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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.

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.

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.

Plants have evolved tightly regulated signaling networks to respond and adapt to environmental perturbations, but the nature of the signaling hub(s) involved have remained an enigma. We have previously established that methylerythritol cyclodiphosphate (MEcPP), a precursor of plastidial isoprenoids and a stress-specific retrograde signaling metabolite, enables cellular readjustments for high-order adaptive functions. Here, we specifically show that MEcPP promotes two Brassicaceae-specific traits, namely endoplasmic reticulum (ER) body formation and induction of indole glucosinolate (IGs) metabolism selectively, via transcriptional regulation of key regulators NAI1 for ER body formation and MYB51/122 for IGs biosynthesis). The specificity of MEcPP is further confirmed by the lack of induction of wound-inducible ER body genes as well as IGs by other altered methylerythritol phosphate pathway enzymes. Genetic analyses revealed MEcPP-mediated COI1-dependent induction of these traits. Moreover, MEcPP signaling integrates the biosynthesis and hydrolysis of IGs through induction of nitrile-specifier protein1 and reduction of the suppressor, ESM1, and production of simple nitriles as the bioactive end product. The findings position the plastidial metabolite, MEcPP, as the initiation hub, transducing signals to adjust the activity of hard-wired gene circuitry to expand phytochemical diversity and alter the associated subcellular structure required for functionality of the secondary metabolites, thereby tailoring plant stress responses.

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.

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.

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.

Summary: 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. [ABSTRACT FROM AUTHOR]

Two medicinal plants, African Locust Bean (Parkia biglobosa) and Lonicera japonica, traditionally used in West Africa and China were investigated against four spoilage microorganisms: Bacillus subtilis, Cronobacter dublinensis, Pantoea agglomerans, and Bacillus sp., isolated from mulberry fruit. The aqueous extract of the P. biglobosa (ALB) was potent with average inhibitory zones against four spoilage microorganisms recorded as (Means ± SD) 6.5 to 19.5 mm as well as 6.5 to 15.5 mm for L. japonica (HSF) at concentrations of 400 mg/ml. The aqueous extracts of P. biglobosa (54%) inhibited decay better than L. japonica (66%). Decay situations of the medicinal plants with 1‐MCP emerged higher than the control group 78%, 72%, and 68% in ALB + 1‐MCP, HSF + 1‐MCP, and deionized water, respectively. Therefore, two plant extracts against spoilage microorganisms isolated from mulberry fruit had good inhibitory activities, but their mixture with 1‐MCP could not preserve the mulberry fruits in limited storage time. Practical applications: Advance in the bacteria control using a natural control like medicinal plants against plant infections was investigated. The present study demonstrated the potentials of African locust bean and L. japonica as a novel biocontrol agent to the extent of preserving the postharvest decay of mulberry fruit in limited storage time at room temperature. The results and findings obtained in this study support the potential application of these two plants in mulberry fruit preservation as well as in traditional therapeutics. In addition, the combination with 1‐MCP also serves as the potential to extend the shelf life of mulberry fruit in order to limit the use of pesticides in postharvest treatments. This study reports for the first time isolated microorganisms from mulberry fruit and the developments of edible coatings utilizing plant extracts from different continents against their activities. [ABSTRACT FROM AUTHOR]

BACKGROUND The silkworm pupae of Bombyx mori L. are considered to be a valuable unsaturated fatty acids (UFAs) resource. Enzymatic transesterification, which can produce structured lipids (SLs), is regarded as a desirable approach to utilize this resource, but requires long reaction time. A method to reduce the reaction time and obtain novel SLs is thus desirable. Hence, microwave irradiation was employed to reduce the reaction time and obtain novel SLs. In addition, high‐performance liquid chromatography with an evaporative light scattering detector was employed to investigate the contribution of the fatty acids (FAs) after microwave irradiation. RESULTS: One hour was required for a 1:5 substrate ratio of silkworm pupae oil (SPO):oleic acid (OA) using a solvent‐free system catalysed by Lipozyme RM IM (7%) at 60 °C with microwave irradiation at 50 W to obtain the novel SLs. The relative content of polyunsaturated FAs glyceride increased from 51.1% to 81.0%, and the relative content of glycerol trioleate (OOO) in SPO obtained in the microwave reactor was twice as high as that in the shaker reactor. In addition, the relative content of OA was increased from 3.9% to 35.8% in total compared with the products from the shaker reactor. Furthermore, the changes of α‐helix relative content of Lipozyme RM IM were observed after the reaction under the microwave irradiation. CONCLUSION: The results indicated that variations in the FA distribution of enzymatic transesterification reactions caused by microwave irradiation could aid the addition of OA and enrichment of UFAs. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: 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 [ABSTRACT FROM AUTHOR]

