Friends in Arms: Flavonoids and the Auxin/Cytokinin Balance in Terrestrialization.

Land plants survive the challenges of new environments by evolving mechanisms that protect them from excess irradiation, nutrient deficiency, and temperature and water availability fluctuations. One such evolved mechanism is the regulation of the shoot/root growth ratio in response to water and nutrient availability by balancing the actions of the hormones auxin and cytokinin. Plant terrestrialization co-occurred with a dramatic expansion in secondary metabolism, particularly with the evolution and establishment of the flavonoid biosynthetic pathway. Flavonoid biosynthesis is responsive to a wide range of stresses, and the numerous synthesized flavonoid species offer two main evolutionary advantages to land plants. First, flavonoids are antioxidants and thus defend plants against those adverse conditions that lead to the overproduction of reactive oxygen species. Second, flavonoids aid in protecting plants against water and nutrient deficiency by modulating root development and establishing symbiotic relations with beneficial soil fungi and bacteria. Here, we review different aspects of the relationships between the auxin/cytokinin module and flavonoids. The current body of knowledge suggests that whereas both auxin and cytokinin regulate flavonoid biosynthesis, flavonoids act to fine-tune only auxin, which in turn regulates cytokinin action. This conclusion agrees with the established master regulatory function of auxin in controlling the shoot/root growth ratio.

Differential oxidative stress responses and tobacco-specific nitrosamine accumulation in two burley varieties.

Tobacco-specific nitrosamines (TSNAs) are carcinogens that accumulate in tobacco leaves during curing, storage, and processing, and their amounts in processed tobacco vary dependent on several intrinsic and extrinsic factors. Here, we assessed the hypothesis that there is a link between reactive oxygen species levels in leaves and TSNA formation during curing. First, we show that burley varieties KT 204LC and NCBH 129LC accumulate TSNAs to different levels but not as a result of a variety-specific abundance of TSNA precursors. Next, we measured the levels of reactive oxygen species, and we show that the variety that accumulates more TSNAs, NCBH 129LC, had significantly higher levels of hydrogen peroxide than KT 204LC. The NCBH 129LC also has more oxidatively damaged and glutathionylated proteins. Finally, we analyzed the antioxidant levels in KT 204LC and NCBH 129LC and their tolerance to oxidative stress. NCBH 129LC contained more of the essential antioxidant glutathione and was more tolerant to the oxidative stress-generating compound paraquat. Collectively, our data suggest that there is indeed a link between foliar oxidative stress parameters and the extent to which TSNAs accumulate in cured tobacco leaves.

Inhibition of Fusarium oxysporum f. sp. nicotianae Growth by Phenylpropanoid Pathway Intermediates.

Fusarium wilt in tobacco caused by the fungus Fusarium oxysporum f. sp. nicotianae is a disease­management challenge worldwide, as there are few effective and environmentally benign chemical agents for its control. This challenge results in substantial losses in both the quality and yield of tobacco products. Based on an in vitro analysis of the effects of different phenylpropanoid intermediates, we found that the early intermediates trans­cinnamic acid and para­coumaric acid effectively inhibit the mycelial growth of F. oxysporum f. sp. nicotianae strain FW316F, whereas the downstream intermediates quercetin and caffeic acid exhibit no fungicidal properties. Therefore, our in vitro screen suggests that trans­cinnamic acid and para­coumaric acid are promising chemical agents and natural lead compounds for the suppression of F. oxysporum f. sp. nicotianae growth.

Cytokinin-induced protein synthesis suppresses growth and osmotic stress tolerance.

Cytokinins control critical aspects of plant development and environmental responses. Perception of cytokinin ultimately leads to the activation of proteins belonging to the type-B Response Regulator family of cytokinin response activators. In Arabidopsis thaliana, ARR1 is one of the most abundantly expressed type-B Response Regulators. We investigated the link between cytokinin signaling, protein synthesis, plant growth and osmotic stress tolerance. We show that the increased cytokinin signaling in ARR1 gain-of-function transgenic lines is associated with increased rates of protein synthesis, which lead to growth inhibition and hypersensitivity to osmotic stress. Cytokinin-induced growth inhibition and osmotic stress hypersensitivity were rescued by treatments with ABA, a hormone known to inhibit protein synthesis. We also demonstrate that cytokinin-induced protein synthesis requires isoforms of the ribosomal protein L4 encoded by the cytokinin-inducible genes RPL4A and RPL4D, and that RPL4 loss-of-function increases osmotic stress tolerance and decreases sensitivity to cytokinin-induced growth inhibition. These findings reveal that an increase in protein synthesis negatively impacts growth and osmotic stress tolerance and explain some of the adverse effects of elevated cytokinin action on plant development and stress physiology.

Metabolomic analyses of the bio-corona formed on TiO2 nanoparticles incubated with plant leaf tissues.

