AF481032 Western Cape Prov. Syntrichia rigescens (Broth. & Geh.) Ochyra MUB 11378 Morocco, High-Atlas AF481037 (Hedw.) Brid. MUB 10887 Morocco, Rif AF480981 200 O. Werner et al.: Phylogenetic relationships of Tortula and related genera

descent ¼ no, COLLAPSE ¼ yes. Parsimony anal- ysis was not expected to swap to completion (see Soltis et al. 1998). Maxtrees was set to 100000. Parsimony analysis were performed with equal weighting of characters and transformations. A bootstrap analysis with 100 replicates was AF481016 performed with the settings as mentioned before, but with MAXTREES set to 10000. The Bayesian approach (Rannala and Yang 1996, Mau and Newton 1997, Mau et al. 1999) is similar to Maximum Likelihood (ML) in that the user postulates a model of evolution and the program searches for the best trees that are consistent with both the model and with the data. However, it differs from ML in that while ML seeks the tree that maximises the probability of observing data given that tree, Bayesian analysis seeks the tree that maximises the probability of the tree given the data and the model for evolution. In essence, this re-scales likelihoods to true probabilities in

La Palma that the sum of the probabilities over all trees is 1.0 with the Bayesian approach, which means that ordinary probability theory can be used to analyze the data (Hall 2001). The program MrBayes (Huelsenbeck and Ronquist, in press) was used to obtain an estimation of the phylogenetic relation- ships under the Bayesian approach. Four charsets were defined representing the codon position of the coding region and the noncoding spacer. A parti- tion (bycodon) in these four charsets was defined. The following settings were used: lset ¼ nst6, rates ¼ sitespec, sitepartition ¼ bycodon. The Mar- kow chain settings were: ngen ¼ 400 000, print- freq ¼ 1000, samplefreq ¼ 100, nchains ¼ 4. The first 100 000 generations were discarded from the further analysis (burnin ¼ 1000) and contype ¼ halfcompat, which is the equivalent of 50% major- ity rule in PAUP*. The clade credibility values as calculated by MrBayes are given in the cladograms.

Results Schiffn. MUB 4933 Spain, Granada AF481009 (Sm.) Lindb. MUB 11352 Spain, Seville AF480990 Mont. MUB 11381The aligned Spain, Albacete sequences have a AF481001 length of 661 bp. (Schimp.) G. Roth MUB 11347 Spain, Malaga AF481030 Hedw. MUB 10951 Morocco, Rif AF481000 (K.F. Schultz) Mont. MUB 2513Spain, Alicante AF481042 Hedw. MUB 12540All Tortula Spain, Canary Islands, s.l. species share a 4 bp deletion compared with Barbula unguiculata. 129 bp of the aligned sequences were variable and 61 of (continued) them parsimony informative. Figures 1 and 2 show the phylograms Table 1 Tortula atrovirens Tortula brevissima Tortula canescens Tortula revolvens Tortula vahliana Tortula subulata obtained under the Bayesian approach and O. Werner et al.: Phylogenetic relationships of Tortula and related genera 201

Fig. 1. Bayesian tree for 36 Pottieae rps4 sequences. The general time-reversible model was assumed. Site specific rate variations were assigned for the three codon positions and the noncoding spacer. The tree is rooted with the sequence of Barbula unguiculata. Numbers indicate the clade credibility values of the nodes. Values below 50% are not shown using the Neighbor-Joining method. In addi- clearly separated from all the taxa supposedly tion, Fig. 2 gives the bootstrap values ob- closely related to Tortula. tained by Maximum Parsimony. All species of Syntrichia form a monophy- Microbryum curvicolle (Hedw.) R. H. letic lineage and are a sister clade to Lepto- Zander, M. davallianum (Sm.) R. H. Zander phascum leptophyllum (Mu¨ ll. Hal.) J. Guerra & and M. rectum (With.) R. H. Zander were M. J. Cano. 202 O. Werner et al.: Phylogenetic relationships of Tortula and related genera

