Skip to main content
SpringerLink
Log in

Medium optimization for efficient somatic embryogenesis and plant regeneration from immature inflorescences and immature scutella of elite cultivars of wheat, barley and tritordeum

  • Published:
Euphytica Aims and scope Submit manuscript

Abstract

Media have been developed for somatic embryogenesis and plant regeneration from immature inflorescences and immature scutella of elite cultivars of wheat, barley and tritordeum. For wheat and tritordeum inflorescences, regeneration from embryogenic calluses induced on medium with picloram was almost twice as efficient as regeneration from cultures induced on 2,4-dichlorophenoxyacetic acid (2,4-D). The addition of zeatin at 5 or 10 mg l−1 to regeneration media had a positive effect on regeneration. For scutella, the highest frequencies of embryogenesis (85%) and regeneration (50%) was obtained using an induction medium containing 2 mg l−1 of 2,4-D and half concentration of aminoacids. The morphogenetic capacities of 19 different cultivars of wheat, barley and tritordeum were compared, and clear differences were found both between explants and genotypes. In wheat, embryogenic capacity from inflorescences (average of 92%) was higher than from immature scutella (average of 62%). However, shoot regeneration from scutella was clearly higher than from inflorescences (averages of 63%, and 18% respectively). Frequencies of regeneration in wheat and barley varied widely among the cultivars tested and in both species no difference was found between spring and winter varieties.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Afsharsterlel, S., E.C.K. Pang, J.S. Brown & J.F. Kollmorgen, 1996. Embryogenic callus induction and plant-regeneration in Triticum tauschii, the diploid D-genome donor for bread wheat (Triticum aestivum). Aust J Bot 44: 489–497.

    Article  Google Scholar 

  • Barcelo, P., A. Vasquez & A. Martin, 1989. Somatic embryogenesis and plant regeneration from tritordeum. Plant Breeding 103: 235–240.

    Article  Google Scholar 

  • Barcelo, P., C. Hagel, D. Becker, A. Martin & H. Lorz, 1994. Transgenic cereal (tritordeum) plants obtained at high efficiency by microprojectile bombardment of inflorescence tissue. Plant J 5: 583–592.

    PubMed  CAS  Google Scholar 

  • Barcelo, P. & P.A. Lazzeri, 1995. Transformation of cereals by microprojectile bombardments of immature inflorescence and scutellum tissues. In: H. Jones (Ed), Methods in Molecular Biology: Plant Gene Transfer and Expression Protocols, 49: 113–123. Humana Press Inc., Totowa, NJ.

    Google Scholar 

  • Becker, D., R. Brettschneider & H. Lorz, 1994. Fertile transgenic wheat from microprojectile bombardment of scutellar tissue. Plant J 5: 299–307.

    Article  PubMed  CAS  Google Scholar 

  • Casas, A.M., A.K. Kononowicz, T.G. Haan, L.Y. Zhang, D.T. Tomes, R.A. Bressan & P.M. Hasegawa, 1997. Transgenic sorghum plants obtained after microprojectile bombardment of immature inflorescences. In Vitro Cell Dev Biol 33: 92–100.

    Google Scholar 

  • Daniel, G., 1993. Anther culture in rye. Improved plant-regeneration using modified MS-media. Plant Breeding 110: 259–261.

    Article  Google Scholar 

  • Denchev, P.D. & B.V. Conger, 1995. In-vitro culture of switchgrass: Influence of 2,4-D and picloram in combination with benzyladenine on callus initiation and regeneration. Plant Cell Tiss Org Cul 40: 43–48.

    Article  CAS  Google Scholar 

  • Dineshkumar, V., P.B. Kirti, J.K.S. Sachan & V.L. Chopra, 1995. Picloram induced somatic embryogenesis in chickpea (Cicer arietinum L.). Plant Sci 109: 207–213.

    Article  CAS  Google Scholar 

  • Elena, E.B. & H.D. Ginzo, 1988. Effect of auxin levels on shoot formation with different embryo tissues from a cultivar and a commercial hybrid of wheat (Triticum aestivum L.). J Plant Physiol 132: 600–603.

