Skip to main content
SpringerLink
Log in

Direct regeneration of haploid or doubled haploid plantlets in cucumber (Cucumis sativus L.) through ovary culture

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

Abstract

This paper reports a simple and effective method of directly producing cucumber plants through unfertilized ovary culture. The paper involves an experiment investigating factorial interactions between TDZ treatment, cold pretreatment, genotypes of cucumber, to improve plant induction. Cold pretreatment was effective in stimulating the ovary. The results showed that cold pretreatment for 4 days, TDZ for 0.06 mg·L−1, the interaction with genotype can be used as an effective strategy to improve the efficiency of gynogenesis. The plant regeneration induction rate was highest (79.3%). In addition, we observed the process of cucumber megasporogenesis and plant regeneration. The plants obtained from ovary culture of cucumber were identified as diploid or haploid by flow cytometry, consistent with the results of chromosome counting. The diploid plants were further identified as pure doubled haploid using simple sequence repeats (SSR). The doubling treatment we used was one of the simplest and most effective methods, completed in a short time (1 h) with a doubling rate of 75%. The acclimation rate for the surviving was 70%. This work provides a basis for promoting haploid breeding in cucumber.

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
Fig. 8
Fig. 9

Similar content being viewed by others

Data availability

The data sets supporting the results of this article are included within the article.

References

  • Bhat JG, Murthy HN (2007) Factors affecting in-vitro gynogenic haploid production in niger (Guizotiaabyssinica (L. f.) Cass.). Plant Growth Regul 52:241–248

    Article  CAS  Google Scholar 

  • Castillo AM, Sánchez-Díaz RA, Vallés MP (2015) Effect of ovary induction on bread wheat anther culture: ovary genotype and developmental stage, and candidate gene association. Front Plant Sci. https://doi.org/10.3389/fpls.2015.00402

  • Chani E, Veilleux RE, Boluarte-Medina T, (2000) Improved androgenesis of interspecific potato and efficiency of SSR markers to identify homozygous regenerants. Plant Cell, Tissue Organ Cult 60(2):101–112

    Article  CAS  Google Scholar 

  • Chen J, Staub J, Qian C, Jiang J, Luo X, Zhuang F (2003) Reproduction and cytogeneticcharacterization of interspecific hybrids from Cucumis hystrix Chakr. × Cucumis sativus L. Theoret Appl Genet 106(4):688–695

    Article  CAS  Google Scholar 

  • Chen JF, Cui L, Malik AA, Mbira KG (2011) In vitro haploid and dihaploid production via unfertilized ovule culture. Plant Cell Tissue Organ Cult 104(3):311–319

    Article  Google Scholar 

  • Claveria E, Garcia-Mas J, Dolcet-Sanjuan R (2005) Optimization of cucumber doubled haploid line production using in vitro rescue of in vivo induced parthenogenic embryos. J Am Soc Hortic Sci 130(4):555–560

    Article  Google Scholar 

  • Diao WP, Jia YY, Song H, Zhang XQ, Lou QF (2009) Efficient embryo induction in cucumber ovary culture and homozygous identification of the regenetants using SSR markers. Sci Hortic 119:246–251

    Article  CAS  Google Scholar 

  • Gémes-Juhász A, Balogh P, Ferenczy A, Kristóf Z (2002) Effect of optimal stage of female gametophyte and heat treatment on in vitro gynogenesis induction in cucumber (Cucumis sativus L). Plant Cell Rep 21(2):105–111

    Article  Google Scholar 

  • Gresshoff PM (1991) Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal Biochem 196(1):80–83

    Article  Google Scholar 

  • Lantos C, Bóna L, Nagy É, Békés F, Pauk J (2018) Induction of in vitro androgenesis in anther and isolated microspore culture of different spelt wheat (Triticum spelta L) genotypes. Plant Cell Tissue Organ Culture 133:385–393 https://dio.org/10.1007/s11240-018-1391-z

  • Hosp J, Tashpulatov A, Roessner U, Barsova E, Katholnigg H, Steinborn R, Melikant B, Lukyanov S, Heberle-Bors E, Touraev A (2007) Transcriptional and metabolic profiles of stress-induced embryogenic tobacco microspores. Plant Mol Biol 63(1):137–149

    Article  CAS  Google Scholar 

  • Li JW, Si SW, Cheng JY, Li JX, Liu JQ (2013) Thidiazuron and silver nitrate enhanced gynogenesis of unfertilized ovule cultures of Cucumis sativus. Biol Plant 57:164–168

    Article  CAS  Google Scholar 

  • Lotfi M, Alan AR, Henning MJ, Jahn MM, Earle ED (2003) Production of haploid and doubled haploid plants of melon (Cucumis melo L) for use in breeding for multiple virus resistance. Plant Cell Rep 21(11):1121–1128

    Article  CAS  Google Scholar 

  • Lou QF, He YH, Cheng CY, Zhang ZH, Li J, Huang SW, Chen JF(2013) Integration of high-resolution physical and genetic mapreveals differential recombination frequency between chromosomes andthe genome assembling quality in cucumber. Plos one 8. htpps://dio.org/10.1371/journal.pone.0062676

  • Loureiro J, Trávníček P, Rauchová J, Urfus T, Vít P, Štech M, Castro S, Suda J (2010) The use of flow cytometry in the biosystematics, ecology and population biology of homoplod plants. Preslia 82(1):3–21