Fig. 2. Neighbor-Joining tree of the 36 rps4 sequences using Kimuras two parameter distances. Numbers on the branches indicate percentage support in 1000 bootstrap replications/percentage support in 100 bootstrap replications using unweighted Maximum Parsimony. Bootstrap values below 50% are not shown

Crossidium laevipilum The´ r. & Trab., Pterygoneurum lamellatum (Lindb.) Jur., C. crassinerve (De Not.) Jur. and C. aberrans Desmatodon latifolius (Hedw.) Brid., and Holz. & E. B. Bartram are in the same clade, Phascum cuspidatum are situated within the but C. squamiferum (Viv.) Jur. is included in Tortula-complex, although their exact place- the clade with Tortula revolvens (Schimp.) ment within this complex remains ambiguous. G. Roth and other species of the same and The Pottia species belonging to the other genera. Crossidium davidai Catches. is Rhynchostegiae group (according to Warn- most closely related to Tortula atrovirens (Sm.) storf 1916) and Protobryum bryoides (Dicks.) Lindb. and Crossidium seriatum (H.A. Crum) J. Guerra & M. J. Cano, both included by Steere is found near Tortula brevissima Zander within Tortula, are situated together Schiffn., T. muralis Hedw. and T. vahliana with Tortula canescens Mont. on a clade (Schultz.) Mont. with high bootstrap support (82% with NJ, O. Werner et al.: Phylogenetic relationships of Tortula and related genera 203

80% with MP, 100% clade credibility value authors (e.g. Brotherus 1924, Mo¨ nkemeyer under Bayesian inference) with Stegonia 1927, Nyholm 1989) to form its own section latifolia (Schwa¨ gr.) Venturi ex Broth. as or subgenus (Mildeella) within Pottia or as an sister group. independent genus, Mildeella bryoides (Dicks.) Our data also support the separation of Limpr. (genus Mildeella Limpr. nom. illeg.) or Protobryum bryoides from the cleistocarpous Protobryum bryoides, due to the presence of a species of the genus Microbryum. cleistocarpous capsule but with a slightly differentiated and non-functional peristome. The Rhynchostegiae species of Pottia, Proto- Discussion bryum bryoides and Tortula canescens form a From our molecular analysis it is clear that the relatively well supported clade, with Stegonia species placed in Microbryum Schimp. by latifolia as a possible sister. Zander (1993) are not closely related with the Stegonia latifolia has been considered as a species considered by this author in Tortula member of Pottia despite its different gameto- sect. Pottia. In this clade are included the clei- phyte, such as the bulbiform , very stocarpous M. curvicolle which was previously concave, nearly circular to very broadly ovate considered by other authors (e.g. Mo¨ nkemeyer or elliptical leaves and the weak costa ending 1927, Smith 1978, Nyholm 1989) to belong below the leaf apex. The well developed peri- to the genus Phascum, the stegocarpous M. stome is similar to that of some Pottia species. davallianum with a conic lid (placed in Pottia Zander (1993) also included in Stegonia a clei- by Warnstorf (1916) as Conostegiae), and the stocarpous species, (S. hyalinotricha [‘‘um’’] cleistocarpous M. rectum, which has a slightly Cardot & The´ r.) R. H. Zander, stating that differentiated lid and considered by other ‘‘this genus, like Tortula, shows a reduction authors to belong to the genera Pottia series in characters’’. Our present (Mo¨ nkemeyer 1927, Smith 1978, Nyholm data suggest a phylogenetic proximity of Ste- 1989) or Phascum (Limpricht 1890). Guerra gonia latifolia to the Rhynchostegiae species of and Cano (2000) suggested that the Conoste- Pottia, but the relatively poor bootstrap giae species of Pottia (including the stegocar- support (54% with NJ, below 50% with MP, pous P. starckeana (Hedw.) Mu¨ ll. Hal. and 51% clade credibility value using Bayesian P. davalliana (Sm.) C.E.O. Jensen) are phylo- inference) preclude a definitive conclusion. genetically closely related with the cleistocar- The Pottia clade might be interpreted as a pous Microbryum, due to the great similarity group in which reduced peristomes evolved. of the morphological features of their gameto- The only Pottia-like species of this clade with a phytes. On the other hand, the rps4 data well developed peristome is Pottia lanceolata, indicate that the Conostegiae group of Pottia with long, lanceolate erect teeth; Protobryum and the other group of species with rostrate bryoides being the only cleistocarpous species operculum (Rhynchostegiae), such as P. inter- with some non-functional peristomial process- media (Turner) Fu¨ rnr., P. lanceolata (Hedw.) es. Tortula canescens, with its long twisted Mu¨ ll. Hal., P. pallida Lindb. and P. truncata peristome and developed basal membrane, (Hedw.) Bruch. & Schimp., belong to two might represent a model of the common different evolutionary lineages. Thus, both the ancestor in this clade. This species appears data presented here and by Zander (1993), within the Rhynchostegiae clade of Pottia but, imply that the old concept of the genus Pottia since the support values within this clade are section or subgenus Eupottia (Conostegiae + low, one cannot be very confident about Rhynchostegiae) cannot be upheld. Zander the exact topology within this clade. This is (1993) included the Rhynchostegiae species of also demonstrated by conflicting topologies Pottia as well as Pottia bryoides within Tortula. obtained by different methods of data analysis The last species has been considered by some (Figs. 1 and 2). 204 O. Werner et al.: Phylogenetic relationships of Tortula and related genera

The analysis supports the recognition at traits of Crossidium are profoundly discussed genus level of Syntrichia. The genus Syntrichia by Delgadillo (1975) and Cano et al. (1993). was established in Bridel (1801) but later its Zander (1993) suggested that Crossidium sect. species were generally considered to belong to Holz. & E.B. Bartram Tortula (De Notaris 1838, Lawton 1971, Smith (C. aberrans Holz. & E.B. Bartram and 1978, Crum and Anderson 1981, Noguchi C. seriatum included in this study) may be 1988, Mishler 1994), probably due to difficul- more closely related with Tortula sect. Pottia ties in separating some species of the two than to Crossidium s. str., through such species genera based on sporophytic characters. Sub- as T. atherodes R.H. Zander (= Phascum sequently Zander (1989, 1993) separated the cuspidatum) and T. protobryoides R.H. Zander two genera based on differences in nerve (= Protobryum bryoides) with their flask- anatomy. Many authors agreed with Zander shaped protuberant ventral costal cells, or in reestablishing Syntrichia as a genus (Du¨ ll Tortula sect. Tortula (forms like T. atrovirens 1992; Ochyra 1992, 1994; Anderson 1997; or T. revolvens included here) with their ventral Gallego et al. 2003, in press). ITS 1 sequence costal pads of vertically elongate cells. data showed that Tortula muralis and Syntri- Crossidium davidai is the only studied chia ruralis (Hedw.) F. Weber & D. Mohr are Crossidium species lacking a hyaline hairpoint only distantly related (Spagnuolo et al. 1999), on the leaves. Crossidium seriatum possesses, but this dataset did not include more species of like C. davidai, 4–6 papillae per cell and short these two genera. filaments, while Crossidium aberrans has 1–2 Leptophascum leptophyllum is a species papillae and also short filaments. In the