In vitro regeneration and genetic transformation of diverse genotypes of chickpea (Cicer arietinum L.)

Yadav, Indu Singh; Singh, N. P.

In vitro regeneration and genetic transformation of diverse genotypes of chickpea (Cicer arietinum L.)

2011

Yadav, Indu Singh | Singh, N. P.

Lack of an efficient in vitro regeneration protocol is a limitingfactor for application of genetic engineering approaches for improvement of chickpea. Although regeneration and genetic transformation system has been optimized in few desi genotypes, there is an urgent need to optimize the regeneration and transformation system in kabuli types as well. Four genotypes of chickpeas (two kabuli and two desi) were screened for shoot induction and multiplication. Genotype L550, which produced highly embryogenic, rapidly growing good-quality shoots capable of regenerating at a high frequency, was selected for transformation experiments. Using a binary vector (pBinAr), frequency of GUS expression was studied. Bombardmentof embryonic axes with gold particles coated with pBinAr at a distance of 9 cm, pressure of 1300psi, and vacuum of 27mm Hg passing through 100μm mesh produced higher transformation frequency. The stable GUS-expressing embryonic axes were multiplied during selection on MS medium containing 50 mg/l kanamycin, incubated at 16/8hr light/dark. Several transformed plants with very strong GUS expression were recovered using the particle gun mediated method. Transformed shoots were confirmed through polymerase chain reaction and Dot blot analysis. These results demonstrated that Cicer arietinum is amenable to particle gun mediated genetic transformation using a binary vector.

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