Uncovering the functional residues of Arabidopsis isoprenoid biosynthesis enzyme HDS.

The methylerythritol phosphate (MEP) pathway is responsible for producing isoprenoids, metabolites with essential functions in the bacterial kingdom and plastid-bearing organisms including plants and Apicomplexa. Additionally, the MEP-pathway intermediate methylerythritol cyclodiphosphate (MEcPP) serves as a plastid-to-nucleus retrograde signal. A suppressor screen of the high MEcPP accumulating mutant plant (ceh1) led to the isolation of 3 revertants (designated Rceh1-3) resulting from independent intragenic substitutions of conserved amino acids in the penultimate MEP-pathway enzyme, hydroxymethylbutenyl diphosphate synthase (HDS). The revertants accumulate varying MEcPP levels, lower than that of ceh1, and exhibit partial or full recovery of MEcPP-mediated phenotypes, including stunted growth and induced expression of stress response genes and the corresponding metabolites. Structural modeling of HDS and ligand docking spatially position the substituted residues at the MEcPP binding pocket and cofactor binding domain of the enzyme. Complementation assays confirm the role of these residues in suppressing the ceh1 mutant phenotypes, albeit to different degrees. In vitro enzyme assays of wild type and HDS variants exhibit differential activities and reveal an unanticipated mismatch between enzyme kinetics and the in vivo MEcPP levels in the corresponding Rceh lines. Additional analyses attribute the mismatch, in part, to the abundance of the first and rate-limiting MEP-pathway enzyme, DXS, and further suggest MEcPP as a rheostat for abundance of the upstream enzyme instrumental in fine-tuning of the pathway flux. Collectively, this study identifies critical residues of a key MEP-pathway enzyme, HDS, valuable for synthetic engineering of isoprenoids, and as potential targets for rational design of antiinfective drugs.

A plastidial retrograde signal potentiates biosynthesis of systemic stress response activators.

Plants employ an array of intricate and hierarchical signaling cascades to perceive and transduce informational cues to synchronize and tailor adaptive responses. Systemic stress response (SSR) is a recognized complex signaling and response network quintessential to plant's local and distal responses to environmental triggers; however, the identity of the initiating signals has remained fragmented. Here, we show that both biotic (aphids and viral pathogens) and abiotic (high light and wounding) stresses induce accumulation of the plastidial-retrograde-signaling metabolite methylerythritol cyclodiphosphate (MEcPP), leading to reduction of the phytohormone auxin and the subsequent decreased expression of the phosphatase PP2C.D1. This enables phosphorylation of mitogen-activated protein kinases 3/6 and the consequential induction of the downstream events ultimately, resulting in biosynthesis of the two SSR priming metabolites pipecolic acid and N-hydroxy-pipecolic acid. This work identifies plastids as a major initiation site, and the plastidial retrograde signal MEcPP as an initiator of a multicomponent signaling cascade potentiating the biosynthesis of SSR activators, in response to biotic and abiotic triggers.

Integrated omics analyses of retrograde signaling mutant delineate interrelated stress-response strata.

To maintain homeostasis in the face of intrinsic and extrinsic insults, cells have evolved elaborate quality control networks to resolve damage at multiple levels. Interorganellar communication is a key requirement for this maintenance, however the underlying mechanisms of this communication have remained an enigma. Here we integrate the outcome of transcriptomic, proteomic, and metabolomics analyses of genotypes including ceh1, a mutant with constitutively elevated levels of both the stress-specific plastidial retrograde signaling metabolite methyl-erythritol cyclodiphosphate (MEcPP) and the defense hormone salicylic acid (SA), as well as the high MEcPP but SA deficient genotype ceh1/eds16, along with corresponding controls. Integration of multi-omic analyses enabled us to delineate the function of MEcPP from SA, and expose the compartmentalized role of this retrograde signaling metabolite in induction of distinct but interdependent signaling cascades instrumental in adaptive responses. Specifically, here we identify strata of MEcPP-sensitive stress-response cascades, among which we focus on selected pathways including organelle-specific regulation of jasmonate biosynthesis; simultaneous induction of synthesis and breakdown of SA; and MEcPP-mediated alteration of cellular redox status in particular glutathione redox balance. Collectively, these integrated multi-omic analyses provided a vehicle to gain an in-depth knowledge of genome-metabolism interactions, and to further probe the extent of these interactions and delineate their functional contributions. Through this approach we were able to pinpoint stress-mediated transcriptional and metabolic signatures and identify the downstream processes modulated by the independent or overlapping functions of MEcPP and SA in adaptive responses.