other with an unclear taxonomic position. For many species included here (C. crassinerve and years it has been considered as a member of C. laevipilum) the papillae are very rare and Tortula (T. rhizophylla (Sakurai) Z. Iwats. & the filaments are longer. Finally, Crossidium K. Saito), although Corley et al. (1981) squamiferum is characterised by smooth cells, mentioned that it ‘‘is completely anomalous up to 10–11 cells long filaments, and thickened in Tortula’’. Zander (1989) included it in the cell walls in leaves and filaments. Interestingly, newly created genus Chenia R. H. Zander. Arts Tortula revolvens, the closest neighbour of and Sollmann (1991) discovered that Phascum Crossidium squamiferum, differs from other leptophyllum Mu¨ ll. Hal. is the earliest name for Tortula s.l. species in its reduced papillosity, this species. Finally Guerra and Cano (2000) while T. brevissima and T. muralis show a created the monotypic genus Leptophascum J. papillosity similar to Crossidium seriatum. The Guerra & M. J. Cano. Based on a cladistic rps4 data suggest that Tortula atrovirens is analysis of morphological data, Zander (1993) closely related to Crossidium davidai (99%, came to the conclusion that it is related to 98% and 100% support with NJ, MP and Syntrichia, a point of view supported by our Bayesian inference), while Tortula brevissima, data in spite of the few shared morphological T. muralis and T. vahliana are closely related to features. Crossidium seriatum (59% and 100% support The distinctive character of the genus with NJ and Bayesian inference, below 50% Crossidium is the presence of supracostal with MP), and T. revolvens might form a filaments, 1–12 cells in length, in the upper monophyletic clade with Crossidium squamife- part of the leaf and forming a pad. This rum, although this is only supported by Baye- character seems to be an adaption to arid sian inference. Our study therefore suggests climates (Frey and Ku¨ rschner 1988, Guerra that Crossidium is polyphyletic. Tortula et al. 1992). Similar morphological adaptions brevissima is morphologically very close to are found in other Pottiaceae, such as Aloina, Crossidium seriatum and, especially in Europe, Aloinella Cardot and some species of Pseudo- it is sometimes difficult to distinguish between crossidium R.S. Williams. The morphological these species. The same is true for the pair of O. Werner et al.: Phylogenetic relationships of Tortula and related genera 205 species formed by Tortula atrovirens and with a cleistocarpous capsule are included in Crossidium davidai, although it should be the genus (P. kozlovii Laz. and P. subsessile borne in mind that the Canarian specimens var. kieneri Habeeb). Frey et al. (1990), of the latter are morphologically closer to Delgadillo (1975) and Zander (1993) discussed Tortula atrovirens than the Australian speci- the possibility that Pterygoneurum might be mens. Thus samples from Northern Africa derived from (ancestors of ) Crossidium. In our with their supracostal filaments 2(3) cells high analysis, the position of P. lamellatum is not have been considered to be xerophytic modi- clearly resolved. With NJ it seems to be closely fications of Tortula atrovirens growing in very related with Tortula revolvens and some species dry conditions (Cano et al., 2002). In these two of Crossidium, but the bootstrap support is cases, a more detailed investigation will be below 50% and the data are conflicting with necessary to delimit the species boundaries. the Bayesian tree. Data on the species with Our data suggest that the filaments on the large and a gymnostomous capsule and ventral side of the costa, which are typical of on those with a cleistocarpous capsule are not species placed