The Glycine soja cytochrome P450 gene GsCYP82C4 confers alkaline tolerance by promoting reactive oxygen species scavenging.

Recent studies have demonstrated the crucial role of Cytochrome P450 enzymes (CYPs) in the production of secondary metabolites, phytohormones and antioxidants in plants. However, their functional characterization specifically under alkaline stress remains elusive. CYP82C4 was the key gene screened from a family of wild soybean CYPs in our previous studies. The aim of this present study was to clone the Glycine soja GsCYP82C4 gene and characterize its functions in Arabidopsis and Glycine max. The results showed that the GsCYP82C4 gene displayed a high expression in different plant tissues at mature stages compared to young stages. Further, higher temporal expression of the GsCYP82C4 gene was noted at 6, 12 and 24 h time points after alkali treatment in leaves compared to roots. In addition, overexpression of GsCYP82C4 improved alkaline stress tolerance in Arabidopsis via increased root lengths and fresh biomass and strengthened the antioxidant defense system via a reduction in superoxide radicals in transgenic lines compared to wild type (WT) and atcyp82c4 mutants. Further, the expression levels of stress-related marker genes were up-regulated in GsCYP82C4 OX lines under alkali stress. The functional analysis of GsCYP82C4 overexpression in soybean displayed better hairy root growth, increased fresh weight, higher antioxidant enzyme activities and reduced lipid peroxidation rates in OX lines compared to the soybean WT (K599) line. In total, our study displayed positive roles of GsCYP82C4 overexpression in both Arabidopsis and Glycine max to alleviate alkaline stress via altering expression abundance of stress responsive genes, stronger roots, higher antioxidant enzyme activities as well as reduced rates of lipid peroxidation and superoxide radicals.

Health risks assessment and source admeasurement of potentially dangerous heavy metals (Cu, Fe, and Ni) in rapidly growing urban settlement.

Environmental contamination is a global challenge that impacts every aspect of ecosystem. The contaminants from anthropogenic or industrial trash continually recirculate into the environment, agricultural land, plants, livestock, and ultimately into humans by way of the food chain. After an increase in human and farmland animal deaths from illnesses due to contaminated drinking water, toxic metal water poisoning has remained a global concern. Diverse environmental and enforcement organisations have attempted to regulate the activities that serve as precursors to these heavy metals which have been proven ineffective. These unnecessary metals have severely hampered most biological processes. The presence of hazardous metals, which are harmful at extremely high levels and have a negative effect on the health of living bodies generally degrades the nutritional value of water. In order to evaluate the heavy metals (Cu, Ni, and Fe) toxicity of groundwater in pri-urban areas, the current study was conducted that have been considered as advance solution to tackle climate change which influence coastal ecosystem. Additionally, the impacts of soil and plant (spinach and brassica) contamination from groundwater were evaluated. The heavy metals were examined in the soil and groundwater samples (Pb, Fe and Ni). While Fe concentrations in water samples were found to be high as 1.978 mg/L as compared to Ni and Cu values low. According to WHO guidelines, the mean value of Fe exceeds the limit value. Similarly, Cu had a higher mean value (0.7 mg/L) in soil samples than other metals (Ni and Fe). In comparison to Ni and Cu, the Fe concentrations in spinach and brassica plants samples are greater, at 17.2 mg/L and 3.22 mg/L, respectively. The possible effects of metal poisoning of groundwater and plants on human health have been assessed using the Hazard Quotient (HQ), Evaluated Daily Intake (EDI), and Incremental Life Time Cancer Risk formulas (ILTCR). When drinking Ni-contaminated water, humans are more at risk of developing cancer (0.0031) than Fe and Cu. Metal concentrations in water and brassica showed substantially more scattered behaviour on the plot and no meaningful relationship, although PCA and masked matrix correlation showed a fair association between Ni and Cu in brassica (r2: 0.46) and Fe and Ni in spinach (r2: 0.31). According to the study's findings, it is anticipated that special management and groundwater monitoring will be needed in the examined area to reduce the health risks related to drinking water that has been contaminated with metals.

GsPKS24, a calcineurin B-like protein-interacting protein kinase gene from Glycine soja, positively regulates tolerance to pH stress and ABA signal transduction.

