Plant amino acid-derived vitamins: biosynthesis and function

Vitamins are essential organic compounds for humans, having lost the ability to de novo synthesize them. Hence, they represent dietary requirements, which are covered by plants as the main dietary source of most vitamins (through food or livestock’s feed). Most vitamins synthesized by plants present amino acids as precursors (B₁, B₂, B₃, B₅, B₇, B₉ and E) and are therefore linked to plant nitrogen metabolism. Amino acids play different roles in their biosynthesis and metabolism, either incorporated into the backbone of the vitamin or as amino, sulfur or one-carbon group donors. There is a high natural variation in vitamin contents in crops and its exploitation through breeding, metabolic engineering and agronomic practices can enhance their nutritional quality. While the underlying biochemical roles of vitamins as cosubstrates or cofactors are usually common for most eukaryotes, the impact of vitamins B and E in metabolism and physiology can be quite different on plants and animals. Here, we first aim at giving an overview of the biosynthesis of amino acid-derived vitamins in plants, with a particular focus on how this knowledge can be exploited to increase vitamin contents in crops. Second, we will focus on the functions of these vitamins in both plants and animals (and humans in particular), to unravel common and specific roles for vitamins in evolutionary distant organisms, in which these amino acid-derived vitamins play, however, an essential role.     

Carotenoid oxygenases: cleave it or leave it

Carotenoid cleavage products (apocarotenoids) are widespread in living organisms and exert key biological functions. In animals, retinoids function as vitamins, visual pigments and signalling molecules. In plants, apocarotenoids play roles as hormones, pigments, flavours, aromas and defence compounds. The first step in their biosynthesis is the oxidative cleavage of a carotenoid catalysed by a non-heme iron oxygenase. A novel family of enzymes, which can cleave different carotenoids at different positions, has been characterized.     

Lipid peroxidation, chloroplastic pigments and antioxidant strategies in Carapa guianensis (Aubl.) subjected to water-deficit and short-term rewetting

The effects of drought on membrane lipids and leaf pigments and the ability of andiroba (Carapa guianensis Aubl.) plants to attenuate oxidative damage through antioxidant enzymes or adjusting carotenoids and glycinebetaine (GB) were examined. Assessments were performed when pre-dawn leaf water potential (Ψ_pd) of water-stressed plants reached −1.35 and −3.21 MPa (15 and 27 days after withholding irrigation) and 12 h after resuming watering (short-term rewetting, day 28). Oxidative damages to lipids were evident on day 15, in which drought caused an increase of 47% in malondialdehyde (MDA) content. On day 27, MDA content did not differ between treatments. The activity of superoxide dismutase remained unchanged over experimental period, while significant increases in the ascorbate peroxidase (APX, 110%) and catalase (CAT, 50%) activities were observed only on day 27. GB content was 62% (day 15) and 112% (day 27) higher in water-stressed plants than in control. Regardless of Ψ_pd, both chlorophyll (Chl) a, Chl b and total carotenoids remained unchanged between well-watered and water-stressed plants, indicating that drought did not result in degradation of leaflet pigments. On day 28, Ψ_pd of water-stressed plants increased near to control plants and both activities of APX and CAT did not differ between treatments. Altogether, adjustments in APX and CAT activity and in the GB content were efficient strategies to prevent expressive oxidative damages in water-stressed andiroba plants.     

Lutein-based plumage coloration in songbirds is a consequence of selective pigment incorporation into feathers

Many birds obtain colorful carotenoid pigments from the diet and deposit them into growing tissues to develop extravagant red, orange or yellow sexual ornaments. In these instances, it is often unclear whether all dietary pigments are used as integumentary colorants or whether certain carotenoids are preferentially excluded or incorporated into tissues. We examined the carotenoid profiles of three New World passerines that display yellow plumage coloration—the yellow warbler (Dendroica petechia), common yellowthroat (Geothlypis trichas) and evening grosbeak (Coccothraustes vespertinus). Using high-performance liquid chromatography, we found that all species used only one carotenoid—lutein—to color their plumage yellow. Analyses of blood carotenoids (which document those pigments taken up from the diet) in two of the species, however, revealed the presence of two dietary xanthophylls—lutein and zeaxanthin—that commonly co-occur in plants and animals. These findings demonstrate post-absorptive selectivity of carotenoid deposition in bird feathers. To learn more about the site of pigment discrimination, we also analyzed the carotenoid composition of lipid fractions from the follicles of immature yellow-pigmented feathers in G. trichas and D. petechia and again detected both lutein and zeaxanthin. This suggests that selective lutein incorporation in feathers is under local control at the maturing feather follicle.     

