Juan Guerra

Molecular phylogeny of Pottiaceae (Musci) based on chloroplast rps4 sequence data

Comparative sequencing of the chloroplast rps4 gene was used to reconstruct the phylogenetic relationships within the family Pottiaceae (Musci). The results confirm that Ephemerum spinulosum, Splachnobryum obtusum, Goniomitrium acuminatum and Cinclidotus fontinaloides are clearly positioned within the Pottiaceae and that Hypodontium dregei is not a member. At subfamily level, the data support the subfamily Pottioideae as being a monophyletic clade. The Trichostomoideae are probably paraphyletic. Neither the subfamily Chionolomideae, represented in this study by Pseudosymblepharis schimperiana, nor the subfamily Erythrophyllopsoideae, represented by both known species, Erythrophyllastrum andinum and Erythrophyllopsis fuscula, are supported by the sequence data. The Timmielloideae should be excluded from the Pottiaceae. The Merceyoideae, represented in this study by Scopelophila cataractae, might form a sister clade to all other Pottiaceae, but their position is not fully resolved. At the genus level, Barbula is clearly polyphyletic since Barbula bolleana and Barbula indica appear in a clade clearly separated from Barbula unguiculata. Pottiopsis caespitosa and Leptobarbula berica are placed within the Trichostomoideae. Likewise, the genera Gymnostomum and Anoectangium are excluded from the Pottioideae and placed within the Trichostomoideae. Leptophascum leptophyllum is closely related with Syntrichia;Aloina is not closely related to Tortula or Crossidium. Evidence of a clade within the Pottioideae, formed of Leptodontium and Triquetrella, is provided.

Direct amplification and NaOH extraction: two rapid and simple methods for preparing bryophyte DNA for polymerase chain reaction (PCR)

Abstract PCR (polymerase chain reaction) has become one of the most important techniques used in molecular systematics. Generally, the methods applied to isolate DNA for PCR amplification depend on multiple steps to isolate and clean the final product. This involves considerable effort and time when many samples have to be processed and poisonous organic solvents are often needed (phenol, chloroform etc.). In this contribution, two very rapid and simple techniques intended for use in higher plants are shown to be useful in bryophyte molecular biology: direct amplification from plant tissues and the NaOH extraction method. In 15 of the 17 investigated bryophytes (two hepatics and 15 mosses) the trn LUAA intron of the chloroplast DNA was successfully amplified by the direct approach, while the NaOH extraction method gave amplifiable DNA in all 17 species. DNA amplified by both methods was successfully used in cycle sequencing.

Preliminary investigation of the systematics of Didymodon (Pottiaceae, Musci) based on nrITS sequence data

Abstract A phylogenetic analysis of species of Didymodon from the Mediterranean area, Macaronesia, and Southwestern and Central Asia is presented. The ITS1, 5.8S rRNA gene, and ITS2 for 30 species have been sequenced, which represent about 25% of the recognized species in the genus. The molecular data confirm the monophyly of Didymodon as clearly separated from Barbula. The transfer of Didymodon giganteus to Geheebia, of D. sinuosus to Oxystegus, and D. australasiae to Trichostomopsis is not supported by the molecular data. The phylogenetic affinities of the species generally do not correspond with the infrageneric classification proposed for the genus based on morphological characteristics. The only monophyletic section is Asteriscium, but only after the inclusion of D. bistratosus.

