Phosphoenolpyruvate Carboxylase Research Articles

Exogenous trehalose differently improves photosynthetic carbon assimilation capacities in maize and wheat under heat stress

ABSTRACT High temperature inhibits the growth and development of maize (Zea mays L.) and wheat (Triticum aestivum L.) seedlings. The current study attempts to elucidate the underlying mechanisms of exogenous trehalose for the improvement of maize and wheat responses to heat stress. Results show that, in maize seedlings, exogenously-supplied trehalose was able to effectively elevate the capacity of photosynthetic carbon assimilation and improve the carbohydrate content via up-regulating the transcript levels of key enzymes including phosphoenol pyruvate carboxylase (PEPC), NADP-malate dehydrogenase (NADP-MDH), NADP-malic enzyme (NADP-ME), pyruvate phosphate dikinase (PPDK) and Ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit. Besides, it was also able to improve C4 pathway enzyme activities, including PEPC, NADP-MDH, NADP-ME and Ribulose-1,5-bisphosphate carboxylase (RUBPCase). In wheat seedlings, exogenously-supplied trehalose promoted photosynthetic carbon assimilation by elevating the activity of RUBPCase. These results show that exogenous trehalose improved photosynthetic carbon assimilation capacities in maize and wheat seedlings under heat stress through different mechanisms.

Polyphenol from Rosaroxburghii Tratt Fruit Ameliorates the Symptoms of Diabetes by Activating the P13K/AKT Insulin Pathway in db/db Mice.

About 4% of the world’s population has type 2 diabetes mellitus (T2DM), and the available hypoglycemic drugs for treating diabetes have some side effects. Therefore, research on the extraction of hypoglycemic components from plants has gradually become popular. This study aimed to investigate the hypoglycemic effects of polyphenol-rich Rosa roxburghii Tratt extract (RP) isolated from Rosa roxburghii Tratt fruit and of four constituents (IRP 1–4 ) isolated from RP on db/db mice. The results indicated that the oral administration of RP and IRP 1–4 could markedly decrease the food intake, water intake, fasting blood glucose (FBG), and serum insulin levels in the db/db mice. Glucose intolerance, insulin resistance, and oxidative stress were ameliorated in the RP and IRP 1–4 groups. Histopathological observation revealed that RP and IRP 1–4 could effectively protect the liver fat against damage and dysfunction. RP and IRP 1–4 also increased the hepatic and muscle glycogen contents by increasing the phosphorylation and reducing the expression of glycogen synthase kinase 3β (GSK3β). The activities of glucokinase (GCK), phosphoenolpyruvate carboxylase (PEPCK), and glucose-6-phosphatase (G6PC) and their respective mRNA expression levels in the liver of db/db mice were simultaneously increased and decreased in the intervention groups. RP and IRP 1–4 significantly increased the expression of phosphatidylinositol 3-kinase (P13K) and the phosphorylation of protein kinase B (AKT). These results indicate that RP and IRP 1–4 exhibit good hypoglycemic effects by activating the P13K/AKT signaling pathway and regulating the expression of FOXO1 and p-GSK3β proteins, controlling hepatic gluconeogenesis and improving hepatic glycogen storage insulin resistance. Therefore, RP and IRP 1–4 could be utilized as the hypoglycemic functional component to alleviate the symptoms of T2DM.

Genome-wide investigation of the ZF-HD gene family in two varieties of alfalfa (Medicago sativa L.) and its expression pattern under alkaline stress

BackgroundZinc finger homeodomain (ZHD) protein is a plant-specific transcription factor and a potential regulator of phosphoenolpyruvate carboxylase (PEPCase)-coding genes, and it also participates in plant growth regulation and abiotic stress responses. To study the function of MsZF-HD genes in the alkaline stress response, this paper assessed biological information and performed transcriptome analysis of the MsZF-HD gene family by using the genomes of two different varieties of alfalfa (XinJiangDa Ye and Zhongmu No. 1).ResultsIn total, 49 and 11 MsZF-HD genes were identified in the two different varieties respectively, including the alleles of XinJiangDa Ye. According to their phylogenetic relationships, the 60 MsZF-HD genes were divided into 5 ZHD subfamilies and 1 MIF subfamily. A total of 88.3% of MsZF-HD genes do not contain introns and are unevenly distributed among the 6 chromosomes of alfalfa. A collinearity analysis indicated that 26 genes of XinJiangDa Ye have no orthologous genes in Zhongmu No. 1, although these genes (such as ZHD-X1–2, ZHD-X3–2 and ZHD-X4–2) have homologous genes in Arabidopsis thaliana, Medicago truncatula and Glycine max. Through RNA-seq and qRT–PCR verification, it was found that MsZF-HD genes are downregulated to participate in the alkaline stress response.ConclusionThe results of this study may lay the foundation for the cloning and functional study of MsZF-HD genes and provide a theoretical basis for revealing the difference between XinJiangDa Ye and Zhongmu No. 1 at the genome level.