SOS2-like protein kinases (PKS/CIPK) family genes are known to be involved in various abiotic stresses in plants. Even though, its functions have been well characterized under salt and drought stresses. The roles of PKS genes associated with alkaline stress response are not fully established yet. In this study, we identified 56 PKS family genes which could be mainly classified into three groups in wild soybean (Glycine soja). PKS family genes transcript profiles revealed different expression patterns under alkali stress. Furthermore, we confirmed the regulatory roles of GsPKS24 in response to NaHCO3, pH and ABA treatments. Overexpression of GsPKS24 enhanced plant tolerance to pH stress in Arabidopsis and soybean hairy roots but conferred suppressed pH tolerance in Arabidopsis atpks mutant. Additionally, Overexpression of GsPKS24 decreased the ABA sensitivity compared to Arabidopsis atpks mutant which displayed more sensitivity towards ABA. Moreover, upregulated expression of stress responsive and ABA signal-related genes were detected in GsPKS24 overexpression lines. In conclusion, we identified the wild soybean PKS family genes, and explored the roles of GsPKS24 in positive response to pH stress tolerance, and in alleviation of ABA sensitivity.

Strigolactone signaling gene from soybean GmMAX2a enhances the drought and salt-alkaline resistance in Arabidopsis via regulating transcriptional profiles of stress-related genes.

Strigolactone (SL) is a new plant hormone, which not only plays an important role in stimulating seed germination, plant branching, and regulating root development, but also plays an important role in the response of plants to abiotic stresses. In this study, the full-length cDNA of a soybean SL signal transduction gene (GmMAX2a) was isolated, cloned and revealed an important role in abiotic stress responses. Tissue-specific expression analysis by qRT-PCR indicated that GmMAX2a was expressed in all tissues of soybean, but highest expression was detected in seedling stems. Moreover, upregulation of GmMAX2a transcript expression under salt, alkali, and drought conditions were noted at different time points in soybean leaves compared to roots. Additionally, histochemical GUS staining studies revealed the deep staining in PGmMAX2a: GUS transgenic lines compared to WT indicating active involvement of GmMAX2a promoter region to stress responses. To further investigate the function of GmMAX2a gene in transgenic Arabidopsis, Petri-plate experiments were performed and GmMAX2a OX lines appeared with longer roots and improved fresh biomass compared to WT plants to NaCl, NaHCO3, and mannitol supplementation. Furthermore, the expression of several stress-related genes such as RD29B, SOS1, NXH1, AtRD22, KIN1, COR15A, RD29A, COR47, H+-APase, NADP-ME, NCED3, and P5CS were significantly high in GmMAX2a OX plants after stress treatment compared to WT plants. In conclusion, GmMAX2a improves soybean tolerance towards abiotic stresses (salt, alkali, and drought). Hence, GmMAX2a can be considered a candidate gene for transgenic breeding against various abiotic stresses in plants.

Health risk assessment and bioaccumulation of potentially toxic metals from water, soil, and forages near coal mines of district Chakwal, Punjab, Pakistan.

