Journal of Plant Biochemistry and Biotechnology

, Volume 21, Issue 1, pp 128–133 | Cite as

Agrobacterium tumefaciens-mediated transformation protocol of Jatropha curcas L. using leaf and hypocotyl segments

  • Pratibha Misra
  • Dibya Dolridge Toppo
  • Manoj Kumar Mishra
  • Syed Saema
  • Gaurav Singh
Short Communication


In the present study, Agrobacterium tumefaciens-mediated transformation protocol has been optimized for Jatropha curcas—an important biofuel plant using leaf segments (LS) and hypocotyl segments (HS) from in vitro-seedlings. Among different accessions used, ‘GJ-Ran-C2’ was selected as the best for transformation. A. tumefaciens strain EHA101, containing the binary vector pIG121Hm was used for transformation, along with the GUS reporter gene containing intron and CaMV 35S as promoter. Various parameters were optimized to get the highest transient transformation expression (TTE). Augmentin® was used in the medium to control A. tumefaciens. Selection of transgenic shoots was done in the presence of kanamycin. Integration of GUS gene was confirmed by PCR analysis, whereas the expression of GUS transgene was confirmed by RT-PCR, using actin gene as internal control. GUS histochemical analysis of explants just after cocultivation showed blue color in approximately all the LS and HS but stable transformation efficiency was 5% in case of LS, whereas 4% in HS.


Actin gene β-glucuronidase GUS histochemical assay Shoot organogenesis 



Cauliflower mosaic virus


Cocultivation medium


β-glucuronidase gene


Hygromycin phosphotransferase gene


Neomycin phosphotransferase gene


Nopaline synthase promoter


Reverse transcription-polymerase chain reaction


Shoot induction medium


Transient transformation expression





The authors are thankful to the Director, National Botanical Research Institute (CSIR), for the facilities provided.


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Copyright information

© Society for Plant Biochemistry and Biotechnology 2011

Authors and Affiliations

  • Pratibha Misra
    • 1
  • Dibya Dolridge Toppo
    • 1
  • Manoj Kumar Mishra
    • 1
  • Syed Saema
    • 1
  • Gaurav Singh
    • 1
  1. 1.National Botanical Research Institute, Council of Scientific and Industrial Research (CSIR)LucknowIndia

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