Skip to main content
SpringerLink
Log in

A protocol for fertility restoration of F1 hybrid derived from Lilium × formolongi ‘Raizan 3’ × Oriental hybrid ‘Sorbonne’

  • Original Article
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

The present study was an attempt to develop an in vitro colchicine chromosome doubling protocol to restore the fertility of an F1 interspecific hybrid in Lilium. Basal scale segments of Lilium × formolongi × Oriental hybrid (FO) bulblets were pre-cultured for three durations (6, 15 and 25 days) and soaked in three colchicine concentrations (0.00, 1.25 and 2.50 mM) for 18, 24 and 36 h. To separate mixoploids, three cycles of adventitious bud induction were performed. The ploidy levels of the surviving plantlets were detected by flow cytometry at 30–31 weeks after induction and were confirmed by chromosome counts. The results indicated that the pre-culture duration, colchicine concentration and exposure time all had significant impacts on the tetraploid induction rate. The preferred procedure was to pre-culture the segments for 6 days and then treat them with 1.25 mM colchicine for 24 h. The morphological traits of rosette leaves were significantly different between the tetraploid and diploid plants. The adult tetraploid plants had considerably longer and wider leaves, larger flowers, and delayed flowering time (8 days later) than did diploid plants. Pollen viability tests and backcross trials of FO hybrids demonstrated fertility restoration at the tetraploid level. This protocol provides a feasible method for inducing fertile tetraploid FO hybrids for further breeding.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Abu-Qaoud H, Skirvin RM, Chevreau E (1990) In vitro separation of chimeral pears into their component genotypes. Euphytica 48:189–196

    Article  Google Scholar 

  • Anderson NO, Younis A, Optiz E (2009) Development of colored, non-vernalization -requiring seed propagated lilies. Acta Hortic 836:193–198

    Article  Google Scholar 

  • Anssour S, Krügel T, Sharbel TF, Saluz HP, Bonaventure G, Baldwin IT (2009) Phenotypic, genetic and genomic consequences of natural and synthetic polyploidization of Nicotiana attenuata and Nicotiana obtusifolia. Ann Bot 103:1207–1217

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bakhshaie M, Babalar M, Mirmasoumi M, Khalighi A (2010) Somatic embryogenesis and plant regeneration of Lilium ledebourii (Baker) Boiss., an endangered species. Plant Cell Tissue Organ Cult 102:229–235

    Article  CAS  Google Scholar 

  • Bakhshaie M, Khosravi S, Azadi P, Bagheri H, Van Tuyl JM (2016) Biotechnological advances in Lilium. Plant Cell Rep 35:1799–1826

    Article  CAS  PubMed  Google Scholar 

  • Barba-Gonzalez R, Ramanna MS, Visser RGF, Van Tuyl JM (2005) Intergenomic recombination in F1 lily hybrids (Lilium) and its significance for genetic variation in the BC1 progenies as revealed by GISH and FISH. Genome 48:884–894

    Article  CAS  PubMed  Google Scholar 

  • Barba-Gonzalez R, Lim KB, Van Tuyl JM (2014) Molecular cytogenetics in Lilium breeding. Acta Hortic 1027:129–142

    Article  Google Scholar 

  • Bi WL, Chen L, Guo L, Pan C, Yin ZF ,Wang QC (2015) Plant regeneration via embryo-like structures: histological observations and genetic stability in regenerants of Lilium spp. J Hortic Sci Biotech 90:626–634

    Article  CAS  Google Scholar 

  • Chandanie MA, Singh SK, Sindhu SS, Singh A, Tomar SMS, Prasad KV (2011) Efficacy of oryzalin as a potent chemical for in vitro induction of polyploids in Asiatic lily (Lilium hybrida L.) var. Polyanna. India J Genet 71:262–268

    CAS  Google Scholar 

  • Dhooghe E, Denis S, Eeckhaut T, Reheul D, Van Labeke MC (2009) In vitro induction of tetraploids in ornamental Ranunculus. Euphytica 168:33–40