Engineering precursor and co-factor supply to enhance D-pantothenic acid production in Bacillus megaterium.

High-yielding chemical and chemo-enzymatic methods of D-pantothenic acid (DPA) synthesis are limited by using poisonous chemicals and DL-pantolactone racemic mixture formation. Alternatively, the safe microbial fermentative route of DPA production was found promising but suffered from low productivity and precursor supplementation. In this study,Bacillus megateriumwas metabolically engineered to produce DPA without precursor supplementation. In order to provide a higher supply of precursor D-pantoic acid, key genes involved in its synthesis are overexpressed, resulting strain was produced 0.53 ± 0.08g/L DPA was attained in shake flasks. Cofactor CH2-THF was found to be vital for DPA biosynthesis and was regenerated through the serine-glycine degradation pathway. Enhanced supply of another precursor, β-alanine was achieved by codon optimization and dosing of the limiting L-asparate-1-decarboxylase (ADC). Co-expression of Pantoate-β-alanine ligase, ADC, phosphoenolpyruvate carboxylase, aspartate aminotransferase and aspartate ammonia-lyase enhanced DPA concentration to 2.56 ± 0.05g/L at shake flasks level. Fed-batch fermentation in a bioreactor with and without the supplementation of β-alanine increased DPA concentration to 19.52 ± 0.26 and 4.78 ± 0.53g/L, respectively. This present study successfully demonstrated a rational approach combining precursor supply engineering with cofactor regeneration for the enhancement of DPA titer in recombinantB. megaterium.

Metabolic engineering of threonine catabolism enables Saccharomyces cerevisiae to produce propionate under aerobic conditions.

Propionate is widely used as a preservative in the food and animal feed industries. Propionate is currently produced by petrochemical processes, and fermentative production of propionate remains challenging. In this study, a synthetic propionate pathway was constructed in the budding yeast Saccharomyces cerevisiae, for propionate production under aerobic conditions. Through expression of tdcB and aldH from Escherichia coli and kivD from Lactococcus lactis, L-threonine was converted to propionate via 2-ketobutyrate and propionaldehyde. The resulting yeast aerobically produced 0.21 g L-1 propionate from glucose in a shake flask. Subsequent overexpression of pathway genes and elimination of competing pathways increased propionate production to 0.37 g L-1 . To further increase propionate production, carbon flux was pulled into the propionate pathway by weakened expression of pyruvate kinase (PYK1), together with overexpression of phosphoenolpyruvate carboxylase (ppc). The final propionate production reached 1.05 g L-1 during fed-batch fermentation in a fermenter. In this work, a yeast cell factory was constructed using synthetic biology and metabolic engineering strategies to enable propionate production under aerobic conditions. Our study demonstrates engineered S. cerevisiae as a promising alternative for the production of propionate and its derivatives.

Novel Galactosyl Moiety-Conjugated Furylchalcones Synthesized Facilely Display Significant Regulatory Effect on Plant Growth.