    CAS  Google Scholar 

  • Fennell, S., N. Bohorova, M. van Ginkel, J. Crossa & D. Hoisington, 1996. Plant regeneration from immature embryos of 48 elite CIMMYT bread wheats. Theor Appl Genet 92: 163–169.

    Article  Google Scholar 

  • Fellers, J.P., A.C. Guenzi & C.M. Taliaferro, 1995. Factors affecting the establishment and maintenance of embryogenic callus and suspension-cultures of wheat (Triticum aestivum L.). Plant Cell Rep 15: 232–237.

    Article  CAS  Google Scholar 

  • He, D.G., Y.M. Yang & K.J. Scott, 1989. The effect of macroelements in the induction of embryogenic callus from immature embryos of wheat (Triticum aestivum L.). Plant Sci 64: 251–258.

    Article  CAS  Google Scholar 

  • Koprek, T., R. Hänsch, A. Nerlich, R.R. Mendel & J. Achulze, 1996. Fertile transgenic barley of different cultivars obtained by adjustment of bombardment conditions to tissue response. Plant Sci 119: 79–91.

    Article  CAS  Google Scholar 

  • Kysely, W., J.R. Myers, P.A. Lazzeri, G.B. Collins & H.J. Jacobsen, 1987. Plant regeneration via somatic embryogenesis in pea (Pisum sativum L.). Plant Cell Rep 6: 305–308.

    Article  CAS  Google Scholar 

  • Lazzeri, P.A., R. Brettschneider, R. Luhrs & H. Lórz, 1991. Stable transformation of barley via PEG-induced direct DNA uptake into protoplasts. Theor Appl Genet 81: 437–444.

    Article  Google Scholar 

  • Lazzeri, P.A. & P.R. Shewry, 1994. Biotechnology of cereals. In: M.P. Tombs (Ed), Biotechnology and Genetic Engineering Reviews, pp. 79–146. Intercept Ltd., Andover.

    Google Scholar 

  • Lehminger-Mertens, R. & H.J. Jacobsen, 1989. Plant regeneration from pea protoplasts via somatic embryogenesis. Plant Cell Rep 8: 379–382.

    Article  Google Scholar 

  • Linacero, R. & A.M. Vazquez, 1990. Somatic embryogenesis from immature inflorescences of rye. Plant Sci 72: 253–258.

    Article  CAS  Google Scholar 

  • Maes, O.C., R.N. Chibbar, K. Caswell, N. Leung & K.K. Kartha, 1996. Somatic embryogenesis from isolated scutella of wheat-effects of physical, physiological and genetic-factors. Plant Sci 121: 75–84.

    Article  Google Scholar 

  • Maddock, S.E., V.A. Lancaster, R. Risiott & J. Franklin, 1983. Plant regeneration from cultured immature embryos and inflorescences of 25 cultivars of wheat (Triticum aestivum). J Exp Bot 34: 915–926.

    Google Scholar 

  • Martin, A. & E. Sanchez-Monge, 1982. Cytology and morphology of the amphiploid Hordeum chilense × Triticum turgidum conv durum. Euphytica 31: 261–267.

    Article  Google Scholar 

  • Mathias, R.J. & E.S. Simpson, 1986. The interaction of genotype and culture medium on the tissue culture responses of wheat (Triticum aestivum L.) callus. Plant Cell Tiss Org Cul 7: 31–37.

    Article  Google Scholar 

  • Murashige, T. & F. Skoog, 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497.

    Article  CAS  Google Scholar 

  • Nehra, N.S., R.N. Chibbar, N. Leung, K. Caswell, C. Mallard, L. Steinhauer, M. Baga & K.K. Kartha, 1994. Self-fertile transgenic wheat plants regenerated from isolated scutellar tissues following microprojectile bombardment with two distinct gene constructs. Plant J 5: 285–297.

    Article  CAS  Google Scholar 

  • Nobre, J., M.R. Davey & P.A. Lazzeri, 1997. Barley scutellum protoplasts. Isolation, culture and plant regeneration. Physiol Plant 98: 868–874.

