Biochemical control systems for small molecule damage in plants.

As a system, plant metabolism is far from perfect: small molecules (metabolites, cofactors, coenzymes, and inorganic molecules) are frequently damaged by unwanted enzymatic or spontaneous reactions. Here, we discuss the emerging principles in small molecule damage biology. We propose that plants evolved at least three distinct systems to control small molecule damage: (i) repair, which returns a damaged molecule to its original state; (ii) scavenging, which converts reactive molecules to harmless products; and (iii) steering, in which the possible formation of a damaged molecule is suppressed. We illustrate the concept of small molecule damage control in plants by describing specific examples for each of these three categories. We highlight interesting insights that we expect future research will provide on those systems, and we discuss promising strategies to discover new small molecule damage-control systems in plants.

D-2-hydroxyglutarate metabolism is linked to photorespiration in the shm1-1 mutant.

The Arabidopsis mutant shm1-1 is defective in mitochondrial serine hydroxymethyltransferase 1 activity and displays a lethal photorespiratory phenotype at ambient CO2 concentration but grows normally at high CO2 . After transferring high CO2 -grown shm1-1 plants to ambient CO2 , the younger leaves remain photosynthetically active while developed leaves display increased yellowing and decreased FV /FM values. Metabolite analysis of plants transferred from high CO2 to ambient air indicates a massive light-dependent (photorespiratory) accumulation of glycine, 2-oxoglutarate (2OG) and D-2-hydroxyglutarate (D-2HG). Amino acid markers of senescence accumulated in ambient air in wild-type and shm1-1 plants maintained in darkness and also build up in shm1-1 in the light. This, together with an enhanced transcription of the senescence marker SAG12 in shm1-1, suggests the initiation of senescence in shm1-1 under photorespiratory conditions. Mitochondrial D-2HG dehydrogenase (D-2HGDH) converts D-2HG into 2OG. In vitro studies indicate that 2OG exerts competitive inhibition on D-2HGDH with a Ki of 1.96 mm. 2OG is therefore a suitable candidate as inhibitor of the in vivo D-2HGDH activity, as 2OG is produced and accumulates in mitochondria. Inhibition of the D-2HGDH by 2OG is likely a mechanism by which D-2HG accumulates in shm1-1, however it cannot be ruled out that D-2HG may also accumulate due to an active senescence programme that is initiated in these plants after transfer to photorespiratory conditions. Thus, a novel interaction of the photorespiratory pathway with cellular processes involving D-2HG has been identified.

Engineering photorespiration: current state and future possibilities.

Reduction of flux through photorespiration has been viewed as a major way to improve crop carbon fixation and yield since the energy-consuming reactions associated with this pathway were discovered. This view has been supported by the biomasses increases observed in model species that expressed artificial bypass reactions to photorespiration. Here, we present an overview about the major current attempts to reduce photorespiratory losses in crop species and provide suggestions for future research priorities.

Non-photosynthetic 'malic enzyme' from maize: a constituvely expressed enzyme that responds to plant defence inducers.

The characterization of a non-photosynthetic isoform of NADP-malic enzyme (NADP-ME) from maize roots, which represents nearly 7% of the total soluble protein of this tissue, was performed. The molecular properties of the purified protein, as well as the kinetic parameters determined, indicate that the NADP-ME isoform present in maize roots differs from the photosynthetic enzyme implicated in the C4 cycle, but is similar, or identical, to the enzyme previously characterized from etiolated maize leaves (Maurino, Drincovich and Andreo, Biochem. Mol. Biol. Int. 38 (1996) 239-250). A full-length ORF encoding a plastidic NADP-ME (almost identical to the maize root NADP-ME, GenBank accession number U39958) was cloned from a root cDNA library as well as isolated by reverse transcription (RT)-PCR using green leaves mRNA as template. These results indicate that root NADP-ME does not constitute a root-specific isoform, but represents a protein with a constitutive pattern of expression in plastids of the C4 plant maize. The amount of NADP-ME measured by activity, western and northern blot was modified when different stress conditions (including treatments with cellulase, fungal elicitors, jasmonate and hypoxic treatment) were applied to maize roots, indicating that the enzyme from maize roots is under transcriptional or post-transcriptional regulation by effectors related to plant defence responses. It is deduced that the induction of housekeeping genes, like non-photosynthetic NADP-ME, whose constitutive role may be the provision of reductive power in non-photosynthetic plastids, is likely to accompany the defence response.

NADP-malic enzyme isoforms in maize leaves.

Two isoforms of NADP-malic enzyme have been characterized in maize leaves. The 72 kDa-form of the protein, present mainly in etiolated maize leaves, has lower specific activity than the 62 kDa-form, which is implicated in C4 metabolism and predominates in green leaves. The larger form of the enzyme has higher Km values for NADP and malate and lower PH optimum. The antibodies raised against the 62 kDa-form of the protein react with the 72 kDa-form. Steady state levels of NADP-malic enzyme, as measured by the amount of protein and activity, increase several-fold when dark-grown maize seedlings are illuminated. This increase in protein is about 13-fold for the 62 kDa-form of the enzyme, while the 72 kDa-form remains practically constant after a transient increase. Northern blot analysis using a specific probe against the 62 kDa-form of the enzyme, reveals the increase of a 2.2 kb mRNA during greening. Southern hybridization analysis with genomic DNA suggests the presence of more than one gene encoding NADP-malic enzyme in maize. In this paper we provide biochemical and inmunological evidence suggesting that both isoforms are closely related and that the 72 kDa-form is also present in low levels in mature green leaves.