Plastid-produced interorgannellar stress signal MEcPP potentiates induction of the unfolded protein response in endoplasmic reticulum.

Cellular homeostasis in response to internal and external stimuli requires a tightly coordinated interorgannellar communication network. We recently identified methylerythritol cyclodiphosphate (MEcPP) as a novel stress-specific retrograde signaling metabolite that accumulates in response to environmental perturbations to relay information from plastids to the nucleus. We now demonstrate, using a combination of transcriptome and proteome profiling approaches, that mutant plants (ceh1) with high endogenous levels of MEcPP display increased transcript and protein levels for a subset of the core unfolded protein response (UPR) genes. The UPR is an adaptive cellular response conserved throughout eukaryotes to stress conditions that perturb the endoplasmic reticulum (ER) homeostasis. Our results suggest that MEcPP directly triggers the UPR. Exogenous treatment with MEcPP induces the rapid and transient induction of both the unspliced and spliced forms of the UPR gene bZIP60. Moreover, compared with the parent background (P), ceh1 mutants are less sensitive to the ER-stress-inducing agent tunicamycin (Tm). P and ceh1 plants treated with Tm display similar UPR transcript profiles, suggesting that although MEcPP accumulation causes partial induction of selected UPR genes, full induction is triggered by accumulation of misfolded proteins. This finding refines our perspective of interorgannellar communication by providing a link between a plastidial retrograde signaling molecule and its targeted ensemble of UPR components in ER.

Control efficacy of Ca-containing foliar fertilizers on bitter pit in bagged 'Fuji' apple and effects on the Ca and N contents of apple fruits and leaves.

BACKGROUND: The preharvest application of Ca-containing foliar fertilizers can reduce the incidence of bitter pit (BP) in apples and improve fruit quality by increasing the Ca content and decreasing both the N content and the N/Ca ratio in fruits. In this study, we aimed to investigate the control efficacy of Ca-containing fertilizers on the incidence of BP and their effects on the Ca and N contents in bagged 'Fuji' apples by spraying foliar fertilizer containing calcium chloride (CaCl2 ), calcium nitrate [Ca(NO3 )2 ] or calcium formate [Ca(HCOO)2 ] at an early stage, five days after full bloom (DAFB) and 40 DAFB, and at a late stage, 80 DAFB and 125 DAFB. RESULTS: The incidences of BP were reduced significantly by 43.2-73.0%, and the efficacy of spraying at an early stage was significantly higher than that of spraying at a late stage. The Ca content of bagged apple fruits increased whereas the N content and N/Ca ratio decreased after spraying Ca-containing foliar fertilizers; however, the Ca content, N content and N/Ca ratio of apple leaves were differentially influenced. CONCLUSION: Foliar fertilizer containing CaCl2, Ca(NO3 )2 or Ca(HCOO)2 can be used at an early stage to control BP in apple and improve the quality of bagged apple fruits. © 2018 Society of Chemical Industry.

Symmetry in Cascade Chirality-Transfer Processes: A Catalytic Atroposelective Direct Arylation Approach to BINOL Derivatives.

Herein we disclose a scalable organocatalytic direct arylation approach for the regio- and atroposelective synthesis of non-C2-symmetric 2,2'-dihydroxy-1,1'-binaphthalenes (BINOLs). In the presence of catalytic amounts of axially chiral phosphoric acids, phenols and naphthols are coupled with iminoquinones via a cascade process that involves sequential aminal formation, sigmatropic rearrangement, and rearomatization to afford enantiomerically enriched BINOL derivatives in good to excellent yields. Our studies suggest that the (local) symmetry of the initially formed aminal intermediate has a dramatic impact on the level of enantioinduction in the final product. Aminals with a plane of symmetry give rise to BINOL derivatives with significantly lower enantiomeric excess than unsymmetrical ones featuring a stereogenic center. Presumably asymmetric induction in the sigmatropic rearrangement step is significantly more challenging than during aminal formation. Sigmatropic rearrangement of the enantiomerically enriched aminal and subsequent rearomatization transfers the central chirality into axial chirality with high fidelity.

