Abstract
Camelina sativa is a promising under-exploited oilseed crop with potential to become a biofuel feedstock. The ability to transform C. sativa would allow for the rapid introduction of novel traits into this emerging crop. We report the development of an Agrobacterium-based floral dip transformation method, requiring no vacuum-infiltration step, with transformation efficiencies up to 0.8%. C. sativa cultivars Ames 26665, “Calena” A3U7761, Ames 1043, and “Celine” were tested using Agrobacterium tumefaciens strains GV3101, EHA105, and At503. Use of all strains and cultivars resulted in transformed plants; however, GV3101 was the only Agrobacterium strain and Ames 1043 the only C. sativa cultivar to yield transformed plants under all conditions tested. Progeny analysis revealed that in approximately 78% of the transformed plants, the transgene segregated as a single locus. Furthermore, a high-throughput, filter paper-based PCR method was developed to screen marker-free transformed plants. Together, these methods will allow for easier introduction of new genes into this promising oilseed crop.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bedbrook JR, Chaleff RS, Falco SC, Mazur BJ, Somerville CR, Yadav NS (1995) Nucleic acid fragment encoding herbicide resistant plant acetolactate synthase. Patent. United States 5378824
Beilstein MA, Al-Shehbaz IA, Kellogg EA (2006) Brassicaceae phylogeny and trichome evolution. Am J Bot 93:607–619
Beilstein MA, Al-Shehbaz IA, Mathews S, Kellogg EA (2008) Brassicaceae phylogeny inferred from phytochrome a and NDHF sequence data: tribes and trichomes revisited. Am J Bot 95:1307–1327
Bent AF (2000) Arabidopsis in planta transformation. Uses, mechanisms, and prospects for transformation of other species. Plant Physiol 124:1540–1547
Chhikara S, Chaudhary D, Yadav M, Sainger M, Jaiwal P (2012) A non-tissue culture approach for developing transgenic Brassica juncea L. plants with Agrobacterium tumefaciens. In Vitro Cell Dev Biol Plant 48:7–14
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743
Curtis IS, Nam HG (2001) Transgenic radish (Raphanus sativus L. longipinnatus Bailey) by floral-dip method—plant development and surfactant are important in optimizing transformation efficiency. Transgenic Res 10:363–371
Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM (1982) Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet 1:561–573
Desfeux C, Clough SJ, Bent AF (2000) Female reproductive tissues are the primary target of Agrobacterium-mediated transformation by the Arabidopsis floral-dip method. Plant Physiol 123:895–904
Falco SC, Famodu OO, Rafalski JA, Ramaker ML, Tarczynski MC, Thorpe C (2006) Plant methionine synthase gene and methods for increasing the methionine content of the seeds of plants. Patent. United States 7026527
Hajdukiewicz P, Svab Z, Maliga P (1994) The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25:989–994
Inan G, Zhang Q, Li P, Wang Z, Cao Z, Zhang H, Zhang C, Quist TM, Goodwin SM, Zhu J, Shi H, Damsz B, Charbaji T, Gong Q, Ma S, Fredricksen M, Galbraith DW, Jenks MA, Rhodes D, Hasegawa PM, Bohnert HJ, Joly RJ, Bressan RA, Zhu JK (2004) Salt cress. A halophyte and cryophyte Arabidopsis relative model system and its applicability to molecular genetic analyses of growth and development of extremophiles. Plant Physiol 135:1718–1737
Kroj T, Savino G, Valon C, Giraudat J, Parcy F (2003) Regulation of storage protein gene expression in Arabidopsis. Development 130:6065–6073
Leonard EC (1998) Camelina oil: alpha-linoleic source. Int News Fats Oils Related Mater 9:830–835
Lu C, Kang J (2008) Generation of transgenic plants of a potential oilseed crop Camelina sativa by Agrobacterium-mediated transformation. Plant Cell Rep 27:273–278
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Putnam D, Budin J, Field L, Breene W (1993) Camelina: a promising low-input oilseed. In: Janick J, Simon J (eds) New crops. Wiley, New York, pp 314–322
Qing CM, Fan L, Lei Y, Bouchez D, Tourneur C, Yan L, Robaglia C (2000) Transformation of pakchoi (Brassica rapa L. ssp. Chinensis) by Agrobacterium infiltration. Mol Breed 6:67–72
Ratcliffe OJ, Riechmann JL, Zhang JZ (2000) INTERFASCICULAR FIBERLESS 1 is the same gene as REVOLUTA. Plant Cell 12:315–317
Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81:8014–8018
Talbert PB, Adler HT, Parks DW, Comai L (1995) The REVOLUTA gene is necessary for apical meristem development and for limiting cell divisions in leaves and stems of Arabidopsis thaliana. Development 121:2723–2735
Wang WC, Menon G, Hansen G (2003) Development of a novel Agrobacterium-mediated transformation method to recover transgenic Brassica napus plants. Plant Cell Rep 22:274–281
Zubr J (1997) Oil-seed crop: camelina sativa. Industrial Crops Products 6:113–119
Acknowledgments
We are grateful to Pioneer Hi-Bred International, Inc. for the chlorsulfuron and for the SAMS–ALS gene cassette, and to Dr. Fernando Guillen-Portal at Sustainable Oils, LLC for “Celine”, Ames 1043, “Calena” A3U7761 and Ames 26665 C. sativa seeds.
Author information
Authors and Affiliations
Corresponding author
Additional information
Editor: J. Forster
X. Liu and J. Brost contributed equally to this work.
Rights and permissions
About this article
Cite this article
Liu, X., Brost, J., Hutcheon, C. et al. Transformation of the oilseed crop Camelina sativa by Agrobacterium-mediated floral dip and simple large-scale screening of transformants. In Vitro Cell.Dev.Biol.-Plant 48, 462–468 (2012). https://doi.org/10.1007/s11627-012-9459-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-012-9459-7