Potentials and limitations of the cross-species transfer of nuclear microsatellite marker in six species belonging to three sections of the genus Populus L.
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The genus Populus is classified into six different sections, and depending on the declaration of hybrids, the number of species varies between 22 and 85. Species within one section, and sometimes between sections, are crossable to each other, resulting in many naturally but also artificially produced hybrids. Morphological attributes for a clone characterisation are often difficult to evaluate when different poplar species or even hybrids are crossed; thus, molecular markers are needed to characterise the different species. Taking advantage of the large microsatellite resource developed for Populus trichocarpa, however, amplification of these microsatellite markers in other Populus species either often fails, or in the case of amplification, unrelated genomic regions are amplified. To meet this obvious problem of the species transferability of microsatellite markers, in total, 305 microsatellite loci, mainly from P. trichocarpa but also few from Populus tremuloides and Populus nigra, were tested for their transferability to diverse genotypes of six species belonging to three sections of the genus Populus. Ultimately, 209 microsatellite loci could be amplified with varying sizes in the different species. The PCR products of selected loci were separated in a polyacrylamide gel and sequenced to assure that the expected loci were derived from the database genome of P. trichocarpa. The present results constitute a large study for microsatellite transferability for Populus species. The documented microsatellite loci can be applied to species-, hybrid- and clone-specific diagnostic approaches or as universal markers for comprehensive ecological studies.
KeywordsPopulus tremula P. nigra Cross-species amplification DNA sequencing Molecular marker Poplar
We like to thank our technical assistants Katrin Groppe for the in silico analyses and Caren Heitmann for the realisation of the PAA gels as well as Dr. Mirko Liesebach, Dr. Georg von Wühlisch and our other colleagues for the collection of different poplar material. Further, we are grateful to Dr. Birgit Kersten (Thünen-Institute of Forest Genetics Grosshansdorf) and Doreen Pahlke (MPI for Molecular Plant Physiology, Potsdam, Germany) for data integration into GabiPD (www.gabipd.org) and sequence submission to the NCBI GenBank database.
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