Water, forages, and soil contamination with potentially toxic metals (PTMs) through anthropogenic activities has become a significant environmental concern. It is crucial to find out the level of PTMs in water, soil, and forages near industrial areas. The PTMs enter the body of living organisms through these sources and have become a potential risk for humans and animals. Therefore, the present study aims at the health risk assessment of PTMs and their accumulation in soil, water, and forages of three tehsils (Kallar Kahar, Choa Saidan Shah, and Chakwal) in district Chakwal. Samples of wastewater, soil, and forages were collected from various sites of district Chakwal. PTMs detected in the present study were cadmium (Cd), chromium (Cr), lead (Pb), zinc (Zn), cobalt (Co), copper (Cu), and nickel (Ni), and their levels were measured through atomic absorption spectrophotometer (AAs GF95 graphite furnace auto sampler). Pollution load index (PLI), bio concentration factor (BCF), soil enrichment factors (EF), daily intake value (DIM), and health risk index (HRI) in sheep, cow, and buffalo were also analyzed. The results revealed that the mean concentration (mg/L) of Cd (0.72-0.91 mg/L), Cr (1.84-2.23 mg/L), Pb (0.95-3.22 mg/L), Co (0.74-2.93 mg/L), Cu (0.84-1.96 mg/L), and Ni (1.39-4.39 mg/L) in wastewater samples was higher than permissible limits set by WHO, NEQS, WWF, USEPA, and Pakistan in all three tehsils of district Chakwal. Similarly, in soil samples, concentrations of Cd (1.21-1.95 mg/kg), Cr (38.1-56.4 mg/kg), and Ni (28.3-55.9 mg/kg) were higher than their respective threshold values. The mean concentration of PTMs in forage samples (Parthenium hysterophorus, Mentha spicata, Justicia adhatoda, Calotropis procera, Xanthium strumarium, Amaranthaceae sp.) showed that maximum values of Cd (5.35-7.55 mg/kg), Cr (5.47-7.51 mg/kg), Pb (30-36 mg/kg), and Ni (12.6-57.5 mg/kg) were beyond their safe limit set for forages. PLI, BCF, and EF were > 1.0 for almost all the PTMs. The DIM and HRI for sheep were less than < 1.0 but for cows and buffalo were > 1.0. The current study showed that soil, water, and forages near coal mines area are contaminated with PTMs which enter the food chain and pose significant harm to humans and animals. In order to prevent their dangerous concentration in the food chain, regular assessment of PTMs present in soil, forages, irrigating water, and food is recommended.

Exogenous strigolactones enhance tolerance in soybean seedlings in response to alkaline stress.

The plant hormone strigolactones (SLs) play crucial roles in regulating plant development and adaptations to abiotic stresses. Even though the functional roles of SLs have been identified in response to abiotic stresses, the function, and mechanism of SLs are not fully established under alkaline stress. In this study, we identified that exogenous SL could improve alkaline tolerance of soybean seedlings, especially when treated with 0.5 µM SL. The application of SL remarkably reduced the malondialdehyde content, hydrogen peroxide content, and increased the activity of antioxidant enzymes under alkaline stress, suggesting that SL improved the alkaline tolerance by regulating the antioxidant defense capacity. The RNA sequencing data showed 530 special differentially expressed genes under SL treatment and alkaline stress, mainly were associated with antioxidant processes and phenylpropanoid biosynthetic pathway. Some transcription factors were also induced by SL under alkaline stress as confirmed by quantitative real-time PCR (qRT-PCR). Furthermore, SL largely increased the Na content in leaves and decreased Na content in roots under alkaline stress, which suggested that SL might promote the transport of Na from the roots to the leaves of the soybean seedlings. Meanwhile, exogenous SL decreased the content of other elements such as K, Mg, Fe, and Cu in leaves or roots under alkaline stress. Collectively, our results suggested a role of SL in regulating antioxidant defense capacity, specific gene expression, and alterations in ionic contents to alleviate harmful effects of alkaline stress in soybean seedlings.

Synthesis and Characterization of an α-Fe2O3-Decorated g-C3N4 Heterostructure for the Photocatalytic Removal of MO.

This study describes the preparation of graphitic carbon nitride (g-C3N4), hematite (α-Fe2O3), and their g-C3N4/α-Fe2O3 heterostructure for the photocatalytic removal of methyl orange (MO) under visible light illumination. The facile hydrothermal approach was utilized for the preparation of the nanomaterials. Powder X-ray diffraction (XRD), Scanning electron microscopy (SEM), Energy dispersive X-ray (EDX), and Brunauer-Emmett-Teller (BET) were carried out to study the physiochemical and optoelectronic properties of all the synthesized photocatalysts. Based on the X-ray photoelectron spectroscopy (XPS) and UV-visible diffuse reflectance (DRS) results, an energy level diagram vs. SHE was established. The acquired results indicated that the nanocomposite exhibited a type-II heterojunction and degraded the MO dye by 97%. The degradation ability of the nanocomposite was higher than that of pristine g-C3N4 (41%) and α-Fe2O3 (30%) photocatalysts under 300 min of light irradiation. The formation of a type-II heterostructure with desirable band alignment and band edge positions for efficient interfacial charge carrier separation along with a larger specific surface area was collectively responsible for the higher photocatalytic efficiency of the g-C3N4/α-Fe2O3 nanocomposite. The mechanism of the nanocomposite was also studied through results obtained from UV-vis and XPS analyses. A reactive species trapping experiment confirmed the involvement of the superoxide radical anion (O2•-) as the key reactive oxygen species for MO removal. The degradation kinetics were also monitored, and the reaction was observed to be pseudo-first order. Moreover, the sustainability of the photocatalyst was also investigated.

