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Summer Research Fellowship Programme of India's Science Academies

To demonstrate the regulation of NAC05 by miR164d using in planta assay

Priyadarshini T.

Tamil Nadu Agricultural University, Lawley Road, Coimbatore 641003

Dr. Neeti Sanan-Mishra

International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110067

Abstract

Plants undergo many adaptive changes at the physiological, biochemical and molecular levels to survive under environmental stress conditions. Regulation of stress related genes is mediated by some transcription factors and regulatory small RNAs. NAC-TFs are the plant specific proteins that have a major role in plant developmental processes and also in stress adaptive responses. They are one of the largest groups of protein family that have been reported in many plant species. Researchers have characterized the involvement of NAC-TFs in abiotic stress conditions such as drought and excess salinity. MicroRNAs (miRNAs), 21-24nt in length are small, endogenous non-coding RNAs that regulate gene expression by gene silencing or by post transcriptional level cleavage. Recent advances in the knowledge of miRNAs led to a way to study about the regulation of genes under diverse conditions. Regulation of NAC-TFs by miRNA164 family has been noticed in some plants. The aim of this project is to demonstrate the interaction between the miR164d and its target GmNAC05 transcript in planta using the model plant Nicotiana benthamiana. To study the interaction, the NAC gene and precursor of miR164d were cloned separately with GFP tag in polylinker site of pCAMBIA1300. GFP (Green Fluorescent Protein) tagged GmNAC05 is to detect the function of the miRNA, how it affects the expression of the protein and tagged miR164d is to differentiate native and introduced miRNA expression in plants. After that in planta assay can be performed by co-infiltration in tobacco.

Keywords: miRNA interaction, cloning, transformation, Agro-infiltration.

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