The expansion of weed infestation has increased the demand on new herbicides. A series of novel galactosyl moiety-conjugated furylchalcones was facilely synthesized in which the furyl group (A ring) was combined with the substituted benzene group (B ring), and a galactosyl moiety was introduced. All these galactosyl furylchalcones were predicted to be phloem-mobile. Most of the galactosyl furylchalcones significantly promoted early seedling growth of sorghum and barnyardgrass under dark conditions, but all of them revealed considerable anti-growth ability on illuminated pot plants; especially, 1-(3'-(4″-O-β-d-galactopyranosyl)furyl)-3-(4″-nitrophenyl)-2-en-1-one (B11) had a better herbicidal activity against rapeseed and Chinese amaranth than haloxyfop-R-methyl. The median efficient concentrations (EC50) of compound B11 against cucumber and wheat were 9.55 and 26.97 mg/L, respectively, also showing a stronger suppressing capacity than 2,4-D. Molecular docking with phosphoenolpyruvate carboxylase protein showed a stable binding conformation in which the galactosyl group interacted with LYS363 and GLU369, the furan ring and carbonyl bound with ARG184, and the crosslink of the nitro group with GLU240 formed a salt bridge. The results demonstrate that galactosyl furylchalcones possess the great potential as new herbicides for weed management, and further evaluations on more weeds are required for practical application.

Dynamic changes and mechanisms of organic acids during black tea manufacturing process

Sourness usually reduces the sensory quality of black tea, and organic acids are considered as one of major contributors. A UPLC-MRM-MS method was successfully developed to analyze organic acids during the processing of black tea made of “Yingshuang” tea variety. The contents of quinic acid and citric acid increased first and then decreased during withering, but didn't significantly changed after the whole processing. Malic acid content obviously dropped in early withering, and then stayed at low level. These three acids at withering stage were regulated by quinate dehydrogenase (QDH), citrate synthase (CS), phosphoenolpyruvate carboxylase (PEPC) and malic enzyme (ME). Gallic acid sharply increased after rolling, resulted from the hydrolysis of epigallocatechin gallate (EGCG) by endogenous tannase. This work revealed the dynamic changes and regulatory mechanisms of organic acids during the black tea manufacturing process, which is helpful to improve the processing technology and quality of black tea. • A UPLC-MRM-MS method was developed to analyze organic acids in black tea. • The contents of quinic acid, citric acid and malic acid changed during withering. • The three acids were regulated by four enzymes including QDH, CS, PEPC and ME. • Gallic acid increased after rolling, due to the hydrolysis of EGCG by tannase. • Fermentation is not the major processing step for organic acid accumulation.

A well-supported nuclear phylogeny of Poaceae and implications for the evolution of C4 photosynthesis

Poaceae (the grasses) includes rice, maize, wheat, and other crops, and is the most economically important angiosperm family. Poaceae is also one of the largest plant families, consisting of over 11 000 species with a global distribution that contributes to diverse ecosystems. Poaceae species are classified into 12 subfamilies, with generally strong phylogenetic support for their monophyly. However, many relationships within subfamilies, among tribes and/or subtribes, remain uncertain. To better resolve the Poaceae phylogeny, we generated 342 transcriptomic and seven genomic datasets; these were combined with other genomic and transcriptomic datasets to provide sequences for 357 Poaceae species in 231 genera, representing 45 tribes and all 12 subfamilies. Over 1200 low-copy nuclear genes were retrieved from these datasets, with several subsets obtained using additional criteria, and used for coalescent analyses to reconstruct a Poaceae phylogeny. Our results strongly support the monophyly of 11 subfamilies; however, the subfamily Puelioideae was separated into two non-sister clades, one for each of the two previously defined tribes, supporting a hypothesis that places each tribe in a separate subfamily. Molecular clock analyses estimated the crown age of Poaceae to be ∼101 million years old. Ancestral character reconstruction of C3/C4 photosynthesis supports the hypothesis of multiple independent origins of C4 photosynthesis. These origins are further supported by phylogenetic analysis of the ppc gene family that encodes the phosphoenolpyruvate carboxylase, which suggests that members of three paralogous subclades (ppc-aL1a, ppc-aL1b, and ppc-B2) were recruited as functional C4ppc genes. This study provides valuable resources and a robust phylogenetic framework for evolutionary analyses of the grass family.

Convergent molecular evolution of phosphoenolpyruvate carboxylase gene family in C4 and crassulacean acid metabolism plants.

