ABSTRACT: Stevia rebaudiana (Bertoni) is known as a natural sweetener plant to produced steviol glycosides. The steviol glycosides biosynthesis is limited in S. rebaudiana plants due to the alteration in the environmental circumstances such as drought, cold, salt and light. These environmental circumstances are a common side-effect in plants affecting the plant growth, metabolism and yield of secondary metabolites. Due to absence of complete genome annotations, the plant metabolites signaling is difficult in order to get the exact enzymatic flow to the product. In this article, we have analyzed the ESTs of S. rebaudiana and predicted their role in plant signaling in term of cis-regulatory elements, their biological function respect to abiotic stress. Further, the predicted abiotic stress responsive factors were also analyzed in order to predict the relevant genes or proteins function in comparison with the genome of Arabidopsis thaliana. Total 5,548 ESTs of stevia were retrieved from NCBI database. EST-contigs assembled from 5393 were 619 contigs and 2,894 singletons elements were identified by assembler program. Due to short expressed sequences related to singletons, it is excluded for further study. Further, retrieved ESTs were resulted in to 619 EST-contigs by using the clustering method. Out of 619, 15 contigs belongs to transcription factor families while 292 contigs, belongs to five enzyme classes. Out of 619 contigs, the 529 contigs showing the correct gene ontology in term of biological process (BP), molecular Functions (MF) and cellular component (CC). Further, these contigs were also screened for metabolic pathways analyses using KEG database. In this, 390 metabolic pathways and 67 involved for signal transduction were identified from 619 contigs. The co-expression analysis was revealed by gene investigators and STRING 10.0 with 0.40 correlations and 0.9 mutual connection. In projected PPI network, the recognized factors (WD40-like protein, MYB-HB like, AP2-EREBP, C2H2, Hap3/NF-YB, bHLH, C2C2-CO-like, CW-Zn, FHA-SMAD, Nin like, SBP3, TIFY, Tc-PD, Znf-B and bIP) belong to plant signaling and MAPK signaling pathways. These TFs introduce as a candidate genes responsive factors may lead to enhanced plant growth and metabolism by overexpression.