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Chloroplast and mitochondrial molecular tests identify European×Japanese larch hybrids

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Abstract

Hybrids between European and Japanese larches combine the properties of both parental species (drought resistance, canker resistance, stem straightness) and exhibit a fast growth rate. They are produced in seed orchards, generally by natural pollination. Seeds are collected and used for afforestation as interspecific hybrids. However, there are no convenient tests to assess the interspecific hybrid proportion. In the present study, we developed diagnostic molecular markers suitable for the individual identification of hybrids, whatever their developmental stage. Our strategy involved testing a combination of maternally inherited markers from the mitochondrial genome (mtDNA) and paternally inherited markers from the chloroplast genome (cpDNA). Hybrids were then identified by the presence of a mitochondrial sequence inherited from one parental species and a chloroplast sequence inherited from the other parental species. To achieve this aim, markers discriminating both parental species were first sought. Amplifications of mitochondrial and chloroplast sequences were performed using specific PCR primers. After testing 33 primer pairs in combination with nine restriction enzymes, we detected one mitochondrial marker, f13 which was amplified in Japanese larch and absent in European larch, and one chloroplast marker, ll-TaqI which showed different restriction patterns depending on the species. A restriction fragment of 601 bp was obtained in Japanese larch while two fragments of 120 bp and 481 bp were observed in European larch. These patterns were found in all 197 individuals tested from the two pure species. These markers were then used for the evaluation of the hybrid proportion in a seed lot produced from seed orchards; this was assessed as between 43% and 53% depending on the parental species. The male and female parental species could be determined for each progeny.

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Acknowledgements

We are grateful to Dominique Scheepers who identified the mtDNA marker and gave us its primer pair sequences. We thank Dominique Jacques for providing us with the seeds from the Halle orchard. We would like to thank Meredith Carter for manuscript correction. We express our thanks to Anne Arcade and Brigitte Demesure for encouragement and valuable discussions. We also thank Gérard Aubard for technical help. This work was supported by the European Union (project LARCH: FAIR-CT98-3354) and the French “Ministère de l’Agriculture et de la Pêche” (DERF grant no. 01.40.12/99).

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Author notes

  1. V. Acheré

    Present address: UMR INRA/UHP 1136, Interactions arbres/micro-organismes, Faculté des Sciences, Université Nancy I, Bd des Aiguillettes, BP 239, 54506, Vandoeuvre Les Nancy, France

  2. P. Faivre Rampant

    Present address: INRA/CNRS, Unité de Recherche en Génomique Végétale, 2 rue Gaston Crémieux, CP 5708, 91057, Evry, Cedex, France

  3. D. Prat

    Present address: EA 3731 Génome et Evolution des Plantes Supérieures, Université Claude Bernard Lyon 1, Bâtiment F.A. Forel, 6 rue Raphaël Dubois, 69622 , Villeurbanne, Cedex, France

Authors and Affiliations

  1. INRA, Unité Amélioration, Génétique et Physiologie Forestières, Avenue de la Pomme de Pin, BP 20619 Ardon, 45166, Olivet, Cedex, France

    V. Acheré, P. Faivre Rampant, L. E. Pâques & D. Prat

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  1. V. Acheré
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Correspondence to D. Prat.

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Communicated by D.B. Neale

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Acheré, V., Faivre Rampant, P., Pâques, L.E. et al. Chloroplast and mitochondrial molecular tests identify European×Japanese larch hybrids. Theor Appl Genet 108, 1643–1649 (2004). https://doi.org/10.1007/s00122-004-1595-y

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