The plastidial retrograde signal methyl erythritol cyclopyrophosphate is a regulator of salicylic acid and jasmonic acid crosstalk.

The exquisite harmony between hormones and their corresponding signaling pathways is central to prioritizing plant responses to simultaneous and/or successive environmental trepidations. The crosstalk between jasmonic acid (JA) and salicylic acid (SA) is an established effective mechanism that optimizes and tailors plant adaptive responses. However, the underlying regulatory modules of this crosstalk are largely unknown. Global transcriptomic analyses of mutant plants (ceh1) with elevated levels of the stress-induced plastidial retrograde signaling metabolite 2-C-methyl-D-erythritol cyclopyrophosphate (MEcPP) revealed robustly induced JA marker genes, expected to be suppressed by the presence of constitutively high SA levels in the mutant background. Analyses of a range of genotypes with varying SA and MEcPP levels established the selective role of MEcPP-mediated signal(s) in induction of JA-responsive genes in the presence of elevated SA. Metabolic profiling revealed the presence of high levels of the JA precursor 12-oxo-phytodienoic acid (OPDA), but near wild type levels of JA in the ceh1 mutant plants. Analyses of coronatine-insensitive 1 (coi1)/ceh1 double mutant plants confirmed that the MEcPP-mediated induction is JA receptor COI1 dependent, potentially through elevated OPDA. These findings identify MEcPP as a previously unrecognized central regulatory module that induces JA-responsive genes in the presence of high SA, thereby staging a multifaceted plant response within the environmental context.

Cytokinins can act as suppressors of nitric oxide in Arabidopsis.

Maintaining nitric oxide (NO) homeostasis is essential for normal plant physiological processes. However, very little is known about the mechanisms of NO modulation in plants. Here, we report a unique mechanism for the catabolism of NO based on the reaction with the plant hormone cytokinin. We screened for NO-insensitive mutants in Arabidopsis and isolated two allelic lines, cnu1-1 and 1-2 (continuous NO-unstressed 1), that were identified as the previously reported altered meristem program 1 (amp1) and as having elevated levels of cytokinins. A double mutant of cnu1-2 and nitric oxide overexpression 1 (nox1) reduced the severity of the phenotypes ascribed to excess NO levels as did treating the nox1 line with trans-zeatin, the predominant form of cytokinin in Arabidopsis. We further showed that peroxinitrite, an active NO derivative, can react with zeatin in vitro, which together with the results in vivo suggests that cytokinins suppress the action of NO most likely through direct interaction between them, leading to the reduction of endogenous NO levels. These results provide insights into NO signaling and regulation of its bioactivity in plants.

Efficient acquisition of high-purity cyanidin-3-O-glucoside from mulberry fruits: An integrated process of ATPS whole-cell transformation and semi-preparative HPLC purification.

As a nutritious fruit, mulberry is an ideal source of high-quality cyanidin-3-O-glucoside (C3G) with various biological activities. However, the difficult separation process of high-purity C3G leads to its high price. To rapidly prepare high-purity C3G, cyanidin-3-O-rutinoside is converted to C3G by direct hydrolysis of rhamnose bond using a whole-cell catalyst containing α-rhamnosidase. Combined with an aqueous two-phase system, a coupling reaction separation system was established. Two monomers were successfully separated by semi-preparative high performance liquid chromatography (semi-preparative HPLC). The conversion of C3G catalyzed by whole-cells in the PEG/Na2SO4 system increased from 47.11 % to 66.56 %, compared with the EtOH/(NH4)2SO4 system, and the whole-cell activity remained above 50 % after five rounds of reuse. Meanwhile, the purity of C3G was increased to 99 % via the semi-preparative HPLC purification and identified by MS. Thus, an integrated process of whole-cell-catalyzed conversion and product peak cutting partition collection provides a novel strategy for efficient biomanufacturing of high-purity C3G.

Reciprocity between a retrograde signal and a putative metalloprotease reconfigures plastidial metabolic and structural states.

