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A Phylogeographic Split in Buxus balearica (Buxaceae) as Evidenced by Nuclear Ribosomal Markers: When ITS Paralogues Are Welcome

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Abstract

Sequences from the ribosomal nuclear internal transcribed spacers (ITS) have been widely used to infer evolutionary hypotheses across a broad range of living organisms. Intraspecific sequence variation is assumed to be absent or negliable in most species, but few detailed studies have been conducted to assess the apportionment of ITS sequence variation within and between plant populations. Buxus balearica was chosen as a model species to assess the levels of infraspecific and intragenomic ITS variation in rare and endangered species occurring in disjunct populations around the Mediterranean basin. Intragenomic polymorphic sites were detected for western and eastern accessions of B. balearica and in two accessions of the sister species B. sempervirens. Overall, 19 different ribotypes were found in B. balearica after sequencing 48 clones, whereas 15 ribotypes were detected in 19 clones of B. sempervirens. The integrity and secondary structure stability of the ribosomal sequences suggest that they are not pseudogenes. The high number of ribotypes recovered through cloning suggested that some sequences could be chimeric or generated in vivo by partial homogenization through gene conversion or unequal crossing-over. Average sequence divergence among B. balearica clones was 0.768%, and the most divergent sequences differed by 1.62%. Available evidence does not suggest that B. balearica paralogues have been obtained from other extant Buxus species through interspecific hybridization. The presence of several ribosomal sequences in box implies that the molecular forces driving the concerted evolution of this multigene family are not fully operational in this genus. Phylogenetic analyses of cloned ITS sequences from B. balearica displayed very poor resolution and only two clades received moderate bootstrap support. Despite the marked intragenomic sequence divergence found, ribosomal data suggest a clear phylogeographic split in B. balearica between western and eastern accessions. The distinct, nonchimeric sequences that are postulated as being present in each biogeographic group suggest that box populations from Anatolia (eastern Mediterranean) are relict.

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Acknowledgments

Jesús Charco helped to locate Moroccan populations, and Mario García and José M. Gutiérrez carried out fieldwork assistance in Morocco and Turkey, respectively. Dr. Gianni Bacchetta sent us the Sardinian samples. Domingo Alcaraz assisted us in several ways. G. Nieto-Feliner and two anonymous referees offered useful suggestions on early versions of the manuscript. Jesús Manzanilla helped with the phylogenetic analyses. The Consejería de Medio Ambiente de la Junta de Andalucía granted us permission to sample in the Andalusian populations. This work was partially funded by projects BOS2001-0610, REN2001-3506-C02-01, and MMA 034/2002. A.L. was supported by a FPU fellowship (Spanish Ministry of Education, MECD).

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  1. Jardí Botánic, Universidad de Valencia, c/ Quart 80, E-46008, Valecia, Spain

    Josep A. Rosselló, Raúl Cosín & Arántzazu Molins

  2. IMEDEA, CSIC, c/ Miguel Marqués 21 07190, E-07190, Esporles, Mallorca, Balearic Islands, Spain

    Amparo Lázaro

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  1. Josep A. Rosselló
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Rosselló, J.A., Lázaro, A., Cosín, R. et al. A Phylogeographic Split in Buxus balearica (Buxaceae) as Evidenced by Nuclear Ribosomal Markers: When ITS Paralogues Are Welcome. J Mol Evol 64, 143–157 (2007). https://doi.org/10.1007/s00239-005-0113-4

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