biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 54:21-32, 2010 | DOI: 10.1007/s10535-010-0004-4

Transformation of chickpea: effect of genotype, explant, Agrobacterium-strain and composition of culture medium

B. Bhattacharjee1, M. Mohan1, S. Nair1,*
1 International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, India

Reproducible and high-frequency transgenic plant regeneration from callus and embryo axes of four different genotypes of chickpea (Cicer arietinum) was achieved after Agrobacterium-mediated transformation. Three different strains of Agrobacterium (EHA105, AGL1 and LBA4404) harboring the binary vector pCAMBIA1301 containing β-glucuronidase (GUS) and hygromycin phosphotransferase (hpt) genes under the control of a CaMV35S promoter were used. The highest number of transgenic plants was obtained from cotyledonary node-derived calli of genotype Pusa-256. A highly efficient rooting was achieved on Murashige and Skoog medium supplemented with indole-3-butyric acid. The stable integration of the gene was confirmed by molecular analyses of the transformed plants. Inheritance of GUS and hpt gene was followed through two generations and they showed the expected 3:1 inheritance.

Keywords: Cicer arietinum; cotyledon- and cotyledonary node-derived calli; β-glucuronidase; hygromycin phosphotransferase
Subjects: acetosyringone; Agrobacterium strains; auxins; cefotaxime; chickpea; Cicer arietinum; cytokinins; ex vitro transfer; β-glucuronidase; hygromycin; in vitro culture, callus; in vitro culture, direct organogenesis; in vitro culture, rooting; polymerase chain reaction (PCR); Southern blot analysis; sucrose; transgenic plants

Received: April 6, 2008; Accepted: October 7, 2008; Published: March 1, 2010  Show citation

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Bhattacharjee, B., Mohan, M., & Nair, S. (2010). Transformation of chickpea: effect of genotype, explant, Agrobacterium-strain and composition of culture medium. Biologia plantarum, 54(1), 21-32. doi: 10.1007/s10535-010-0004-4
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