Arch. Biol. Sci., Be�gr�de, 61 (1), 85-91, 2009 DOI:10.2298/ABS0901085S GENETIC DIVERSITY AND PHYLOGEOGRAPHY OF THE RARE RIPARIAN MOSS DICHELYMA CAPILLACEUM (WITH.) MYR. INFERRED FROM TRNL-F PLASTID DNA SEQUENCES M. SaboVljeVIć1 and J.-P. Frahm2 1Institute of Botany and Jevremovac Botanical Garden, Faculty of Biology, University of Belgrade, 11000 Be�gr�de, Serbi� 2AG Bryologie, Nees Institut für Biodiversität der Pflanzen, RFW Universität Bonn, D-53115 Bo��, Germany Abstract — The ge�etic relatio�ships of Dichelyma capillaceum (With.) Myr. �re studied from ch�oroplast seque�ces of the tr�L-� regio�. O� the b�sis of the mo�ecular d�t� obt�i�ed, the German populatio� can be co�sidered to be derived from Scandinavian (Swedish) r�ther than North American populatio�s. To judge from the ge�etic distances betwee� the Swedish and German populatio�s, the sep�r�tio� must h�ve occurred � �o�g time �go. Key words: Mosses, Dichelyma capillaceum, ge�etic relatio�ship, phy�ogeogr�phy, re�ict�ess Udc582.32:581.9:575.2 INTRODUCTION There h�ve bee� �o reports of European fossi� or subfossi� fi�ds of D. capillaceum (e.g., Jovet-Ast, The dioecious moss Dichelyma capillaceum (With.) 1967; Dickso�, 1973), and �t �e�st �o Qu�ternary Myr. (�o���ti����nalace�e) h�s � very sc�ttered �mphi- subfossi�s of this species h�ve bee� fou�d i� North Atlantic range (�ig. 1), m�inly i�c�udi�g the �orth- Americ� either (Miller, 1980; Jansse�s, 1983). e�ster� p�rt of North Americ� and Scandinavi� (Crum and A�derso�, 1981; Ireland, 1989; Nyho�m, Dichelyma capillaceum is � yellowish-brow� to 1960; Toivo�e�, 1972). I� Europe, most of the gree� medium-sized p�euroc�rpous �qu�tic moss populatio�s (19) �re situ�ted i� Souther� Swede�, growi�g i� small tufts up to 10 cm �o�g. Le�ves but they �re o� the dec�i�e there (ECCB, 1995; �re erect-spre�di�g, s�ight�y falc�te, seco�d, and Hylander, 1998). E�sewhere, � very small �umber of lanceolate. The �erve is excurre�t i� � �o�g �rist�te populatio�s �re k�ow� from �inland, Poland, and poi�t �bout ½ the �e�gth of the �e�f. C�psu�es �re extreme�y r�re and immersed. The species is almost K�re�i� i� Russi� (Kotirant� et al., 1998). This moss without exceptio� steri�e i� Europe, and it is sus- cannot be fou�d i� Esto�i�, and populatio�s k�ow� pected to h�ve �ost its �bi�ity to produce c�psu�es from De�m�rk, Italy, and �rance (all but o�e) �re due to dec�i�i�g h�bit�t quality and freque�cies of �ssumed to be exti�ct (Allorge and Jovet-Ast, 1948; male plants (Hede�äs et al., 1996). Curre�t spre�d- ECCB, 1995). No d�t� o� rece�t populatio�s from i�g occurred �sexually with fr�gme�ts of you�g Poland and K�re�i� �re �v�ilab�e. E�ch populatio� branches. No specialized veget�tive dispersal u�its from Germany and the o�e from �rance seem to �re k�ow� i� D. capillaceum, but plants can be be still prese�t. The records from the Netherlands, dispersed veget�tive�y, either �t � �imited �ocality Greece, and Sici�y d�te from 1760, 1851, and 1888, through e�o�g�tio� of sto�o�iferous shoots th�t �re respective�y (Touw, 1989; Presto�, 1984; Di� et al., sometimes see� firm�y �tt�ched to the substr�te or 1987), but �o�e of these h�s bee� verified. Thus, the withi� the w�tercourse by fr�gme�t�tio� (Toivo�e�, �ctual ce�ters of distributio� of this r�re and e�dan- 1972). There is �o evide�ce of �o�g-distance disper- gered species �re �oc�ted i� �orthe�ster� North sal, and the m�i� dispersal vector seems to be stre�m Americ� and Norther� Europe. w�ter. 85 86 M. SABOVLJEVIć and J.-P. �RAHM Fig. 1. The world distribution of Dichelyma capillaceum. The moss i�h�bits rip�rian h�bit�ts, growi�g o� and St�pper, 1998). Eve� though the species is cited tree b�rk and rocks. �s exc�usive�y steri�e over almost all of its prese�t range i� Europe, o�e herb�rium specime� from I� Europe, this species is tre�ted �s vu��er- 1923 be�rs sporophytes. �b�e and is i�c�uded i� Appe�dix I of the Ber� Co�ve�tio� and i� A��ex 2 of the EC H�bit�ts and A populatio� w�s rece�t�y discovered �g�i� i� Species Directive. Germany. However, it is �ot c�e�r if it sett�ed the Dichelyma capillaceum w�s fou�d i� Germany prese�t h�bit�t �s � resu�t of �o�g-distance dispersal for the first time i� � small village �e�r the tow� of from North American or Scandinavian relatives or Brüh� �t the begi��i�g of the 20th ce�tury (Br�sch, w�s prese�t there for � �o�g time alre�dy. Co�sideri�g 1923). The �ocality is c�. 