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Horizontal genome transfer as an asexual path to the formation of new species

Nature volume 511, pages 232–235 (2014)Cite this article

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Abstract

Allopolyploidization, the combination of the genomes from two different species, has been a major source of evolutionary innovation and a driver of speciation and environmental adaptation1,2,3,4. In plants, it has also contributed greatly to crop domestication, as the superior properties of many modern crop plants were conferred by ancient allopolyploidization events5,6. It is generally thought that allopolyploidization occurred through hybridization events between species, accompanied or followed by genome duplication6,7. Although many allopolyploids arose from closely related species (congeners), there are also allopolyploid species that were formed from more distantly related progenitor species belonging to different genera or even different tribes8. Here we have examined the possibility that allopolyploidization can also occur by asexual mechanisms. We show that upon grafting—a mechanism of plant–plant interaction that is widespread in nature—entire nuclear genomes can be transferred between plant cells. We provide direct evidence for this process resulting in speciation by creating a new allopolyploid plant species from a herbaceous species and a woody species in the nightshade family. The new species is fertile and produces fertile progeny. Our data highlight natural grafting as a potential asexual mechanism of speciation and also provide a method for the generation of novel allopolyploid crop species.

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Figure 1: Grafting and experimental selection for horizontal transfer of nuclear DNA between tobacco plants.
Figure 2: Grafting and horizontal transfer of nuclear DNA between different species.
Figure 3: Presence of nuclear genes from both Nicotiana tabacum and Nicotiana glauca in Nicotiana tabauca and their Mendelian inheritance.
Figure 4: Phenotype and karyotype of Nicotiana tabauca plants.

References

  1. Comai, L. The advantages and disadvantages of being polyploid. Nature Rev. Genet. 6, 836–846 (2005)

    Article  CAS  Google Scholar 

  2. Sobel, J. M., Chen, G. F., Watt, L. R. & Schemske, D. W. The biology of speciation. Evolution 64, 295–315 (2010)

    Article  Google Scholar 

  3. Madlung, A. Polyploidy and its effect on evolutionary success: old questions revisited with new tools. Heredity 110, 99–104 (2013)

    Article  CAS  Google Scholar 

  4. Leitch, A. R. & Leitch, I. J. Genomic plasticity and the diversity of polyploid plants. Science 320, 481–483 (2008)

    Article  ADS  CAS  Google Scholar 

  5. Paterson, A. H. Polyploidy, evolutionary opportunity, and crop adaptation. Genetica 123, 191–196 (2005)

    Article  CAS  Google Scholar 

  6. Hegarty, M. J. & Hiscock, S. J. Genomic clues to the evolutionary success of polyploid plants. Curr. Biol. 18, R435–R444 (2008)

    Article  CAS  Google Scholar 

  7. Soltis, P. S. & Soltis, D. E. The role of hybridization in plant speciation. Annu. Rev. Plant Biol. 60, 561–588 (2009)

    Article  CAS  Google Scholar 

  8. Joly, S., Heenan, P. B. & Lockhart, P. J. A Pleistocene inter-tribal allopolyploidization event precedes the species radiation of Pachycladon (Brassicaceae) in New Zealand. Mol. Phylogenet. Evol. 51, 365–372 (2009)

    Article  CAS  Google Scholar 

  9. Seidel, C. F. Ueber Verwachsungen von Stämmen und Zweigen von Holzgewächsen und ihren Einfluss auf das Dickenwachsthum der betreffenden Theile. Naturwiss. Ges. Isis Dresden Sitzber. 161–168 (1879)

  10. Küster, E. Über Stammverwachsungen. Jahrb. Wiss. Bot. 33, 487–512 (1899)

    Google Scholar 

  11. Beddie, A. D. Natural root grafts in New Zealand trees. Transact. Proc. R. Soc. New Zeal. 71, 199–203 (1942)

    Google Scholar 

  12. Larson, P. R. The vascular cambium: development and structure. Springer Series in Wood Science 1–725 (Springer-Verlag, 1994)