in the genus Crossidium, have available (the assays with P. ovatum were not evolved from ancestors close to the genus successful). Therefore, a final conclusion can- Tortula, as proposed by Chen (1941), Delga- not be drawn with the present data. Whatever dillo (1975) and Cano et al. (1993), however the case, it seems that morphological adap- not once, but several times. According to the tions such as filaments or lamellae on the rps4 data, Crossidium aberrans is placed near ventral surface of the costa, developed inde- C. crassinerve and C. laevipilum, although pendently at various times within the Pottioi- C. aberrans is more similar to some species of deae. Tortula because of the short supracostal fila- The rps4 sequence data clearly place Des- ments. Therefore this species might represent a matodon latifolius and Phascum cuspidatum model for the transition between Tortula and within Tortula s.l. excluding Syntrichia. Both the species of Crossidium with long supracostal are situated in somewhat isolated clades. filaments and smooth or slightly papillose Phascum cuspidatum might be related with laminal cells. Tortula subulata, but the support is below 50% Similar adaptions to those of Crossidium with both NJ and MP, while Bayesian infer- are present in the genus Pterygoneurum, con- ence gives a clade credibility value of 85%. sisting of lamellae on the ventral surface of the Definition of the genus Tortula remains costa. Nevertheless sporophytic characters are unresolved. At present it comprises a high very heterogeneous within the genus. For number of species with a heterogeneous com- instance, P. ovatum (Hedw.) Dix., P. sampaia- bination of morphological characters in the num (A. M. L. Guim.) A. M. L. Guim. and , and only a deeper study includ- P. subsessile (Brid.) Jur. have gymnostomous ing as many species as possible and using capsules and large spores (25–57 (60) lm). The additional molecular markers will help resolve last species has an immersed capsule, while the problem. P. lamellatum has an exerted one, a peristome The molecular analysis carried out suggests with a basal, sometimes fragile membrane, and that the evolution within Pottiaceae is charac- peristome teeth ranging from short to moder- terised by a high degree of parallel and ately long and slightly twisted; furthermore, convergent evolution. Characters such as clei- this species possesses smaller spores stocarpous capsules, hairy filaments on leaves (15–25(30) lm) than the rest of the species of or costa, probably have evolved various times Pterygoneurum. The spores of P. lamellatum and independently as a response to environ- are more similar in size and morphology to mental pressures. This leads to a situation those present in the genera Crossidium and where it is extremely difficult to reconstruct Tortula (Carrio´ n et al. 1995). Also, some taxa phylogenetic relationships within this family 206 O. Werner et al.: Phylogenetic relationships of Tortula and related genera based on morphological data alone. Due to Cano M. J., Guerra J., Ros R. M. (1993) A revision the theoretically almost unlimited number of of the genus Crossidium (Pottiaceae) with characters, molecular analysis combined with the description of the new genus Microcrossi- classical morphological technique should dium. Plant Syst. Evol. 188: 213–235. contribute to a better understanding of the Cano M. J., Ros R. M., Gallego M. T., Jime´ nez phylogeny of this important family of . J. A., Guerra J. (2002) Contribution to the flora of Morocco: the Anti-Atlas The authors thank the deceased Theo Arts for catalogue. Cryptogamie, Bryol. 