Abstract
Amplified fragment length polymorphism (AFLP) fingerprinting and three different plastidic DNA regions (rpl16, rps16, atpF-atpH) were used to investigate species identity in the genus Wolffiella. For this purpose, clones (67 in total) belonging to all ten species were selected. Almost all the species were represented by more than one clone. The fingerprinting technique, AFLP, clearly distinguished the species, W. caudata, W. gladiata, W. neotropica, W. rotunda, and W. welwitschii. Apart from confirming the molecular identity of these five species, the plastidic markers could delineate two additional species, W. hyalina and W. denticulata, although the conclusion concerning the latter is restricted by the availability of only one clone. The efficiency of the plastid-derived markers in identifying the number of species-specific clusters followed the sequence rps16 > rpl16 > atpF-atpH. The species W. lingulata, W. oblonga, and W. repanda could not be identified by any of the molecular methods presented here, but could be strictly defined on a morphological basis. In several clones, high amounts of genetic admixtures between different species were detected. Further, simulation studies demonstrated that these clones are genetic hybrids. This might be one of the obstacles in molecular identification of species in the genus Wolffiella.
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Acknowledgements
We thank the late Elias Landolt (1926–2013) for his essential contribution at the beginning of the present project and for providing majority of the clones. We also thank Ms Katja Kuehdorf for experimental support in the initial experiments. KSS is thankful to Science and Engineering Research Board, Government of India, Fast Track Young Scientist project for the support.
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Online Resource 1. 0/1 matrix of AFLP investigations of clones of Wolffiella.
Online Resource 2. Alignment of sequences related to Fig. 5a.
Online Resource 3. Alignment of sequences related to Fig. 5b.
Online Resource 4. Alignment of sequences related to Fig. 5c.
Online Resource 5. Alignment of sequences related to Fig. 6.
Online Resource 6. Cluster assignment of Wolffiella clones according to the ten independent runs from Structure for K = 9; run with highest lnPD highlighted in bold red.
Online Resource 7. Correlations between the maximum intraspecific distances and the minimum interspecific distances (K2P) for the regions rpl16, rps16, and atpF-atpH as well as all possible combinations.
Online Resource 8. Number of haplotypes per plastidic marker and for all possible combinations of the three markers for the sequenced Wolffiella clones. In addition, the species specificity of the haplotypes is indicated in an overview table.
Online Resource 9. Recalculated UPGMA tree based on atpF-atpH sequences from all 21 clones of Wolffiella investigated by Wang et al. (2010). In comparison with the published tree (Fig. 3 in Wang et al. 2010), also clones of W. denticulata 8221 and W. hyalina 8640 were included and the sequence of W. rotunda 9072 was revised.
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Bog, M., Landrock, M.F., Drefahl, D. et al. Fingerprinting by amplified fragment length polymorphism (AFLP) and barcoding by three plastidic markers in the genus Wolffiella Hegelm. Plant Syst Evol 304, 373–386 (2018). https://doi.org/10.1007/s00606-017-1482-z
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DOI: https://doi.org/10.1007/s00606-017-1482-z