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Direct somatic embryogenesis from pericycle cells of broccoli (Brassica oleracea L. var. italica) root explants

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Abstract

Cotyledon, hypocotyl or root explants of 7-day-old broccoli seedlings were cultured on Murashige and Skoog (MS) agar or liquid medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). The frequency of direct somatic embryo formation was 100% when root explants were cultured in liquid medium. Histological analysis indicated that somatic embryos were initiated directly from the pericycle cell layers of root explants as early as 1 day after liquid culture. Genotype did not affect the frequency of somatic embryo formation or the number of somatic embryos per explant. All broccoli genotypes examined had 100% somatic embryo induction efficiency, and the number of somatic embryos per 0.8 mm root segment ranged from 22.9 in ‘Luhui’ to 26.0 in ‘Haizi’. The number of normally developed somatic embryos in culture increased with increasing 2,4-D concentration. Plantlet regeneration frequency was the highest (73.3%) when germinated plantlets were transferred to 1/2 strength MS agar medium containing 1.0 mg l−1 6-benzyladenine (BA). When regenerated plantlets were transferred to a greenhouse, approximately 75% survived and there were no morphological differences between regenerated plants and seed-derived controls. The protocols established in this study will benefit large-scale vegetative propagation and transformation-based genetic improvement of broccoli.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BA:

6-Benzyladenine

GA3 :

Gibberellic acid

KN:

Kinetin

LRM:

Lateral root meristem

MS:

Murashige and Skoog

NAA:

α-Naphthaleneacetic Acid

PGRs:

Plant growth regulators

SAM:

Shoot apical meristem

TDZ:

Thidiazuron

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Acknowledgments

This research was supported by the National Natural Science Foundation of China (No. 30671701).

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Authors and Affiliations

  1. Key Laboratory of Forest Improvement and Biotechnology, Ministry of Education, Northeast Forestry University, 150040, Harbin, China

    J. L. Yang & Cheng Hao Li

  2. Bioherb Research Institute, Kangwon National University, Chuncheon, 200-701, South Korea

    E. S. Seong, M. J. Kim, W. H. Kang & Chang Yeon Yu

  3. Department of Horticulture, College of Agriculture and Life Sciences, Kangwon National University, Chuncheon, 200-701, South Korea

    W. H. Kang

  4. Department of Applied Life Science, Konkuk University, Seoul, 143-701, South Korea

    B. K. Ghimire

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  1. J. L. Yang
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  2. E. S. Seong
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  3. M. J. Kim
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Correspondence to Chang Yeon Yu or Cheng Hao Li.

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J. L. Yang and E. S. Seong contributed equally to this work.

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Yang, J.L., Seong, E.S., Kim, M.J. et al. Direct somatic embryogenesis from pericycle cells of broccoli (Brassica oleracea L. var. italica) root explants. Plant Cell Tiss Organ Cult 100, 49–58 (2010). https://doi.org/10.1007/s11240-009-9615-x

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