23: 249–262. sending some specimens as gift. Also thanks to Carrio´ n J. S., Cano M. J., Guerra J. (1995) Lloyd Stark for the loan of some American morphology in the moss genus Pterygoneurum Crossidium seriatum specimens and the financial Jur. (Pottiaceae). Nova Hedwigia 61: 481–496. support of the Spanish DGI (Project BOS2000- Chen P.-C. (1941) Studien u¨ ber die ostasiatischen 0296-C03-01). Lars Hedena¨ s and an other anony- Arten der Pottiaceae, I-II. Hedwigia 80: 1–76; mous reviewer made useful comments on an early 141–322. version of the manuscript. Corley M. F. V., Crundwell A. C., Du¨ ll R., Hill M. O., Smith A. J. E. (1981) Mosses of Europe References and the Azores; an annotated list of species, with synonyms from the recent literature. J. Bryol. 11: Agnew S., Vondra´ cˇ ek M. (1975) A Moss Flora of 609–689. Iraq. Feddes Repert. 86: 341–489. Crum H., Anderson L. E. (1981) Mosses of Eastern Anderson L. E. (1997) Syntrichia magilliana North America Vol. 1 & 2. Columbia University (Pottiaceae), a new species from South Africa. Press, New York. J. Hattori Bot. Lab. 82: 15–18. De Notaris G. (1838) Specimen de Tortulis italicis. Arts T., Sollmann P. (1991) Remarks on Phascum Mem. Reale Accad. Sci. Torino 40: 281–326. leptophyllum C. Mu¨ ll., an earlier name for Delgadillo C. M. (1975) Taxonomic revision of Tortula rhizophylla (Sak.) Iwats. & K. Saito. Aloina, Aloinella and Crossidium (Musci). Bry- Lindbergia 17: 20–27. ologist 78: 245–303. Bayes T. (1763) An essay towards solving a Du¨ ll R. (1992) Distribution of the European and problem in the doctrine of chances. Philos. Macaronesian mosses (Bryophytina), part I. Trans. 53: 370–418. Reprinted in: Pearson E.S., Bryol. Beitr. 4: 1–113. Kendall M. G. (eds.) (1970) Studies in the Fitch W. M. (1971) Toward defining the course of history of statistics and probability. Charles evolution: minimum change for a specific tree Griffin, London, pp. 131–153. topology. Syst. Zool. 20: 406–416. Blockeel T. L. (1990) The genus Par., Frey W., Herrnstadt I., Ku¨ rschner H. (1990) a note on the affinities of Tortula brevis White- Pterygoneurum crossidioides (Pottiaceae, Musci), house & Newton and T. stanfordensis Steere. a new species to the desert flora of the Dead Sea J. Bryol. 16: 187–192. area. Nova Hedwigia 50: 239–244. Bridel S. E. (1801) Animadversiones in Muscolo- Frey W., Ku¨ rschner H. (1988) Re-evaluation of giae Recentiorum Tomum secundum, ab ipso Crossidium geheebii (Broth.) Broth. (Pottiaceae) auctore propositae. J. Bot. (Schrader) 1800: 268– from Sinai, a xerothermic Pangaean element. 299. J. Bryol 15: 123–126. Brotherus V. F. (1924) Musci (Laubmoose). In: Gallego M. T., Cano M. J., Ros R. M., Guerra J. Engler A., Prantl K. (eds.) Die natu¨ rlichen (2003) An overview of Syntrichia ruralis complex Pflanzenfamilien 10, edn 2. Duncker & Humblot, (Pottiaceae: Musci) in the Mediterranean region Berlin. and neighbouring areas. Bot. J. Linn. Soc. 138: Buck W. R., Goffinet B., Shaw A. J. (2000) Testing 209–224. morphological concepts of orders of pleurocar- Gallego M. T., Cano M. J., Ros R. M., Guerra J. pous mosses (Bryophyta) using phylogenetic (2002) New taxonomic data on a circum-Teth- reconstructions based on trnL-trnF and rps4 yan group of Syntrichia (Pottiaceae, Bryophyta): sequences. Mol. Phyl. Evol. 16: 180–198. the S. caninervis complex. Syst. Bot. (in press). O. Werner et al.: Phylogenetic relationships of Tortula and related genera 207

