Germin and germin-like protein is an important family of extracellular plant glycoproteins providing resistance against biotic and abiotic stresses. Though various enzymatic activities have been assigned to these proteins but still their function is not fully understood. In the present study, 1228bp upstream promoter region of Oryza sativa root germin-like protein 1 (OsRGLP1) gene was characterized by computational as well as in vivo analysis. Computational analysis of OsRGLP1 gene promoter sequence was also investigated. Transcriptional factor binding sites (TFBS) analysis revealed a total of 190 matches of 46 different TFs families. The most frequent (Yeast TATA box binding protein (YTBP) (19 copies), Arabidopsis homeobox protein (AHBP) (15 copies) and Vertebrate TATA box binding protein (VTBP) (14 copies) and unique cis-regulatory elements were also investigated. In-silico functional analysis revealed three modules in which DOFF_OPAQ_03 was the most frequent. Interaction of three long stretches of promoter (-408 to -378, -537 to -513 and -708 to -638) with TBP (TATA box binding protein) was analyzed using Haddock webserver suggesting that Adenine, Thymine, Serine and Lysine were the most active residues. For in vivo studies, OsRGLP1 gene promoter was ligated upstream to GUS reporter gene and transformed into Nicotiana tabacum using agrobacterium mediated transformation. In transgenic plants, GUS expression was observed in root, shoot and leaves. Strong GUS activity was noticed in leaf veins, epidermal hair, stomata guard cells, cell wall, stem cortex, petiole stem junction, root and root hair etc. Moreover, wound inducibility was analyzed in root, shoot and leaves at different time interval (10, 30 min, 1, 2, 3, 4, 5 hours), which showed linear association. Wound inducible property of this promoter confirmed its involvement in various biotic and mechanical stresses which can be used for the production of biologically important crop species in deriving tissue specific expression at the site of injury/wound.
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