Journal of Plant Research

, Volume 132, Issue 4, pp 461–471 | Cite as

Nicotiana suaveolens accessions with different ploidy levels exhibit different reproductive isolation mechanisms in interspecific crosses with Nicotiana tabacum

  • Hai He
  • Takahiro Iizuka
  • Maho Maekawa
  • Kumi Sadahisa
  • Toshinobu Morikawa
  • Masanori Yanase
  • Shuji Yokoi
  • Masayuki Oda
  • Takahiro TezukaEmail author
Regular Paper


Reproductive isolation, including prezygotic and postzygotic barriers, is a mechanism that separates species. Many species in the Nicotiana section Suaveolentes exhibit reproductive isolation in crosses with Nicotiana tabacum. In this study, we investigated whether the chromosome numbers and ploidy levels of eight Nicotiana suaveolens accessions are related to the reproductive isolation after crosses with N. tabacum by flow cytometry and chromosome analyses. Additionally, the internal transcribed spacer (ITS) regions of the eight N. suaveolens accessions were sequenced and compared with the previously reported sequences of 22 Suaveolentes species to elucidate the phylogenetic relationships in the section Suaveolentes. We revealed that four N. suaveolens accessions comprised 64 chromosomes, while the other four accessions carried 32 chromosomes. Depending on the ploidy levels of N. suaveolens, several types of reproductive isolation were observed after crosses with N. tabacum, including decreases in the number of capsules and the germination rates of hybrid seeds, as well as hybrid lethality and abscission of enlarged ovaries at 12–17 days after pollination. A phylogenetic analysis involving ITS sequences divided the eight N. suaveolens accessions into three distinct clades. Based on the results, we confirmed that N. suaveolens accessions vary regarding ploidy levels and reproductive isolation mechanisms in crosses with N. tabacum. These accessions will be very useful for revealing and characterizing the reproductive isolation mechanisms in interspecific crosses and their relationships with ploidy levels.


Internal transcribed spacer region Interspecific hybridization Phylogenetics Polyploidy Reproductive isolation Tobacco 



This study was partly supported by JSPS KAKENHI Grants (JP20880024 and JP25870627) from the Japan Society for the Promotion of Science.

Supplementary material

10265_2019_1114_MOESM1_ESM.pdf (784 kb)
Supplementary material 1 (PDF 784 kb)