Nature,Source and Function of Pigments in Tardigrades: In Vivo Raman Imaging of Carotenoids in Echiniscus blumi

Tardigrades are microscopic aquatic animals with remarkable abilities to withstand harsh physical conditions such as dehydration or exposure to harmful highly energetic radiation. The mechanisms responsible for such robustness are presently little known, but protection against oxidative stresses is thought to play a role. Despite the fact that many tardigrade species are variously pigmented, scarce information is available about this characteristic. By applying Raman micro-spectroscopy on living specimens, pigments in the tardigrade Echiniscus blumi are identified as carotenoids, and their distribution within the animal body is visualized. The dietary origin of these pigments is demonstrated, as well as their presence in the eggs and in eye-spots of these animals, together with their absence in the outer layer of the animal (i.e., cuticle and epidermis). Using in-vivo semi-quantitative Raman micro-spectroscopy, a decrease in carotenoid content is detected after inducing oxidative stress, demonstrating that this approach can be used for studying the role of carotenoids in oxidative stress-related processes in tardigrades. This approach could be thus used in further investigations to test several hypotheses concerning the function of these carotenoids in tardigrades as photo-protective pigments against ionizing radiations or as antioxidants defending these organisms against the oxidative stress occurring during desiccation processes.     

Enrichment of some B-vitamins in plants with application of organic fertilizers

A review of the literature showed that plants grown with organic fertilizers often contain higher concentrations of vitamins B₁ (thiamin) and B₁₂ (cyanocobalamin) as compared with plants grown with inorganic fertilizers. Since plant roots were recently shown to be able to absorb B₁ and B₁₂, it was thus suspected that organic fertilizers (such as manure of diverse sources or sewage sludges which often contain relatively high concentrations of several vitamins) introduce additional vitamins into the soil which in turn leads to increased vitamins in the plants. This possibility was studied by measuring the B₁₂ content in the seeds of soybean and barley and in the leaves of spinach plants grown in soils amended with pure B₁₂ or cow dung (which is naturally rich in B₁₂). The addition of pure B₁₂ or cow dung did not alter the B₁₂ content in the soybean seeds but significantly increased that in the barley kernels and in the spinach leaves. For example, the addition of cow dung at the rate of 10 g kg^–1 increased the B₁₂ content in barley kernels by more than threefold (from 2.6 to 9.1 ng g^–1 DW) and in spinach leaves by close to twofold (from 6.9 to 17.8 ng g^–1 DW). Long-term addition of organic fertilizers to the soil also significantly increased the soil content of this vitamin. Since plants cannot synthesize B₁₂ and thus plant foods are normally fully devoid of (or have very low concentrations of) this vitamin, the finding that plants grown with organic fertilizers may contain relatively higher concentrations of this vitamin may have nutritional consequences in that the consumption of these plants by humans would inadvertently increase their intake of this vitamin. This may be of special benefit to people living by choice or by necessity on strict vegetarian diets who are known to be in danger of B₁₂ deficiency.     

Carotenoid production in <Emphasis Type="Italic">Bacillus subtilis</Emphasis> achieved by metabolic engineering

The carotenoid synthetic genes, crtM and crtN, derived from Staphylococcus aureus, were introduced into B. subtilis, resulting in yellow pigmentation. Absorption maxima of pigments and MALDI-TOF mass spectrometry demonstrated that the pigmented strain accumulated two C₃₀ carotenoids, 4,4′-diapolycopene and 4,4′-diaponeurosporene. A survival test using H₂O₂ revealed that the pigmented strain was more resistant to oxidative stress than the strain harboring an empty-vector. These findings indicate that B. subtilis can produce carotenoids, and the strain accumulating the carotenoids, CarotenoBacillus, will become a basal host for production of C₃₀ carotenoids and evaluation of their antioxidative effects.     