    Google Scholar 

  • Luo XD, Dai LF, Qian CT, Chen JF (2007) Cytological studies on male and female gametophytes development in an interspecific hybrid F1 from Cucumis hystrixChakr× C sativus L. Scientia Agricultura Sinica 40(2):338–344

    Google Scholar 

  • Malik AA, Cui L, Zhang SX, Chen JF (2011) Efficiency of SSR markers for determining the origin of melon plantlets derived through unfertilized ovary culture. Horticult Sci 38:27–34

    Article  CAS  Google Scholar 

  • Maraschin SF, de Priester W, Spaink HP, Wang M (2005) Androgenic switch: an example of plant embryogenesis from the male gametophyte perspective. J Exp Bot 56(417):1711–1726

    Article  CAS  Google Scholar 

  • Metwally EI, Moustafa SA, El-Sawy BI, Haroun SA, Shalaby TA (1998) Production of haploid plants from in vitro culture of unpollinated ovules of Cucurbita pepo. Plant Cell Tissue Organ Cult 52(3):117–121

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

    Article  CAS  Google Scholar 

  • Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8(19):4321–4326

    Article  CAS  Google Scholar 

  • Pazuki A, Aflaki F, Gürel S, Ergül A, Gürel E (2018) Production of doubled haploids in sugar beet (Beta vulgaris) an efficient method by a multivariate experiment. Plant Cell Tissue Organ Cult 132:85–97. https://doi.org/10.1007/s11240-017-1313-5

    Article  CAS  Google Scholar 

  • Perera PI, Perera L, Hocher V, Verdeil JL, Yakandawala DMD, Weerakoon LK (2008) Use of SSR markers to determine the anther-derived homozygous lines in coconut. Plant Cell Rep 27(11):1697–1703

    Article  CAS  Google Scholar 

  • Shalaby TA (2007) Factors affecting haploid induction through in vitro gynogenesis in summer squash (Cucurbita pepo L). Sci Hortic 115(1):1–6

    Article  Google Scholar 

  • Plapung P, Khamsukdee S, Potapohn N, Smitamana P (2014) Screening for cucumber mosaic resistant lines from the ovule culture derived double haploid cucumbers. Am J Agric Biol Sci 9:261–269

    Article  Google Scholar 

  • Popova T, Grozeva S, Todorova V, Stankova G, Anachkov N, Rodeva V (2016) Effects of low temperature, genotype and culture media on in vitro androgenic answer of pepper (Capsicum annuum L). Acta Physiol Plant 38:273. https://doi.org/10.1007/s11738-016-2294-4

    Article  CAS  Google Scholar 

  • Tantasawat PA, Sorntip A, Poolsawat O, Chaowiset W, Pornbungkerd P (2015) Evaluation of factors affecting embryo-like structure and callus formation in unpollinated ovary culture of cucumber (Cucumis sativus L). Int J Agric Biol 17(3):613–618

    Article  CAS  Google Scholar 

  • Tugce O, Volkan G, Ahmet NO (2017) Cucumber gynogenesis: effects of 8 different media on embryo and plant formation. Int J Agric Innovat Res 2:418–421

    Google Scholar 

  • Wang Y, Gu XF, Zhang SP, Miao H (2015) Studies on haploid plant induction via in vitro unfertilized ovule culture of cucumber. Acta Horticulturae Sinica 42:2174–2182

    CAS  Google Scholar 

  • Yadegari R (2004) Female gametophyte development. The Plant Cell Online 16((suppl_1)):S133–S141

    Article  CAS  Google Scholar 

  • Yetisir H, Sari N (2003) A new method for haploid muskmelon ( Cucumis melo L.) dihaploidization. Sci Hortic 98:277–283

    Article  Google Scholar 

Download references

Acknowledgements

The authors would like to acknowledge the Dr. Tan (Institute of Horticulture, Guizhou Academy of Agricultural Sciences, Guiyang, China) for critical reading of the manuscript and Springer Nature Author Services for writing services. This study was funded by the financial support from National Natural Science Foundation of China (No. 31460515), Jiangsu Agricultural Innovation of New Cultivars (No. PZCZ201719), Guizhou Science and Technology Support Plan (No. 20192259, Guizhou Construction of Genetics and Breeding Laboratory of Vegetable Industry System (NO. 2019-0102) and Special Fund for Agro-Scientific Research of Guizhou Academy of Agricultural Sciences (No. 2015-19).

Author information

Author notes

  1. Ying Deng and Bing Tang have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China

    Ying Deng, Xia Zhou, Lu Zhang & Jinfeng Chen

  2. Institute of Horticulture, Guizhou Academy of Agricultural Sciences, Guiyang, 55006, China

    Ying Deng, Bing Tang, Wenyuan Fu & Lian Tao

Authors
  1. Ying Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bing Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xia Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wenyuan Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lian Tao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lu Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jinfeng Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

The research and design of were guided by JFC. Data collection and analysis were performed by YD, BT, XZ, WYF, LT and LZ. The first draft of the manuscript was written by YD and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Jinfeng Chen.

Ethics declarations

Conflicts of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by Jose M. Segui-Simarro.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Deng, Y., Tang, B., Zhou, X. et al. Direct regeneration of haploid or doubled haploid plantlets in cucumber (Cucumis sativus L.) through ovary culture. Plant Cell Tiss Organ Cult 142, 253–268 (2020). https://doi.org/10.1007/s11240-020-01839-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-020-01839-w

Keywords

Search

Navigation