Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 135, Issue 3, pp 407–417 | Cite as

Improved chromosome doubling of parthenogenetic haploid plants of cucumber (Cucumis sativus L.) using colchicine, trifluralin, and oryzalin

  • Hamed Ebrahimzadeh
  • Hassan SoltanlooEmail author
  • Mehran E. ShariatpanahiEmail author
  • Ali Eskandari
  • Seiede Sanaz Ramezanpour
Original Article


An effective chromosome doubling protocol was established in essential garden crop of cucumber (Cucumis sativus L.) Cv. Hi Power. The different concentrations of colchicine (0, 250, 500, 750, and 1500 mg/L), oryzalin (0, 5, 15, 25, 50, 75, and 150 mg/L) and trifluralin (0, 5, 15, 25, 50, 75, and 150 mg/L) were applied on parthenogenesis-induced haploid nodal and shoot tip explants of cucumber for 18 and 38 h in three independent factorial experiments. Increasing concentrations of applied antimitotic agents led to the significant reduction in the survival rate of both shoot tip and nodal explants, especially in longer exposure duration. Three ploidy levels including haploid, mixoploid, and doubled haploid were regenerated form both explant types treated with colchicine, oryzalin, and trifluralin. Flow cytometry analysis proved successful chromosome doubling of haploid plants. Based on the results obtained, the highest number of regenerated doubled haploid plants (92.31%) and fruit set (86.21%) were related to immersion of nodal explants in 50 mg/L oryzalin for 18 h. The highest doubled haploid regeneration for colchicine and trifluralin antimitotic agents were 58.33 and 83.33%, respectively. The leaf size of doubled haploid plants was larger than their correspond haploids. The optimized chromosome doubling protocol would be applicable for doubled haploid production in garden crops of Cucurbitaceae family, which is recalcitrant to the spontaneous doubling, and also for in vitro polyploidy induction studies.


Artificial genome duplication Double haploids Antimitotic agent Embryos rescue 



This research was supported by grants from Agricultural Biotechnology Research Institute of Iran (ABRII) Project No. 12-05-05-9451-94001.

Author Contributions

HE contributed in the practical process of parthenogenesis induction, regeneration of plants, chromosome doubling, cytological and flow cytometer analysis. HS and MES (corresponding authors) contributed in the supervising of the whole process. AE contributed in the pollen irradiation experiments. SSR contributed in data analysis.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Hamed Ebrahimzadeh
    • 1
    • 2
  • Hassan Soltanloo
    • 1
    Email author
  • Mehran E. Shariatpanahi
    • 2
    Email author
  • Ali Eskandari
    • 3
  • Seiede Sanaz Ramezanpour
    • 1
  1. 1.Department of Plant Breeding and BiotechnologyGorgan University of Agricultural Sciences and Natural ResourcesGorganIran
  2. 2.Department of Tissue and Cell Culture, Agricultural Biotechnology Research Institute of Iran (ABRII)Agricultural Research, Education and Extension Organization (AREEO)KarajIran
  3. 3.Nuclear Agriculture Research School- NSTRIKarajIran

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