Figure 5

(A) Melting-curve analysis of one representative HAP gene with ILP marker in quantitative RT-PCR assay using the cDNA-pools of seedlings and two seed developmental stages of IR 64 (at least two biological replicates) produces single peak as desired, confirming the efficacy of ILP marker to amplify single gene-specific PCR product of accurate fragment size. (B) Hierarchical cluster display illustrated the differential expression profile of four HAP genes with ISM and ILP markers in seedling (SL) and two seed developmental stages (S1–S2: early cell division and S3–S5: late maturation) of one rice accession IR 64. The blue, black and yellow colour scale (mentioned at the top) signify the low, medium and high level of average log signal expression values of genes in various tissues/stages, respectively. The expression values across diverse tissues/development stages of accession were normalized using an endogenous control Actin1 in RT-PCR assay. The differential expression profiling of genes in two seed developmental stages of IR 64 was compared with their respective vegetative seedling tissue by assigning the gene expression in this tissue as a reference calibrator 1. The detail structural and functional annotation four rice HAP genes with markers are mentioned in the Tables S1 and S2. The genes with marker and tissues/stages used for expression profiling are indicated on the top and right side of an expression map, respectively.