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Establishment of a simple and efficient system for somatic embryo induction via ovule culture in Arabidopsis thaliana

  • Cell Biology and Morphogenesis
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Abstract

We established a simple and effective system to induce somatic embryos in Arabidopsis via ovule culture. Agar-solidified B5 basic medium supplemented with 10 μ M 2,4-dichlorophenoxyacetic acid was used for callus induction. Ovules at all developmental stages were tested, and among these, ovules older than 48 h after anthesis could be successfully induced to form embryogenic calli at high frequencies (42–82%). Structural and molecular probe analyses confirmed that the embryogenic calli were derived from embryos in the ovules. These calli were then easily induced to generate somatic embryos at frequencies of 63–95%. Subculture of the somatic embryos onto 1/2 strength MS medium resulted in their direct conversion into plants. The regenerants appeared morphologically normal and were fertile. This method provides a useful alternative tool to create sufficient numbers of somatic embryos for the study of biochemical and molecular mechanisms of embryogenesis, especially to recover early defective embryos in some mutations for cell-biological analyses.

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Abbreviations

CIM:

Callus-induction medium

EC:

Embryogenic calli

HAA:

Hours after anthesis

SE:

Somatic embryo

2,4-D:

2,4-dichlorophenoxyacetic acid

References

  • Berleth T (1998) Experimental approaches to Arabidopsis embryogenesis. Plant Physiol Biochem 36:69–82

    Article  CAS  Google Scholar 

  • Berleth T, Chatfield C (2002) Embryogenesis: pattern formation from a single cell. In: Somerville CR, Meyerowitz EM (eds) The Arabidopsis Book. American Society of Plant Biologists, Rockville, MD

    Google Scholar 

  • Chen JG, Ullah H, Young JC, Sussman MR, Jones AM (2001) ABP1 is required for organized cell elongation and division in Arabidopsis embryogenesis. Genes Dev 15:902–911

    Article  PubMed  CAS  Google Scholar 

  • Gaj MD (2001) Direct somatic embryogenesis as a rapid and efficient system for in vitro regeneration of Arabidopsis thaliana. Plant Cell Tiss Org 64:39–46

    Article  Google Scholar 

  • Gambog OL, Miller RA, Ojima K (1968) Nutrient requirement of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  Google Scholar 

  • Hadfi K, Speth V, Neuhaus G (1998) Auxin—induced developmental patterns in Brassica juncea embryos. Development 125:879–887

    PubMed  CAS  Google Scholar 

  • Higashi K, Shiota H, Kamada H (1998) Patterns of expression of the genes for glutamine synthetase isoforms during somatic and zygotic emryogenesis in carrot. Plant Cell Physiol 39:418–424

    PubMed  CAS  Google Scholar 

  • Hoagland DR, Arnon DJ (1938) The Water culture method for growing plants without soil. Univ Calif Agr Expt Sta Cire, 347p

  • Ikeda-Iwai M, Satoh S, Kamada H (2002) Establishment of a reproducible tissue culture system for induction Arabidopsis somatic embryos. J Exp Bot 53:1575–1580

    Article  PubMed  CAS  Google Scholar 

  • Jürgens G, Mayer U (1994) Embryos: color atlas of development. In: Bard JBL (ed) Arabidopsis. Wolfe Publishers, London, pp 7–21

    Google Scholar 

  • Jürgens G, Mayer U, Ruiz RAT, Berleth T (1991) Genetic analysis of pattern formation in the Arabidopsis embryo. Development Suppl 1:27–38

    Google Scholar 

  • Laux T, Jürgens G (1997) Embryogenesis: a new start in life. Plant Cell 9:989–1000

    Article  PubMed  CAS  Google Scholar 

  • Liu CM, Xu ZH, Chua NH (1993) Auxin polar transport is essential for the establishment of bilateral symmetry during early plant embryogenesis. Plant Cell 5:621–630

    Article  PubMed  CAS  Google Scholar 

  • Luo Y, Koop HU (1997) Somatic embryogenesis in cultured immature zygotic embryos and leaf protoplasts from Arabidopsis thaliana ecotypes. Planta 202:387–396

    Article  PubMed  CAS  Google Scholar 

  • Malamy JE, Benfey PN (1997). Organization and cell differentiation in lateral roots of Arabidopsis thaliana. Development 124:33–44

    PubMed  CAS  Google Scholar 

  • Mansfield SG, Briarty LG (1991) Early embbryogensis in arabidopsis thaliana: II The developing embryo. Can J Bot 69:461–476

    Google Scholar 

  • Meinke DW (1991) Perspectives on genetic analysis of plant Embryogenesis. Plant Cell 3:857–866