  1. Baldwin BG (1993) Molecular phylogenetics of Calycadenia (Compositae) based on ITS sequences of nuclear ribosomal DNA: chromosomal and morphological evolution reexamined. Am J Bot 80:222–238CrossRefGoogle Scholar
  2. Bushell C, Spielman M, Scott RJ (2003) The basis of natural and artificial postzygotic hybridization barriers in Arabidopsis species. Plant Cell 15:1430–1442CrossRefPubMedPubMedCentralGoogle Scholar
  3. Chase MW, Knapp S, Cox AV et al (2003) Molecular systematics, GISH and the origin of hybrid taxa in Nicotiana (Solanaceae). Ann Bot 92:107–127CrossRefPubMedPubMedCentralGoogle Scholar
  4. Clarkson JJ, Knapp S, Garcia VF, Olmstead RG, Leitch AR, Chase MW (2004) Phylogenetic relationships in Nicotiana (Solanaceae) inferred from multiple plastid DNA regions. Mol Phylogenet Evol 33:75–90CrossRefPubMedGoogle Scholar
  5. Clarkson JJ, Kelly LJ, Leitch AR, Knapp S, Chase MW (2010) Nuclear glutamine synthetase evolution in Nicotiana: phylogenetics and the origins of allotetraploid and homoploid (diploid) hybrids. Mol Phylogenet Evol 55:99–112CrossRefPubMedGoogle Scholar
  6. Dickinson GR, Lee DJ, Wallace HM (2012) The influence of pre- and post-zygotic barriers on interspecific Corymbia hybridization. Ann Bot 109:1215–1226CrossRefPubMedPubMedCentralGoogle Scholar
  7. Doležel J, Bartos J (2005) Plant DNA flow cytometry and estimation of nuclear genome size. Ann Bot 95:99–110CrossRefPubMedPubMedCentralGoogle Scholar
  8. Goodspeed TH (1954) The genus Nicotiana. Chronica Botanica Company, WalthamGoogle Scholar
  9. Guo J, Xu X, Li W et al (2016) Overcoming inter-subspecific hybrid sterility in rice by developing indica-compatible japonica lines. Sci Rep 6:26878CrossRefPubMedPubMedCentralGoogle Scholar
  10. Ichitani K, Namigoshi K, Sato M et al (2007) Fine mapping and allelic dosage effect of Hwc1, a complementary hybrid weakness gene in rice. Theor Appl Genet 114:1407–1415CrossRefPubMedGoogle Scholar
  11. Inoue E, Marubashi W, Niwa M (1994) Simple method for overcoming the lethality observed in the hybrid between Nicotiana suaveolens and N. tabacum. Breed Sci 44:333–336Google Scholar
  12. Ishikawa R, Ohnishi T, Kinoshita Y, Eiguchi M, Kurata N, Kinoshita T (2011) Rice interspecies hybrids show precocious or delayed developmental transitions in the endosperm without change to the rate of syncytial nuclear division. Plant J 65:798–806CrossRefPubMedGoogle Scholar
  13. Japan Tobacco Inc (1994) The genus Nicotiana illustrated. Japan Tobacco Inc, TokyoGoogle Scholar
  14. Kradolfer D, Wolff P, Jiang H, Siretskiy A, Köhler C (2013) An imprinted gene underlies postzygotic reproductive isolation in Arabidopsis thaliana. Dev Cell 26:525–535CrossRefPubMedGoogle Scholar
  15. Kuboyama T, Saito T, Matsumoto T et al (2009) Fine mapping of HWC2, a complementary hybrid weakness gene, and haplotype analysis around the locus in rice. Rice 2:93–103CrossRefGoogle Scholar
  16. Ladiges PY, Marks CE, Nelson G (2011) Biogeography of Nicotiana section Suaveolentes (Solanaceae) reveals geographical tracks in arid Australia. J Biogeogr 38:2066–2077CrossRefGoogle Scholar
  17. Leitch IJ, Hanson L, Lim KY et al (2008) The ups and downs of genome size evolution in polyploid species of Nicotiana (Solanaceae). Ann Bot 101:805–814CrossRefPubMedPubMedCentralGoogle Scholar
  18. Lewis RS, Nicholson JS (2007) Aspects of the evolution of Nicotiana tabacum L. and the status of the United States Nicotiana Germplasm Collection. Genet Resour Crop Evol 54:727–740CrossRefGoogle Scholar
  19. Li Z, Pinson SRM, Paterson AH, Park WD, Stansel JW (1997) Genetics of hybrid sterility and hybrid breakdown in an intersubspecific rice (Oryza sativa L.) population. Genetics 145:1139–1148PubMedPubMedCentralGoogle Scholar
  20. Manabe T, Marubashi W, Onozawa Y (1989) Temperature-dependent conditional lethality in interspecific hybrids between Nicotiana suaveolens Lehm. and N. tabacum L. In: Proceedings of the 6th international congress of SABRAO, pp 459–462Google Scholar
  21. Marubashi W, Onosato K (2002) Q chromosome controls the lethality of interspecific hybrids between Nicotiana tabacum and N. suaveolens. Breed Sci 52:137–142CrossRefGoogle Scholar
  22. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  23. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325CrossRefPubMedPubMedCentralGoogle Scholar
  24. Nei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, New YorkGoogle Scholar