    Article  CAS  Google Scholar 

  • Dhooghe E, Van Laere K, Eeckhaut T, Leus L, Van Huylenbroeck J (2011) Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tissue Organ Cult 104:359–373

    Article  Google Scholar 

  • Doležel J, Greilhuber J, Suda J (2007) Estimation of nuclear DNA content in plants using flow cytometry. Nat Protoc 2:2233–2244

    Article  PubMed  Google Scholar 

  • Gomes SSL, Saldanha CW, Neves CS, Trevizani M, Raposo NRB, Notini MM, Santos MDO, Campos JMS, Otoni WC, Viccini LF (2014) Karyotype, genome size, and in vitro chromosome doubling of Pfaffia glomerata (Spreng.) Pedersen. Plant Cell Tissue Organ Cult 118:45–56

    Google Scholar 

  • Han DS, Niimi Y (2008) Fertility restoring of interspecific hybrids between Lilium nobilissimum and L. regale by chromosome doubling. Acta Hortic 766:421–426

    Article  Google Scholar 

  • Hu FR (2007) The wild and artificial germplasm assessment by cytogenetics and mass propagation system establishment in Lilium. PhD thesis, Nanjing Forestry University, Beijing

  • Lim KB, Van Tuyl JM (2002) Identification of parental chromosomes and detection of ribosomal DNA sequences in interspecific hybrids of Lilium revealed by multicolor in situ hybridization. Acta Hortic 570:403–408

    Article  CAS  Google Scholar 

  • Lim KB, Chung JD, Van Kronenburg BCE, Ramanna MS, Hans de Jong J, Van Tuyl JM (2000) Introgression of Lilium rubellum Baker chromosomes into L. longiflorum Thunb.: a genome painting study of the F1 hybrid, BC1 and BC2 progenies. Chromosome Res 8:119–125

    Article  CAS  PubMed  Google Scholar 

  • Lim KB, Wennekes J, Jong JHd, Jacobsen E, Van Tuyl JM (2001) Karyotype analysis of Lilium longiflorum and Lilium rubellum by chromosome banding and fluorescence in situ hybridisation. Genome 44:911–918

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Ning Y, Zhou H, Wang F (2003) The bulblet morphogenesis of Lilium × formolongi in scale propagation. Acta Hortic 30:229–231

    Google Scholar 

  • Okazaki K (1996) Lilium species native to Japan, and breeding and production of Lilium in Japan. Acta Hortic 414:81–92

    Article  Google Scholar 

  • Omran A, Mohammad BN (2008) Polyploidization effect in two diploid cotton (Gossypium herbaceum L. and G. arboreum L.) species by colchicine treatments. African. J Biotechnol 7:102–108

    CAS  Google Scholar 

  • Regalado JJ, Carmona-Martín E, Castro P, Moreno R, Gil J, Encina CL (2015) Study of the somaclonal variation produced by different methods of polyploidization in Asparagus officinalis L. Plant Cell Tissue Organ Cult 122:31–44

    Article  CAS  Google Scholar 

  • Rhee HK, Kim KS (2008) Interspecific hybridization and polyploidization in lily breeding. Acta Hortic 766:441–445

    Article  Google Scholar 

  • Rose JB, Kubba J, Tobutt KR (2000) Chromosome doubling in sterile Syringa vulgaris × S. pinnatifolia hybrids by in vitro culture of nodal explants. Plant Cell Tiss Organ Cult 63:127–132

    Article  Google Scholar 

  • Roy AT, Leggett G, Koutoulis A (2001) In vitro tetraploid induction and generation of tetraploids from mixoploids in hop (Humulus lupulus L.). Plant Cell Rep 20:489–495

    Article  CAS  Google Scholar 

  • Schwarzacher T, Heslop-Harrison JS (2000) Reprobing of preparations. In: Kingston F (ed) Practical in situ hybridization, BIOS Scientific Publishers Limited. Oxford, UK, p 110

