2.3. Molecular Characterization of Selected Wheat Genotypes
The genes related to pigment-binding proteins were selected to examine the different expression in four genotypes of wheat which was grown in normal condition 100% field capacity and drought stress condition 80% field capacity and 60% field capacity. Total 6 genes were studied and specific primers for genes encoding LHCI and LHCII were designed based on published expressed sequence tags. The primers were: TaLhca1 - F 5’ CAACCTGCCGACCATCCTG-3’ and TaLhca1- R 5’CAGCCGCCCGTTCTTGAT-3’ and TaLhca2 - F 5’CCCCAACCGCAAGAACC-3’ andTaLhca2- R 5’CCGACGAAGGCGAGCAT-3’, and TaLhca3 - F 5’CCTCACCAGCCTCAAGTTCC-3’, and TaLhca3- R 5’CGCACGCTCACGTTTCC-3’, and TaLhcb1 - F 5’GGAGAACACACAATACACC-3’ and TaLhcb1- R 5’CCCATTATGTGTGCAGTTC-3’ and TaLhcb4 - F 5’AAAGGCCGAGGAGGACAA-3’ and TaLhcb4- R 5’ CCACCGACCACTTAAGAGG-3’ and TaLhcb6 - F TCAGCGACCTCACCGTCA-3’ and TaLhcb6- R 5’CCCCAAAGAAGTCACGGACA-3’, and amplification of the T. aestivum tubulin gene using primerTubulin- F 5’ACCGCCAGCTCTTCCACCCT- 3’ and Tubulin- R 5 TCACTGGGGCATAGGAGGAA- 3’ exhibiting constitutive expression was used as a positive control, from which a linear relationship between the amount of RNA used for amplification and the amount of cDNA fragment amplified, as well as the quality of both extracted RNA and RT–PCRs were determined.