The soybean GmNFY-B1 transcription factor positively regulates flowering in transgenic Arabidopsis.

Nuclear Factor Y (NF-Y) gene family regulates numbers of flowering processes. Two independent transgenic Arabidopsis lines overexpressing (OX) GmNFY-B1 and GmNFYB1-GR (GmNFYB1 fused with the glucocorticoid receptor) were used to investigate the function of NFY-B1 in flowering. Furthermore, GmNFYB1-GR lines were chemically treated with dexamethasone (Dex, synthetic steroid hormone), cycloheximide (Cyc, an inhibitor of protein biosynthesis), and ethanol to examine their effects on different flowering related marker genes. Our results indicated that the transgenic lines produced longer hypocotyl lengths and had fewer numbers of rosette leaves compared to the wild-type and nf-yb1 mutant plants under both long and short-day (LD and SD) conditions. The qRT-PCR assays revealed that transcript levels of all flowering time regulating genes, i.e. SOC, FLC, FT, TSF, LFY, GI2, AGL, and FCA showed higher transcript abundance in lines OX GmNFYB1-GR. However, FT and GI genes showed higher transcript levels under Dex and Dex/Cyc treatments compared to Cyc and ethanol. Additionally, 24 differentially expressed genes were identified and verified through RNA-seq and RT-qPCR in GmNF-YB1-GR lines under Cyc and Dex/Cyc treatments from which 14 genes were up-regulated and 10 were down-regulated. These genes are involved in regulatory functions of circadian rhythm, regulation of flower development in photoperiodic, and GA pathways. The overexpression of GmNF-YB1 and GmNF-YB1-GR promote flowering through the higher expression of flowering-related genes. Further GmNF-YB1 and its attachment with the GR receptor can regulate its target genes under Dex/Cyc treatment and might act as flowering inducer under LD and SD conditions.

A novel AP2/ERF family transcription factor from Glycine soja, GsERF71, is a DNA binding protein that positively regulates alkaline stress tolerance in Arabidopsis.

KEY MESSAGE: Here we first found that GsERF71, an ERF factor from wild soybean could increase plant alkaline stress tolerance by up-regulating H+-ATPase and by modifing the accumulation of Auxin. Alkaline soils are widely distributed all over the world and greatly limit plant growth and development. In our previous transcriptome analyses, we have identified several ERF (ethylene-responsive factor) genes that responded strongly to bicarbonate stress in the roots of wild soybean G07256 (Glycine soja). In this study, we cloned and functionally characterized one of the genes, GsERF71. When expressed in epidermal cells of onion, GsERF71 localized to the nucleus. It can activate the reporters in yeast cells, and the C-terminus of 170 amino acids is essential for its transactivation activity. Yeast one-hybrid and EMSA assays indicated that GsERF71 specifically binds to the cis-acting elements of the GCC-box, suggesting that GsERF71 may participate in the regulation of transcription of the relevant biotic and abiotic stress-related genes. Furthermore, transgenic Arabidopsis plants overexpressing GsERF71 showed significantly higher tolerance to bicarbonate stress generated by NaHCO3 or KHCO3 than the wild type (WT) plants, i.e., the transgenic plants had greener leaves, longer roots, higher total chlorophyll contents and lower MDA contents. qRT-PCR and rhizosphere acidification assays indicated that the expression level and activity of H+-ATPase (AHA2) were enhanced in the transgenic plants under alkaline stress. Further analysis indicated that the expression of auxin biosynthetic genes and IAA contents were altered to a lower extent in the roots of transgenic plants than WT plants under alkaline stress in a short-term. Together, our data suggest that GsERF71 enhances the tolerance to alkaline stress by up-regulating the expression levels of H+-ATPase and by modifying auxin accumulation in transgenic plants.

Synthesis, characterization and biological evaluation of novel chalcone sulfonamide hybrids as potent intestinal alkaline phosphatase inhibitors.

