Sorghum is the major grain crop in semi arid tropics. Until recently the genetic improvement of sorghum for abiotic stresses has been carried out through traditional plant breeding methods. Non-availability of efficient transformation techniques is one of the limitations for the application of biotechnology for genetic improvement of this crop. Present investigation was aimed at developing transgenic sorghum with glyoxalase I gene through micro projectile mediated gene transfer.
The gly I gene which is 784 bp length, was mobilized along with CaMV35S promoter and terminator from pRT100 vector. The whole casette was cleaved by HindIII restriction enzyme and cloned into pCAMBIA 1300 binary vector having hygromycin resistance for plant selection. Sorghum shoot apices isolated from 3 days old seedlings were used as explant to induce calli on MS medium containing 2,4-D and kinetin. Thirteen days old calli were bombarded with gly I gene construct. The putative transgenics were selected and maintained till rooting on hygromycin selection pressure. PCR verification of putative transgenics was done using nptII, hpt and gly I primers. The amplification products obtained were 1050 bp, 350 and 558 bp, respectively, confirming the presence of nptII, hpt and gly I genes. |
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