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Not AvailableMicroRNAs are small noncoding regulatory RNAs which control gene expression by mRNA degradation or translational repression. They are significant molecular players regulating important biological processes such as developmental timing and stress response. We report here the discovery of miRNAs derived from ribosomal DNA using the small RNA datasets of 16 deep sequencing libraries of rice. Twelve putative miRNAs were identified based on highly stringent criteria of novel miRNA prediction. Surprisingly, 10 putative miRNAs (mi_7403, mi_8435, mi_12675, mi_4266, mi_4758, mi_4218, mi_8200, mi_4644, mi_14291, mi_16235) originated from rDNA of rice chromosome 9. Expression analysis of putative miRNAs and their target genes in heat tolerant and susceptible rice cultivars in control and high temperature treated seedlings revealed differential regulation of rDNA derived miRNAs. This is the first report of rDNA derived miRNAs in rice which indicates their role in gene regulation during high temperature stress in plants. Further studies in this area will open new research challenges and opportunities to broaden our knowledge on gene regulation mechanisms.Not Availabl