Interorganellar cooperation maintained via exquisitely controlled retrograde-signaling pathways is an evolutionary necessity for maintenance of cellular homeostasis. This signaling feature has therefore attracted much research attention aimed at improving understanding of the nature of these communication signals, how the signals are sensed, and ultimately the mechanism by which they integrate targeted processes that collectively culminate in organellar cooperativity. The answers to these questions will provide insight into how retrograde-signal-mediated regulatory mechanisms are recruited and which biological processes are targeted, and will advance our understanding of how organisms balance metabolic investments in growth against adaptation to environmental stress. This review summarizes the present understanding of the nature and the functional complexity of retrograde signals as integrators of interorganellar communication and orchestrators of plant development, and offers a perspective on the future of this critical and dynamic area of research. [ABSTRACT FROM AUTHOR]

The pupae is one of the most valuable components of the silkworm, a by-product of silk reeling. Employing them as a raw material to create valued products will help to recycle and reuse silkworm pupae, as well as contribute to the food industry's long-term development. Silkworm pupae protein (SPP) and pupae oil (SPO) are high in nutritional and bioactive components that have a variety of applications in the food industry as well as health-promoting characteristics. In this review, we first elaborate on the modification, nutritional, and functional properties of silkworm pupae in terms of requirements for protein and lipids processing as well as current extraction technology. In the subsequent sections, we summarize the modification techniques including the science and technology involved in attaining fatty acid components of SPO. Finally, we focus on recent studies related to the utilization of SPP and SPO subject to their inclusion in foods for their health benefits. The key to silkworm pupae product design may be to fully exploit and utilize the inherent characteristics of the pupae to produce functional peptides and structural lipids with high nutritional content, which may necessitate further improvements in the bioactive and functional properties. Designing more ecologically friendly, low-cost, and efficient processes for producing protein and oil from silkworm pupae is also a trend in technology. This may further provide new insights into the mechanisms behind modifying silkworm pupae as food and its health advantages. [Display omitted] • Modifications and functions of silkworm pupae protein (SPP) and oil (SPO) were reviewed. • Modification of SPP and SPO reveal valuable nutritional properties. • Inclusion of SPP and SPO in food and feed applications were summarized. • Peptide composition in silkworm pupae aid in valuable biological activities. • Further research to compare SPP and SPO with standard proteins and oils is recommended. [ABSTRACT FROM AUTHOR]

The reaction can be scaled up. [ABSTRACT FROM AUTHOR]

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. Unlabelled Image • Two genes of rhaB1 and EGFP were firstly co-expressed by a bi-promoter expression vector. • Crude rhaB1-EGFP possessed higher thermal and pH stabilities than those of crude rhaB1. • Expression vector activity could be quantitatively detected by measuring fluorescence intensity. • Affinity of rhaB1-EGFP to substrate was not significantly reduced confirmed by SPR assay. • A labeled and recycling whole-cell catalyst was developed for flavonoids transformation. [ABSTRACT FROM AUTHOR]

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 H 2 O 2 , 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.

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 ( F v / F m ), actual photochemical efficiency ( Φ PSⅡ ), photochemical quenching coefficient ( q P ) 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 Q A to Q B . Metamitron treatment increased W k , and significantly decreased ψ o , RC / CS m , and PI abs . 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.

Boron is an essential mineral nutrient for higher plant growth and development. However, excessive amounts of boron can be toxic. Here, we report on the characterization of an Arabidopsis mutant, shb1 ( sensitive to high-level of boron 1 ), which exhibits hypersensitivity to excessive boron in roots. Positional cloning demonstrated that the shb1 mutant bears a point mutation in a gene encoding a heme oxygenase 1 (HO1) corresponding to the HY1 gene involved in photomorphogenesis. The transcription level of the SHB1/HY1 gene in roots is up-regulated under excessive boron stimulation. Either overexpressing SHB1/HY1 or applying the HO1 inducer hematin reduces boron accumulation in roots and confers high boron tolerance. Furthermore, carbon monoxide and bilirubin, catalytic products of HO1, partially rescue the boron toxicity-induced inhibition of primary root growth in shb1 . Additionally, the mRNA level of BOR4 , a boron efflux transporter, is reduced in shb1 roots with high levels of boron supplementation, and hematin cannot relieve the boron toxicity-induced root inhibition in bor4 mutants. Taken together, our study reveals that HO1 acts via its catalytic by-products to promote tolerance of excessive boron by up-regulating the transcription of the BOR4 gene and therefore promoting the exclusion of excessive boron in root cells.

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.