STUDYBlackwell OF Science, LtdOxford, UKBOJBotanical Journal of the Linnean Society0024-4074The Linnean Society of London, 2005? 2005 TORTUL149? A SUBULATA COMPLEX333350 M.Original J. CANO Article ET AL. Botanical Journal of the Linnean Society, 2005, 149, 333–350. With 7 figures A morphometric and molecular study in Tortula subulata complex (Pottiaceae, Bryophyta) MARÍA J. CANO*, OLAF WERNER and JUAN GUERRA Departamento de Biología Vegetal (Botánica), Facultad de Biología, Universidad de Murcia, E-30100, Spain Received March 2005; accepted for publication June 2005 Specimens belonging to Tortula subulata complex (T. inermis, T. mucronifolia and T. subulata) were analysed using a combination of morphometric methods based on quantitative characters [principal component analysis (PCA), dis- criminant analysis (DA); 76 samples)] and molecular methods (ITS1 – 5.8S rRNA gene and ITS2; 47 samples) to assess patterns of morphological and molecular differentiation within this complex of taxa. The study shows that four species can be recognized: T. mucronifolia, T. subulata, T. inermis and T. subulata var. angustata with bistratose border, which is elevated to the species rank as T. schimperi nom. nov. The most valuable quantitative characters for identification of these species are the strata number of the marginal laminal cells, the ratio of middle marginal laminal cell width/middle marginal laminal cell length, basal membrane of peristome length, middle laminal cell width and papillae number on the middle laminal cells. The internal transcribed spacer (ITS) data suggest that this group of taxa is not monophyletic, T. mucronifolia being close to Protobryum bryoides (= Tortula protobryoides). The remaining species seem to be a monophyletic group, T. schimperi being the sister group of the clade composed by T. inermis and T. subulata. T. subulata is considered to be of high morphological variability, for which ITS sequences did not resolve the internal relationships. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society, 2005, 149, 333–350. ADDITIONAL KEYWORDS: bryophytes – ITS sequence – molecular systematics – taxonomy. INTRODUCTION not been monographed or critically revised for any wide region; the only floristic treatments are included The genus Tortula Hedw. represents one of the most in a diverse range of flora. Outstanding treatments complex and diverse genera in terms of morphological are those of Steere (1937) for the flora of North Amer- variation in the family Pottiaceae. Its taxonomic cir- ica and north Mexico, Lawton (1971) for the Pacific cumscriptions have been controversial during the last north-west of North America, Nyholm (1989) for Scan- two centuries, and there has been no consensus about dinavia and Finland, Mishler (1994) for Mexico, and which species or even other genera should be included Smith (2004) for Great Britain and Ireland. According in it. Zander (1989, 1993), in his new classification of to the treatment by Zander (1993), the genus Tortula the genera of Pottiaceae, recognized genera such as includes 141 species (Crosby et al., 1999). Chenia R. H. Zander, Dolotortula R. H. Zander, From the morphological study carried out on Hennediella Paris, Hilpertia R. H. Zander, Sageno- Tortula for the Flora Briofítica Ibérica (Cano, 2004), tortula R. H. Zander, Stonea R. H. Zander or Syntri- we identified many taxonomic problems in the species chia Brid. as segregates of Tortula and included taxa related to Tortula subulata. These difficulties are the that traditionally were placed in other genera such as result of overlapping between the characters usually some species of Pottia Ehrh. ex Fürnr., Phascum used and the inability to fit some forms found to any Hedw. (e.g. Phascum cuspidatum Hedw.) and the taxa described. This assemblage of taxa does not cur- genus Desmatodon Brid. With the exception of the rently have any taxonomic status, but it shares a com- treatment of Brotherus (1924), the genus Tortula has bination of morphological characters that differentiate it from the remaining of taxa of the genus Tortula. *Corresponding author. E-mail: [email protected] These include large leaves, (2.2)2.5–5.5 mm in length, © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society, 2005, 149, 333–350 333 334 M. J. CANO ET AL. without hair-point (generally mucronate or apiculate) which he tried to classify all the variation observed. and with a large capsule with twisted peristome from Consequently, he described new species, and accepted a high tessellated basal membrane similar to those a great number of varieties and forms besides those present in the genus Syntrichia. According to our pre- previously described, recognizing three sections, vious morphological study (Cano, 2004), this