Phosphoenolpyruvate carboxylase (PEPC), as the key enzyme in initial carbon fixation of C4and crassulacean acid mechanism (CAM) pathways, was thought to undergo convergent adaptive changes resulting in the convergent evolution of C4 and CAM photosynthesis in vascular plants. However, the integral evolutionary history and convergence of PEPC in plants remain poorly understood. In the present study, we identified the members of PEPC gene family across green plants with seventeen genomic datasets, found ten conserved motifs and modeled three-dimensional protein structures of 90 plant-type PEPC genes. After reconstructing PEPC gene family tree and reconciled with species tree, we found PEPC genes underwent 71 gene duplication events and 16 gene loss events, which might result from whole-genome duplication events in plants. Based on the phylogenetic tree of the PEPC gene family, we detected four convergent evolution sites of PEPC in C4 species but none in CAM species. The PEPC gene family was ubiquitous and highly conservative in green plants. After originating from gene duplication of ancestral C3-PEPC, C4-PEPC isoforms underwent convergent molecular substitution that might facilitate the convergent evolution of C4 photosynthesis in Angiosperms. However, there was no evidence for convergent molecular evolution of PEPC genes between CAM plants. Our findings help to understand the origin and convergent evolution of C4 and CAM plants and shed light on the adaptation of plants in dry, hot environments.

γ-Aminobutyric acid regulates mitochondrial energy metabolism and organic acids metabolism in apples during postharvest ripening

“Golden Delicious” apples were treated with 1.0 mmol L−1 γ-aminobutyric acid (GABA) to evaluate the changes of mitochondrial energy metabolism and organic acids metabolism during ripening. Results showed that GABA dipping enhanced malic, citric, succinic, and tartaric acids contents in apple fruit. GABA treatment enhanced phosphoenolpyruvate carboxylase (PEPC), NADP-dependent isocitrate dehydrogenase (NADP-IDH), cytoplasm aconitase (Cyt-ACO), citrate synthase (CS), and cytosolic NAD-dependent malate dehydrogenase (NAD-MDH) activities, but decreased cytosolic NAD phosphate-dependent malic enzyme (NADP-ME) activity. GABA also enhanced the expressions of MdPEPC, MdNADP-IDH, MdCyt-ACO, MdNAD-MDH and MdCS, and decreased MdNADP-ME expression in apples. Moreover, higher level of energy charge, adenosine triphosphate (ATP) content, H+-ATPase, cytochrome C oxidase (CCO), succinic dehydrogenase (SDH), and Ca2+-ATPase activities were recorded in GABA-treated fruit. Additionally, GABA up-regulated MdSDH, MdCCO, MdH+-ATPase, and MdCa2+-ATPase expressions of apples. In conclusion, GABA could regulate the key gene expression and enzyme activity in mitochondrial energy metabolism and organic acids metabolism during ripening to maintain freshness of apple fruit.

Partial gene sequencing of phosphoenol pyruvate carboxylase (PEPc) gene from Saccharum spp. hybrid CoLk 94184

Phosphoenol pyruvate carboxylase (PEPc), a key enzyme in photosynthesis, an important carboxylase for plant cell metabolism, fixes CO2 in mesophyll cells to form dicarboxylic acids, and being more specific for inorganic carbon, improves the photosynthetic rate of C4 plants. In the present study, partial gene sequencing of PEPc was done using Saccharum spp. hybrid, CoLk 94184 of sugarcane, one of the most popular variety released by ICAR-IISR, Lucknow. Utilizing PEPc of different cultivars of sugarcane species, the primers were designed for PEPc gene and the first ever nucleotide sequence was determined, especially for Saccharum spp. hybrid CoLk 94184 (accession no: MW766370.1). Sequence based phylogenetic tree showed close relatedness between the PEPc sequences for various Saccharum cultivars, thus pointing to little genetic change, while that for sorghum and maize were relatively distantly related. Results may help in understanding the photosynthetic process using sugarcane hybrid specific genes

Effects of elevated air temperature coupling with soil drought on carbohydrate metabolism and oil synthesis during cottonseed development.