Guerra J., Cano M. J. (2000) A taxonomic contri- Nyholm E. (1989) Illustrated flora of Nordic bution on the European cleistocarpous species of mosses. Fasc. 2. Pottiaceae-Splachnaceae-Schis- Pottiaceae (Musci). J. Bryol. 22: 91–97. tostegaceae. Nordic Bryological Society, Copen- Guerra J., Martı´ nez-Sa´ nchez J. J., Ros R. M. (1992) hagen. On the degree of adaptation of the moss flora and Ochyra R. (1992) New combinations in Syntrichia vegetation in gypsiferous zones of the south-east and Warnstorfia (Musci). Frag. Flor. Geobot. Iberian Peninsula. J. Bryol 17: 133–142. 37: 211–214. Hall G. H. (2001) Phylogenetic trees made easy. Ochyra R. (1994) Nomenclatural notes on mosses: Sinauer Associates Inc., Sunderland, Massachu- 3. Syntrichia virescens (Pottiaceae). Frag. Flor. setts. Geobot. 39: 311–314. Hall T. A. (1999) BioEdit: a user-friendly biological Rannala B., Yang Z. H. (1996) Probability distri- sequence alignment editor and analysis program bution of molecular evolutionary trees: A new for Windows 95/98/NT. Nucl. Acids Symp. Ser. method of phylogenetic inference. J. Molec. 41: 95–98. Evol. 43: 304–311. Huelsenbeck J. P., Ronquist F. R. (2002) MrBayes: Saitou N., Nei M. (1987) The neighbor-joining Bayesian inference of phylogeny. Biometrics (in method: a new method for reconstructing press). phylogenetic trees. Mol. Biol. Evol. 4: 406–425. La Farge K., Mishler B. D., Wheeler J. A., Wall Smith A. J. E. (1978) The Moss Flora of Britain D. P., Johannes K., Schaffer S., Shaw J. A. and Ireland. Cambridge University Press, Cam- (2000) Phylogenetic relationships within the bridge. haplolepideous mosses. Bryologist 103: 257–276. Soltis D. E., Soltis P. S., Mort M. E., Chase M. W., Lawton E. (1971) The moss flora of the Pacific Savolainen V., Hoot S. B., Morton C. M. (1998) Nothwest. The Hattori Botanical Laboratory, Inferring complex phylogenies using parsimony: Nichinan. an empirical approach using three large DNA Limpricht K. G. (1890) Die Laubmoose Deutsch- sets for angiosperms. Syst. Biol. 47: 32–42. lands, O¨ sterreichs und der Schweiz. 1. Abt.: Spagnuolo V., Caputo P., Cozzolino S., Castaldo Sphagnaceae, Andreaeaceae, Archidiaceae, Bryi- R., de Luca P. (1999) Patterns of relationships neae. In: Rabenhorst’s Kryptogamenflora von in Trichostomoideae (Pottiaceae, Musci). Plant Deutschland, O¨ sterreich und der Schweiz, 2. Syst. Evol. 216: 69–79. Aufl., Bd. 4, Eduard Kummer, Leipzig. Swofford D. L. (1998) PAUP* Phylogenetic anal- Mau B., Newton M. (1997) Phylogenetic inference ysis using parsimony (*and other methods). for binary data on dendrograms using Markow Version 4. Sinauer Associates, Sunderland, chain Monte Carlo. J. Comp. Graph. Stat. 6: Massachusetts. 122–131. Wareham, W. T. (1972) Pottia. In: Grout A. J. (ed.) Mau B., Newton M., Larget B. (1999) Bayesian Moss flora of North America, north of Mexico, phylogenetic inference via Markow chain Monte I. Hafner Publishing Company, New York, Carlo methods. Biometrics 55: 1–12. pp. 197–208. Mishler B. D. (1994) Tortula. In: Sharp A. J., Crum Warnstorf C. (1916) Pottia-Studien. Hedwigia 58: H., Eckel P. M. (eds.) The moss flora of Mexico, 35–152. vol. 1. The New York Botanical Garden, New Zander R. H. (1989) Seven new genera in Pottia- York, pp. 319–350. ceae (Musci) and a lectotype for Syntrichia. Mo¨ nkemeyer W. (1927) Die Laubmoose Europas. Phytologia 65: 424–436. In: Rabenhorst L. (ed.) Kryptogamenflora von Zander R. H. (1993) Genera of the Pottiaceae: Deutschland, O¨ sterreich und der Schweiz, vol. 4 mosses of harsh environments. Bull. Buffalo Soc. (Erga¨ nzungsband). Akademische Verlagsgesell- Nat. Sci. 32: 1–378. schaft, Leipzig. Nadot S., Bajon R., Lejeune B. (1994) The chlo- roplast gene rps4 as a tool for the study of Address of the authors: O. Werner, R. M. Ros, Poaceae phylogeny. Plant Syst. Evol. 191: 27–38. M. J. Cano, J. Guerra (E-mail: [email protected]) Noguchi A. (1988) Illustrated moss flora of Japan. Departamento de Biologı´ a Vegetal (Bota´ nica), Part 2. The Hattori Botanical Laboratory, Facultad de Biologı´ a, Universidad de Murcia, Hiroshima. Campus de Espinardo, E-30100-Murcia, Spain.