Agrobacterium mediated transformation of chickpea (Cicer arietinum L.) embryo axes

Embryo axes of four accessions of chickpea (Cicer arietinum L.) were treate d with Agrobacterium tumefaciens strains C58C1/GV2260 carrying the plasmid p35SGUSINT and EHA101 harbouring the plasmid pIBGUS. In both vectors the GU S gene is interrupted by an intron, After inoculation shoot formation was p romoted on MS medium containing 0.5 mg/l BAP under a selection pressure of 100 mg/l kanamycin or 10 mg/l phosphinothricin, depending on the construct used for transformation. Expression of the chimeric GUS gene was confirmed by histochemical localization of GUS activity in regenerated shoots. Resist ant shoots were grafted onto 5-day-old dark-grown seedlings, and mature pla nts could be recovered. T-DNA integration was confirmed by Southern analysi s by random selection of putative transformants, The analysis of 4 plantlet s of the T1 progeny revealed that none of them was GUS-positive, whereas th e presence of the nptII gene could be detected by polymerase chain reaction .