EFFECTS OF CERTAIN LIMITING CONDITIONS ON THE SYNTHESIS OF B VITAMINS BY YEAST

In yeast crops which were grown in the presence of various inhibitors, there was considerable variation in content of the various B vitamins. A higher degree of parallelism in variation in content was found to exist between thiamine and niacin than between any other pair of vitamins; this has been interpreted as indicating that the predominant functions of these two vitamins are their established rôles in fermentation. Values for inositol indicate that it may be involved in fermentation processes, but this is not the case for other members of the B complex. Biotin appears to be unique since in no case did the biotin content of yeast grown in the presence of an inhibitor fall below that of the control yeast. There was some evidence of synthesis of biotin, or a material with biotin activity, in the presence of certain inhibitors, the most striking instance being with sulfaguanidine. An exogenous supply of biotin was essential for extensive proliferation of F. B. yeast, and yeast grown in a medium to which biotin was the only added vitamin contained the B vitamins in amounts very similar to those found in the control yeast, the most marked differences being in increased vitamin B₆ and p-aminobenzoic acid contents. In the absence of biotin, significant amounts of all of the B vitamins except biotin were synthesized, both in the presence and absence of certain other members of the B complex. The addition of thiamine, pyridoxine, inositol, and β-alanine to the culture medium caused a reduction in the amounts of vitamin B₆ and p-aminobenzoic acid synthesized. F. B. yeast was able to grow in a xylose medium only when certain of the B vitamins were present, and even then growth was limited. Evidence was obtained for some synthesis of all of the vitamins investigated except biotin and vitamin B₆. The most significant differences in vitamin content between galac yeast and the parent F. B. strain were in folic acid and vitamin B₆, the former being considerably reduced in amount, the latter being increased.     

Effects of carotenoid sequestration on a caterpillar's cryptic coloration and susceptibility to predation

Carotenoids are used for many functions by animals, including combining with other pigments to produce aposematic and cryptic coloration. Carotenoids in combination with blue pigments are responsible for green coloration in many caterpillars, and thus carotenoid sequestration may reduce their contrast against a green foliage background. We tested the hypothesis that carotenoid sequestration reduces contrast and enhances survival by rearing Trichoplusia ni Hübner (Lepidoptera: Noctuidae) on Brassica oleracea L. var. Acephala (Brassicaceae) leaves and exposing them to predators. We found that carotenoids derived from the host plant are partially excreted, along with chlorophyll, but also sequestered in hemolymph. Larvae that were given plants that provided carotenoids showed less contrast against their host plants within 1 day compared to larvae that were not provided with carotenoids. Last, both short‐term field observations and laboratory trials of larvae caged with predatory Podisus maculiventris Say (Hemiptera: Pentatomidae) nymphs showed that survival of carotenoid‐sequestering larvae was higher compared to larvae that did not sequester. These results suggest that carotenoid sequestration may be an important adaptive strategy that reduces susceptibility to natural enemies that hunt by sight. Further research that examines the mechanisms by which carotenoids are absorbed and modified will lend insights into the evolution of carotenoids functioning as passive defensive compounds.     

Contributions of pterin and carotenoid pigments to dewlap coloration in two anole species

Animals can acquire bright coloration using a variety of pigmentary and microstructural mechanisms. Reptiles and amphibians are known to use two types of pigments - pterins and carotenoids - to generate their spectrum of colorful red, orange, and yellow hues. Because both pigment classes can confer all of these hues, the relative importance of pterins versus carotenoids in creating these different colors is not always apparent. We studied the carotenoid and pterin content of red and yellow dewlap regions in two neotropical anole species - the brown anole (Norops sagrei) and the ground anole (N. humilis). Pterins (likely drosopterins) and carotenoids (likely xanthophylls) were present in all tissues from all individuals. Pterins were more enriched in the lateral (red) region, and carotenoids more enriched in the midline (yellow) region in N. humilis, but pterins and carotenoids were found in similar concentrations among lateral and midline regions in N. sagrei. These patterns indicate that both carotenoid and pterin pigments are responsible for producing color in the dichromatic dewlaps of these two species, and that in these two species the two pigments interact differently to produce the observed colors.     