BACKGROUND: The surface of a nanoparticle adsorbs molecules from its surroundings with a specific affinity determined by the chemical and physical properties of the nanomaterial. When a nanoparticle is exposed to a biological system, the adsorbed molecules form a dynamic and specific surface layer called a bio-corona. The present study aimed to identify the metabolites that form the bio-corona around anatase TiO2 nanoparticles incubated with leaves of the model plant Arabidopsis thaliana. RESULTS: We used an untargeted metabolomics approach and compared the metabolites isolated from wild-type plants with plants deficient in a class of polyphenolic compounds called flavonoids. CONCLUSIONS: These analyses showed that TiO2 nanoparticle coronas are enriched for flavonoids and lipids and that these metabolite classes compete with each other for binding the nanoparticle surface.

Anatase TiO2 Nanoparticles Induce Autophagy and Chloroplast Degradation in Thale Cress (Arabidopsis thaliana).

The extensive use of TiO2 nanoparticles and their subsequent release into the environment have posed an important question about the effects of this nanomaterial on ecosystems. Here, we analyzed the link between the damaging effects of reactive oxygen species generated by TiO2 nanoparticles and autophagy, a housekeeping mechanism that removes damaged cellular constituents. We show that TiO2 nanoparticles induce autophagy in the plant model system Arabidopsis thaliana and that autophagy is an important mechanism for managing TiO2 nanoparticle-induced oxidative stress. Additionally, we find that TiO2 nanoparticles induce oxidative stress predominantly in chloroplasts and that this chloroplastic stress is mitigated by autophagy. Collectively, our results suggest that photosynthetic organisms are particularly susceptible to TiO2 nanoparticle toxicity.

Antagonistic activity of auxin and cytokinin in shoot and root organs.

The hormones auxin and cytokinin are essential for plant growth and development. Because of the central importance of root and shoot apical meristems in plant growth, auxin/cytokinin interactions have been predominantly analyzed in relation to apical meristem formation and function. In contrast, the auxin/cytokinin interactions during organ growth have remained largely unexplored. Here, we show that a specific interaction between auxin and cytokinin operates in both the root and the shoot where it serves as an additional determinant of plant development. We found that auxin at low concentrations limits the action of cytokinin. An increase in cytokinin level counteracts this inhibitory effect and leads to an inhibition of auxin signaling. At higher concentrations of both hormones, these antagonistic interactions between cytokinin and auxin are absent. Thus, our results reveal a bidirectional and asymmetrical interaction of auxin and cytokinin beyond the bounds of apical meristems. The relation is bidirectional in that both hormones exert inhibitory effects on each other's signaling mechanisms. However, this relation is also asymmetrical because under controlled growth conditions, auxin present in nontreated plants suppresses cytokinin signaling, whereas the reverse is not the case.

Modulation of auxin and cytokinin responses by early steps of the phenylpropanoid pathway.

BACKGROUND: The phenylpropanoid pathway is responsible for the synthesis of numerous compounds important for plant growth and responses to the environment. In the first committed step of phenylpropanoid biosynthesis, the enzyme phenylalanine ammonia-lyase (PAL) deaminates L-phenylalanine into trans-cinnamic acid that is then converted into p-coumaric acid by cinnamate-4-hydroxylase (C4H). Recent studies showed that the Kelch repeat F-box (KFB) protein family of ubiquitin ligases control phenylpropanoid biosynthesis by promoting the proteolysis of PAL. However, this ubiquitin ligase family, alternatively named Kiss Me Deadly (KMD), was also implicated in cytokinin signaling as it was shown to promote the degradation of type-B ARRs, including the key response activator ARR1. Considering that ubiquitin ligases typically have narrow target specificity, this dual targeting of structurally and functionally unrelated proteins appeared unusual. RESULTS: Here we show that the KFBs indeed target PAL but not ARR1. Moreover, we show that changes in early phenylpropanoid biosynthesis alter cytokinin sensitivity - as reported earlier - but that the previously documented cytokinin growth response changes are primarily the result of altered auxin signaling. We found that reduced PAL accumulation decreased, whereas the loss of C4H function increased the strength of the auxin response. The combined loss of function of both enzymes led to a decrease in auxin sensitivity, indicating that metabolic events upstream of C4H control auxin sensitivity. This auxin/phenylpropanoid interaction impacts both shoot and root development and revealed an auxin-dependent stimulatory effect of trans-cinnamic acid feeding on leaf expansion and thus biomass accumulation. CONCLUSIONS: Collectively, our results show that auxin-regulated plant growth is fine-tuned by early steps in phenylpropanoid biosynthesis and suggest that metabolites accumulating upstream of the C4H step impact the auxin response mechanism.

Cytokinin signaling promotes differential stability of type-B ARRs.

Cytokinins control key aspects of plant growth, including shoot and root meristem development and the timing of senescence of leaves and stems. Cytokinin perception triggers a 2-component signaling mechanism that ultimately leads to phosphorylation-dependent activation of a class of transcriptional regulators called type-B ARRs (RRBs). We have recently shown that the stability of the RRB family member ARR1 is increased in response to elevated cytokinin concentrations. In contrast, cytokinin decreases the stability of the closely related RRB member ARR2. The molecular mechanism governing the differential stability regulation of these 2 closely related RRBs remains unknown.