    Article  Google Scholar 

  • Ozias-Akins, P. & I.K. Vasil, 1982. Plant regeneration from cultured immature embryos and inflorescences of Triticum aestivum L. (wheat): evidence for somatic embryogenesis. Protoplasma 110: 95–105.

    Article  Google Scholar 

  • Parrot, W.A., S.A. Merkle & E.G. Williams, 1991. Somatic embryogenesis: potential for use in propagation and gene transfer system. In: D.R. Murray (Ed), Advanced Methods in Plant Breeding and Biotechnology, pp. 158–200. CAB International, Wallinford.

    Google Scholar 

  • Ranch, J.P., L. Ogelsby & A.C. Zielinsky, 1986. Plant regeneration from tissue cultures of soybean by somatic embryogenesis. In: I.K. Vasil (Ed), Cell Culture and Somatic Cell Genetics of Plants, pp. 97–110. Academic Press Inc., New York.

    Google Scholar 

  • Redway, F.A., V. Vasil, D. Lu & I.K. Vasil, 1990. Identification of callus types for long-term maintenance and regeneration from commercial cultivars of wheat (Triticum aestivum L.). Theor Appl Genet 79: 609–617.

    Article  Google Scholar 

  • Rhodes, C.A., C.E. Green & R.L. Phillips, 1986. Factors affecting culture initiation from maize tassels. Plant Sci 46: 225–232.

    Article  Google Scholar 

  • Rout, J.R. & W.J. Lucas, 1996. Characterization and manipulation of embryogenic response from in-vitro-cultured immature inflorescences of rice (Oryza sativa L.). Planta 198: 127–138.

    Article  CAS  Google Scholar 

  • Songstad, D.D., W.L. Petersen & C.L. Armstrong, 1992. Establishment of friable embryogenic (type II) callus from immature tassels of Zea mays (Poaceae). Am J Bot 79: 761–764.

    Article  Google Scholar 

  • Takumi, S. & T. Shimada, 1997. Variation in transformation frequencies among 6 common wheat cultivars through particle bombardment of scutellar tissues. Genes Genet Syst 72: 63–69.

    Article  PubMed  CAS  Google Scholar 

  • Thomas, M.R. & K.J. Scott, 1985. Plant regeneration by somatic embryogenesis from callus initiated from immature embryos and immature inflorescences of Hordeum vulgare. J Plant Physiol 121: 159–169.

    CAS  Google Scholar 

  • Vasil, I.K., 1987. Developing cell and tissue culture systems for the improvement of cereal and grass crops. J Plant Physiol 128: 193–218.

    Google Scholar 

  • Vasil, V., V. Srivastava, A.M. Castillo, M.E. From & I.K. Vasil, 1993. Rapid production of transgenic wheat plants by direct bombardment of cultured embryos. Bio/Technology 11: 1553–1558.

    Article  Google Scholar 

  • Wan, Y. & P.G. Lemaux, 1994. Generation of large numbers of independently transformed fertile barley plants. Plant Physiol 104: 37–48.

    PubMed  CAS  Google Scholar 

  • Weeks, J.T., O.D. Anderson & A.E. Blechl, 1993. Rapid production of multiple independent lines of fertile transgenic wheat (Triticum aestivum L.). Plant Physiol 102: 1077–1084.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. IACR-Rothamsted, Harpenden Herts, AL5 2JQ, U.K.

    Francisco Barro, Paul A. Lazzeri & Pilar Barceló

  2. Instituto de Agricultura Sostenible, Apdo 4084, 14080-, Córdoba, Spain

    Antonio Martin

Authors
  1. Francisco Barro
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Antonio Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paul A. Lazzeri
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pilar Barceló
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Barro, F., Martin, A., Lazzeri, P.A. et al. Medium optimization for efficient somatic embryogenesis and plant regeneration from immature inflorescences and immature scutella of elite cultivars of wheat, barley and tritordeum. Euphytica 108, 161–167 (1999). https://doi.org/10.1023/A:1003676830857

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1003676830857

Search

Navigation