Alkaline phosphatase (AP) and ecto-5'-nucleotidase (e5'NT) belong to same family that hydrolyze the extracellular nucleotides and ensure the bioavailability of nucleotides and nucleosides at purinergic receptors. During pathophysiological conditions, the over expression of AP and e5'NT lead to an increased production of adenosine that enhance tumor proliferation, invasiveness, neoangiogenesis and disrupts the body antitumor response. As both enzymes are abundantly expressed in above mentioned conditions, therefore it is of great interest to synthesize and develop potent inhibitors of these enzymes that augment the antitumor therapy. Herein we reported the synthesis and biological activity of a new series of chalcone-sulfonamide hybrids (4a-j). These derivatives were then evaluated for their inhibitory potential against two members of ecto-nucleotidase family, e5'NT (human and rat) and APs isozyme (intestinal and tissue nonspecific). Only six derivatives were found to inhibit both human and rat e5'NT enzymes. Compounds 4e and 4d showed maximum inhibition of human and rat e5'NT with an IC50±SEM=0.26±0.01 and 0.33±0.004µM, respectively. Moreover, on APs, these derivatives were identified as the selective inhibitors of calf intestinal AP (c-IAP). The derivative 4a exhibited maximum inhibition of c-IAP with an IC50±SEM=0.12±0.02µM. In conclusion, these chalcone-sulfonamide hybrids exhibited dual inhibition of both family of isozymes but was more selective towards c-IAP enzyme.

GsERF6, an ethylene-responsive factor from Glycine soja, mediates the regulation of plant bicarbonate tolerance in Arabidopsis.

MAIN CONCLUSION: This is an original study focus on ERF gene response to alkaline stress. GsERF6 functions as transcription factor and significantly enhanced plant tolerance to bicarbonate (HCO 3 (-) ) in transgenic Arabidopsis . Alkaline stress is one of the most harmful, but little studied environmental factors, which negatively affects plant growth, development and yield. The cause of alkaline stress is mainly due to the damaging consequence of high concentration of the bicarbonate ion, high-pH, and osmotic shock to plants. The AP2/ERF family genes encode plant-specific transcription factors involved in diverse environmental stresses. However, little is known about their physiological functions, especially in alkaline stress responses. In this study, we functionally characterized a novel ERF subfamily gene, GsERF6 from alkaline-tolerant wild soybean (Glycine soja). In wild soybean, GsERF6 was rapidly induced by NaHCO3 treatment, and its overexpression in Arabidopsis enhanced transgenic plant tolerance to NaHCO3 challenge. Interestingly, GsERF6 transgenic lines also displayed increased tolerance to KHCO3 treatment, but not to high pH stress, implicating that GsERF6 may participate specifically in bicarbonate stress responses. We also found that GsERF6 overexpression up-regulated the transcription levels of bicarbonate-stress-inducible genes such as NADP-ME, H (+)-Ppase and H (+)-ATPase, as well as downstream stress-tolerant genes such as RD29A, COR47 and KINI. GsERF6 overexpression and NaHCO3 stress also altered the expression patterns of plant hormone synthesis and hormone-responsive genes. Conjointly, our results suggested that GsERF6 is a positive regulator of plant alkaline stress by increasing bicarbonate ionic resistance specifically, providing a new insight into the regulation of gene expression under alkaline conditions.

Exogenous 6-BA inhibited hypocotyl elongation under darkness in Picea crassifolia Kom revealed by transcriptome profiling.