Reconfiguration of the plastidial proteome in response to environmental cues is central to tailoring adaptive responses. To define the underlying mechanisms and consequences of these reconfigurations, we performed a suppressor screen, using a mutant (ceh1) accumulating high levels of a plastidial retrograde signaling metabolite, MEcPP. We isolated a revertant partially suppressing the dwarf stature and high salicylic acid of ceh1 and identified the mutation in a putative plastidial metalloprotease (VIR3). Biochemical analyses showed increased VIR3 levels in ceh1, accompanied by reduced abundance of VIR3-target enzymes, ascorbate peroxidase, and glyceraldehyde 3-phophate dehydrogenase B. These proteomic shifts elicited increased H2O2, salicylic acid, and MEcPP levels, as well as stromule formation. High light recapitulated VIR3-associated reconfiguration of plastidial metabolic and structural states. These results establish a link between a plastidial stress-inducible retrograde signaling metabolite and a putative metalloprotease and reveal how the reciprocity between the two components modulates plastidial metabolic and structural states, shaping adaptive responses.

Effects of metamitron on fruit set and photosynthetic biological characteristics of apples.

With dwarfing interstock Fuji apples as the test materials and water treatment as the control (CK), we examined the fruit thinning effect and its influences on leaves' photosynthesis by spraying 200, 300, and 400 mg·L-1 metamitron during the young fruit period to solve artificial fruit thinning problems (time-consuming, much labor, and low efficiency). The results showed that metamitron application could significantly reduce the inflorescence and flowers' fruit-setting rate by 16.5%-22.8% and 50.9%-53.9%, respectively. The treatment of 300 mg·L-1 metamitron had the strongest fruit thinning effect, with a single fruit rate of 46.6% and a double fruit rate of 18.3%. As a photosynthesis inhibitor, metamitron application reduced the chlorophyll content of leaves and strongly affected photosynthesis. The inhibitory effect on chlorophyll content disappeared after 15 days of the treatment, while that on the net photosynthetic rate disappeared gradually after 11 days of the treatment. The application of metamitron significantly reduced the maximum quantum yield of PSⅡ reaction center (Fv/Fm), actual photochemical efficiency (ΦPSⅡ), photochemical quenching coefficient (qP) and non-photochemical quenching coefficient (NPQ), with such inhibitory effect having been lasted for 15 days. OJIP analysis showed that metamitron caused damage to the apple leaves' oxygen-evolving complex, especially limiting the transfer of electrons in the PSⅡ reaction center from QA to QB. Metamitron treatment increased Wk, and significantly decreased ψo, RC/CSm, and PIabs. Besides, 300 mg·L-1 metamitron had the most significant effect. Our results showed that metamitron destroyed the structure of the PSⅡ reaction center of apple leaves and hindered the transfer of electrons from the donor to the receptor of PSⅡ. Consequently, the photosynthetic rate was affected, and the young fruits fell off due to the lack of accumulation of photosynthetic products.

Chiral Phosphoric Acid Catalyzed Asymmetric Desymmetrization of para-Quinamines with Isocyanates: Access to Functionalized Imidazolidin-2-one Derivatives.

The development of enantioselective desymmetrization of para-quinamines with isocyanates catalyzed by chiral phosphoric acid is reported. The strategy provides concise access to functionalized imidazolidin-2-one derivatives in high yields and enantioselectivities under mild reaction conditions. Remarkably, this reaction could be performed on a gram scale using 5 mol % catalyst loading and the chiral imidazolidin-2-one derivatives could be easily transformed into valuable scaffolds without disturbing the enantiopurity, demonstrating the synthetic utility of this protocol.

Nutritional targeting modification of silkworm pupae oil catalyzed by a smart hydrogel immobilized lipase.

To prepare a nutritional supplement using silkworm pupae oil (SPO) as a feedstock, a microfluidic reactor with a smart hydrogel immobilized lipase was first constructed to reduce the relative content of palmitic acid at sn-1,3 and improve the nutritional function. The effects of flow rate, reaction temperature, and substrate molar ratio were investigated. In vitro digestion and pH-stat models were employed to analyze the digestion feature after the modification of SPO, while HPLC-ELSD, zeta potential, DSC, and TGA were used to evaluate the nutritional function. The relative content of "OOO" and "OPO" type triglycerides was increased by 49.48% and 107.67%, and that of palmitic acid at sn-1,3 was decreased by 49.61% in 10 s. After the verification of the in vitro digestion model, the fatty acid release rate of the modified SPO was significantly improved by 22.07%, indicating the nutritional function improvement of SPO. Therefore, the nutritional function of SPO has been improved successfully by the application of a microchannel reactor with photo-immobilized lipase, which could set a reference for the utilization of insect oil resources.

