Agrobacterium-mediated transformation of Camelina sativa for production of transgenic plants
Camelina sativa (C. sativa), an oilseed species rich in poly-unsaturated fatty acids, has gained great importance as an industrial oil platform crop in recent years. Despite the potential benefits of C. sativa for bioenergy applications, limited research has been conducted to improve its agronomic qualities. Hence, a simple and efficient technique for production of transgenic C. sativa plants is warranted. In the present study, shoot apical meristems of two C. sativa cultivars (Pl650159 and Pl650161) were transformed with Agrobacterium strain ‘EHA 105’ harboring the enhanced green fluorescent protein (EGFP) and neomycin phosphotransferase II (nptII) genes. After two days of co-cultivation in the dark, explants were transferred to selection medium. Transgenic shoots were identified on the basis of green fluorescence and kanamycin resistance. Shoots were then rooted and transferred to potting mix soil for acclimatization. This protocol describes an efficient method to generate transgenic C. sativa plants in as little as 4 weeks.
1. Kumar A, Kumar K, Kaushik N, Sharma S, Mishra S (2010) Renewable energy in India: current status and future potentials. Renew Sust Energ Rev 14: 2434-2442.
2. Shonnard DR, Williams L, Kalnes TN (2010) Camelina‐derived jet fuel and diesel: Sustainable advanced biofuels. Environ Prog Sust Energ 29: 382-392.
3. Gugel RK, Falk KC (2006) Agronomic and seed quality evaluation of Camelina sativa in western Canada. Can J Plant Sci 86: 1047-1058.
4. Soriano NU, Narani A (2012) Evaluation of biodiesel derived from Camelina sativa oil. J Am Oil Chem Soc 89: 917-923.
5. Frohlich A, Rice B (2005) Evaluation of Camelina sativa oil as a feedstock for biodiesel production. Ind Crop Prod 21: 25-31.
6. Bernardo A, Howard HR, O'Connell A, Nichol R, Ryan J, Rice B, Leahy J (2003) Camelina oil as a fuel for diesel transport engines. Ind Crop Prod 17:191-197.
7. Narasimhulu SB, Kirti PB, Bhatt SR, Prakash S, Chopra VL (1994) Intergeneric protoplast fusion between Brassica carinata and Camelina sativa. Plant Cell Rep 13: 657-660.
8. Sigareva MA, Earle ED (1999) Camalexin induction in intertribal somatic hybrids between Camelina sativa and rapid-cycling Brassica oleracea. Theor Appl Genet 98: 164-170.
9. Tattersall A, Millam S (1998) Establishment and in vitro regeneration studies of the potential oil crop species Camelina sativa. Plant cell Tiss Org 55: 147-150.
10. 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.
11. Liu X, Brost J, Hutcheon C, Guilfoil R, Wilson AK, Leung S, De Rocher J (2012) Transformation of the oilseed crop Camelina sativa by Agrobacterium-mediated floral dip and simple large-scale screening of transformants. In Vitro Cell Dev Pl 48: 462-468.
12. Li J, Tan X, Zhu F, Guo J (2010) A rapid and simple method for Brassica napus floral-dip transformation and selection of transgenic plantlets. Int J Biol 2: 127-131.
13. Barlass M, Skene KGM (1978) In vitro propagation of grapevine (Vitis vinifera L.) from fragmented shoot apices. Vitis 17: 335-340.
14. Dhekney SA, Li ZT, Dutt M, Gray DJ (2012) Initiation and transformation of grapevine embryogenic cultures. In Transgenic Plants 1: 215-225.
15. Dutt M, Grosser JW (2009) Evaluation of parameters affecting Agrobacterium-mediated transformation of citrus. Plant Cell Tissue Organ Cult 98:331–340.