Hypocotyl elongation is an important process in plant growth and development, and is under hormonal regulatory signaling pathways. In our study, exogenous 6-BA significantly inhibited Picea crassifolia hypocotyl elongation more than ethylene in the dark, indicating the existence of different regulatory strategies in conifers, therefore, the P. crassifolia transcriptome was studied to explore the responsive genes and their regulatory pathways for exogenous N6-benzyladenine (6-BA) inhibition of hypocotyl elongation using RNA-Sequencing approach. We present the first transcriptome assembly of P. crassifolia obtained from 24.38 Gb clean data. With lowly-expressed and short contigs excluded, the assembly contains roughly 130,612 unigenes with an N50 length of 1,278 bp. Differential expression analysis found 3,629 differentially expressed genes (DEGs) and found that the differential expression fold of genes was mainly concentrated between 2 and 8 (1 ≤ log2FoldChange ≤ 3). Functional annotation showed that the GO term with the highest number of enriched genes (83 unigenes) was the shoot system development (GO: 0048367) and the KEGG category, plant hormone signal transduction (ko04075), was enriched 30 unigenes. Further analysis revealed that several cytokinin dehydrogenase genes (PcCTD1, PcCTD3 and PcCTD6) catabolized cytokinins, while xyloglucan endotransglucosylase hydrolase gene (PcXTH31), WALLS ARE THIN 1-like gene (PcWAT1-1) and Small auxin-induced gene (PcSAUR15) were strongly repressed thus synergistically completing the inhibition of hypocotyl elongation in P. crassifolia. Besides, PcbHLH149, PcMYB44 and PcERF14 were predicted to be potential core TFs that may form a multi-layered regulatory network with the above proteins for the regulation of hypocotyl growth.

Experimental Correlation of the Role of Synthesized Biochar on Thermal, Morphological, and Crystalline Properties of Coagulation Processed Poly(1,4-phenylene sulfide) Nanocomposites.

This work aimed to study the thermal and crystalline properties of poly (1,4-phenylene sulfide)@carbon char nanocomposites. Coagulation-processed nanocomposites of polyphenylene sulfide were prepared using the synthesized mesoporous nanocarbon of coconut shells as reinforcement. The mesoporous reinforcement was synthesized using a facile carbonization method. The investigation of the properties of nanocarbon was completed using SAP, XRD, and FESEM analysis. The research was further propagated via the synthesis of nanocomposites through the addition of characterized nanofiller into poly (1,4-phenylene sulfide) at five different combinations. The coagulation method was utilized for the nanocomposite formation. The obtained nanocomposite was analyzed using FTIR, TGA, DSC, and FESEM analysis. The BET surface area and average pore volume of the bio-carbon prepared from coconut shell residue were calculated to be 1517 m2/g and 2.51 nm, respectively. The addition of nanocarbon to poly (1,4-phenylene sulfide) led to an increase in thermal stability and crystallinity up to 6% loading of the filler. The lowest glass transition temperature was achieved at 6% doping of the filler into the polymer matrix. It was established that the thermal, morphological, and crystalline properties were tailored by synthesizing their nanocomposites with the mesoporous bio-nanocarbon obtained from coconut shells. There is a decline in the glass transition temperature from 126 °C to 117 °C using 6% filler. The measured crystallinity was decreased continuously, with the mixing of the filler exhibiting the incorporation of flexibility in the polymer. So, the loading of the filler into poly (1,4-phenylene sulfide) can be optimized to enhance its thermoplastic properties for surface applications.

Study of synergistic effects induced by novel base composites on heavy metals removal and pathogen inactivation.

The green-collar strategies for nanomaterial synthesis with novel structural competencies have received significant attention in nanotechnology owing to their potential benefits. The utilization of silica nanoparticles for wastewater treatment through heavy metal ions remediation is the focal point of the present study. With this intent, silica was extracted from bagasse ash by the sol-gel method and modified using chitosan. Chemical and physical characteristics of silica(S), silica/Chitosan (SCs), were reckoned through X-ray Powder Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), and Scanning Electron Microscopy (SEM) and the efficiency of synthesized biomaterials for removing heavy metal ions. Cadmium and Lead from wastewater was evaluated by conducting closed batch experiments. Isotherm and kinetics models were applied to understand the adsorption mechanism. Results of heavy metal ions removal showed that the S possesses the highest removal efficiency of 88% for cadmium. Equilibrium was established within 56 min following a Langmuir isotherm model and pseudo-second-order reaction. The synthesized biomaterials were also tested against the fungal (Aspergillus Niger) and bacterial strains (Escherichia coli and Staphylococcus aureus) to determine their antimicrobial properties Maximum inhibition of 26 mm was shown by SCs for E.coli. Synthesized samples were not so effective for A.niger. The high adsorption potential of silica nanoparticles reveals their potential to treat wastewater containing inorganic pollutants like calcium and lead released from the sugar industry firsthand, thereby building a circular economy by controlling the pollution from source to sink. The synthesized silica nanoparticles and silica/chitosan biomaterials demonstrated high adsorption potential for heavy metal ions, making them promising candidates for integration into Algal Membrane Bioreactors to enhance wastewater treatment efficiency and remove toxic pollutants. Their multifunctional properties, including antimicrobial activity, also offer potential for improving microbial control within AMBRs, ensuring a more effective and sustainable wastewater treatment process.

