Plant Tissue Cult. 9(2) : 89-95, 1999 (December)

Genotype-dependent Embryogenesis, Organogenesis and Agrobacterium-mediated Transformation in Pigeonpea (Cajanus cajan L.)

C. H. Venu, U Pavan, T. Jaya Sree, R. V. Ramana, C. Cheralu¹ and A. Sadanandam

Plant Cell and Tissue Culture Laboratory, Department of Botany, Kakatiya University, Warangal-506 009, India

Key words: Pigeonpea, Genotypes, Transformation, Kanamycin

Abastract

Differential morphogenic responses of seven genotypes of pigeonpea (Cajanus cajan L.) was studied. Immature cotyledons were evaluated for embryogenesis and immature embryonal axes for organogenesis. Somatic embryos were induced directly from the immature cotyledons with 2, 4-D (9.0 ?M) or 5-T (7.8 ?M) in combination with BA (4.4 ?M) or Kn (4.6 ?M). Multiple shoots were obtained from embryonal axes in all the seven genotypes in response to 5?M BA. The regeneration protocol developed was employed in Agrobacterium- mediated transformation of pigeonpea. The Agrobacterium tumefaciens strain LBA 4404 carrying a binary plasmid vector system containing neomycin phosphotransferase (NPT II) gene as the selectable marker and ?-glucuronidase (GUS) as a reporter gene was used. Positive histochemical staining for GUS expression after four weeks in culture gave preliminary evidence for transformation.

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