New national and regional bryophyte records, 41

Dicranum spurium has been recorded for the first time in Croatia. In neighbouring countries it is known from Italy, Slovenia, Hungary, Bosnia and Herzegovina, and Montenegro (Cortini Pedrotti, 1992 ; Sabovljevic´ et al., 2008 ; Papp et al., 2010). In Hungary, it is treated as VU (vulnerable) (Papp et al., 2010), in Montenegro as DD (data-deficient) (Sabovljevic´ et al., 2004), and further in the Balkans as EN (endangered) in Romania (Stefanut¸ & Goia, 2012). Dicranum spurium was found on Papuk Mountain (northeastern Croatia). Ninety-five per cent of Papuk Mountain is covered by forest vegetation, the yearly mean temperature is between 8 and 11C, and annual precipitation varies between 800 and 1300 mm. Almost the whole of the mountain is protected as a Nature Park. The locality in Svinjarevac where D. spurium occurred is beech forest (Fagus sylvatica L.), unique in Croatia, with a dense cover of Sphagnum quinquefarium (Braithw.) Warnst. on the forest floor. The bedrock is quartzite and gneiss, sloping between 35 and 45, and exposed to the northwest. This Fagus-Sphagnum quinquefarium forest was spread over 2.4 ha., with a poorly developed layer of herbaceous plants. Sphagnum platyphyllum was recorded during an extensive survey of spruce forests throughout Croatia. It was found in Stirovaca, a plateau in the central part of Velebit Mountain. The locality for this first record of S. platyphyllum for Croatia is just near the border of the Northern Velebit National Park (The whole Velebit Mountain is protected as a Nature Park). According to Dull et al. (1999), Sabovljevic (2006), and Sabovljevic et al. (2008), the species has not been recorded from Croatia, although it is known to occur in neighbouring countries: Italy, Slovenia, Hungary, Bosnia and Herzegovina, Montenegro, and Serbia (Cortini Pedrotti, 1992 ; Sabovljevic et al., 2008 ; Papp et al., 2010). In many of these countries, it is red listed: as VU (vulnerable) in Hungary, Slovenia, and Serbia (Martincicc, 1992 ; Sabovljevic et al., 2004 ; Papp et al., 2010) and further in the Balkans as NT (near threatened) in Romania (Stefanut & Goia, 2012). This part of Velebit Mountain is characterized by cool and long winters, with 1900 mm average annual precipitation and an average annual temperature of 3.5C. The natural vegetation consists of beech-fir and spruce forests and some of the forest stands are still primeval. Sphagnum platyphyllum occurred in a wet spruce (Picea abies (L.) H.Karst.) forest, where it grew in dense carpets covering the forest floor and the banks of shallow depressions (ca 100 m2) filled with water. The collection site in Stirovaca is the only known locality for this unique spruce forest community with peat mosses in Croatia. Sphagnum teres was found at two localities in Croatia, in or near the National Park Plitvicka jezera lakes, but in different habitats within the belt of beech-fir forests. The climate is moderately temperate, with 1500 mm annual precipitation and an average temperature of 7.9C. This is the first record of this species for Croatia, although in the neighbouring countries, it is known from Italy, Slovenia, Hungary and Serbia, and further to the south-east, in FYR Macedonia, Bulgaria and Greece (Papp et al., 2010 ; Sabovljevic et al., 2008). S. teres has been recently recorded as new for Albania (Marka & Sabovljevic, 2011), Bosnia- Herzegovina (Sabovljevic´ et al., 2010) and Montenegro (Martincic, 2006), and is red listed as CR (critically endangered) in Hungary (Papp et al., 2010), and as VU (vulnerable) in Serbia (Sabovljevic et al., 2004). The first locality, Ljeskovacˇke bare, is within the National Park Plitvicka jezera lakes. Here, the species occurred in transitional peat bog belonging to the community Drosero-Caricetum echinatae and occupying an area of ca 1 ha., S. teres was the dominant Sphagnum species. Sphagnum squarrosum Crome, S. palustre L., S. centrale C.E.O.Jensen and S. flexuosum Dozy & Molk. were also relatively abundant. The bog is wet throughout the year, but mostly without open water. Depending on the amount of precipitation, it can be flooded during winter and spring. The bog is not shaded ; it is completely exposed to the sun, but partially overgrown by Molinia coerulea Moench, which is mowed in the management of the Park. Some other species characteristic for this site were Drosera rotundifolia L., Carex echinata Murray, C. lasiocarpa Ehrh., and Menyanthes trifoliata L. The second locality, Vrhovinsko polje, is situated near the eastern border of Plitvicˇka jezera lakes National Park. It is a typical karst field, with the lowest, central part periodically flooded during winter and early spring. It is covered with grassland vegetation of different communities reflecting gradients of water and soil pH. The flora was very rich. In the lowest part of the field were several shallow depressions with a perimeter of 10–20 m overgrown by sedges, mostly Carex vesicaria L. On the edges of these depressions were scattered peat moss hummocks of Sphagnum palustre L., S. capillifolium (Ehrh.) Hedw., and S. subnitens Russow & Warnst. Among them, S. teres (Schimp.) Angstr. also occured, but in much lower abundance.