  13. Bock, R. The give-and-take of DNA: horizontal gene transfer in plants. Trends Plant Sci. 15, 11–22 (2010)

    Article  CAS  Google Scholar 

  14. Mudge, K., Janick, J., Scofield, S. & Goldschmidt, E. E. A history of grafting. Hortic. Rev. (Am. Soc. Hortic. Sci.) 35, 437–493 (2009)

    Article  Google Scholar 

  15. Stegemann, S. & Bock, R. Exchange of genetic material between cells in plant tissue grafts. Science 324, 649–651 (2009)

    Article  ADS  CAS  Google Scholar 

  16. Stegemann, S., Keuthe, M., Greiner, S. & Bock, R. Horizontal transfer of chloroplast genomes between plant species. Proc. Natl Acad. Sci. USA 109, 2434–2438 (2012)

    Article  ADS  CAS  Google Scholar 

  17. Thyssen, G., Svab, Z. & Maliga, P. Cell-to-cell movement of plastids in plants. Proc. Natl Acad. Sci. USA 109, 2439–2443 (2012)

    Article  ADS  CAS  Google Scholar 

  18. Zhang, W. C., Yan, W. M. & Lou, C. H. Intercellular movement of protoplasm in vivo in developing endosperm of wheat caryopses. Protoplasma 153, 193–203 (1990)

    Article  Google Scholar 

  19. Zhang, W.-C. Progress in research on intercellular movement of protoplasm in higher plants. Acta Bot. Sin. 44, 1068–1074 (2002)

    CAS  Google Scholar 

  20. Mursalimov, S. R., Sidorchuk, Y. V. & Deineko, E. V. New insights into cytomixis: specific cellular features and prevalence in higher plants. Planta 238, 415–423 (2013)

    Article  CAS  Google Scholar 

  21. Melaragno, J. E., Mehrotra, B. & Coleman, A. W. Relationship between endopolyploidy and cell size in epidermal tissue of Arabidopsis. Plant Cell 5, 1661–1668 (1993)

    Article  Google Scholar 

  22. Madlung, A. et al. Genomic changes in synthetic Arabidopsis polyploids. Plant J. 41, 221–230 (2005)

    Article  CAS  Google Scholar 

  23. Pontes, O. et al. Chromosomal locus rearrangements are a rapid response to formation of the allotetraploid Arabidopsis suecica genome. Proc. Natl Acad. Sci. USA 101, 18240–18245 (2004)

    Article  ADS  CAS  Google Scholar 

  24. Chester, M. et al. Extensive chromosomal variation in a recently formed natural allopolyploid species, Tragopogon miscellus (Asteraceae). Proc. Natl Acad. Sci. USA 109, 1176–1181 (2012)

    Article  ADS  CAS  Google Scholar 

  25. Xiong, Z., Gaeta, R. T. & Pires, J. C. Homoeologous shuffling and chromosome compensation maintain genome balance in resynthesized allopolyploid Brassica napus. Proc. Natl Acad. Sci. USA 108, 7908–7913 (2011)

    Article  ADS  CAS  Google Scholar 

  26. Yu, H.-S. & Russell, S. D. Occurrence of mitochondria in the nuclei of tobacco sperm cells. Plant Cell 6, 1477–1484 (1994)

    Article  CAS  Google Scholar 

  27. Shepard, J. F., Bidney, D., Barsby, T. & Kemble, R. Genetic transfer in plants through interspecific protoplast fusion. Science 219, 683–688 (1983)

    Article  ADS  CAS  Google Scholar 

  28. Evans, D. A., Wetter, L. R. & Gamborg, O. L. Somatic hybrid plants of Nicotiana glauca and Nicotiana tabacum obtained by protoplast fusion. Physiol. Plant. 48, 225–230 (1980)

    Article  CAS  Google Scholar 

  29. Trojak-Goluch, A. & Berbec, A. Cytological investigations of the interspecific hybrids of Nicotiana tabacum L. × N. glauca Grah. J. Appl. Genet. 44, 45–54 (2003)

