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Effect of plant growth regulator combination and culture period on in vitro regeneration of spinach (Spinacia oleracea L.)

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Abstract

The objective of this study was to develop an efficient system for the regeneration of spinach plants (Spinacia oleracea L.) by investigating the factors influencing callus and shoot induction. All plant growth regulator (PGR) combinations tested induced callus with high frequency (73–100 %), and the combination of 5 μM α-naphthaleneacetic acid (NAA), 10 μM 6-benzyladenine (BA) and 0.1 μM gibberellic acid (GA3) had the most significant effect on callus growth in term of weight (120.98 ± 22.56 mg). A high auxin-containing medium induced competent callus for shoot formation, while high cytokinin-containing media enhanced callus growth and made callus incompetent for shoot regeneration. Longer periods of callus induction in a high auxin-containing medium were required to form competent callus and led to a high regeneration capacity. The PGR combination shift from a high auxin to cytokinin ratio (ACR) to a low ACR resulted in highly efficient regeneration. Among the regeneration systems tested, the combination of 10 μM NAA and 0.3 μM GA3 for callus induction for 6 weeks followed by 2 μM NAA and 5 μM BA resulted in the highest plant regeneration frequency (83.33 ± 6.43 %) and the highest number of plantlets per explant (7.93 ± 1.24). Somatic embryos at cotyledonary stage and plantlets were transferred to PGR-free medium to establish whole plants. Regenerated female plants grew well to maturity in the greenhouse (77.17 ± 9.80 %) and produced seeds (175.21 ± 28.01 firm seeds per plant).

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Acknowledgments

This work was supported by the grants from Priority Research Centers Program (2011-0030748) and Mid-career Researcher Program (2011-0027519) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology, and the Next-Generation BioGreen 21 Program (SSAC, PJ008122; TAGC, PJ009094), Rural Development Administration, Republic of Korea.

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Authors and Affiliations

  1. College of Applied Life Science (SARI), Jeju National University, Jeju, 690-756, Republic of Korea

    Quyen Van Nguyen, Hyeon Jin Sun, Kyung Hwan Boo, Doseung Lee, Ji-Hyun Lee, Hyo Yeon Lee, Key-Zung Riu & Dong-Sun Lee

  2. Subtropical Horticulture Research Institute, Jeju National University, Jeju, 690-756, Republic of Korea

    Hyeon Jin Sun, Kyung Hwan Boo, Doseung Lee, Pyung Ok Lim, Hyo Yeon Lee, Key-Zung Riu & Dong-Sun Lee

  3. Department of Science Education, Jeju National University, Jeju, 690-756, Republic of Korea

    Pyung Ok Lim

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  1. Quyen Van Nguyen
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  2. Hyeon Jin Sun
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  3. Kyung Hwan Boo
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  4. Doseung Lee
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  5. Ji-Hyun Lee
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  6. Pyung Ok Lim
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  7. Hyo Yeon Lee
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  8. Key-Zung Riu
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  9. Dong-Sun Lee
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Corresponding authors

Correspondence to Key-Zung Riu or Dong-Sun Lee.

Additional information

Q. V. Nguyen, H. J. Sun and K. H. Boo contributed equally to this work.

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Nguyen, Q.V., Sun, H.J., Boo, K.H. et al. Effect of plant growth regulator combination and culture period on in vitro regeneration of spinach (Spinacia oleracea L.). Plant Biotechnol Rep 7, 99–108 (2013). https://doi.org/10.1007/s11816-012-0242-3

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  • DOI: https://doi.org/10.1007/s11816-012-0242-3

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