    Google Scholar 

  • Takamura T, Lim KB, Van Tuyl JM (2002) Effect of a new compound on the mitotic polyploidization of Lilium longiflorum and Oriental hybrid lilies. Acta Hortic 572:37–40

    Article  CAS  Google Scholar 

  • Takayama S, Misawa M (1980) Differentiation in Lilium bulbscales grown in vitro. Effects of activated charcoal, physiological age of bulbs and sucrose concentration on differentiation and scale leaf formation in vitro. Physiol Plant 48:121–125

    Article  CAS  Google Scholar 

  • Van Tuyl JM, Lim KB (2003) Interspecific hybridisation and polyploidisation as Tools in ornamental plant breeding. Acta Hortic 612:13–22

    Article  Google Scholar 

  • Van Tuyl JM, Van Holsteijn HCM (1996) Lily breeding research in the Netherlands. Acta Hortic 414:35–45

    Article  Google Scholar 

  • Van Tuyl JM, Van-Holsteijn HMC, Kwakkenbos AAM (1990) Research on polyploidy in interspecific hybridization of lily. Acta Hortic 266:323–330

    Article  Google Scholar 

  • Van Tuyl JM, Meijer B, Van Diën MP (1992) The use of oryzalin as an alternative for colchicine in in vitro chromosome doubling of Lilium and Nerine. Acta Hortic 325:625–630

    Article  Google Scholar 

  • Van Tuyl JM, Van Dijken A, Chi HS, Lim KB, Villemoes S, Van Kronenburg BCE (2000) Breakthroughs in interspecific hybridization of lily. Acta Hortic 508:83–90

    Article  Google Scholar 

  • Xu CP, Huang Z, Liao T, Li Y, Kang XY (2016) In vitro tetraploid plants regeneration from leaf explants of multiple genotypes in Populus. Plant Cell Tissue Organ Cult 125:1–9

    Article  CAS  Google Scholar 

  • Yan HJ, Xiong Y, Zhang HY, He ML (2016) In vitro induction and morphological characteristics of octoploid plants in Pogostemon cablin. Breeding Sci 66:169–174

    Article  Google Scholar 

  • Yin ZF, Zhao B, Bi WL, Chen L, Wang QC (2013) Direct shoot regeneration from basal leaf segments of Lilium and assessment of genetic stability in regenerants by ISSR and AFLP markers. In Vitro Cell Dev Biol- Plant 49:333–342

    Article  CAS  Google Scholar 

  • Zhang J, Zhang M, Deng XX (2007) Obtaining autotetraploids in vitro at a high frequency in Citrus sinensis. Plant Cell Tissue Organ Cult 89:211–216

    Article  Google Scholar 

  • Zhou SJ (2007) Intergenomic recombination and introgression breeding in Longiflorum × Asiatic lilies. Ph D thesis, Wageningen University, The Netherlands

Download references

Acknowledgements

The work was supported by grants from the National Forestry Public Welfare Industry Research Project (201204609) and the National Natural Science Foundation of China (31470106).

Author contributions

The authors have made the following declarations regarding their contributions: GXJ, XQZ conceived and designed the experiments; XQZ, QZC performed the experiments; and XQZ analyzed the data and wrote the manuscript.

Author information

Authors and Affiliations

  1. Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, National Engineering Research Center for Floriculture, Beijing Laboratory of Urban and Rural Ecological Environment and College of Landscape Architecture, Beijing Forestry University, Beijing, 100083, China

    Xiqing Zhang, Qinzheng Cao & Guixia Jia

Authors
  1. Xiqing Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qinzheng Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Guixia Jia
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guixia Jia.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, X., Cao, Q. & Jia, G. A protocol for fertility restoration of F1 hybrid derived from Lilium × formolongi ‘Raizan 3’ × Oriental hybrid ‘Sorbonne’. Plant Cell Tiss Organ Cult 129, 375–386 (2017). https://doi.org/10.1007/s11240-017-1184-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-017-1184-9

Keywords

Search

Navigation