Cotton, as the fifth-largest oilseed crop, often faces the coupling stress of heat and drought. Still, the effects of combined stress on cottonseed oil synthesis and its closely related carbon metabolism are poorly investigated. To this end, experiments were conducted with two cultivars (Sumian 15 and PHY370WR) under two temperature regimes: ambient temperature (AT) and elevated temperature (ET, which was 2.5°C-2.7°C higher than AT) and three water regimes: optimum soil moisture (soil relative water content [SRWC] at 75% ± 5%), and drought (SD) including SRWC 60% ± 5% and SRWC 45% ± 5%, during 2016-2018. Results showed that ET plus SD decreased cottonseed kernel yield, seed index, kernel weight, and kernel percentage more than either single stress. The content of hexoses, the carbon skeleton source for oil synthesis, was decreased by ET while increased by SD. The combined stress increased the hexose content by increasing the activities of sucrose synthase (SuSy, EC 2.4.1.13) and invertase (Inv, EC 3.2.1.26) and upregulating GhSuSy expression; however, hexose content under combined stress was lower than that under SD alone. Increased oil content under SD was attributed to the high phosphoenolpyruvate carboxylase (PEPCase, EC 4.1.1.31), acetyl-CoA carboxylase (ACCase, EC 6.4.1.2), and diacylglycerol acyltransferase (DGAT, EC 2.3.1.20) activities, whereas the opposite effects were seen under ET. Under combined stress, although ACCase activity decreased, PEPCase and DGAT activities, and GhPEPC-1 and GhDGAT-1 expression upregulated, enhancing carbon flow into oil metabolism and triacylglycerol synthesis, ultimately generating higher oil content.

Responses to Aluminum and Cadmium of a Rnai Sorghum Line with Decreased Levels of Phosphoenolpyruvate Carboxylase 3 (Ppc3)

Responses to Aluminum and Cadmium of a Rnai Sorghum Line with Decreased Levels of Phosphoenolpyruvate Carboxylase 3 (Ppc3)

Genome-Wide In Silico Analysis and Expression Profiling of Phosphoenolpyruvate Carboxylase Genes in Loquat, Apple, Peach, Strawberry and Pear

Phosphoenolpyruvate carboxylase (PEPC) genes have multiple potential roles in plant metabolism such as regulation and accumulation of organic acids in fruits, movement of guard cells and stress tolerance, etc. However, the systematic identification and characterization of PEPC genes in Rosaceae species i.e., loquat, apple, peach, strawberry, and pear are yet to be performed. In present study, 27 putative PEPC genes (loquat 4, apple 6, peach 3, strawberry 9, and pear 5) were identified. To further investigate the role of those PEPC genes, comprehensive bioinformatics and expression analysis were performed. In bioinformatic analysis, the physiochemical properties, conserved domains, gene structure, conserved motif, phylogenetic and syntenic analysis of PEPC genes were performed. The result revealed that the PEPcase superfamily domain was conserved in all examined PEPC proteins. Most of the PEPC proteins were predicted to be localized in cytonuclear. Genomic structural and motif analysis showed that the exon and motif number of each PEPC gene ranged dramatically, from 8 to 20, and 7 to 10, respectively. Syntenic analysis indicated that the segmental or whole-genome duplication played a vital role in extension of PEPC gene family in Rosacea species. The Ka and Ks values of duplicated genes depicted that PEPC genes have undergone a strong purifying selection. Furthermore, the expression analysis of PEPC genes in root, mature leaf, stem, full-bloom flower, and ripened fruit of loquat, apple, peach, strawberry, and pear was performed. Some genes were differentially expressed in aforementioned plant tissues, signifying their role in plant metabolism. This study provides the first genome-wide identification, characterization, and expression profiling of PEPC gene family in Rosaceae species, and provides the foundation for further functional analysis.

Metabolic profiles in C3, C3-C4 intermediate, C4-like, and C4 species in the genus Flaveria.

C4 photosynthesis concentrates CO2 around Rubisco in the bundle sheath, favouring carboxylation over oxygenation and decreasing photorespiration. This complex trait evolved independently in >60 angiosperm lineages. Its evolution can be investigated in genera such as Flaveria (Asteraceae) that contain species representing intermediate stages between C3 and C4 photosynthesis. Previous studies have indicated that the first major change in metabolism probably involved relocation of glycine decarboxylase and photorespiratory CO2 release to the bundle sheath and establishment of intercellular shuttles to maintain nitrogen stoichiometry. This was followed by selection for a CO2-concentrating cycle between phosphoenolpyruvate carboxylase in the mesophyll and decarboxylases in the bundle sheath, and relocation of Rubisco to the latter. We have profiled 52 metabolites in nine Flaveria species and analysed 13CO2 labelling patterns for four species. Our results point to operation of multiple shuttles, including movement of aspartate in C3–C4 intermediates and a switch towards a malate/pyruvate shuttle in C4-like species. The malate/pyruvate shuttle increases from C4-like to complete C4 species, accompanied by a rise in ancillary organic acid pools. Our findings support current models and uncover further modifications of metabolism along the evolutionary path to C4 photosynthesis in the genus Flaveria.