Correction: Nutritional targeting modification of silkworm pupae oil catalyzed by a smart hydrogel immobilized lipase.

Correction for 'Nutritional targeting modification of silkworm pupae oil catalyzed by a smart hydrogel immobilized lipase' by Jin-Zheng Wang et al., Food Funct., 2021, 12, 6240-6253, DOI: 10.1039/D1FO00913C.

An alternative solution for α-linolenic acid supplements: in vitro digestive properties of silkworm pupae oil in a pH-stat system.

α-Linolenic acid (ALA) is recognised to have a regulatory effect on cardiovascular diseases. Due to the low bioavailability of linseed oil (LINO), which is the most common ALA supplement, it is necessary to find a replacement for ALA supplements that is more easily accepted by the human body. The content of ALA in silkworm pupae oil (SPO) is 32.60 ± 0.67%, and SPO can be substituted as a dietary lipid to meet the demand of the human body. In the present study, a pH-stat system was used to investigate the release degree of free fatty acids (FFAs) from SPO and construct a first-order kinetic model. Digestion experiments in vitro with different lipids showed that the maximum release FFA levels were SPO > SO (soybean oil) > LO (lard oil) > MSO (mulberry seed oil) > LINO, and the first-order kinetic apparent rate constants were LINO > SPO > LO > SO > MSO. Triacylglycerol (TAG) and fatty acid composition are the decisive factors in determining the level of lipid digestion. Therefore, the maximum level of FFAs released from SPO (84.34 ± 1.37%) was much higher than that of LINO (49.78 ± 0.52%) when the hydrolysis rates were 0.2114 s-1 and 0.2249 s-1, respectively. In addition, the smaller emulsion droplet size (609.24 ± 43.46 nm) and weaker surface charge (-17.93 ± 0.42 mV) also resulted in higher levels of SPO under in vitro digestion conditions. Meanwhile, due to low melting and crystallisation temperature, SPO is quickly absorbed by the human body. Overall, SPO can be used as a new alternative for ALA supplements based on its superior digestive properties.

A novel nanoparticle loaded with methyl caffeate and caffeic acid phenethyl ester against Ralstonia solanacearum-a plant pathogenic bacteria.

Developing a novel agent and understanding the interaction model between multipolymer nanoparticles and bacteria could be worthwhile to induce the protection of crops with the prevalence of frequent hazards because of the use of pesticides and chemical resistance. Unlike metal nanoparticles, multipolymer nanoparticles have bacteriostatic properties against Ralstonia solanacearum that can trigger bacterial wilt by infecting the plant. Therefore, a novel poly(lactic-co-glycolic acid) nanoparticle containing caffeic acid phenethyl ester (CAPE) and methyl caffeate (MC) was prepared with the sustained-release property (for 10 d at pH 6.5); here, 50% of the cumulative release rate was achieved. It was observed that the cytomembrane of R. solanacearum was jeopardized by the nanoparticle by the creation of large holes on the bacterial surface. The nanoparticle has an approximate EC50 value of 0.285 mg mL-1 with active pharmaceutical ingredients (APIs), while the drug dosage could be reduced by 2/3. Furthermore, to reveal the possible mechanism of interaction between the multipolymer nanoparticles and bacteria, a formidable inhibition effect was observed; the pathogenicity-related genes, namely, phcA, phcB, pehC, egl, pilT, and polA, of R. solanacearum were downregulated by 1/2, 1/42, 1/13, 1/6, 1/2, and 1/8, respectively, showing significant effects on the major virulence-related genes. Hence, a novel nanoparticle with excellent antibacterial and sustained-release properties has been prepared, possessing the potential to replace chemical pesticides and serve as a new control strategy for mulberry blight disease.

Dual promoter strategy enhances co-expression of α-L-rhamnosidase and enhanced fluorescent protein for whole-cell catalysis and bioresource valorization.