Genomic analysis of Chryseobacterium indologenes and conformational dynamics of the selected DD-peptidase.

Chrysobacterium indologenes is an emerging MDR pathogen that belongs to the family Flavobacteriaceae. The genome of the C. indologenes, isolated from the nephrotic patient, was sequenced through Illumina MiSeq. The pangenomics of available 56 C. indologenes strains using BPGA revealed an open pangenome (n=5553 CDS), core genome (2141), and accessory genome (2013). The CEG/DEG database identified 662 essential genes that drastically reduced to 68 genes after non-homology analyses towards human and gut microbiome. Further filtering the data for other drug target prioritizing parameters resulted in 32 putative targets. Keeping in view the crucial role played in cell wall biosynthesis, dacB was selected as the final target that encodes D-alanyl-d-alanine carboxypeptidase/endopeptidase (DD-peptidase). The 3D structure of dacB was modelled and rendered to docking analyses against two compound libraries of African plants (n=6842) and Tibetan medicines (n=52). The ADMET profiling exhibited the physicochemical properties of final compounds. The MD simulations showed the stability of inhibitor-DD-peptidase complex and interactions in terms of RMSD, RMSF, binding free energy calculation and H-bonding. We propose that the novel compounds Leptopene and ZINC95486338 from our findings might be potent DD-peptidase inhibitors that could aid in the development of new antibiotic-resistant therapy for the emerging MDR C. indologenes.

Priming with copper-chitosan nanoparticles elicit tolerance against PEG-induced hyperosmotic stress and salinity in wheat.

In this study Cu-chitosan nanoparticles (Cu-CNP) have been employed as eco-friendly and safer priming agents to induce salt and PEG-induced hyperosmotic stress tolerance in wheat seedlings. Seed priming is a facile on-farm stress management technique that requires a little amount of priming agent and minimizes the eco-toxicological effects on soil fertility. The wheat seeds were primed with 0.12% and 0.16% Cu-CNP for eight hours and were allowed to germinate under normal, PEG-induced hyperosmotic stress (15% PEG-6000  - 3.0 Mpa) and salt stress (150 mM). For comparison, non-primed and hydro-primed seeds were also allowed to germinate as control under the same conditions. The biochemical analyses suggested the priming treatments enhanced the POD activity under salt stress but it was decreased under PEG-induced hyperosmotic stress. Priming with 0.12% Cu-CNP induced a significant increase in CAT while the opposite effect was observed in 0.16% treated seedling under stress and non-stress conditions. Both priming treatments did not allow the over-expression of SOD under both stress conditions. The total phenolic contents were also decreased significantly under all conditions. Except for priming with 0.16% Cu-CNP under PEG-induced hyperosmotic stress, a suppression in MDA was observed under both stress conditions. Surprisingly, the Cu-CNP priming induced a significant increase in ß-carotenoids, total carotenoids, chlorophyll a, b and total chlorophyll under normal and stress conditions. In conclusion, the controlled expression of enzymatic antioxidants, low contents of non-enzymatic antioxidants and suppression of MDA mirror the stress mitigating role of Cu-CNP against PEG-induced hyperosmotic stress and salinity. The stress-insulating potential has also been reinforced by the enhanced production of plant and photosynthetic pigments. All these priming-induced biochemical changes produced positive effects on growth and germinating parameters in wheat seedlings under PEG-induced hyperosmotic stress as well as salinity.

Anti-Wear and Anti-Erosive Properties of Polymers and Their Hybrid Composites: A Critical Review of Findings and Needs.

