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Cross-species amplification of mitochondrial DNA sequence-tagged-site markers in conifers: the nature of polymorphism and variation within and among species in Picea

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Abstract

Primers previously developed to amplify specific non-coding regions of the mitochondrial genome in Angiosperms, and new primers for additional non-coding mtDNA regions, were tested for their ability to direct DNA amplification in 12 conifer taxa and to detect sequence-tagged-site (STS) polymorphisms within and among eight species in Picea. Out of 12 primer pairs, nine were successful at amplifying mtDNA in most of the taxa surveyed. In conifers, indels and substitutions were observed for several loci, allowing them to distinguish between families, genera and, in some cases, between species within genera. In Picea, interspecific polymorphism was detected for four loci, while intraspecific variation was observed for three of the mtDNA regions studied. One of these (SSU rRNA V1 region) exhibited indel polymorphisms, and the two others (nad1 intron b/c and nad5 intron1) revealed restriction differences after digestion with Sau3AI (PCR-RFLP). A fourth locus, the nad4L-orf25 intergenic region, showed a multibanding pattern for most of the spruce species, suggesting a possible gene duplication. Maternal inheritance, expected for mtDNA in conifers, was observed for all polymorphic markers except the intergenic region nad4L-orf25. Pooling of the variation observed with the remaining three markers resulted in two to six different mtDNA haplotypes within the different species of Picea. Evidence for intra-genomic recombination was observed in at least two taxa. Thus, these mitotypes are likely to be more informative than single-locus haplotypes. They should be particularly useful for the study of biogeography and the dynamics of hybrid zones.

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Acknowledgements

We thank S. Plante and N.O. Tremblay (C.R.B.F., Univ. Laval) for valuable help in the laboratory, S. Boisclair and S. Senneville (C.R.B.F., Univ. Laval) for help with the collection of samples, and two anonymous reviewers for constructive comments and suggestions to improve the manuscript. This research was supported by grants from Fonds Québécois de la Recherche sur la Nature et les Technologies (F.Q.R.N.T.), the National Sciences and Engineering Research Council of Canada (N.S.E.R.C.), and the Canadian Forest Service (C.F.S.) to J. Bousquet, J. Beaulieu and N. Isabel, and an international fellowship from the Québec Ministry of Education to J.P. J.-C.

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Authors and Affiliations

  1. Chaire de recherche du Canada en génomique forestière et environnementale and Centre de recherche en biologie forestière, Pavillon Charles-Eugène-Marchand, Université Laval, Sainte-Foy Québec, Canada, G1K 7P4

    J. P. Jaramillo-Correa, J. Bousquet, J. Beaulieu, N. Isabel & M. Bouillé

  2. Natural Resources Canada, Canadian Forest Service – Laurentian Forestry Centre, 1055 du P.E.P.S., P.O. Box 3800, Sainte-Foy Québec, Canada, G1V 4C7

    J. Beaulieu & N. Isabel

  3. Direction de la recherche forestière, Ministère des ressources naturelles du Québec, 2700, rue Einstein, Sainte-Foy, Québec, Canada, G1P 3W8

    M. Perron

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  1. J. P. Jaramillo-Correa
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Correspondence to J. Bousquet.

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Communicated by D.B. Neale

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Jaramillo-Correa, J.P., Bousquet, J., Beaulieu, J. et al. Cross-species amplification of mitochondrial DNA sequence-tagged-site markers in conifers: the nature of polymorphism and variation within and among species in Picea. Theor Appl Genet 106, 1353–1367 (2003). https://doi.org/10.1007/s00122-002-1174-z

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