    PubMed  Google Scholar 

  30. Greiner, S. & Bock, R. Tuning a ménage à trois: co-evolution and co-adaptation of nuclear and organellar genomes in plants. Bioessays 35, 354–365 (2013)

    Article  CAS  Google Scholar 

  31. Murashige, T. & Skoog, F. A revised medium for rapid growth and bio assays with tobacco tissue culture. Physiol. Plant. 15, 473–497 (1962)

    Article  CAS  Google Scholar 

  32. Galbraith, D. W. et al. Rapid flow cytometric analysis of the cell cycle in intact plant tissues. Science 220, 1049–1051 (1983)

    Article  ADS  CAS  Google Scholar 

  33. Doležel, J., Greilhuber, J. & Suda, J. Estimation of nuclear DNA content in plants using flow cytometry. Nature Protocols 2, 2233–2244 (2007)

    Article  Google Scholar 

  34. Doyle, J. J. & Doyle, J. L. Isolation of plant DNA from fresh tissue. Focus 12, 13–15 (1990)

    Google Scholar 

  35. Moon, H. S., Nicholson, J. S. & Lewis, R. S. Use of transferable Nicotiana tabacum L. microsatellite markers for investigating genetic diversity in the genus Nicotiana. Genome 51, 547–559 (2008)

    Article  CAS  Google Scholar 

  36. Shinozaki, K. et al. The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J. 5, 2043–2049 (1986)

    Article  CAS  Google Scholar 

  37. Sugiyama, Y. et al. The complete nucleotide sequence and multipartite organization of the tobacco mitochondrial genome: comparative analysis of mitochondrial genomes in higher plants. Mol. Genet. Genomics 272, 603–615 (2005)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank the MPI-MP Green Team for help with plant transformation, K. Köhl for providing the hpt vector and S. Ruf (MPI-MP) for discussions. We are grateful to J. Fuchs (IPK Gatersleben) for advice on flow cytometry measurements. This research was financed by the Max Planck Society.

Author information

Author notes

  1. Ignacia Fuentes and Sandra Stegemann: These authors contributed equally to this work.

Authors and Affiliations

  1. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, D-14476 Potsdam-Golm, Germany,

    Ignacia Fuentes, Sandra Stegemann, Daniel Karcher & Ralph Bock

  2. Department of Molecular Biology, Institute of Biotechnology, John Paul II Catholic University of Lublin, Konstantynow 1I, 20-708 Lublin, Poland,

    Hieronim Golczyk

Authors
  1. Ignacia Fuentes
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  2. Sandra Stegemann
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  3. Hieronim Golczyk
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  4. Daniel Karcher
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  5. Ralph Bock
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Contributions

I.F., S.S. and H.G. performed the experiments. All authors participated in data evaluation and experimental design. R.B. conceived the study, and wrote the manuscript.

Corresponding author

Correspondence to Ralph Bock.

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The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Autotetraploidy and chromosome loss in NGT plants.

a, Chromosome preparations of mitotic cells. Along with preparations from each of the two graft partners (Nt-kan:yfp and Nt-hyg), four examples of mitotic cells from four individual progeny plants of the self-pollinated line NGT-2 are shown. Chromosomes are visualized by hot tissue hydrolysis in HCl and staining with toluidine blue. Chromosome numbers are given in parentheses. Chr, chromosomes. b, Chromosome counts for individual seedlings from the two graft partners and the selfed line NGT-2. Mitotic cells from the root tips were analysed. The total number of metaphases investigated was 32 for Nt-kan:yfp (blue), 35 for Nt-hyg (purple) and 86 for NGT-2 (pink). c, Absolute genome size determination by flow cytometry. Leaf samples were mixed with tomato leaves (Solanum lycopersicum, Sl) that served as internal standard, nuclei were isolated and the relative fluorescence intensity of propidium iodide (PI) was measured. Each peak corresponds to a population of nuclei. For each sample, the ratio between the peak of the analysed tobacco line and the peak of the internal standard (4C tomato nuclei) was calculated to determine the absolute genome size. Sample 1: Nt-hyg; sample 2: Nt-kan:yfp; sample 3: seedling from a cross between NGT-1 and wild type; sample 4: seedling from a self-pollinated NGT-2 plant.