Metabolic recovery of the Antarctic liverwort <Emphasis Type="Italic">Cephaloziella varians</Emphasis> during spring snowmelt

We measured the responses of pigments and chlorophyll a fluorescence parameters of the Antarctic leafy liverwort Cephaloziella varians to snowmelt during austral spring 2005 at Rothera Point on the western Antarctic Peninsula. Although no changes to the concentrations of UV-B photoprotective pigments were detected during snowmelt, chlorophyll and carotenoid concentrations and maximum photosystem (PS)II yield (F _v /F _m) were respectively 88, 60 and 144% higher in the tissues of the liverwort that had recently emerged from snow than in those under a 10 cm depth of snow. A laboratory experiment similarly showed that effective PSII yield increased rapidly within the first 45 min after plants sampled from under snow were removed to an illuminated growth cabinet. The pigmentation and PSII yields of plants during snowmelt were also compared with those of plants in January, during the middle of the growing season at Rothera Point. During snowmelt, plants had lower F _v /F _m values, chlorophyll a/b ratios and concentrations of UV-B photoprotective pigments and carotenoids than during mid-season, suggesting that although there is some recovery of PSII activity and increases in concentrations of photosynthetic pigments during snowmelt, the metabolism of C. varians is restricted during this period.     

Photosynthetic Pigments of Tomato Plants under Conditions of Biotic Stress and Effects of Furostanol Glycosides

The adaptogenic effect of furostanol glycosides (FG) on biosynthesis of photosynthetic pigments in tomato plants (Lycopersicon esculentum Mill.) was studied under conditions of biotic stress caused by root-knot nematode (Meloidogyne incognita Kofoid et White). Treatment of plants with 5 × 10^–4 M FG was accompanied by an increase in the rate of biosynthesis of pigments (particularly, chlorophyll b and carotenoids), which was observed against the background of a decrease in the relative contribution of -carotene and an increase in the relative contribution of pigments of the violaxanthin cycle (VXC) to the overall pool of carotenoids. It was suggested that FG stimulated phytoimmunity by shifting metabolism of carotenoids toward enhanced biosynthesis of VXC pigments. These pigments play a protective role and facilitate stabilization of the photosynthetic apparatus, which is particularly important under stress conditions.     

Response of two Antarctic bryophytes to stratospheric ozone depletion

We report a study which measured changes to the radiative environment arising from stratospheric O₃ depletion at Rothera Point on the western Antarctic Peninsula (67°S, 68°W) and subsequent associations between these changes and the pigmentation and maximum quantum yield of photochemistry (F_v/F_m) of two Antarctic bryophytes, the liverwort Cephaloziella varians and the moss Sanionia uncinata. We found a strong relationship between O₃ column depth and the ratio of UV‐B to PAR irradiance (F_UV‐B/F_PAR) recorded at ground level. Weaker, but significant, associations were also found between O₃ column depth and noon irradiances and daily doses of unweighted and biologically effective UV‐B radiation received at ground level. Regression analyses indicated that F_UV‐B/F_PAR and daily dose of unweighted UV‐B were best predictors for concentrations of total carotenoids and UV‐B screening pigments extracted from bryophyte tissues. Concentrations of these pigments were loosely but significantly positively associated with O₃‐dependent irradiance parameters. HPLC analyses of carotenoids also suggested that both species increased the synthesis of neoxanthin during periods of O₃ depletion. Violaxanthin, lutein, zeaxanthin and b,bββ‐carotene concentrations were also apparently influenced by O₃ reduction, but not consistently across both bryophyte species. Concentrations of chlorophylls a and b were apparently unaffected by O₃ depletion. No direct associations between F_v/F_m and O₃‐dependent irradiance parameters were found. However stepwise multiple regression analyses suggested that the production of UV‐B screening pigments conferred protection from elevated F_UV‐B/F_PAR on F_v/F_m in both species and that carotenoids conferred protection on F_v/F_m in Sanionia. Our data suggest that changes to the radiative environment associated with stratospheric O₃ depletion influence the pigmentation of two Antarctic bryophytes, but that F_v/F_m is unaffected, at least in part because of rapid synthesis of protective pigments.     

