Skip to main content
SpringerLink
Log in

Dehydrating immature embryo split apices and rehydrating withAgrobacterium tumefaciens: A new method for genetically transforming recalcitrant sunflower

  • Commentary
  • Published:
Plant Molecular Biology Reporter Aims and scope Submit manuscript

Abstract

Split embryonic axes of 21-day-old immature sunflower (Helianthus annuus L. var. RHA266) embryos were bombarded, vacuum infiltrated, or dehydrated before being infected byAgrobacterium tumefaciens strain EHA 105, bearing a binary vector carrying the selectable markerNPTII and the scorable markeruidA gene. RHA266 was found to be particularly sensitive to in vitro culture and recalcitrant to organogenesis. Of the 3 methods used, only dehydration and rehydration withAgrobacterium gave rise to transient GUS expression in shoot apices and stably transformed plants as confirmed by Southern blotting analysis.

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

Abbreviations

BAP:

6-Benzylaminopurine

MS:

Murashige-Skoog

NAA:

naphthaleneacetic acid

NPTII :

neomycin phosphotransferase gene

OD:

optical density

T-DNA:

transferred DNA

uidA (GUS):

β-glucuronidase

X-Gluc:

5-bromo-4-chloro-3-indoyl-β-D-glucuronide

References

  • Alibert B, Lucas O, Le Gall V, Kallerhoff J, and Alibert G (1999) Pectolytic enzyme treatment of sunflower explants prior to wounding and cocultivation withAgrobacterium tumefaciens, enhances efficiency of transient β-glucuronidase expression. Physiol Plant 106: 232–237.

    Article  CAS  Google Scholar 

  • Bechtold N, Ellis J, and Pelletier G (1993)In planta Agrobacterium mediated gene transfer by infiltration of adultArabidopsis thaliana plants. CR Acad Sci Vie 316: 1194–1196.

    CAS  Google Scholar 

  • Bidney D, Scelonge C, Martich J, Burrus M, Sims L, and Huffman G (1992) Microprojectile bombardment of plant tissues increases transformation frequency byAgrobacterium tumefaciens. Plant Mol Biol 18: 301–313.

    Article  PubMed  CAS  Google Scholar 

  • Burrus M, Molinier J, Himber C, Hunold R, Bronner R, Rousselin P, and Hahne G (1996)Agrobacterium-mediated transformation of sunflower (Helianthus annuus L.) shoot apices: transformation patterns. Mol Breeding 2: 329–328.

    Article  CAS  Google Scholar 

  • Everett NP, Robinson KEP, and Mascarenhas D (1987) Genetic engineering of sunflower (Helianthus annuus L.). Biotechnology 5: 1201–1204.

    Article  CAS  Google Scholar 

  • Fischer C, Klethi P, and Hahne G (1992) Protoplasts from cotyledon and hypocotyl of sunflower (Helianthus annuus L.): shoot regeneration and seed production. Plant Cell Rep 11: 632–636.

    Article  Google Scholar 

  • Flores Berrios E, Sarrafi A, Fabre F, Alibert G, and Getzbittel L (2000a) Genotypic variation and chromosomal location of QTLs for somatic embryogenesis revealed by epidermal layers culture of recombinant inbread lines in the sunflower (Helianthus annuus L.). Theor Appl Genet 101: 1307–1312.

    Article  Google Scholar 

  • Flores Berrios E, Gentzbittel L, Kayyal H, Alibert G, and Sarrafi A (2000b) AFLP mapping of QTLs for in vitro organogenesis traits using recombinant inbred lines in sunflower (Helianthus annuus L.). Theor Appl Genet 101: 1299–1306.

    Article  Google Scholar 

  • Fulton TM, Chunwongse J, and Tanksley SD (1995) Microprep protocol for extraction of DNA from tomato and other herbaceous plants. Plant Mol Biol 13: 207–209.

    Article  CAS  Google Scholar 

  • Grayburn WS, and Vick BA (1995) Transformation of sunflower (Helianthus annuus L.) following wounding with glass beads. Plant Cell Rep 14: 285–289.

    Article  CAS  Google Scholar 

  • Ishida Y, Saito H, Ohta S, Hiei Y, Komari T, and Kumashiro T (1996) High efficiency transformation of maize (Zea mays L.) mediated byAgrobacterium tumefaciens. Biotechnology 14: 745–750.

    Article  CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimaeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405.

    Article  CAS  Google Scholar 

  • Knittel N, Gruber V, Hahne G, and Lénée P (1994) Transformation of sunflower (Helianthuus annuus L.): a reliable protocol. Plant Cell Rep 14: 81–86.