Extended Data Figure 2 Meiotic chromosome missegregation in autopolyploid NGT tobacco lines.

a–c,Young flower buds were fixed and stained, either with aceto-orceine (a, b) or with DAPI (c). The anthers were collected and the pollen mother cells (PMC) were examined under the microscope. Two plants from the F1 generation of self-pollinated NGT plants were used. a, Four representative PMCs of a wild-type tobacco plant. Scale bar, 50 µm. b, Six PMC from NGT plants. The upper three cells are from line NGT-2, the lower three from line NGT-3. Scale bar, 20 µm. c, Two PMCs from line NGT-3 stained with DAPI. Scale bar, 5 µm. Mis-segregating chromosomes are indicated by arrowheads in b and c.

Extended Data Figure 3 Phenotypes of NGT progeny plants.

a, b, Wild-type tobacco (Nt-wt), the two transgenic graft partners (Nt-hyg and Nt-kan:yfp) and the second generation of three NGT lines (NGT-1, NGT-2 and NGT-3) were grown under greenhouse conditions. A total of 21 NGT progeny plants were investigated, 14 of them resulted from self-pollinated lines (NGT-2 and NGT-3) and the remaining 7 from the cross of the NGT-1 line with a wild-type plant (which was male sterile and could not be selfed). Pictures were taken 30 (a) and 45 (b) days after sowing.

Extended Data Figure 4 Phenotypes of Nicotiana tabauca progeny plants.

a, b, Wild-type tobacco (Nt-wt), the two transgenic graft partners (Nt-hyg and Ng-kan) and the F1 generation of an N. tabauca line were raised from seeds and grown under greenhouse conditions. Pictures were taken 28 (a) and 47 (b) days after sowing.

Extended Data Figure 5 Detection of allopolyploid and aberrant karyotypes in Nicotiana tabauca F1 progeny plants.

a, Two examples of DAPI-stained metaphases with fewer than 72 chromosomes. b, As an alternative to the DAPI staining shown in Fig. 4c and in panel a, a method based on hot tissue hydrolysis in HCl and staining with toluidine blue was used (see Methods). Shown is a Nicotiana tabauca F1 plant with the full allopolyploid chromosome set of 72 chromosomes.

Extended Data Figure 6 Molecular analysis of four polymorphic regions in the plastid genome of Nicotiana tabauca lines.

a, Physical map of the Nicotiana tabacum plastid genome showing the four plastid polymorphic regions analysed (pt1, pt2, pt3 and pt4). pt1: polymorphism amplified with oligonucleotides Ppt1F and Ppt1R (Extended Data Table 2) resulting in a 211 bp fragment in N. tabacum cv. SNN and a 203 bp fragment in N. glauca; pt2: polymorphism amplified with oligonucleotides Ppt2F and Ppt2R resulting in a 221 bp fragment in N. tabacum cv. SNN and a 229 bp fragment in N. glauca; pt3: polymorphic region of ∼3 kb (containing altogether 25 polymorphisms) amplified using the primer pairs Ppt3-1F + Ppt3-1R, Ppt3-2F + Ppt3-2R and Ppt3-3F + Ppt3-3R; pt4: polymorphic region of ∼3 kb (containing altogether 32 polymorphisms) amplified with the primer pairs Ppt4-1F + Ppt4-1R, Ppt4-2F + Ppt4-2R and Ppt4-3F + Ppt4-3R. The polymorphisms were selected based on published sequence information of the plastid genomes of N. tabacum and N. glauca16,36. b, Overview of the plant lines and the polymorphic regions analysed. Nicotiana tabacum sequence is represented with Nt on yellow background, Nicotiana glauca sequence is represented with Ng on pink background. c, A 4% agarose gel showing the size difference of the PCR fragments for the pt1 and pt2 polymorphisms between N. tabacum, N. glauca and five N. tabauca lines.