Effect of glutamate on Pyricularia oryzae infection of rice monitored by changes in photosynthetic parameters and antioxidant metabolism

Considering the importance of blast caused by Pyricularia oryzae in the decrease of rice yield worldwide, this study aimed to assess the photosynthetic performance [leaf gas exchange and chlorophyll (Chl) a fluorescence parameters as well as the photosynthetic pigments concentration], the activities of antioxidant enzymes [ascorbate peroxidase, catalase (CAT), peroxidase (POX), superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione-S-transferase] and concentrations of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) in the leaves of rice plants non-supplied (−Glu) or supplied (+Glu) with glutamate (Glu) and non-infected or infected by P. oryzae. Blast severity was reduced in the leaves of +Glu plants. On the infected leaves of +Glu plants, the values for internal CO₂ concentration were lower while the values for net carbon assimilation rate, stomatal conductance as well as for the concentrations of Chl a, Chl b and carotenoids were higher in comparison to infected leaves of −Glu plants. The functionality of the photosynthetic apparatus was preserved in the infected leaves of +Glu plants. The activities of CAT, GPX, GR, POX and SOD increased in the infected leaves of both −Glu and +Glu plants compared to their non-inoculated counterparts, but their activities were lower for +Glu plants. The lower activity of these antioxidative enzymes was triggered by the reduced hydrogen peroxide concentration in the infected leaves of +Glu plants resulting in lower MDA concentration. It can be concluded that photosynthesis was less impaired in infected plants supplied with glutamate due to the lower biochemical constraints for CO₂ fixation. Moreover, there was a need for lower activity of reactive oxygen species scavenging enzymes in infected leaves of plants supplied with glutamate due to the lower oxidative stress imposed by P. oryzae infection.     

Diversity and evolution of photosynthetic antenna systems in green plants

Photosynthetic antenna systems are mainly involved in the absorption of light energy required for photosynthesis. The typical green plants arrange chlorophylls a and b and carotenoids, including lutein and 9′‐cis neoxanthin, in their antenna systems; such antenna systems have prospered on earth. Therefore, these antenna systems should be highly evolved and should adapt to the photoenvironments in which plants grow. However, little information is available on the diversity and evolution of antenna systems in green plants as a whole. To approach this, the present study focused on the antenna systems in the Prasinophyceae, an assemblage of early diverging lineages of green plants and analyzed their photosynthetic pigments in detail. In the present study, various novel blue–green light‐absorbing siphonaxanthin series were detected in the early diverging species of the Prasinophyceae and the distribution of these carotenoids was revealed. Additionally, to clarify the evolution of antenna systems in the Ulvophyceae, a highly developed green algal group that specializes in inhabiting various aquatic environments, members of the Cladophorales belonging to this class were selected and their carotenoid compositions were determined to compare them with the molecular phylogenetic tree constructed on the basis of the 18S rRNA gene sequences of the Cladophorales. In this review, these data will be summarized and the remarkable variation of photosynthetic pigments will be presented. A possible scenario detailing the evolution of antenna systems in green plants will be elucidated.     

Photosynthetic efficiency of marine plants

Multicellular marine plants were collected from their natural habitats and the quantum efficiency of their photosynthesis was determined in the laboratory in five narrow wave length bands in the visible spectrum. The results along with estimates of the relative absorption by the various plastid pigments show a fairly uniform efficiency of 0.08 molecules O₂ per absorbed quantum for (a) chlorophyll of one flowering plant, green algae, and brown algae, (b) fucoxanthol and other carotenoids of brown algae, and (c) the phycobilin pigments phycocyanin and phycoerythrin of red algae. The carotenoids of green algae are sometimes less efficient while those of red algae are largely or entirely inactive. Chlorophyll a of red algae is about one-half as efficient (_o2 = 0.04) as either the phycobilins, or the chlorophyll of most other plants. These results as well as those of high intensity and of fluorescence experiments are consistent with a mechanism in which about half the chlorophyll is inactive while the other half is fully active and is an intermediate in phycoerythrin- and phycocyanin-sensitized photosynthesis.     

Is carotenoid ornamentation linked to the inner mitochondria membrane potential? A hypothesis for the maintenance of signal honesty

Several mechanistic hypotheses have been proposed for how carotenoid pigmentation of integumentary structures can serve as an honest signal of individual quality. These hypotheses are founded on proposed links between carotenoids, immuno responsiveness, and oxidative stress, but an absence of biochemical information on the oxidative pathways of carotenoids has limited the sophistication of such hypotheses. Based on published evidence, we propose that the oxidation of carotenoids for the purpose of ornamentation in birds and reptiles is coupled to the inner mitochondria membrane. We predict that several carotenoid oxidation reactions yielding ornamental pigments occur on the inner mitochondrial membrane. Three of these reactions are proposed to occur within the ubiquinone biosynthesis cluster known as the Coq cluster consisting of approximately a dozen Coq members, tightly integrated and intimately associated with Complex I and III of the electron transport system. Ubiquinone and highly oxidized ornamental carotenoids share a stereochemically-conserved binding region suggesting that these two molecules may have shared similar pathways in the past. Carotenoids and ubiquinones may cooperate as redox participants in anti-radical reactions or independently in helping to maintain membrane or supra-complex stabilization during times of high-energy demand. Under this hypothesis, oxidation of carotenoids is coupled to the inner mitochondria membrane potential such that ornamental coloration reflects the efficiency of cellular respiration.     

