Myo-inositol oxygenase (MIOX), the only catabolic enzyme of the inositol pathway, catalyzes conversion of myo-inositol to D-GlcA (glucuronic acid). The present study encompasses bioinformatic analysis of MIOX gene across phylogenetically related plant lineages and representative animal groups. Comparative motif analysis of the MIOX gene(s) across various plant groups suggested existence of abiotic- stress related cis-acting elements such as, DRE, MYB, MYC, STRE, MeJa among others. A detailed analysis revealed a single isoform of MIOX gene, located in chromosome 6 of indica rice (Oryza sativa) with an open reading frame of 938 bp coding for 308 amino acids producing a protein of ~ 35 kD. Secondary structure prediction of the protein gave the predicted number of 144 alpha helices and 154 random coils. The three-dimensional structure suggested it to be a monomeric protein with a single domain. Bacterial overexpression of the protein, purification and enzyme assay showed optimal catalytic activity at pH 7.5-8 at an optimal temperature of 37 °C with Michaelis constant of 40.92 mM. The range of Km was determined as 22.74-28.7 mM and the range of Vmax was calculated as 3.51-3.6 µM/min, respectively. Four salt-tolerant and salt-sensitive rice cultivars displayed differential gene expression of OsMIOX at different time points in different tissues under salinity and drought stress as observed from qRT-PCR data, microarray results and protein expression profile in immunoblot analysis. Gel volumetric analysis confirmed a very high expression of MIOX in roots and leaves on 7th day following germination. Microarray data showed high expression of MIOX at all developmental stages including seedling growth and reproduction. These data suggest that OsMIOX might have a role to play in rice abiotic stress responses mediated through the myo-inositol oxidation pathway.
Supplementary information: The online version contains supplementary material available at 10.1007/s12298-023-01340-6.
Keywords: Cis-acting elements; Enzyme kinetics; Expression analysis; Motif analysis; Myo-inositol oxygenase; Oryza sativa; Salinity stress.
© Prof. H.S. Srivastava Foundation for Science and Society 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.