1Department of Genetics and Plant Breeding, College of Agriculture, University of Agricultural Sciences, Dharwad-580 005 Karnataka, India.
2Institute of Agribiotechnology, University of Agriculture Sciences, Dharwad-580005, Karnataka, India.
A gene transfer system that ensured recovery of whole plant transformation was developed for Chickpea (Cicer arietinum L.). Embryo axes of germinating seeds with one of the cotyledons removed were pricked with a sterile needle at the cotyledonary node and infected by gentile agitation for 10 min in a suspension of Agrobacterium tumefeciens. In another method; embryos were pricked with needle dipped in agro-inoculum following a 48 h co-cultivation and decontamination with cefotaxime, they were sown in vermiculate: soil (in 1:1 proportion) mixture, moistened with water to allow germination to progress. Later, the seedlings were transferred to soil in pots where they grew into normal healthy plants in the greenhouse. The leaf paint assay with kanamycin was used for early determination of transformants, whereas PCR analysis of T1 plant DNA was used to confirm integration of genes. The combined results indicated that dipping injured embryos in inoculum for 10 min produced 5 transgenic plants in T1 generation out of 100 embryos treated in To generation. Second method, ie.. pricking with needle dipped in inoculum gave 2 transgenic plants in T1 generation out of 100 embryos treated in To generation. The strategy, in principle, should be applicable to all cultivars and genotypes of chickpea, which are susceptible to A. tumefaciens infection.
Chickpea (Cicer arietinum L.), transformation, non-tissue culture method, and embryo axes, Agrobacterium