complex namely section Vulgatae (including T. subulata with of species could be represented by three species: six varieties and one form plus T. serrulata Warnst., Tortula inermis (Brid.) Mont., Tortula mucronifolia nom. illeg. = T. crenulata Warnst., with two varieties Schwägr. and T. subulata, although numerous and two forms), section Intermediae (including Tortula infraspecific taxa have been described of T. subulata. graeffii with two varieties and one form plus Tortula Thus, Zander (1993) includes in T. subulata two sub- bürgeneri Loeske) and sect. Levifoliae (which included species, 13 varieties and three forms, although the T. mucronifolia with three varieties and six forms). number according to van der Wijk, Margadant & The current infrageneric classification of Zander Florschütz (1969) is close to 30. In recent floral (1993) of this complex of species is also rather confus- descriptions (Nyholm, 1989; Cortini-Pedrotti, 2001; ing, because Tortula inermis is classified in the Smith, 2004), only three infraspecific taxa of Tortula genus Syntrichia [S. inermis (Brid.) Bruch], Tortula subulata are usually recognized [T. subulata var. subulata is included in Tortula sect. Tortula, and angustata (Schimp.) Kindb., Tortula subulata var. gra- T. mucronifolia is included in Tortula sect. Pottia effii Warnst. and T. subulata var. subinermis (Bruch & (Rchb.) Kindb., which Ochyra, Zarnowiec & Bednarek- Schimp.) Wilson]. Most authors separate T. subulata Ochyra (2003) renamed as Tortula sect. Cuneifoliae var. angustata from other varieties of T. subulata (Schimp.) Ochyra, because Pottia does not seem to be based on its lingulate-lanceolate leaves, crenulated the oldest available name for this group, Barbula sect. near the apex, acuminate, strongly developed border Cuneifoliae Schimp. being the priority. extending almost to the apex and upper and middle Tortula inermis, placed in the genus Syntrichia by cells 12–16 mm wide, with conspicuous papillae (Cort- Zander (1993), was considered in Tortula by Gallego ini-Pedrotti, 2001; Smith, 2004). Only Nyholm (1989) (2002) and Cano & Gallego (2003) after the study of distinguished this taxon by its bistratose leaf border, the type material and numerous specimens from the which is also drawn in Dixon (1924). Tortula subulata Mediterranean basin. This was based on possession var. subinermis is characterized by obtuse leaf apex, of a differentiated dorsal costal epidermis, a semicir- mucronate, poorly developed border ceasing at about cular shape in the ventral stereid band and a yellow mid-leaf, and upper and middle cells 12–16 mm wide, to orange KOH reaction of the upper laminal cells. In with conspicuous papillae, and Tortula subulata var. contrast, Syntrichia shows an undifferentiated dorsal graeffii by its acute or acuminate leaf apex, apiculate, costal epidermis, a lunate ventral stereid band and with poorly developed border from base to upper third, red KOH reaction. In addition, a molecular study, and upper and middle cells 16–28 mm, with inconspic- based on chloroplast gene rps4 and the rps4–trnS uous papillae. T. inermis var. submarginata Schiffn., a spacer, showed the inclusion of T. inermis in the variety with laminal marginal cells and leaf shape genus Tortula, and that T. subulata and T. inermis closer to T. inermis but with recurvature of the leaf form a well-supported sister clade to the remaining margin and habit of the plant when dry closer to Tortula species included in that study (Werner et al., T. subulata, has been recorded in some Mediterranean 2003). countries (Düll, 1992; Cano, 2004). The objectives of this paper are to evaluate (1) All of these taxa show a Holarctic distribution, the best quantitative taxonomic characters, (2) the with a punctuated disjunction in New Zealand recognizable species within this group based both on (T. mucronifolia). quantitative and on qualitative characters, and (3) Historically, the Tortula subulata complex has been monophyly and proper circumscription of this group of included in different genera and suprageneric taxa. species and the internal relationship between them Some authors have included this complex with the based on internal transcribed spacer (ITS) sequences. species presently recognized in the genus Syntrichia, e.g. Schimper (1876) [Barbula subgen. Syntrichia (Brid.) Schimp., Barbula sect. Subulatae Bruch & MATERIAL AND METHODS Schimp.], or Mönkemeyer (1927) [Syntrichia sect. Zygotrichia (Brid.) Mönk.]. Others have included it in MORPHOMETRIC ANALYSIS the genus Tortula, e.g. Brotherus (1924) [Tortula sect. A total of 76 herbarium specimens (Appendix) were Zygotrichia (Brid.) Mitt.] or more recently Corley et al. studied to span the morphological variability and geo- (1981) [Tortula sect. Tortula Broth.].