Isoprenoid Metabolism and Engineering in Glandular Trichomes of Lamiaceae

The isoprenoids play important ecological and physiological roles in plants. They also have a tremendous impact on human lives as food additives, medicines, and industrial raw materials, among others. Though some isoprenoids are highly abundant in nature, plants produce many at extremely low levels. Glandular trichomes (GT), which cover the aerial parts of more than 25% of vascular plants, have been considered as natural biofactories for the mass production of rare industrially important isoprenoids. In several plant genera (e.g., Lavandula and Mentha), GTs produce and store large quantities of the low molecular weight isoprenoids, in particular mono- and sesquiterpenes, as essential oil constituents. Within each trichome, a group of secretory cells is specialized to strongly and specifically express isoprenoid biosynthetic genes, and to synthesize and deposit copious amounts of terpenoids into the trichome’s storage reservoir. Despite the abundance of certain metabolites in essential oils and defensive resins, plants, particularly those lacking glandular trichomes, accumulate small quantities of many of the biologically active and industrially important isoprenoids. Therefore, there is a pressing need for technologies to enable the mass production of such metabolites, and to help meet the ever-increasing demand for plant-based bioproducts, including medicines and renewable materials. Considerable contemporary research has focused on engineering isoprenoid metabolism in GTs, with the goal of utilizing them as natural biofactories for the production of valuable phytochemicals. In this review, we summarize recent advances related to the engineering of isoprenoid biosynthetic pathways in glandular trichomes.

Occurrence, Evolution and Specificities of Iron-Sulfur Proteins and Maturation Factors in Chloroplasts from Algae

Iron-containing proteins, including iron-sulfur (Fe-S) proteins, are essential for numerous electron transfer and metabolic reactions. They are present in most subcellular compartments. In plastids, in addition to sustaining the linear and cyclic photosynthetic electron transfer chains, Fe-S proteins participate in carbon, nitrogen, and sulfur assimilation, tetrapyrrole and isoprenoid metabolism, and lipoic acid and thiamine synthesis. The synthesis of Fe-S clusters, their trafficking, and their insertion into chloroplastic proteins necessitate the so-called sulfur mobilization (SUF) protein machinery. In the first part, we describe the molecular mechanisms that allow Fe-S cluster synthesis and insertion into acceptor proteins by the SUF machinery and analyze the occurrence of the SUF components in microalgae, focusing in particular on the green alga Chlamydomonas reinhardtii. In the second part, we describe chloroplastic Fe-S protein-dependent pathways that are specific to Chlamydomonas or for which Chlamydomonas presents specificities compared to terrestrial plants, putting notable emphasis on the contribution of Fe-S proteins to chlorophyll synthesis in the dark and to the fermentative metabolism. The occurrence and evolutionary conservation of these enzymes and pathways have been analyzed in all supergroups of microalgae performing oxygenic photosynthesis.

The mevalonate pathway contributes to monoterpene production in peppermint

ABSTRACTPeppermint produces monoterpenes which are of great commercial value in different traditional and modern pharmaceutical and cosmetic industries. In the classical view, monoterpenes are synthesized via the plastidic 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, while the cytosolic mevalonate (MVA) pathway produces sesquiterpenes. Interactions between both pathways have been documented in several other plant species, however, a quantitative understanding of the metabolic network involved in monoterpene biosynthesis is still lacking. Isotopic tracer analysis, steady state 13C metabolic flux analysis (MFA) and pathway inhibition studies were applied in this study to quantify metabolic fluxes of primary and isoprenoid metabolism of peppermint glandular trichomes (GT). Our results offer new insights into peppermint GT metabolism by confirming and quantifying the crosstalk between the two isoprenoid pathways towards monoterpene biosynthesis. In addition, a quantitative description of precursor pathways involved in isoprenoid metabolism is given. While glycolysis was shown to provide precursors for the MVA pathway, the oxidative bypass of glycolysis fueled the MEP pathway, indicating prominent roles for the oxidative branch of the pentose phosphate pathway and RuBisCO. This study reveals the potential of 13C-MFA to ascertain previously unquantified metabolic routes of the trichomes and thus advancing insights on metabolic engineering of this organ.