Erosion caused by the repeated impact of particles on the surface of a substance is a common wear method resulting in the gradual and continual loss of affected objects. It is a crucial problem in several modern industries because the surfaces of various products and materials are frequently subjected to destructively erosive situations. Polymers and their hybrid materials are suitable, in powdered form, for use as coatings in several different applications. This review paper aims to provide extensive information on the erosion behaviors of thermoset and thermoplastic neat resin and their hybrid material composites. Specific attention is paid to the influence of the properties of selected materials and to impingement parameters such as the incident angle of the erodent, the impact velocity of the erodent, the nature of the erodent, and the erosion mechanism. The review further extends the information available about the erosion techniques and numerical simulation methods used for wear studies of surfaces. An investigation was carried out to allow researchers to explore the available selection of materials and methods in terms of the conditions and parameters necessary to meet current and future needs and challenges, in technologically advanced industries, relating to the protection of surfaces. During the review, which was conducted on the findings in the literature of the past fifty years, it was noted that the thermoplastic nature of composites is a key component in determining their anti-wear properties; moreover, composites with lower glass transition, higher ductility, and greater crystallinity provide better protection against erosion in advanced surface applications.

Cloning and in silico characterization of an abiotic stress-inducible U-box domain-containing protein gene GsPUB8 from Glycine soja.

The ubiquitination pathway is involved in the posttranslational modification of cellular proteins. However, the role of E3 ubiquitin ligase family proteins under abiotic stress conditions remains unclear, particularly in soybean. The core objective of the current study was to isolate and functionally characterize the GsPUB8 protein gene from wild soybean (Glycine soja) by using a homologous cloning method to investigate its abiotic stress responses. The GsPUB8 is a 40,562 Da molecular weight protein with 373 amino acid residues. The sequence alignment revealed the presence of U-box domain while the phylogenetic analysis showed an abundance of PUB8 proteins in both monocot and dicot plants. Analysis of gene structure predicted the absence of introns along with the presence of one exon. Furthermore, the activity of the GsPUB8 protein was anticipated in the plasma membrane and its expression was persuaded with NaCl, ABA, PEG6000, and NaHCO3 treatments with considerably higher manifestation in roots than leaves although, expressed in both vegetative and reproductive parts of G. soja. GsPUB8 protein showed 54% and 32% sequence identity to U-box domain containing 8 and 12 proteins from Arabidopsis thaliana and Oryza sativa subsp. japonica, respectively. GsPUB8 exhibited relatively higher expression under saline and drought stress particularly in roots. Whereas, the 3D model of GsPUB8 protein was generated using the SWISS-MODEL. This study can be used to manipulate the GsPUB8 protein or GsPUB8 gene for transformation purposes and its functional characterization under abiotic stress conditions.

Genome-wide identification and characterization of NHL gene family in response to alkaline stress, ABA and MEJA treatments in wild soybean (Glycine soja).

Background: NDR1/HIN1-like (NHL) family genes are known to be involved in pathogen induced plant responses to biotic stress. Even though the NHL family genes have been identified and characterized in plant defense responses in some plants, the roles of these genes associated with the plant abiotic stress tolerance in wild soybean is not fully established yet, especially in response to alkaline stress. Methods: We identified the potential NHL family genes by using the Hidden Markov model and wild soybean genome. The maximum-likelihood phylogenetic tree and conserved motifs were generated by using the MEME online server and MEGA 7.0 software, respectively. Furthermore, the syntenic analysis was generated with Circos-0.69. Then we used the PlantCARE online software to predict and analyze the regulatory cis-acting elements in promoter regions. Hierarchical clustering trees was generated using TM4: MeV4.9 software. Additionally, the expression levels of NHL family genes under alkaline stress, ABA and MEJA treatment were identified by qRT-PCR. Results: In this study, we identified 59 potential NHL family genes in wild soybean. We identified that wild soybean NHL family genes could be mainly classified into five groups as well as exist with conserved motifs. Syntenic analysis of NHL family genes revealed genes location on 18 chromosomes and presence of 65 pairs of duplication genes. Moreover, NHL family genes consisted of a variety of putative hormone-related and abiotic stress responsive elements, where numbers of methyl jasmonate (MeJA) and abscisic acid (ABA) responsive elements were significantly larger than other elements. We confirmed the regulatory roles of NHL family genes in response to alkaline stress, ABA and MEJA treatment. In conclusion, we identified and provided valuable information on the wild soybean NHL family genes, and established a foundation to further explore the potential roles of NHL family genes in crosstalk with MeJA or ABA signal transduction mechanisms under alkaline stress.