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Kinetic and regulatory properties of plant ADP-glucose pyrophosphorylase genetically modified by heterologous expression of potato upreg mutants in vitro and in vivo

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Abstract

In this study, the uses of the mutated genes, upreg1 and upreg2, encoding upregulated ADP-glucose pyrophosphorylase (AGPase) large subunits with increased enzymatic activity, to improve crop yield productivity was evaluated in vitro and in planta. For in vitro examination, wild type and upregs were co-expressed with three different AGPase small subunit genes from potato and perilla to produce nine AGPase isoforms. In kinetic experiments, 3-Phosphoglycerate increased the V max and decreased the K M for the recombinant AGPase. Regardless of the specific small subunit, Upreg-type AGPases had much larger increases in enzymatic activity with concomitant decreases in values as compared to the wild type enzyme. Transformation of lettuce with the upreg1 gene altered the regulatory properties of leaf AGPase. AGPases from transgenic lettuce showed greater 3-PGA activation and lower Pi inhibition than was observed for wild type AGPase. Fresh weights of the aerial parts of transgenic plants were larger than non-transgenic controls. Based on these results, upreg mutant genes could be used for the genetic improvement of plant AGPases other than potato and effectively increase crop yield productivity.

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Abbreviations

ADP-Glc:

ADP-glucose

AGPase:

ADP-glucose pyrophosphorylase

Glc-1-P:

Glucose-1-phosphate

3-PGA:

3-Phosphoglycerate

Pi:

Orthophosphate

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Acknowledgment

This research was supported by National Institute of agricultural Biotechnology, RDA (05-5-11-22-3).

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Authors and Affiliations

  1. Biosafety Division, National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon, 441-707, South Korea

    Si-Myung Lee, Tae-Hun Ryu & Donghern Kim

  2. Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, College of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, South Korea

    Su-Il Kim

  3. Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164-6340, USA

    Thomas Okita

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  1. Si-Myung Lee
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  2. Tae-Hun Ryu
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Correspondence to Donghern Kim.

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Lee, SM., Ryu, TH., Kim, SI. et al. Kinetic and regulatory properties of plant ADP-glucose pyrophosphorylase genetically modified by heterologous expression of potato upreg mutants in vitro and in vivo. Plant Cell Tiss Organ Cult 96, 161–170 (2009). https://doi.org/10.1007/s11240-008-9472-z

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