Melatonin Enhances Drought Tolerance by Regulating Leaf Stomatal Behavior, Carbon and Nitrogen Metabolism, and Related Gene Expression in Maize Plants

It is commonly known that exogenously applied melatonin can alleviate the impact of drought stress, but the mechanism used by melatonin to regulate stomatal behavior and carbon (C) and nitrogen (N) metabolism to increase drought resistance remains elusive. Herein, our aim was to investigate the influence of exogenous melatonin on the regulation of C and N metabolism in maize plants under water deficit. In this study, we analyzed stomatal behavior, the key components of C and N metabolism, and the gene expression and activity of enzymes involved in the C and N metabolism in maize plants. The results showed that the application of melatonin (100 μM) significantly increased maize growth and sustained the opening of stomata, and secondarily increased the photosynthetic capacity in maize. Under drought stress, foliar application of melatonin induced the gene transcription and activities of sucrose phosphate synthetase, ADP-glucose pyrophosphorylase, phosphoenolpyruvate carboxylase, and citrate synthase, resulting in the enhancement of sucrose and starch synthesis and the tricarboxylic acid (TCA) cycle. This enhancement in sugar biosynthesis and the TCA cycle might lead to stronger N assimilation. As anticipated, NO3– reduction and NH4+ assimilation were also strengthened after melatonin treatment under drought stress. An increase was observed in some key enzymatic activities and transcription involved in nitrogen metabolism, such as that of nitrate reductase, nitrite reductase, glutamate synthase, and glutamine synthetase, in melatonin-treated, drought-stressed maize. Moreover, melatonin attenuated the drought-induced damage by reducing protein degradation and increasing the level of proline. Conclusively, our results indicate that exogenous melatonin enhances drought tolerance in maize via promoting stomatal opening and regulating C and N metabolism and related gene expression.

Comparing the Effects of N and P Deficiency on Physiology and Growth for Fast- and Slow-Growing Provenances of Fraxinus mandshurica

With the continuous increase in atmospheric carbon dioxide emissions, nitrogen (N) and phosphorus (P) as mineral elements increasingly restrict plant growth. To explore the effect of deficiency of P and N on growth and physiology, Fraxinus mandshurica (hereafter “F. mandshurica”) Rupr. annual seedlings of Wuchang (WC) provenance with fast growth and Dailing (DL) provenance with slow growth were treated with complete nutrition or starvation of N (N-), P (P-) or both elements (NP-). Although P- and N- increased the use efficiency of P (PUE) and N (NUE), respectively, they reduced the leaf area, chlorophyll content and activities of N assimilation enzymes (NR, GS, GOGAT), which decreased the dry weight and P or N amount. The free amino acid content and activities of Phosphoenolpyruvate carboxylase (PEPC) and acid phosphatase enzymes were reduced by N-. The transcript levels of NRT2.1, NRT2.4, NRT2.5, NRT2.7, AVT1, AAP3, NIA2, PHT1-3, PHT1-4 and PHT2-1 in roots were increased, but those of NRT2.1, NRT2.4, NRT2.5, PHT1-3, PHT1-4, PHT2-1 and AAP3 in leaves were reduced by P-. WC was significantly greater than DL under P- in dry weight, C amount, N amount, leaf area, PUE, NUE, which related to greater chlorophyll content, PEPC enzyme activity, N assimilation enzyme activities, and transcript levels of N and P transporter genes in roots and foliage, indicating a greater ability of WC to absorb, transport and utilize N and P under P-. WC was also greater than DL under N- in terms of the above indicators except the transcript levels of N and P assimilation genes, but most of the indicators did not reach a significant level, indicating that WC might be more tolerant to N- than DL, which requires further verification. In summary, WC was identified as a P-efficient provenance, as the growth rate was greater for the genetic type with high than low tolerance to P-.