  25. Otto F (1990) DAPI staining of fixed cells for high-resolution flow cytometry of nuclear DNA. In: Darzynkiewickz Z, Crissman HA (eds) Methods in cell biology. Academic Press, San Diego, pp 105–110Google Scholar
  26. Purdie RW, Symon DE, Haegi L (1982) Solanaceae. Flora of Australia 29:1–208Google Scholar
  27. Rebernig CA, Lafon-Placette C, Hatorangan MR, Slotte T, Köhler C (2015) Non-reciprocal interspecies hybridization barriers in the Capsella genus are established in the endosperm. PLoS Genet 11:e1005295CrossRefPubMedPubMedCentralGoogle Scholar
  28. Rieseberg LH, Blackman BK (2010) Speciation genes in plants. Ann Bot 106:439–455CrossRefPubMedPubMedCentralGoogle Scholar
  29. Scott RJ, Spielman M, Bailey J, Dickinson HG (1998) Parent-of-origin effects on seed development in Arabidopsis thaliana. Development 125:3329–3341Google Scholar
  30. Sekine D, Ohnishi T, Furuumi H et al (2013) Dissection of two major components of the post-zygotic hybridization barrier in rice endosperm. Plant J 76:792–799CrossRefPubMedGoogle Scholar
  31. Stebbins GL (1966) Reproductive isolation and the origin of species. Processes of organic evolution. Prentice-Hall, Upper Saddle River, pp 85–112Google Scholar
  32. Sun Y, Skinner DZ, Liang GH, Hulbert SH (1994) Phylogenetic analysis of Sorghum and related taxa using internal transcribed spacers of nuclear ribosomal DNA. Theor Appl Genet 89:26–32CrossRefGoogle Scholar
  33. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729CrossRefPubMedPubMedCentralGoogle Scholar
  34. Tezuka T (2012) Hybrid lethality in the genus Nicotiana. In: Mworia JK (ed) Botany. InTech, Rijeka, pp 191–210Google Scholar
  35. Tezuka T (2013) Hybrid lethality in Nicotiana: a review with special attention to interspecific crosses between species in sect. Suaveolentes and N. tabacum. In: Wallner F (ed) Herbaceous plants: cultivation methods, grazing and environmental impacts. Nova Science Publishers, New York, pp 69–94Google Scholar
  36. Tezuka T, Marubashi W (2004) Apoptotic cell death observed during the expression of hybrid lethality in interspecific hybrids between Nicotiana tabacum and N. suaveolens. Breed Sci 54:59–66CrossRefGoogle Scholar
  37. Tezuka T, Marubashi W (2006) Hybrid lethality in interspecific hybrids between Nicotiana tabacum and N. suaveolens: evidence that the Q chromosome causes hybrid lethality based on Q-chromosome-specific DNA markers. Theor Appl Genet 112:1172–1178CrossRefPubMedGoogle Scholar
  38. Tezuka T, Marubashi W (2012) Genes in S and T subgenomes are responsible for hybrid lethality in interspecific hybrids between Nicotiana tabacum and Nicotiana occidentalis. PLoS One 7:e36204CrossRefPubMedPubMedCentralGoogle Scholar
  39. Tezuka T, Kuboyama T, Matsuda T, Marubashi W (2010) Seven of eight species in Nicotiana section Suaveolentes have common factors leading to hybrid lethality in crosses with Nicotiana tabacum. Ann Bot 106:267–276CrossRefPubMedPubMedCentralGoogle Scholar
  40. Tezuka T, Matsuo C, Iizuka T, Oda M, Marubashi W (2012) Identification of Nicotiana tabacum linkage group corresponding to the Q chromosome gene(s) involved in hybrid lethality. PLoS One 7:e37822CrossRefPubMedPubMedCentralGoogle Scholar
  41. USDA ARS National Genetic Resources Program (2010) Germplasm Resources Information Network—(GRIN) [Online Database]. National Germplasm Resources Laboratory, Beltsville, Maryland. Accessed Apr 2015
  42. Wheeler HM (1935) Studies in Nicotiana. II. A taxonomic survey of the Australasian species. Univ Calif Publ Bot 18:45–68Google Scholar
  43. Yamada T, Marubashi W, Niwa M (1999) Detection of four lethality types in interspecific crosses among Nicotiana species through the use of three rescue methods for lethality. Breed Sci 49:203–210CrossRefGoogle Scholar
  44. Yamada T, Marubashi W, Niwa M (2000) Apoptotic cell death induces temperature-sensitive lethality in hybrid seedlings and calli derived from the cross of Nicotiana suaveolens × N. tabacum. Planta 211:614–622CrossRefPubMedGoogle Scholar

Copyright information

© The Botanical Society of Japan and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Graduate School of Life and Environmental SciencesOsaka Prefecture UniversitySakaiJapan
  2. 2.School of Life and Environmental SciencesOsaka Prefecture UniversitySakaiJapan
  3. 3.Education and Research Field, College of Life, Environment, and Advanced SciencesOsaka Prefecture UniversitySakaiJapan

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