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A large decrease of cytosolic triosephosphate isomerase in transgenic potato roots affects the distribution of carbon in primary metabolism

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Abstract

Triosephosphate isomerase (TPI, EC 5.3.1.1) catalyzes the interconversion of dihydroxyacetone-P and glyceraldehyde 3-P in the glycolytic pathway. A constitutively expressed antisense construct for cytosolic TPI was introduced into potato (Solanum tuberosum) using Agrobacterium rhizogenes to examine the metabolic effects of a reduction in cytosolic TPI in roots. We obtained a population of transgenic root clones displaying ~36 to 100 % of the TPI activity found in control clones carrying an empty binary vector. Ion exchange chromatography and immunoblot analysis showed that the antisense strategy significantly decreased the cytosolic TPI isoform, while levels of plastidial TPI activity remained apparently unaffected. Transgenic roots were characterized with respect to the activity of glycolytic enzymes, their metabolite contents and carbon fluxes. Metabolite profiling of sugars, organic acids, amino acids and lipids showed elevated levels of sucrose, glucose, fructose, fumarate, isocitrate, 4-aminobutyrate, alanine, glycine, aromatic amino acids and saturated long chain fatty acids in roots containing the lowest TPI activity. Labelings with 14C-glucose, 14C-sucrose and 14C-acetate indicated that a reduction of cytosolic TPI activity in roots increased carbon metabolism through the pentose phosphate pathway, O2 uptake and catabolism of sucrose to CO2, and capacity for lipid synthesis. These results demonstrate that a large reduction of cytosolic TPI alters the distribution of carbon in plant primary metabolism.

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Abbreviations

cTPI:

Cytosolic triosephosphate isomerase

DGDG:

Digalactosyldiacylglycerol

DHAP:

Dihydroxyacetone phosphate

FAA:

Free amino acid

Fru:

Fructose

Fru1,6P:

Fructose-1,6-bisphosphate

Fru6P:

Fructose-6-phosphate

GAP:

Glyceraldehyde-3-phosphate

Glc:

Glucose

Glc1P:

Glucose-1-phosphate

Glc6P:

Glucose-6-phosphate

Glc6PDH:

Glucose-6-phosphate dehydrogenase

MGDG:

Monogalactosyldiacylglycerol

PEP:

Phosphoenolpyruvate

PPP:

Pentose phosphate pathway

pTPI:

Plastidial triosephosphate isomerase

Suc:

Sucrose

TCA:

Tricarboxylic acid

TPI:

Triosephosphate isomerase

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Acknowledgments

This work was supported by a Discovery grant from the Natural Sciences and Engineering Research Council of Canada (NSERC) to J.R. S.D. was supported by an NSERC postdoctoral fellowship. J.J. was supported by an NSERC undergraduate summer research award. N.P. was supported in part by a Development Fellowship from the Association of Commonwealth Universities.

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Authors and Affiliations

  1. IRBV, Université de Montréal, 4101 rue Sherbrooke est, Montréal, QC, H1X 2B2, Canada

    Sonia Dorion, Audrey Clendenning, Julie Jeukens, Nanhi Parveen & Jean Rivoal

  2. Instituto de la Grasa, CSIC, Av. Padre García Tejero, 4, 41012, Seville, Spain

    Joaquín J. Salas & Enrique Martínez Force

  3. Department of Biology, Lakehead University, 955 Oliver Road, Thunder Bay, ON, P7B 5E1, Canada

    Andrea A. Haner & R. David Law

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  1. Sonia Dorion
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  2. Audrey Clendenning
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Correspondence to Jean Rivoal.

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Dorion, S., Clendenning, A., Jeukens, J. et al. A large decrease of cytosolic triosephosphate isomerase in transgenic potato roots affects the distribution of carbon in primary metabolism. Planta 236, 1177–1190 (2012). https://doi.org/10.1007/s00425-012-1675-1

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