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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 139, Issue 3, pp 479–492 | Cite as

Embryogenesis efficiency and genetic stability of Dianthus caryophyllus embryos in response to different light spectra and plant growth regulators

  • Mostafa Aalifar
  • Mostafa ArabEmail author
  • Sasan AliniaeifardEmail author
  • Shirin Dianati
  • Mahboobeh Zare Mehrjerdi
  • Erik Limpens
  • Margrethe Serek
Original Article
  • 156 Downloads

Abstract

Carnation is an important cut flower with industrial and medicinal applications. To establish an efficient protocol without somaclonal variation for micropropagation of Dianthus caryophyllus, direct and indirect somatic embryogenesis (DSE and ISE) were investigated under six different light spectra (white, red, green, blue, red + blue and far red + red) and four combinations of different plant growth regulators (PGRs) never tested so far for carnation. The best results were achieved with 2,4-dichlorophenoxyacetic acid (2,4-D) + N-(2-chloro-4-pyridyl)-N′-phenylurea (4-CPPU) for ISE and picloram + 4-CPPU or naphthoxyacetic acid (NOA) + 6-benzylaminopurine (BAP) for DSE. The DSE method was faster (3 weeks compared to 8 weeks) and easier (no subculturing compared to two rounds of subculture with ISE methods) but the percentage of somatic embryos in the ISE method was higher compared to the DSE method. Our results showed that the highest DSE, formation of embryogenic callus, embryo maturation (generation of globular, heart and torpedo shapes) and ISE rate was observed in carnation explants exposed to blue light (450–495 nm). In contrast, green (495–570 nm), red (610–700) and far red (710–730 nm) lights caused negative effects on embryogenesis compared to white light controls (380–750 nm). For the first time, genetic stability of regenerated carnation plants was estimated using inter-simple sequence repeat (ISSR) markers. The amplified products showed 75 distinct and scorable bands, and regenerants [plants obtained by primary (PSE) and secondary SE (SSE)] were completely identical to the mother plant. Similarly, flow cytometric analysis confirmed that somatic embryo-derived plants had on average 1.53 pg nuclear DNA (2C), and all plants maintained their ploidy. In conclusion, obtained embryos under blue light were big in size and torpedo-shaped and their germination was highest compared to other light spectra. Moreover, blue light was effective for direct and indirect somatic embryogenesis in carnation without induction of somaclonal variation.

Key message

An effective protocol through application of phytohormones is introduced. Blue light can be used to improve in vitro propagation of carnation by somatic embryogenesis. Genetic stability of regenerated carnation plants was confirmed using inter-simple sequence repeat (ISSR) markers.

Keywords

Light spectrum ISSR marker Embryogenesis Carnation 

Notes

Acknowledgements

LED lights were provided by Iran grow light company (http://www.Irangrowlight.ir). The authors wish to thank Dr Christian Gehl, Faculty of Natural Sciences, Institute of Horticultural Production Systems, Floriculture, Leibniz University Hannover, Germany, for his thoughtful and inspirational comments. We would like to thank Iran National Science Foundation (INSF) (grant number 96006991) and University of Tehran for their supports.

Author contributions

MF, MA and SA performed most in vitro culture experiments and together with SD, MZ, and MF contributed to in vitro initiation experiments; MF and MZ performed the genetic analyses by ISSR markers; MA conceived and designed the experiments and together with MF, SA, EL, MS and MZ contributed to data interpretation. MF, EL, and MS contributed mostly to manuscript elaboration and other authors contributed to its revision.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Mostafa Aalifar
    • 1
  • Mostafa Arab
    • 1
    Email author
  • Sasan Aliniaeifard
    • 1
    Email author
  • Shirin Dianati
    • 1
  • Mahboobeh Zare Mehrjerdi
    • 1
  • Erik Limpens
    • 2
  • Margrethe Serek
    • 3
  1. 1.Department of Horticulture, Aburaihan CampusUniversity of TehranPakdashtIran
  2. 2.Laboratory of Molecular Biology, Department of Plant ScienceWageningen UniversityWageningenThe Netherlands
  3. 3.Faculty of Natural Sciences, Institute of Horticultural Production Systems, FloricultureLeibniz University HannoverHannoverGermany

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