1500 km �s the crow f�ies th�t the species h�s �ot bee� see� for dec�des with �w�y from the m�i� European range i� Scandinavi� sporophytes �cross its range and �o prop�gu�es for (Swede�). �e�d (1958) cited the �ocality �g�i� with- �o�g-range spre�di�g �re k�ow�, we here ex�mi�e out co�firmi�g the existe�ce of the populatio�s and the questio� of relatio�ships with represe�t�tives i� me�tio�ed two other records for E�ster� Germany the two prese�t ce�ters of its distributio�. (S�gan and Westprußie�), t�ke� from Mö�kemeyer (1927). Se�rches to fi�d German populatio�s f�i�ed Phy�ogeogr�phical methods h�ve m�de it pos- over the ye�rs, and Düll (1980) st�ted th�t the spe- sib�e to e�ucid�te the effects of large-scale histori- cies is impossib�e to fi�d due to landsc�pe change cal eve�ts (such �s P�eistoce�e c�im�te change) o� and finally co�sidered it exti�ct (Düll, 1994). The the distributio� and subdivisio� of biot� and put species w�s re-fou�d i� the steri�e st�ge i� 1997, i�to co�text the ro�e of reproductive bio�ogy i� the more than 70 ye�rs �fter previous records (�r�hm ge�etic structuri�g of species. GENETIC DIVERSITY AND PHYLOGEOGRAPHY OF DICHELYMA CAPILLACEUM (WITH.) MYR. 87 MATERIAL AND METHODS Table 1. Specimens used in this study, with GenBank accession numbers. Ch�oroplast m�rkers �re ofte� successfully �pp�ied Species Origi� i� plant phy�ogeogr�phy rese�rch �ot only bec�use Swede�, Vöxjö, they �re better co�served, but also bec�use ge�es of Dichelyma capillaceum �eg. K. Hylander ch�oroplast RNAs h�ve sp�cers and i�tro�s th�t �re Dichelyma capillaceum Swede�, Sandvike�, usefu� for studyi�g differe�ces �mo�g both popula- �eg. K. Hylander tio�s and ge�er�. Wide�y used is the high�y v�ri�b�e Dichelyma capillaceum Swede�, Närke, �eg. L. Hede�äs USA, Missouri, sp�cer positio�ed betwee� �ysi�e-tRNA-Exo� trnL Dichelyma capillaceum �eg. ����������B. Summers ��an�������������d C. D. Scott (UAA-3’) and the tRNA ge�e for phe�ylanali�e trn� USA, New H�mpshire, (GAA) (e.g., T�ber�et et al., 1991; Ke�ch�er, 2000). A Dichelyma capillaceum �eg. B. Alle� previous survey demo�str�ted th�t this fr�gme�t is Dichelyma capillaceum USA, M�i�e, �ike�y to exhibit an i�form�tive �mou�t of diversity �eg. ����B. A��lle� Germany, Brüh�, (Voge� et al., 1996). It h�s also bee� show� th�t the Dichelyma capillaceum �eg. S�bov�jević and �r�hm ch�oroplast is m�ternally (i.e., u�ip�re�tally) i�her- Dichelyma falcatum USA, from Ge�Bank�� A�191505 ited i� most plants (Voge� et al., 1998). Dichelyma falcatum USA, from Ge�Bank�� A�191506 We extr�cted DNA from herb�rium specime�s Dichelyma uncinatum USA, from Ge�Bank A�191504 usi�g the 2x CTAB (hex�decy�trimethylammo�ium bromide) method �s described by S�bov�jević et al. (2005). RESULTS Amp�ific�tio� of the trnL-F regio� (T�ber�et et The resu�ts obt�i�ed i� PAUP analyses c�e�r�y show al., 1991) w�s c�rried out usi�g the forw�rd primer th�t the German populatio� is distant from all C and the reverse primer � s�ight�y modified for the other tested populatio�s. Si�ce the species is bryophytes �ccordi�g to Meiß�er et al. (1998) and quite r�re and i� danger of exti�ctio�, three North Sugiur� et al. (2003). European populatio�s and three from the �orthe�st- er� p�rt of North Americ� were chose� for comp�ri- The alig�me�t of seque�ces w�s cre�ted manu- so� with the German o�e. Additionally, the trnL-� ally with the alig�me�t editor A�ig� 32 (H�pper�e, seque�ces of two specime�s of Dichelyma falcatum 2003), and PAUP4.0b10 (Swoford, 2002) w�s used and o�e of D.