Extended Data Figure 7 Molecular analysis of five mitochondrial genome polymorphisms in Nicotiana tabauca lines by sequencing of amplified PCR products.

a, Physical map of the Nicotiana tabacum mitochondrial genome showing the five mitochondrial polymorphic regions analysed (mt1, mt2, mt3, mt4 and mt5). mt1: polymorphic region of 693 bp (containing 5 polymorphisms) amplified with oligonucleotides Pmt1F and Pmt1R (Extended Data Table 2); mt2: polymorphism (SNP) amplified with oligonucleotides Pmt2F and Pmt2R; mt3: polymorphism (SNP) amplified with oligonucleotides Pmt3F and Pmt3R; mt4: polymorphism (SNP) amplified with oligonucleotides Pmt4F and Pmt4R; mt5: polymorphic region amplified with oligonucleotides Pmt5F and Pmt5R and resulting in a 298 bp fragment in N. tabacum and a 288 bp fragment in N. glauca. The polymorphisms were selected based on published sequence information of the mitochondrial genomes of N. tabacum and N. glauca16,37. b, Overview of the plant lines and the polymorphic regions analysed. Nicotiana tabacum sequence is represented with Nt on yellow background, Nicotiana glauca sequence is represented with Ng on dark pink background. Heteroplasmy (that is, detectability of both the N. tabacum and the N. glauca sequence in an N. tabauca plant) is indicated by Nt/Ng on light pink background. A and B denote two different plants from the same N. tabauca line.

Extended Data Figure 8 Detection of mitochondrial heteroplasmy in N. tabauca plants.

a, Example of sequences amplified from the mt1 polymorphic region in Nt-hyg, Ng-kan and in an N. tabauca (Ntca) line. The mt1 polymorphic region was amplified with the oligonucleotide combination Pmt1F and Pmt1R (Extended Data Table 2) resulting in a 693 bp fragment (corresponding to nucleotide positions 36,746 to 37,438 in the Nicotiana tabacum mitochondrial genome, accession number: NC_006581.1). The fragment contains five single nucleotide polymorphisms (SNPs) that are denoted with an arrow in the sequence chromatograms and the nucleotide position in the N. tabacum mitochondrial genome. A mixed nucleotide position indicating heteroplasmy is denoted by two letters above each other. b, A 4% agarose gel analysing the mt4 polymorphism in N. tabacum, N. glauca and nine N. tabauca plants (representing five different lines). A 310 bp fragment was amplified by PCR and digested with the two restriction enzymes NcoI and BsaBI. Digestion of the PCR product amplified from the N. tabacum mitochondrial genome yields three restriction fragments (25 bp, 185 bp and 100 bp), whereas digestion of the PCR product amplified from the N. glauca mitochondrial genome results in two fragments (25 bp and 285 bp). This difference between the two species is due to a T to G substitution in N. glauca (relative to N. tabacum; position in the N. tabacum mitochondrial genome: 305,402), resulting in the loss of a BsaBI site. The 25 bp restriction fragment is not detectable in the gel because of its small size. Note that the low level of heteroplasmy detected by restriction-fragment length polymorphism (RFLP) analysis was not reliably detectable by DNA sequencing (compare with Extended Data Fig. 7b). A and B denote two different plants from the same N. tabauca line.

Extended Data Table 1 Segregation ratios of three autopolyploid NGT lines
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Extended Data Table 2 List of oligonucleotides used in this study
Full size table

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Fuentes, I., Stegemann, S., Golczyk, H. et al. Horizontal genome transfer as an asexual path to the formation of new species. Nature 511, 232–235 (2014). https://doi.org/10.1038/nature13291

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