African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12278

Full Length Research Paper

Agrobacterium tumefaciens-mediated transformation of biofuel plant Jatropha curcas using kanamycin selection

Jingli Pan1, Qiantang Fu2 and Zeng-Fu Xu1, 2*
1State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, Guangdong, China. 2Laboratory of Molecular Breeding of Energy Plants, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming 650223, Yunnan, China.
Email: [email protected]

  •  Accepted: 26 August 2010
  •  Published: 27 September 2010

Abstract

Establishment of an efficient transformation system is a prerequisite for genetic improvement of Jatropha curcas, a promising biodiesel feedstock plant, by transgenic approach. In this study an efficient Agrobacterium-mediated transformation protocol using cotyledon explants from J. curcas seeds was developed. The integration and expression of the transgenes in the putatively transformed J. curcas plants was confirmed by polymerase chain reaction (PCR), Southern blot analysis and β-glucuronidase (GUS) activity staining. In the protocol used, the critical step for successful transformation is that the selective agent kanamycin (20 mg/L) is not included in the callus-inducing medium within the first 4 weeks after co-cultivation with Agrobacterium.   

 

Keywords: Agrobacterium-mediated transformation, biofuel, Jatropha curcas, kanamycin, physic nut, plant regeneration, transgenic plants.

Abbreviation

BA, 6-Benzylaminopurine; CaMV, cauliflower mosaic virus; GUS,β-glucuronidase; IBA, indole-3-butyric acid; MS, Murashige and Skoog; NPT II,neomycin phosphotransferase II; PCR, polymerase chain reaction; YEP, yeast extract peptoneCIM, callus-inducing mediumSIM, shoot-inducing medium; RM,rooting mediumCTAB, cetyl trimethylammonium bromide.