Quantitative Multilevel Analysis of Central Metabolism in Developing Oilseeds of Oilseed Rape during in Vitro Culture

authored by

Jörg Schwender, Inga Hebbelmann, Nicolas Heinzel, Tatjana Hildebrandt, Alistair Rogers, Dhiraj Naik, Matthias Klapperstück, Hans Peter Braun, Falk Schreiber, Peter Denolf, Ljudmilla Borisjuk, Hardy Rolletschek

Abstract

Seeds provide the basis for many food, feed, and fuel products. Continued increases in seed yield, composition, and quality require an improved understanding of how the developing seed converts carbon and nitrogen supplies into storage. Current knowledge of this process is often based on the premise that transcriptional regulation directly translates via enzyme concentration into flux. In an attempt to highlight metabolic control, we explore genotypic differences in carbon partitioning for in vitro cultured developing embryos of oilseed rape (Brassica napus). We determined biomass composition as well as 79 net fluxes, the levels of 77 metabolites, and 26 enzyme activities with specific focus on central metabolism in nine selected germplasm accessions. Overall, we observed a tradeoff between the biomass component fractions of lipid and starch. With increasing lipid content over the spectrum of genotypes, plastidic fatty acid synthesis and glycolytic flux increased concomitantly, while glycolytic intermediates decreased. The lipid/starch tradeoff was not reflected at the proteome level, pointing to the significance of (posttranslational) metabolic control. Enzyme activity/flux and metabolite/flux correlations suggest that plastidic pyruvate kinase exerts flux control and that the lipid/starch tradeoff is most likely mediated by allosteric feedback regulation of phosphofructokinase and ADP-glucose pyrophosphorylase. Quantitative data were also used to calculate in vivo mass action ratios, reaction equilibria, and metabolite turnover times. Compounds like cyclic 39,59-AMP and sucrose-6-phosphate were identified to potentially be involved in so far unknown mechanisms of metabolic control. This study provides a rich source of quantitative data for those studying central metabolism.

Organisation(s)

Institute of Plant Genetics

External Organisation(s)

Brookhaven National Laboratory (BNL)
Indian Institute of Advanced Research
Monash University
Martin Luther University Halle-Wittenberg
Bayer Corporation - USA
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)

Type

Article

Journal

Plant physiology

Volume

168

Pages

828-848

No. of pages

21

ISSN

0032-0889

Publication date

06.07.2015

Publication status

Published

Peer reviewed

Yes

ASJC Scopus subject areas

Physiology, Genetics, Plant Science

Electronic version(s)

https://doi.org/10.1104/pp.15.00385 (Access: Open)
https://doi.org/10.15488/11683 (Access: Open)