Altering potato isoprenoid metabolism increases biomass and induces early flowering

Isoprenoids constitute the largest class of plant natural products and have diverse biological functions including in plant growth and development. In potato (Solanum tuberosum), the regulatory mechanism underlying the biosynthesis of isoprenoids through the mevalonate pathway is unclear. We assessed the role of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) homologs in potato development and in the metabolic regulation of isoprenoid biosynthesis by generating transgenic lines with down-regulated expression (RNAi-hmgr) or overexpression (OE) of one (StHMGR1 or StHMGR3) or two genes, HMGR and farnesyl diphosphate synthase (FPS; StHMGR1/StFPS1 or StHMGR3/StFPS1). Levels of sterols, steroidal glycoalkaloids (SGAs), and plastidial isoprenoids were elevated in the OE-HMGR1, OE-HMGR1/FPS1, and OE-HMGR3/FPS1 lines, and these plants exhibited early flowering, increased stem height, increased biomass, and increased total tuber weight. However, OE-HMGR3 lines showed dwarfism and had the highest sterol amounts, but without an increase in SGA levels, supporting a rate-limiting role for HMGR3 in the accumulation of sterols. Potato RNAi-hmgr lines showed inhibited growth and reduced cytosolic isoprenoid levels. We also determined the relative importance of transcriptional control at regulatory points of isoprenoid precursor biosynthesis by assessing gene–metabolite correlations. These findings provide novel insights into specific end-products of the sterol pathway and could be important for crop yield and bioenergy crops.

Plastoglobular protein 18 is involved in chloroplast function and thylakoid formation

Plastoglobules are lipoprotein particles that are found in different types of plastids. They contain a very specific and specialized set of lipids and proteins. Plastoglobules are highly dynamic in size and shape, and are therefore thought to participate in adaptation processes during either abiotic or biotic stresses or transitions between developmental stages. They are suggested to function in thylakoid biogenesis, isoprenoid metabolism, and chlorophyll degradation. While several plastoglobular proteins contain identifiable domains, others provide no structural clues to their function. In this study, we investigate the role of plastoglobular protein 18 (PG18), which is conserved from cyanobacteria to higher plants. Analysis of a PG18 loss-of-function mutant in Arabidopsis thaliana demonstrated that PG18 plays an important role in thylakoid formation; the loss of PG18 results in impaired accumulation, assembly, and function of thylakoid membrane complexes. Interestingly, the mutant accumulated less chlorophyll and carotenoids, whereas xanthophyll cycle pigments were increased. Accumulation of photosynthetic complexes is similarly affected in both a Synechocystis and an Arabidopsis PG18 mutant. However, the ultrastructure of cyanobacterial thylakoids is not compromised by the lack of PG18, probably due to its less complex architecture.

Antioxidant Balance and Regulation in Tomato Genotypes of Different Color

Antioxidants, antioxidant capacity, and the expression of isoprenoid metabolism–related genes and two pigmentation-related transcription factors were studied in four native and four hybrid tomato (Solanum lycopersicum) genotypes with different-colored fruit. Red fruit genotypes were associated with greater lycopene, β-carotene, lipophilic antioxidant capacity, and greater chromoplast-specific lycopene β-cyclase (CYC-B) transcript levels. Orange fruit genotypes had greater concentrations of tocopherols and greater transcript levels of homogentisate phytyl transferase (VTE-2), 1-deoxy-D-xylulose phosphate synthase (DXS), and 4-hydroxyphenylpyruvate dioxygenase (HPPD). The yellow fruit genotype was greater in total polyphenol and hydrophilic antioxidant capacity with greater expression of geranylgeranyl reductase (GGDR), phytol kinase (VTE-5), phytoene synthase (PSY) 2, lycopene β-cyclase (LCY-B), SlNAC1, and SINAC4. Greater levels of individual antioxidants were associated with specific coloration of tomato fruit. Moreover, the negative correlations between the expression of PSY1 and VTE-5, and between lycopene and chlorophyll, suggest a balance between carotenoids, tocopherols, and chlorophylls. The results of this study support either the direct commercialization of tomatoes with different color fruit or use of their genotypes in breeding programs to increase antioxidant levels among existing cultivars.