Figure 1
Diagrammatic representation of gene constructs and molecular evaluation of transgenic rice lines. (a) (I) Schematic representation of T-DNA construct harboring OsCHI11 gene under the green tissue-specific promoter P D54O-544 used for rice transformation. (II) Diagrammatic representation of T-DNA construct AtNPR1 gene placed under the control of maize green tissue-specific PEPC promoter. (III) Diagrammatic representation of the T-DNA construct used to transform mature embryogenic calli of jaldi-13 rice variety. OsCHI11 and AtNPR1 genes under the control of the rice green tissue-specific promoter (P D54O–544 ) and maize green tissue-specific promoter (PEPC), respectively. (b) (I) PCR analysis of T2 transgenic rice plants with partial gene-specific (OsCHI11) primers which amplified 490 bp product. (II) PCR analysis of T2 transgenic rice plants with partial gene-specific (AtNPR1) primers which amplified 1.7Kbp product. (III) PCR based screening of T2 transgenic and non-transformed WT performed with partial gene specific primer (OsCHI11) which amplified 490 bp product. (IV) PCR analysis performed with partial gene specific primer (AtNPR1) showing amplification of 1.7Kbp product. PC-Positive control and NC- Negative control. (c) (I) Southern blot analysis of T2 transgenic plants (C): genomic DNA digested with SalI restriction enzyme and probed with 1.1 kbp HPT gene fragment. (II) Southern blot analysis T2 transgenic plants (N): genomic DNA digested by EcoRI restriction enzyme and probed with 1.1 kbp HPT gene fragment. (III) Southern hybridization analysis of T2 transgenic plants (C-N): genomic DNA was digested with SalI restriction enzyme and probed with 1.1 kbp HPT (hygromycin phosphotransferase) gene fragment. WT represents wild type.