Fortifying nursery soil-less media with cyanobacteria for enhancing the growth of tomato

Priming nursery media to develop robust nurseries is a major objective of horticulturists for improving yields and quality of produce, and use of cyanobacteria, with both plant growth promoting and biofertilizing properties, can be valuable for generating robust nurseries of tomato. The potential of cyanobacteria as supplements to different soilless media combinations involving perlite, cocopeat and vermiculite (P,C,V) in different combinations was evaluated in terms of their ability to enhance the growth of tomato, from seed to fruiting stage. Agriculturally beneficial cyanobacterial cultures - Anabaena laxa, Calothrix elenkinii, and biofilm- Anabaena torulosa-Trichoderma viride (An-Tr) were used to augment these media and evaluated in Pro-Trays, pot and polyhouse beds respectively. Significant differences were recorded in terms of plant and media parameters. A positive correlation (r = 0.55, 0.78) between PEP carboxylase and Carbonic anhydrase enzyme activities was observed in Pro-Tray and polyhouse experiments. The performance of C:V + C. elenkinii, P:C:V + An-Tr, PM (Potting mix; C:V:P, 3:1:1) + A. laxa in terms of leaf, soil and nutrient data suggested their superiority among all other treatments. An enhancement of 14.70% and 36.55% in fruit and flower count was observed in PM + A. laxa over control. Among all the media evaluated, PM was most responsive to augmentation using any of the cultures, highlighting it as an effective nursery medium. A. laxa proved most superior among the cultures evaluated. Future research is focused on the upscaling the use of A. laxa as a nursery medium supplement for enhancing growth and productivity of tomato.

Flux balance analysis for overproduction of organic acids by Synechocystis sp. PCC 6803 under dark anoxic condition

Cyanobacteria have recently been considered as potential organisms for metabolic engineering with an objective to improve its ability to synthesize biofuel precursors and other value-added products. A modified version of the genome-scale metabolic model of Synechocystis sp. PCC 6803 model organism was used to theoretically demonstrate the flux distribution in the central carbon metabolism towards optimal accumulation of organic acids, namely succinate, acetate, lactate and malate. We further validate the model using experimental data reported by Hasunuma et al. Met. Engg. Comm., 2016, 3, 130–141 for organic acid synthesis under dark anoxic conditions from glycogen accumulated during photoautotrophic growth conditions by sequestering atmospheric CO2. We show that key limiting factors are redox balance causing gluconeogenesis, availability of CO2 and channeling of carbon flux to TCA cycle, in addition to the rate of breakdown of glycogen. Sodium bicarbonate supplementation enhances succinate production flux by eliminating the CO2 limitations for the phosphoenolpyruvate (PEP) to oxaloacetate biochemical reaction catalyzed by PEP carboxylase. A combined strategy of supplementation with NaHCO3 and increased temperature can only offer ~25% of the theoretical maximum succinate production. We suggest possible metabolic pathway interventional approaches to improve the succinate productivity.

Down-Regulation of Phosphoenolpyruvate Carboxylase Kinase in Grapevine Cell Cultures and Leaves Is Linked to Enhanced Resveratrol Biosynthesis.

In grapevine, trans-Resveratrol (tR) is produced as a defence mechanism against stress or infection. tR is also considered to be important for human health, which increases its interest to the scientific community. Transcriptomic analysis in grapevine cell cultures treated with the defence response elicitor methyl-β-cyclodextrin (CD) revealed that both copies of PHOSPHOENOLPYRUVATE CARBOXYLASE KINASE (PPCK) were down-regulated significantly. A role for PPCK in the defence response pathway has not been proposed previously. We therefore analysed the control of PPCK transcript levels in grapevine cell cultures and leaves elicited with CD. Moreover, phosphoenolpyruvate carboxylase (PPC), stilbene synthase (STS), and the transcription factors MYB14 and WRKY24, which are involved in the activation of STS transcription, were also analysed by RT-qPCR. The results revealed that under CD elicitation conditions PPCK down-regulation, increased stilbene production and loss of PPC activity occurs in both tissues. Moreover, STS transcripts were co-induced with MYB14 and WRKY24 in cell cultures and leaves. These genes have not previously been reported to respond to CD in grape leaves. Our findings thus support the hypothesis that PPCK is involved in diverting metabolism towards stilbene biosynthesis, both for in vitro cell culture and whole leaves. We thus provide new evidence for PEP being redirected between primary and secondary metabolism to support tR production and the stress response.