    Article  CAS  Google Scholar 

  • Laparra H, Burrus M, Hunold R, Damm B, Bravo-Angel A, Bronner R, and Hahne G (1995) Expression of foreign genes in sunflower (Helianthus annuus L.)-evaluation of three gene transfer methods. Euphytica 85: 63–74.

    Article  CAS  Google Scholar 

  • Lucas O, Kallerhoff J, and Alibert G (2000) Production of stable transgenic sunflowers (Helianthus annuus L.) from wounded immature embryos by particle bombardment and co-cultivation withAgrobacterium tumefaciens. Mol Breeding 6: 479–487.

    Article  CAS  Google Scholar 

  • Malone-Schoneberg JB, Scelonge CJ, Burrus M, and Bidney D (1994) Stable transformation sunflower usingAgrobacterium and split embryonic axis explants. Plant Sci 103: 199–207.

    Article  CAS  Google Scholar 

  • Moore GA, Jacono CC, Neidigh JL, Lawrence SD, and Cline K (1992)Agrobacterium-mediated transformation ofCitrus stem segments and regeneration of transgenic plants. Plant Cell Rep 11: 238–242.

    Article  CAS  Google Scholar 

  • Mouzeyar S, Roeckel-Drevet P, Gentzbittel L, Phillipon J, Tourviele De labrouhe D, Vear F, and Nicolas P (1995) RFLP and RAPD mapping of the sunflower P11 locus for resistance toPlasmopara halstedii race 1. Theor Appl Genet 91: 733–737.

    Article  CAS  Google Scholar 

  • Müller A, Iser M, and Hess D (2001) Stable transformation of sunflower (Helianthus annuus L.) using a non-meristematic regeneration protocol and green fluorescent protein as a vital marker. Transgenic Res 10: 435–444.

    Article  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Paul MH, and Barthou H (1994) Technique simplifiée et non stérile pour la culture d'embryons immatures de tournesol (Helianthus annuus). Agronomie 14: 281–284.

    Article  Google Scholar 

  • Potrykus I (1990) Gene transfer to cereals: an assessment. Biotechnology 8: 535–542.

    Article  CAS  Google Scholar 

  • Rao KS, and Rohini VK (1999)Agrobacterium-mediated transformation of sunflower (Helianthus annuus L.): a simple protocol. Ann Bot 83: 347–354.

    Article  CAS  Google Scholar 

  • Sanford JC, Smith FD, and Russell JA (1993) Optimising the biolistic process for different biological applications. Meth Enzymol 217: 483–509.

    Article  PubMed  CAS  Google Scholar 

  • Schrammeijer B, Sijmons PC, van den Elzen JM, and Hoekema A (1990) Meristem transformation of sunflower viaAgrobacterium. Plant Cell Rep 9: 55–60.

    Article  CAS  Google Scholar 

  • Ulian EC, Smith RH, Gould JH, and McKnight TD (1988) Transformation of plants via the shoot apex. In Vitro Cell Dev Biol 24: 951–954.

    Article  Google Scholar 

  • Vancanneyt G, Schmidt R, O'Connor-Sanchez A, Willmitzer L, and Rocha-Sosa M (1990). Construction of an intron-containing marker gene: splicing of the intron in transgenic plants and its use in monitoring early events inAgrobacterium-mediated plant transformation. Mol Gen Genet 220: 245–250.

    Article  PubMed  CAS  Google Scholar 

  • Vear F, Gentzbittel L, Phillipon J, Mouzeyar S, Mestries E, Roeckel-Drevet P, Tourvielle de Labrouhe D, and Nicolas P (1997) The genetics of resistance to five races of downy mildew (Plasmopara halstedii) in sunflower (Helianthus annuus L.). Theor Appl Genet 95: 584–589.

    Article  Google Scholar 

  • Zapata C, Park SG, El-Zik KM, and Smith RH (1999) Transformation of a Texas cotton cultivar by using Agrobacterium and the shoot apex. Theor Appl Genet 98: 252–256.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Biotechnologie et d'Amélioration des Plantes (BAP), INP-ENSAT, Pôle de Biotechnologie Végétale, 18 Chemin de Borde Rouge, BP107, 31326, Auzeville Castanet Tolosan, France

    Tarek Hewezi, Annie Perrault, Gilbert Alibert & Jean Kallerhoff

Authors
  1. Tarek Hewezi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Annie Perrault
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gilbert Alibert
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jean Kallerhoff
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jean Kallerhoff.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hewezi, T., Perrault, A., Alibert, G. et al. Dehydrating immature embryo split apices and rehydrating withAgrobacterium tumefaciens: A new method for genetically transforming recalcitrant sunflower. Plant Mol Biol Rep 20, 335–345 (2002). https://doi.org/10.1007/BF02772121

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02772121

Key words

Search

Navigation