Genetic variations and migration pathway of Juniperus thurifera L. (Cupressaceae) in the western Mediterranean region

Random amplified polymorphic DNA (RAPD) analysis and length variations in the chloroplast trnL-trnF intergenic spacer were used to analyze the geographic structure of Juniperus thurifera L. populations in Spain and North Africa. The chloroplast DNA analysis indicated three size classes of the spacer, but was uninformative of any geographic pattern. In contrast, the RAPD data based on 127 polymorphous bands clearly showed the existence of three population groups: (a) a group comprising two populations from northeastern Spain and two from southern Spain, (b) a group comprising the rest of the Spanish populations, and (c) a group which consisted of the two Moroccan populations. Analysis of molecular variance (AMOVA) showed that 24% of the variation is found between groups of populations, 15% between populations within groups, and 61% within populations. The results are consistent with a hypothetical migration of J. thurifera starting from central Europe, passing through France and Spain, and finally reaching...

Molecular data Confirm the Presence of Anacolia menziesii (Bartramiaceae, Musci) in Southern Europe and Its Separation from Anacolia webbii

Abstract ISSR (Inter-Simple Sequence Repeat) fingerprint data and nrITS sequences confirm the presence of Anacolia menziesii in Europe. The species is more variable genetically in North America than in Europe. The data show only minor differentiation between the North American and European populations of A. menziesii. Anacolia webbii is morphologically and genetically very similar to Anacolia menziesii, but can be separated from Anacolia menziesii with high confidence based on the ITS and ISSR data. Long distance dispersal is the most likely explanation for the disjunction of Anacolia menziesii between North America and Europe.

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

Abstract. According to chloroplast rps4 sequence data the genus Syntrichia forms a monophyletic clade clearly separated from Tortula, while Pottia is shown to be polyphyletic and the Rhynchostegiae species (with rostrate lid) are very similar to Tortula. Crossidium is probably polyphyletic. The close affinity of Desmatodon, Stegonia, Pterygoneurum and some of the species of Phascum with Tortula is confirmed by the molecular data.

Contribution to the bryophyte flora of Morocco: terricolous and saxicolous bryophytes of the Jbel Bouhalla

Abstract The terricolous and saxicolous bryophytes of the Jbel Bouhalla (Rif Cordillera, Morocco) have been studied. This mountainous system, characterized by basic geology, contains the largest and best conserved forest of Abies pinsapo subsp. maroccana in Northern Africa. The catalogue is composed of 121 taxa, of which 108 are mosses and 13 liverworts. Of these, nine are new to the African continent (Acaulon mediterraneum, Claopodium whippleanum, Gymnostomum lanceolatum, Hedwigia stellata, Orthotrichum cupulatum var. baldacci, Schistidium brunnescens subsp. griseum, S. crassipilum, Scorpiurium sendtneri, Seligeria acutifolia), Eurhynchium schleicheri is new for mainland Africa, and seven are new records for Morocco (Barbula enderesii, Bryum dunense, Campyliadelphus chrysophyllus, Fissidens dubius, Hedwigia ciliata var. leucophaea, Pleuridium acuminatum, Pseudoleskeella catenulata).

Contribution to the bryophyte flora of Morocco: the Jbel Toubkal

Abstract The bryophyte flora of the Jbel Toubkal, included in the Toukbal National Park (Morocco), is catalogued. Only 12 bryophytes had been reported previously from this mountain, the highest in northern Africa. As a result of this study, the catalogue consists of 101 taxa (95 mosses and six liverworts), of which nine species are new for Africa (Amblyodon dealbatus, Brachythecium collinum, Grimmia mollis, Hyocomium armoricum, Pseudoleskeella tectorum, Schistidium cinclidodonteum, Syntrichia bolanderi, Syntrichia calcicola, Tortula guepinii), three species are newly reported for northern Africa (Bryum klinggraeffii, Bryum muehlenbeckii, Schistidium rivulare) and another six are new for Morocco (Bryum mildeanum, Cirriphyllum crassinervium, Fissidens rivularis, Plagiochila porelloides, Rhynchostegiella teneriffae and Weissia levieri).

A revision of the moss genusCrossidium (Pottiaceae) with the description of the new genusMicrocrossidium

A world revision of the genusCrossidiumJur. recognizes 11 species, which are described and discussed in the context of important taxonomic characters. An identification key is provided.C. asirenseFrey & Kürschner is reduced to synonymy withC. davidaiCatcheside, and the geographical range ofC. laevipilumTher. & Trab. is extended to Europe. Phylogenetic trends are interpreted on the basis of two main evolutionary lines associated with the presence or absence of hyaline hair-points on the leaves. A twelfth species is transferred toMicrocrossidiumGuerra & Cano, gen. nov., asM. apiculatum (Magill)Guerra & Cano, comb. nova, because of differences in stem anatomy, peristome configuration, and spore morphology.