Subcellular localization of proteins ofOryza sativaL. in the model tobacco and tomato plants

Abstract

The cellular localization and molecular interactions are indicative of functions of a protein. The development of a simple and efficient method for subcellular localization of a protein is indispensable to elucidate gene function in plants. In this study, we assessed the feasibility of Agrobacterium-mediated transformation (agroinfiltration) of tobacco and tomato leaf tissue to follow intracellular targeting of proteins from rice fused to green fluorescent protein (GFP). For this, a simple in planta assay for subcellular localization of rice proteins in the heterologous host systems of tobacco and tomato leaf via transient transformation was developed. We have tested the applicability of this method by expressing GFP fusions of the putative antiphagocytic protein 1 (APP1) (OsAPP, LOC_Os03g56930) and ZOS3-18 - C2H2 zinc-finger protein (OsZF1, LOC_Os03g55540) from Oryza sativa L. subsp. japonica in tobacco and tomato leaf tissues. Our results demonstrate the suitability of GFP as a reporter in gene expression studies in tomato cv MicroTom. The use of GFP-fused proteins from rice for subcellular targeting in the heterologous hosts of tobacco and tomato plant systems has been confirmed.

Transient Transformation of Red Algal Cells: Breakthrough Toward Genetic Transformation of Marine Crop Porphyra Species

“…However, living cell imaging to monitor gene expression is possible using fluorescent proteins without additional substrates and cofactors (Ehrhardt, 2003). Indeed, fluorescent proteins are usually employed as useful tools to visualize protein localization in living cells in land plants (Buschmann et al, 2010;Kokkirala et al, 2010;Meng et al, 2010;Pribat et al, 2010). We first examined the usability of fluorescent proteins as reporters in P. yezoensis cells, since the expression of fluorescent protein has not yet been demonstrated in red algal cells.…”

Comprehensive Expression Profiling of Rice Tetraspanin Genes Reveals Diverse Roles During Development and Abiotic Stress

“…In silico prediction of subcellular localization suggested that majority of OsTET proteins are targeted to plasma membrane, with two exceptions, of OsTET6 and OsTET7 genes that were predicted to reside in chloroplast, and endoplasmic reticulum, respectively. Transient expression of genes is a reliable, simple, and rapid approach for determining the subcellular localization of several plant proteins ( Kokkirala et al, 2010 ). We employed model plant system N. benthamiana (tobacco) leaves for agroinfiltration of several OsTET-YFP fusion constructs.…”

Heterologous Expression of a Glycine soja C2H2 Zinc Finger Gene Improves Aluminum Tolerance in Arabidopsis

“…We transformed the plasmids into the Agrobacterium tumefaciens strain GV3101 by the heat shock method. Then, the Agrobacterium tumefaciens with plasmids was injected into the young leaves of 4-week-old tobacco plants in accordance with a previously described method [64]. After 48 h, we observed and photographed the lower epidermal cells of the leaves with a confocal laser scanning microscope (Leica, Germany).…”