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LNA mediated in situ hybridization of miR171 and miR397a in leaf and ambient root tissues revealed expressional homogeneity in response to shoot heat shock in Arabidopsis thaliana
Authors:Barku M Mahale  Bashasab Fakrudin  Sudipta Ghosh  P U Krishnaraj
Institution:1. Department of Biotechnology, University of Agricultural Sciences, Dharwad, 580005, Karnataka, India
Abstract:MicroRNAs (miRNAs) are endogenous non-protein coding RNA molecules of approximately 21 nucleotides in length capable of modulating gene expression in animals and plants. The role of miRNA based gene regulation has been proved in several pathways including in plant growth, development and stress response. In this study miR171 and miR397a were tested for their expression pattern under different heat shock regimes in shoot and root tissues of Arabidopsis thaliana using Locked Nucleic Acid (LNA) mediated in situ hybridization. With an increase in temperature across 35 °C, 40 °C and 45 °C there was a corresponding increased up-regulation of miR171 in leaf tissues compared to ambient temperature. Similarly, an unambiguous elevated expression of miR171 within increase in duration of exposure at each temperature regime across 1 h, 2 h and 3 h was noticed in comparison to ambient control leaf tissue. On the other hand, miR397a, which expressed at ambient control conditions, got down-regulated both with increase in heat and exposure regime in leaf tissues. Both miRNAs expressed in control ambient root tissues. Maintaining the root zone temperature at ambient conditions, upon imposing heat shock regime to shoot system, miR171 recorded corresponding increased up-regulation as indicated by the intensity of in situ hybridization, while miR397a got down-regulated. Given the differential homogeneity in expression pattern of both miRNA in leaf and root tissues experiencing heat shock regimes, possibilities of movement of heat shock induced signals to root tissues seem to be obvious.
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