In vivo NMR for ¹³C Metabolic Flux Analysis.

The use of in vivo NMR within the framework of Metabolic Flux Analysis in plants is presented. In vivo NMR allows to visualize the active metabolic network, to determine metabolic and isotopic steady state and to measure metabolic fluxes which are not necessarily accessible by isotopic steady state (stationary) Metabolic Flux Analysis. The kinetic data can be used as input for dynamic (nonstationary) Metabolic Flux Analysis. Both 1D and 2D NMR methods are employed.

Potassium deficiency induces the biosynthesis of oxylipins and glucosinolates in Arabidopsis thaliana.

BACKGROUND: Mineral fertilization and pest control are essential and costly requirements for modern crop production. The two measures go hand in hand because plant mineral status affects plant susceptibility to pests and vice versa. Nutrient deficiency triggers specific responses in plants that optimize nutrient acquisition and reprogram metabolism. K-deficient plants illustrate these strategies by inducing high-affinity K-uptake and adjusting primary metabolism. Whether and how K deficient plants also alter their secondary metabolism for nutrient management and defense is not known. RESULTS: Here we show that K-deficient plants contain higher levels of the phytohormone jasmonic acid (JA), hydroxy-12-oxo-octadecadienoic acids (HODs) and 12-oxo-phytodienoic acid (OPDA) than K-sufficient plants. Up-regulation of the 13-LOX pathway in response to low K was evident in increased transcript levels of several biosynthetic enzymes. Indole and aliphatic glucosinolates accumulated in response to K-deficiency in a manner that was respectively dependent or independent on signaling through Coronatine-Insensitive 1 (COI1). Transcript and glucosinolate profiles of K-deficient plants resembled those of herbivore attacked plants. CONCLUSIONS: Based on our results we propose that under K-deficiency plants produce oxylipins and glucosinolates to enhance their defense potential against herbivorous insects and create reversible storage for excess S and N.

Absorption, metabolism and excretion of Choladi green tea flavan-3-ols by humans.

Ten healthy human subjects consumed 500 mL of Choladi green tea, containing 648 mumol of flavan-3-ols after which plasma and urine were collected over a 24 h period and analysed by HPLC-MS. Plasma contained a total of ten metabolites, in the form of O-methylated, sulphated and glucuronide conjugates of (epi)catechin and (epi)gallocatechin, with 29-126 nM peak plasma concentrations (C(max)) occurring 1.6-2.3 h after ingestion, indicative of absorption in the small intestine. Plasma also contained unmetabolised (-)-epigallocatechin-3-gallate and (-)-epicatechin-3-gallate with respective C(max) values of 55 and 25 nM. Urine excreted 0-24 h after consumption of green tea contained 15 metabolites of (epi)catechin and (epi)gallocatechin, but (-)-epigallocatechin-3-gallate and (-)-epicatechin-3-gallate were not detected. Overall flavan-3-ol metabolite excretion was equivalent to 8.1% of intake, however, urinary (epi)gallocatechin metabolites corresponded to 11.4% of (epi)gallocatechin ingestion while (epi)catechin metabolites were detected in amounts equivalent to 28.5% of (epi)catechin intake. These findings imply that (epi)catechins are highly bioavailable, being absorbed and excreted to a much greater extent than most other flavonoids. It is also evident that flavan-3-ol metabolites are rapidly turned over in the circulatory system and as a consequence C(max) values are not an accurate quantitative indicator of the extent to which absorption occurs.

The effect of potassium nutrition on pest and disease resistance in plants.

Providing a fast growing world population with sufficient food while preserving ecological and energy resources of our planet is one of the biggest challenges in this century. Optimized management of chemical fertilizers and pesticides will be essential for achieving sustainability of intensive farming and requires both empirical data from field trials and advanced fundamental understanding of the molecular processes controlling plant growth. Genes involved in plant responses to nutrient deficiency and pathogen/herbivore attack have been identified, but we are lacking information about the cross-talk between signalling pathways when plants are exposed to a combination of abiotic and biotic stress factors. The focus of this review is on the relationship between the potassium status of plants and their susceptibility to pathogens and herbivorous insects. We combine field evidence on potassium-disease interaction with existing knowledge on metabolic and physiological factors that could explain such interaction, and present new data on metabolite profiles and hormonal pathways from the model plant Arabidopsis thaliana. The latter provides evidence that facilitated entry and development of pathogens or insects in(to) potassium-deficient plants as a result of physical and metabolic changes is counteracted by an increased defence. A genetic approach should now be applied to establish a causal relationship between disease susceptibility on the one hand and individual enzymatic and signal components on the other. Once identified, these can be used to design agricultural strategies that support the nutritional status of the crops while exploiting their inherent potential for defence.

In vivo 13C NMR determines metabolic fluxes and steady state in linseed embryos.

The dynamics of developing linseed embryo metabolism was investigated using (13)C-labelling experiments where the real-time kinetics of label incorporation into metabolites was monitored in situ using in vivo NMR. The approach took advantage of the occurrence in this plant tissue of large metabolite pools - such as sucrose or lipids - to provide direct and quantitative measurement of the evolution of the labelling state within central metabolism. As a pre-requisite for the use of steady state flux measurements it was shown that isotopic steady state was reached within 3 h at the level of central intermediates whereas it took a further 6h for the sucrose pool. Complete isotopic and metabolic steady state took 18 h to be reached. The data collected during the transient state where label was equilibrated but the metabolic steady state was incomplete, enabled the rates of lipid and sucrose synthesis to be measured in situ on the same sample. This approach is suitable to get a direct assessment of metabolic time-scales within living plant tissues and provides a valuable complement to steady state flux determinations.