Developing circular economy is the only way to improve the efficiency of resource utilization. Whole-cell catalysis is an effective method to recycle enzymes, improve catalytic efficiency, and reduce production costs. The enzyme, α-L-rhamnosidase has considerable application prospects in the field of biocatalysis as it can hydrolyze a variety of α-L rhamnoses. In the present study, the genes for α-L-rhamnosidase (rhaB1) and enhanced fluorescent protein (EGFP) were co-expressed using a bi-promoter expression vector pRSFDuet1 and their enzymatic properties were evaluated. To our knowledge, this study has established an effective rhamnosidase-fluorescent indicator and whole-cell catalytic system for the first time. Moreover, we analyzed the change in the activity of the crude rhaB1-EGFP as well as its whole-cell during the biocatalysis process using fluorescence intensity. Recombinant rhaB1-EGFP as a product which contains rhaB1 and EGFP showed higher thermal stability, pH stability, and conversion efficiency than rhaB1, and its optimum temperature for rutin catalysis was ideal for industrial applications. Moreover, under the optimal conditions of a rutin concentration of 0.05 g/L, pH of 6.0, temperature of 40 °C, a yield of 92.5% was obtained. Furthermore, we demonstrated the relationship between the fluorescence intensity and enzyme activity. This study established a highly efficient whole-cell catalytic system whose activity can be evaluated by fluorescence intensity, providing a reference for enzyme recycling.

Microfluidic tools for lipid production and modification: a review.

Microfluidics has great potential as an efficient tool for a large range of applications in industry. The ability of such devices to deal with an extremely small amount of fluid has additional benefits, including superlatively fast and efficient mass and heat transfer. These characteristics of microfluidics have attracted an enormous amount of interest in their use as a novel tool for lipid production and modification. In addition, lipid resources have a close relationship with energy resources, and lipids are an alternative renewable energy source. Here, recent advances in the application of microfluidics for lipid production and modification, especially in the discovery, culturing, harvesting, separating, and monitoring of lipid-producing microorganisms, will be reviewed. Other applications of microfluidics, such as the modification of lipids from microorganisms, will also be discussed. The novel microfluidic tools in this review will be useful in applications to improve lipid production and modification in the future.

ER: the Silk Road of interorganellar communication.

Cellular adaptive responses arise from an array of spatially and temporally distinct biochemical interactions that modulate biological processes and reorganize subcellular structures tailored to the nature of stimulus. As such, cells have evolved elegantly and tightly regulated mechanisms to enable interorganellar communication in part through the dynamic readjustment of physical distance enabling the tethering between two closely apposed membranous organelles and thus formation of Membrane Contact Sites (MCSs). MCSs are dynamic and ubiquitous interorganellar structures that serve as regulatory interfaces to facilitate transmission of signals and to integrate synthesis of metabolic pathways such as lipids required for upholding cellular homeostasis in response to environmental and developmental inputs. Endoplasmic reticulum (ER) is the most copious endomembrane system that extend throughout the cell, and functions in production, processing, and transport of proteins and lipids, as well as in intracellular signaling. Reminiscent of the ancient Silk Road, ER connection to other membranous organelles via MCSs alters cellular landscape and serves as nexus for coordinating exchange of metabolites such as lipids, ions such as Ca2+, and other small molecules involved in maintaining cellular integrity under prevailing conditions. Delineating the molecular organization of the tethering complexes, molecular action of exchanged molecules and hence the nature of information transmitted will afford insight into underlying basis of interorganellar communication and shed light on the evolutionarily conserved function of ER as the ancient trans-kingdom Silk Road trafficking vital metabolites via the non-vesicular pathway.

Interplay of the two ancient metabolites auxin and MEcPP regulates adaptive growth.

The ancient morphoregulatory hormone auxin dynamically realigns dedicated cellular processes that shape plant growth under prevailing environmental conditions. However, the nature of the stress-responsive signal altering auxin homeostasis remains elusive. Here we establish that the evolutionarily conserved plastidial retrograde signaling metabolite methylerythritol cyclodiphosphate (MEcPP) controls adaptive growth by dual transcriptional and post-translational regulatory inputs that modulate auxin levels and distribution patterns in response to stress. We demonstrate that in vivo accumulation or exogenous application of MEcPP alters the expression of two auxin reporters, DR5:GFP and DII-VENUS, and reduces the abundance of the auxin-efflux carrier PIN-FORMED1 (PIN1) at the plasma membrane. However, pharmacological intervention with clathrin-mediated endocytosis blocks the PIN1 reduction. This study provides insight into the interplay between these two indispensable signaling metabolites by establishing the mode of MEcPP action in altering auxin homeostasis, and as such, positioning plastidial function as the primary driver of adaptive growth.