Effects of root morphology, respiration and carboxylate exudation on carbon economy in two non-mycorrhizal lupines under phosphorus deficiency.

Under phosphorus (P) deficiency, Lupinus albus develops cluster roots that allow efficient P acquisition, while L. angustifolius without cluster roots also grows well. Both species are non-mycorrhizal. We quantitatively examined the carbon budgets to investigate the different strategies of these species. Biomass allocation, respiratory rates, protein amounts and carboxylate exudation rates were examined in hydroponically-grown plants treated with low (1 µM; P1) or high (100 µM; P100) P. At P1, L. albus formed cluster roots, and L. angustifolius increased biomass allocation to the roots. The respiratory rates of the roots were faster in L. albus than in L. angustifolius. The protein amounts of the non-phosphorylating alternative oxidase and uncoupling protein were greater in the cluster roots of L. albus at P1 than in the roots at P100, but similar between the P treatments in L. angustifolius roots. At P1, L. albus exuded carboxylates at a faster rate than L. angustifolius. The carbon budgets at P1 were surprisingly similar between the two species, which is attributed to the contrasting root growth and development strategies. L. albus developed cluster roots with rapid respiratory and carboxylate exudation rates, while L. angustifolius developed a larger root system with slow respiratory and exudation rates.

Phosphorus toxicity disrupts Rubisco activation and reactive oxygen species defence systems by phytic acid accumulation in leaves.

Phosphorus (P) is an essential mineral nutrient for plants. Nevertheless, excessive P accumulation in leaf mesophyll cells causes necrotic symptoms in land plants; this phenomenon is termed P toxicity. However, the detailed mechanisms underlying P toxicity in plants have not yet been elucidated. This study aimed to investigate the molecular mechanism of P toxicity in rice. We found that under excessive inorganic P (Pi) application, Rubisco activation decreased and photosynthesis was inhibited, leading to lipid peroxidation. Although the defence systems against reactive oxygen species accumulation were activated under excessive Pi application conditions, the Cu/Zn-type superoxide dismutase activities were inhibited. A metabolic analysis revealed that excessive Pi application led to an increase in the cytosolic sugar phosphate concentration and the activation of phytic acid synthesis. These conditions induced mRNA expression of genes that are activated under metal-deficient conditions, although metals did accumulate. These results suggest that P toxicity is triggered by the attenuation of both photosynthesis and metal availability within cells mediated by phytic acid accumulation. Here, we discuss the whole phenomenon of P toxicity, beginning from the accumulation of Pi within cells to death in land plants.

Comparison of the response to phosphorus deficiency in two lupin species, Lupinus albus and L. angustifolius, with contrasting root morphology.

White lupin (Lupinus albus) produces cluster roots, an adaptation to low soil phosphorus (P). Cluster roots exude large levels of P-solubilizing compounds such as citrate and malate. In contrast, narrow leaf lupin (L. angustifolius) is closely related to L. albus, but does not produce cluster roots. To examine the different strategies for P acquisition, we compared the growth, biomass allocation, respiratory properties and construction cost between L. albus and L. angustifolius under P-deficient conditions. Both Lupinus species were grown in hydroponic culture with 1 or 100 µM P. Under the P-deficient regime, L. albus produced cluster roots with little change in biomass allocation, while L. angustifolius significantly increased biomass allocation to roots. The rate of cyanide-resistant SHAM (salicylhydroxamic acid)-sensitive respiration was high in cluster roots and very low in roots of L. angustifolius. These results suggest a low alternative oxidase (AOX) activity in L. angustifolius roots, and thus, ATP would be produced efficiently in L. angustifolius roots. The construction cost was highest in cluster roots and lowest in L. angustifolius roots. This study shows that under P deficiency, L. albus produces high-cost cluster roots to increase the P availability, while L. angustifolius produces large quantities of low-cost roots to enhance P uptake.

Systematic exploration of thioredoxin target proteins in plant mitochondria.

The thioredoxin (Trx) system is known to play a pivotal role in cellular redox regulation, but its target proteins in plant mitochondria remain largely uncharacterized. In this study, we systemically screened Trx target candidates in plant mitochondria. Mitochondrial protein extracts were prepared from Arabidopsis shoots, spinach leaves and potato tubers, and then subfractionated into soluble matrix and insoluble membrane fractions. Protein extracts were loaded onto an affinity column immobilizing Arabidopsis mitochondria-localized o-type Trx mutant protein, in which one of two internal cysteines at the active site was substituted by serine. Proteins forming mixed-disulfide intermediates with the mutated Trx were identified by proteomic approaches. This procedure allowed the determination of 101 Trx target candidate proteins involved in a broad spectrum of mitochondrial processes. Furthermore, biochemical assay revealed that one of the potential Trx target proteins, alternative oxidase, is actually redox regulated by Trx. This study provides insights into the regulatory mechanism of diverse functions in mitochondrial biology that are mediated through the Trx system.

Responses of spinach leaf mitochondria to low N availability.

Low N availability induces carbohydrate accumulation in leaf cells, which often causes suppression of photosynthesis. Under low N supply, excess carbohydrates would be preferentially respired by the non-phosphorylating pathways, such as the alternative oxidase (AOX) and uncoupling protein (UCP), which would suppress the excessive increase in the ratio of C to N (C/N ratio). In leaves, however, responses of these pathways to the low N stress are still unknown. We examined the mitochondrial respiratory pathways in spinach leaves grown at three different N availabilities to clarify whether the respiratory pathways change depending on the N availabilities. With the decrease in N availability, leaf respiratory rates per leaf area decreased, but the rates on the leaf N basis were comparable. Using fumarase activities of whole leaf extracts and isolated mitochondria, we estimated mitochondrial protein contents per leaf N. The contents increased with the decrease in the N availability, that is, at the low N availability, N was preferentially invested into mitochondria. On the mitochondrial protein basis, capacities of cytochrome pathway (CP) and cytochrome c oxidase (COX) were comparable regardless of the N availabilities, whereas both AOX capacity and the amounts of AOX protein increased with the decrease in the N availability. Some enzymes of tricarboxylic acid (TCA) cycle, especially NAD-dependent malic enzyme (NAD-ME), showed higher capacities under lower N. On the other hand, amounts of UCP did not differ amongst the N availabilities. These results indicated that, under low N stress, AOX will be preferentially up-regulated and will efficiently consume excess carbohydrates, which leads to suppressing the rise in the C/N ratio to a moderate level.