Int.J.Curr.Microbiol.App.Sci (2017) 6(5): 476-480 476 Original Research Article https://doi.org/10.20546/ijcmas.2017.605.055 Evaluation of Endogenous Gene Validation: An Important Contrivance for Differential Gene Study in Pearl Millet Poonam Kanani* and Y.M. Shukla Department of Agricultural Biotechnology, Anand Agricultural University, Anand-388 110, India *Corresponding author ABSTRACT Introduction Pearl millet (Pennisetum glaucum L.) is a monocot, diploid (2n=2x=14) cereal crop plant belongs to family Poaceae, sub family Penicedae, also known as bajra. It is one of most important cereals growing worldwide in tropical semi arid regions mainly as food and forage crop in Africa and Asia, and It is the sixth most important crop after rice, wheat, maize, barley and sorghum, nutritionally rich in zinc, iron, and high antioxidants. It has haploid genome size of 2450 Mbp and has DNA content of 1C = 2.36 pg. (Budak et al., 2003, Malik, 2015). Plants being sessile in nature are enforced to face different environmental conditions in time course of their life span. The stresses such as drought, salinity and temperature: heat or cold are major abiotic and biotic stresses Biotic stress include four major diseases are downy mildew (Sclerospora graminicola), ergot (Claviceps fusiformis), smut (Tolysporium penicillariae) and rust (Puccinia substriata var. indica) which develop depending on the weather conditions and genotype are serious restraint for growth and productivity of pearlmillet (Neya and Kabore, 1987; Thakur et al., 2008). All the processes related to physiological, morphological and biochemical changes are International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 5 (2017) pp. 476-480 Journal homepage: http://www.ijcmas.com Nowadays selection and validation of endogenous gene is commonly being studied worldwide for differential gene study. And it is also considered as inevitable for gene validation studies. Endogenous gene expression varies profoundly with every cell and tissue with even mere deviations at nominal levels too, thus a need of normalization. So far, the most refined and steadfast method is preferably Quantitative Real-Time PCR (qRT-PCR) for precise quantification of gene expression to understand explicit gene functions at various development phases and beneath diverse environmental conditions that plant encounters during its life. The data normalization to find most stable gene are obtained through different software programs, such as geNorm, Norm finder and Best keeper. Pearl millet [Pennisetum glaucum (L.) R. Br.] a major grain and forage crop, frequently encounter various stresses both biotic and abiotic, and therefore more chances of studying genes responsible for stress adaption. Thus insight into evaluation of endogenous gene (such as actin, tubulin, GAPDH, ubiquitin etc.) validation studies can prove to target more exact molecular mechanism for stress tolerance. Keywords qRT-PCR, normalization, gene expression, validation, endogenous gene. Accepted: 04 April 2017 Available Online: 10 May 2017 Article Info