Abstract
Key message
Co-suppressed MIPS2 transgenic lines allow bypass of the embryo lethal phenotype of the previously published triple knock-out and demonstrate the effects of MIPS on later stages of development.
Abstract
Regulation of inositol production is of interest broadly for its effects on plant growth and development. The enzyme l-myo-inositol 1-phosphate synthase (MIPS, also known as IPS) isomerizes d-glucose-6-P to d-inositol 3-P, and this is the rate-limiting step in inositol production. In Arabidopsis thaliana, the MIPS enzyme is encoded by three different genes, (AtMIPS1, AtMIPS2 and AtMIPS3), each of which has been shown to produce proteins with biochemically similar properties but differential expression patterns. Here, we report phenotypic and biochemical effects of MIPS co-suppression. We show that some plants engineered to overexpress MIPS2 in fact show reduced expression of AtMIPS1, AtMIPS2 and AtMIPS3, and show altered vegetative phenotype, reduced size and root length, and delayed flowering. Additionally, these plants show reduced inositol, increased glucose levels, and alteration of other metabolites. Our results suggest that the three AtMIPS genes work together to impact the overall synthesis of myo-inositol and overall inositol homeostasis.
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Acknowledgements
We acknowledge the students of BIOL 300 at Emory & Henry College for help with MIPS transformants and Janet Donahue for assistance with myo-inositol measurements. This work was supported by awards from the Appalachian Colleges Association (C.M.F.) and National Science Foundation Division of Molecular and Cellular Biosciences 1052034 (J.Y. and G.G.).
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CMF conducted phenotypic analysis and most qPCR. JYY and GEG prepared the transgenic lines and conducted the metabolite analysis as well as western blot and complementation analysis. EAH conducted qPCR of vegetative tissue.
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Fleet, C.M., Yen, J.Y., Hill, E.A. et al. Co-suppression of AtMIPS demonstrates cooperation of MIPS1, MIPS2 and MIPS3 in maintaining myo-inositol synthesis. Plant Mol Biol 97, 253–263 (2018). https://doi.org/10.1007/s11103-018-0737-6
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DOI: https://doi.org/10.1007/s11103-018-0737-6