    Article  PubMed  CAS  Google Scholar 

  • Meinke DW (1995) Molecular genetics of plant embryogenesis. Ann Rev Plant Physiol Plant Mol Biol 46:369–394

    Article  CAS  Google Scholar 

  • Mordhorst AP, Toonen MAJ, de Vries SC (1997) Plant embryogenesis. Crit Rev Plant Sci 16:535–576

    Google Scholar 

  • Mordhorst AP, Voerman KJ, Hartog MV, Meijer EA, VanWent J, Koornneef M, de Vries SC (1998) Somatic embryogenesis in Arabidopsis thaliana is facilitated by mutation in genes repressing meristematic cell divisions. Genetics 149:549–563

    PubMed  CAS  Google Scholar 

  • Müller AJ (1961) Zur chrakterisierung der bluten und infloreszenzen von Arabidopsis thaliana (L.) Heynh. Kulturpflanze 9:364–393

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:437–497

    Article  Google Scholar 

  • O’Neill CM, Matthias RJ (1993) Regeneration of plants from protoplasts of Arabidopsis thaliana L. cv. Columbia (C24) via direct embryogenesis. J Exp Bot 44:1579–1585

    Google Scholar 

  • Pillon E, Teriz M, Baldan B, Mariani P, Schiavo FL (1996) A protocol for obtaining embryogenic cell lines from Arabidopsis. Plant J 9:573–577

    Article  PubMed  CAS  Google Scholar 

  • Raghavan V (2002) Induction of vivipary in Arabidopsis by silique culture: Implications for seed dormancy and germination. Am J Bot 89:766–776

    Google Scholar 

  • Sangwan RS, Bourgeois Y, Duboid F, Sangwan NBS (1992) In vitro regeneration of Arabidopsis thaliana of cultured zygotic embryos and analysis of regenerants. J Plant Physiol 140:588–595

    CAS  Google Scholar 

  • Sauer M, Friml J (2004) In vitro culture of Arabidopsis embryos within their ovules. Plant J 40:835–843

    Article  PubMed  Google Scholar 

  • Schneider T, Dinkins R, Robinson K, Shellhammer J, Meinke DW (1989) An embryo-lethal mutant of Arabidopsis thaliana is a biotin auxotroph. Dev Biol 131:161–167

    PubMed  CAS  Google Scholar 

  • Shellhammer J, Meinke DW (1990) Arrested embryos from the bio1 auxotroph of Arabidopsis thaliana contain reduced levels of biotin. Plant Physiol 93:1162–1167

    Article  PubMed  CAS  Google Scholar 

  • Shiota H, Satoh R, Watabe K, Harada H, Kamade H (1998) C-ABI3, the carrot homologue of the Arabidopsis ABI3, is expressed during both zygotic and somatic embryogenesis and functions in the regulation of embryo-specific ABA-inducible genes. Plant Cell Physiol 39:1184–1193

    PubMed  CAS  Google Scholar 

  • Shiota H, Tachikawa K, Watabe K, Kamada H (1999) Successful long-term preservation of abscisic acid-treated and desiccated carrot somatic embryos. Plant Cell Rep 18:749–753

    Article  CAS  Google Scholar 

  • Wu Y, Haberland G, Zhou C, Koop HU (1992) Somatic embryogenesis, formation of morphogenetic callus and normal development in zygotic embryos of Arabidopsis thaliana in vitro. Protoplasma 169:89–96

    Article  Google Scholar 

  • Zimmerman JL (1993) Somatic embryogenesis: a model for early development in higher plant. Plant Cell 5:1411–1423

    Article  PubMed  Google Scholar 

Download references

Acknowledgement

We thank Professor Tom J. Guilfoyle for his kindness in providing us DR5::GUS (Columbia ecotype). This research is supported by the National Natural Science Foundation of China (30370743, 90408002), National Outstanding Youth Science Fund (30225006), and “PCSIRT.”

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Authors and Affiliations

  1. Key Laboratory of MOE for Plant Development Biology, College of Life Science, Wuhan University, Wuhan, 430072, P.R. China

    Jun Wei, Xin-Ran Li & Meng-Xiang Sun

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  1. Jun Wei
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  2. Xin-Ran Li
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  3. Meng-Xiang Sun
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Correspondence to Meng-Xiang Sun.

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Communicated by P. Lakshmanan

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Wei, J., Li, XR. & Sun, MX. Establishment of a simple and efficient system for somatic embryo induction via ovule culture in Arabidopsis thaliana . Plant Cell Rep 25, 1275–1280 (2006). https://doi.org/10.1007/s00299-006-0166-x

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  • DOI: https://doi.org/10.1007/s00299-006-0166-x

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