Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

A Simplified Floral Dip Method for Transformation of Brassica napus and B. carinata

Abstract

A floral dip method of genetic transformation has been developed in two different species of Brassica namely B. napus cv Elect and i Pusa Gaurav. Binary vector containing B. carinata Zinc Finger transcription factor (BcZF1) gene under the transcriptional control of abiotic stress-inducible Late Embryogenesis Abundant 1 (i) promoter was transformed into Agrobacterium tumefaciens strain GV-3101. This strain was used for transformation of Brassica spp via Agrobacterium-mediated floral dip method. Agrobacterium cells cultured in Yeast Extract Mannitol (YEM) were harvested and resuspended in half MS solution containing 5% sucrose and 0.05% Silwet L-77. The transformation was performed by dipping inflorescence at the initial blossom stage. Seeds from these plants were harvested and screened by germinating them on MS medium supplemented with kanamycin (25 mg l−1). The kanamycin resistant putative transgenic plants were further confirmed by PCR and RT PCR using nptll gene-specific primers. The results showed that the Agrobacterium-mediated floral-dip transformation can be a successful strategy to develop transgenic B. napus and B. carinata in a short time without undergoing cumbersome vacuum infiltration or tissue culture steps.

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

References

  1. 1

    Feldmann KA & Marks MD, Mol Gen Genet, 208 (1987) 1.

  2. 2

    Bechtold N, Ellis J & Pelletier G, C R Acad Sci Paris Life Sci, 316 (1993) 1194.

  3. 3

    Trujillo MM, Briones VL, Ponce JLC, Estrella LH, Plant Mol Biol Rep, 22 (2004) 63.

  4. 4

    Trieu AT, Burleigh SH, Kardailsky IV, Maldonado-Mendoza IE, Versaw WK, Blaylock LA, Shin H, Chiou TJ, Katagi H, Dewbre GR, Weigel D & Harrison MJ, Plant J, 22 (2000) 531.

  5. 5

    Qing CM, Fan L, Lei Y, Bouchez D, Tourneur C, Yan L & Robaglia C, Mol Breed, 6 (2000) 67.

  6. 6

    Liu F, Cao MQ, Yao L, Li Y, Robaglia C & Tourneur C, Acta Hort, 467 (1998) 187.

  7. 7

    Wang WC, Menon G & Hansen G, Plant Cell Rep, 22 (2003) 274.

  8. 8

    Wang T, Wu W, Zhang C, Nowack LM, Liu Z & Thompson JE, Physiol Plant, 124 (2005) 493.

  9. 9

    Lu C & Kang J, Plant Cell Rep, 27 (2008) 273.

  10. 10

    Clough SJ & Bent AF, Plant J, 16 (1998) 735.

  11. 11

    Bechtold N, Jaudeau B, Jolivet S, Maba B, Vezon D, Voisin R & Pelletier G, Genetics, 155 (2000) 1875.

  12. 12

    Chung MH, Chen MK, & Pan SM, Transgenic Res, 9 (2000) 471.

  13. 13

    Desfeux C, Clough SJ & Bent AF, Plant Physiol, 123 (2000) 895.

  14. 14

    Zhang X, Henriques R, Lin SS, Niu QW & Chua NH, Nature Protocols, 1 (2006) 641.

  15. 15

    Curtis IS & Nam HG, Transgenic Res, 10 (2001) 363.

  16. 16

    Bent AF, Plant Physiol, 124 (2000) 1540.

  17. 17

    Doyle JJ & Doyle JL, Focus, 12 (1990) 13.

Download references

Author information

Correspondence to Kailash C. Bansa.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Verma, S.S., Chinnusamy, V. & Bansa, K.C. A Simplified Floral Dip Method for Transformation of Brassica napus and B. carinata . J. Plant Biochem. Biotechnol. 17, 197–200 (2008). https://doi.org/10.1007/BF03263286

Download citation

Key words

  • Brassica napus
  • B. carinata
  • floral-dip transformation
  • transgenic