Columnar cacti as sources of energy and protein for frugivorous bats in a semi‐arid ecosystem

Columnar cacti constitute the dominant elements in the vegetation structure of arid and semi‐arid New World ecosystems representing a plethora of food resources for vertebrate consumers. Previous stable isotope analysis in Central Mexico showed that columnar cacti are of low importance to build tissue for frugivorous bats. We used carbon stable isotope analysis of whole blood and breath samples collected from four species of frugivorous bats (Sturnira parvidens, Sturnira ludovici, Artibeus jamaicensis, and Artibeus intermedius) to reconstruct the importance of cactus plants in their diet. Breath samples were collected within 10 min (B₁₀) of bat capture and ~12 h after capture (B₇₂₀), representing the oxidation of recently ingested food and of body reserves, respectively. We expected that bats relied primarily on non‐cactus food to construct tissues and fuel oxidative metabolism. Non‐cactus food strongly predominated for tissue building, whereas oxidative metabolism was supported by a moderate preponderance of non‐cactus food for B₁₀ samples, and a moderate preponderance of cactus food or an equal contribution of both sources for B₇₂₀ samples. Artibeus and Surnira species appear to cover a narrow part of the diet with cactus food, confirming that the incorporation of nutrients derived from these plants is not generalized among vertebrate consumers.     

Issue Information ‐ Referees List

Rapid developments in remote‐sensing of vegetation and high‐throughput precision plant phenotyping promise a range of real‐life applications using leaf optical properties for non‐destructive assessment of plant performance. Use of leaf optical properties for assessing plant performance requires the ability to use photosynthetic pigments as proxies for physiological properties and the ability to detect these pigments fast, reliably and at low cost. We describe a simple and cost‐effective protocol for the rapid analysis of chlorophylls, carotenoids and tocopherols using high‐performance liquid chromatography (HPLC). Many existing methods are based on the expensive solvent acetonitrile, take a long time or do not include lutein epoxide and α‐carotene. We aimed to develop an HPLC method which separates all major chlorophylls and carotenoids as well as lutein epoxide, α‐carotene and α‐tocopherol. Using a C₃₀‐column and a mobile phase with a gradient of methanol, methyl‐tert‐butyl‐ether (MTBE) and water, our method separates the above pigments and isoprenoids within 28 min. The broad applicability of our method is demonstrated using samples from various plant species and tissue types, e.g. leaves of Arabidopsis and avocado plants, several deciduous and conifer tree species, various crops, stems of parasitic dodder, fruit of tomato, roots of carrots and Chlorella algae. In comparison to previous methods, our method is very affordable, fast and versatile and can be used to analyze all major photosynthetic pigments that contribute to changes in leaf optical properties and which are of interest in most ecophysiological studies.     

The Pseudoenzyme PDX1.2 Boosts Vitamin B6 Biosynthesis under Heat and Oxidative Stress in Arabidopsis

Vitamin B₆ is an indispensable compound for survival, well known as a cofactor for numerous central metabolic enzymes and more recently for playing a role in several stress responses, particularly in association with oxidative stress. Regulatory aspects for the use of the vitamin in these roles are not known. Here we show that certain plants carry a pseudoenzyme (PDX1.2), which is involved in regulating vitamin B₆ biosynthesis de novo under stress conditions. Specifically, we demonstrate that Arabidopsis PDX1.2 enhances the activity of its catalytic paralogs by forming a heterododecameric complex. PDX1.2 is strongly induced by heat as well as singlet oxygen stress, concomitant with an enhancement of vitamin B₆ production. Analysis of pdx1.2 knockdown lines demonstrates that boosting vitamin B₆ content is dependent on PDX1.2, revealing that this pseudoenzyme acts as a positive regulator of vitamin B₆ biosynthesis during such stress conditions in plants.