MOLECULAR EVALUATION OF SOME SALINITY TOLERANT GENOTYPES OF SORGHUM USING RT-PCR

Abstract

This study was carried out in the Plant Transformation and Biopharmaceuticals Laboratory (PTBL), Plant Molecular Biology Department, Agricultural Genetic Engineering Research Institute (AGERI), Agricultural Research Center (ARC), Giza, Egypt during the period from 2014 to 2020. It aimed to fingerprint of 20 sorghum genotypes using ISSR markers and analyze their related relationship using UPGMA and PCoA, morphologically evaluation to the 20 genotypes after exposure to tested concentration of salt stress (172.5 mM) to pick up the two most tolerant and sensitive genotypes. Also evaluate the most two tolerant and sensitive genotypes morphologically, analyze the recorded data statistically using one way ANOVA and LSD and molecularly using RT-PCR to study the gene expression of the salinity tolerance gene SbBADH1 and SbBADH2. Twenty sorghum (Sorghum bicolor (L.) Monech) genotypes were assessed using 10 ISSR primers. The used primers generated polymorphic and scorable amplification fragments among the tested sorghum genotypes with a 93 % of polymorphism. All tested ISSR primers yielded amplified products and germinated 152 alleles (average of 15.2 bands/primer). Out of the 10 used primers, eight primers obtained 24 unique markers among the 20 studied sorghum genotypes. Out of these eight primers, only three obtained four markers which distinguished between all A and all B genotypes. However, molecular characterization revealed polymorphism percentage of a 100% for ISSR markers, indicating a high level of polymorphism among the studied sorghum genotypes.