Evaluation of housekeeping genes for gene expression study in cluster bean under drought stress

Abstract

For reliable gene expression results, normalization of realtime PCR data is required against a control gene, which exhibits uniform expression in living entity during various development stages and under different environmental situations. Among various stresses, drought is one of the limiting constraints in growth and yield of cluster bean (Cyamopsis tetragono/oba). Gene expression study through Real-Time PCR technique is the most common for accurate quantification of genes. However, the pre-requisite for this technique is the normalization of target gene expression, which is not available in case of cluster bean during drought stress. Thus, an experiment was setup to evaluate endogenous gene expression in two cluster bean genotypes viz., Pusa Navbahar and IC369860. A total of 44 housekeeping genes from different legumes were analyzed in cluster bean resulting in EF1a, EF1B, UBQ10, 18SrRNA, 25SrRNA, ACT1, ACT11, IF4a, ADH3, and LEC with successful amplification and analyzed through GeNorm and Normfinder algorithm. It was concluded that among ten endogenous genes, all nine housekeeping genes were more or less influenced by drought stress in cluster bean exceptACT1. Therefore, ACT1 endogenous gene can further be used to normalized the expression of drought responsive genes in RT-PCR.