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

• Published in: Plant biotechnology reports Vol. 7; no. 1; pp. 99 - 108
• Main Authors: Nguyen, Quyen Van, Sun, Hyeon Jin, Boo, Kyung Hwan, Lee, Doseung, Lee, Ji-Hyun, Lim, Pyung Ok, Lee, Hyo Yeon, Riu, Key-Zung, Lee, Dong-Sun
• Format: Journal Article
• Language: English
• Published: Japan Springer-Verlag 2013; Springer Japan; Springer Nature B.V; 한국식물생명공학회
• Subjects: Acids; Agricultural biotechnology; Agriculture; benzyladenine; Biomedical and Life Sciences; Biotechnology; callus; Cell Biology; cytokinins; Embryos; gibberellic acid; Growth regulators; in vitro regeneration; Life Sciences; naphthaleneacetic acid; Original Article; Plant Biochemistry; Plant growth; Plant propagation; Plant Sciences; plantlets; Science education; Seeds; shoots; somatic embryos; Spinach; Spinacia oleracea; Studies; Sucrose; Vegetables; 농학; South Korea; Spinach; Plant regeneration; Plant growth regulator combination; Somatic embryogenesis; Culture period
• ISSN: 1863-5466; 1863-5474
• DOI: 10.1007/s11816-012-0242-3

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).