Abstract
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.
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Abbreviations
- CRE:
-
Cis Regulatory element
- DRE:
-
Dehydration responsive element
- MYB:
-
Myeloblastosis
- MYC:
-
Master regulator of cell cycle entry and proliferative metabolism
- STRE:
-
Stress responsive element
- MeJa:
-
Methyl jasmonate
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Acknowledgements
The work is supported by grants from Department of Biotechnology, GOI (BT/AB/05/02/2007-III dt.21/09/2010 and BT/IN/NWO/17/ALM dated 02.09.2015) awarded to ALM, while an INSA Senior Scientist. The research is also supported by grants from, Department of Biotechnology to SR [WBDBT Sanction No.237(Sanc.)/BT(Estt.)/RD-28/2016 dated 29-03-2017]. SA is funded by Govt. of India University Grants Commission-Rajiv Gandhi National Fellowship (2011-12/RGNF-SC-WES-13271). PD is funded by Department of Science and Technology-SERB research grant (YSS/2015/001872).
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ALM, SA and SR designed the research. SA, TA and PD conducted the experiments. SA, TA, PD and SR prepared the draft manuscript. ALM finalized the manuscript with input and approval from all the authors.
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Adak, S., Agarwal, T., Das, P. et al. Characterization of myo-inositol oxygenase from rice (OsMIOX): influence of salinity stress in different indica rice cultivars. Physiol Mol Biol Plants 29, 927–945 (2023). https://doi.org/10.1007/s12298-023-01340-6
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DOI: https://doi.org/10.1007/s12298-023-01340-6