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Enzymatic and antioxidative responses are key defense mechanisms in plants following pathogen invasion, collectively known as induced systemic resistance (ISR). Alternaria sp., a well-known soil-borne pathogen, causes blight diseases in various crops. This study investigates the defence response in lentil plants through the treatment-induced application of two multipotent pesticide degrading plant growth-promoting rhizobacteria (PGPR), Bacillus cereus and Bacillus safensis, to mitigate the destructive effects of Alternaria. Both bacterial strains were applied in different carrier-based bioformulations via soil drenching. We assessed the modulation of defense-related enzymes by various combinational treatments with the Alternaria pathogen. The in vitro production of antimicrobial compounds was analyzed using GC-MS to confirm their pathogen-suppressive capabilities. Field trials showed a positive correlation between treatments and improvements in yield and growth index (GI). The highest (180%) enzymatic induction of phenylalanine ammonia lyase (PAL) followed by catalase (CAT)(100%) and polyphenol oxidase (PPO) (54%), was observed in treatments with B. cereus alone or in combination with B. safensis, in presence of Alternaria, in respect to the control. In vitro analysis revealed the production of antimicrobial compounds, including benzoic acid derivatives, cyclotetrasiloxanes, hexacosane, chlorpyrifos, and phthalates, which may contribute to pathogen suppression. Our findings demonstrate that these biocontrol agents (BCAs) not only stimulate the plant's enzymatic defense system but also enhance growth, seed yield and produce several antimicrobial compounds in vitro. Thus, pesticide-tolerant PGPR, used in this study, exhibit both disease control and plant growth-promoting properties, offering promising applications in sustainable agriculture.

Background Purple acid phosphatases, an important sub-class of metallo-phosphoesterase, play a key role in the uptake and homeostasis of phosphorus in plants, especially under phosphorus limited conditions. Phosphorus deficiency in legumes leads to an impaired growth, poor nodulation and a significant decline in yield potential. Results In this study, 22 putative purple acid phosphatase genes in Lens culinaris ( LcPAP s) were identified, distributed across seven chromosomes with chromosome 4 harbouring the highest number. In-silico analysis revealed their gene structures, physico-chemical properties, structures of the PAP proteins, and predicted sub-cellular localization. Most of the LcPAPs were found to possess a signal peptide suggesting their involvement in the secretory pathway, whereas LcPAP3 lacking any signal peptide was uniquely localized in the nucleus. Considering molecular weights, the LcPAPs clustered into three groups and were classified in five out of six clades in a phylogenetic tree constructed with PAPs from soyabean, rice and Arabidopsis . Synteny analysis of LcPAPs revealed the highest conservation with soyabean followed by Arabidopsis and least with rice, consistent with the lineage divergence of dicots and monocots that occurred million years ago. Promoter analysis of the LcPAP s revealed the presence of multiple cis elements associated with stress responsive transcription factors, including PHR1, a key regulator of phosphorus starvation response. Docking studies of the protein with ligands like para-nitrophenyl phosphate (pNPP) and phytic acid revealed the involvement of the conserved motifs and different covalent bonds in protein–ligand interaction. Gene expression analysis under differential phosphorus regimes revealed root specific upregulation of LcPAP20 , while constitutive upregulation in both root and shoot tissues of LcPAP4 annotated with oxidoreductase and phosphatase enzyme activity under low as well as high phosphorus conditions as compared to control. In contrast, LcPAP9 predicted to possess dual phosphatase-phytase activity was highly upregulated in the shoots under phosphorus deficient condition as compared to phosphorus sufficiency. Conclusions These findings provide foundational insights into the functional diversification of LcPAPs on an intricate molecular level, and might pave the way of identifying key regulatory genes of lentil involved in phosphorus homeostasis. Further in-depth bioinformatics study on LcPAPs with their two potential interactors has unravelled their potential role and binding efficacies.

Nitrogen doped carbon quantum dots (NCQDs) were synthesized via hydrothermal route. The NCQDs are thermally and optically stable with high flouresence yield. For the synthesis of NCQDs, citric acid and urea was taken as carbon and nitrogen sources, respectively. The Transmission Electron Microscopy (TEM) of these quantum dots revealed nearly spherical shape and average size of 1.5 nm, which was calculated using Image J software. The quantum dots were also well-characterized using spectroscopic techniques such as FTIR, UV-Visible absorption and fluorescence. These synthesized and characterized dots were utilized for selective detection of lactose in Milli Q water. The bioprobe provide a wide linear range varying from (10.00–77.41) μM with limit of detection 11.36 μM and sensitivity equal to (0.0065 ± 0.0002) μM−1.[Images not available. See PDF.]Graphical Abstract

Copper-containing amine oxidase (CuAO) has been proposed to play a role in H2O2 production in plant cell walls during cell development and in response to pathogen attack. We have compared the localisation of CuAO in pea (Pisum sativum L.), lentil (Lens culinaris M.) and chick pea (Cicer arietinum L.) grown under different light conditions, using both immuno- and histochemical techniques. The enzyme was detected by indirect immunofluorescence in the cell walls of parenchyma tissues of etiolated pea and lentil plants and was particularly abundant at intercellular spaces. Upon deetiolation, CuAO largely disappeared from cortical cell walls except in the region of intercellular spaces. In the apical internode of light-grown seedlings, CuAO occurred mainly in cortical cell walls and, to some extent, in cell walls of xylem vessels. In both the elongation zone and mature regions of roots, CuAO was restricted to cortical cell walls and some cell junctions close to the meristem. Extensin epitopes co-localised to intercellular spaces of the cortex in de-etiolated pea, indicating that CuAO may have a role in cell wall strengthening at intercellular spaces. In chick pea, the localisation of the enzyme varied between different cultivars that have differing susceptibility to the fungus Ascochyta rabiei. In a susceptible cultivar Calia, immunogold labelling localised CuAO to cell walls of the cortex, as in lentil and pea, while in a resistant cultivar Sultano, it was most abundant in xylem vessels and, in light-grown plants, in the epidermis. These expression patterns are discussed with regard to the possible functions of amine oxidase in cell growth, cell differentiation and pathogen resistance.

The thermal stability of copper/quinone containing amine oxidases from Euphorbia characias latex (ELAO) and lentil seedlings (LSAO) was measured in 100 mM potassium phosphate buffer (pH 7.0) following changes in absorbance at 292 nm. ELAO was shown to be about 10°C more stable than LSAO. The dissociative thermal inactivation of ELAO was studied using putrescine as substrate at different temperatures in the range 47-70°C, and a \"conformational lock\" was developed using the theory pertaining to oligomeric enzyme. Moreover ELAO was shown to be more stable towards denaturants than LSAO, as confirmed by dodecyl trimethylammonium bromide denaturation curves. A comparison of the numbers of contact sites in inter-subunits of ELAO relative to LSAO led us to conclude that the higher stability of ELAO to temperature and towards denaturants was due to the presence of larger number of contact sites in the conformational lock of the enzyme. This study also gives a putative common mechanism for thermal inactivation of amine oxidases and explains the importance of C-terminal conserved amino acids residues in this class of enzymes.[PUBLICATION ABSTRACT]

Copper/TPQ amine oxidases from mammalian and plant sources have shown many differences in substrate specificity and molecular properties. In this work the activity of lentil seedling amine oxidase was followed at various temperatures in 100 mM potassium phosphate buffer, pH 7, using benzylamine as substrate. The discontinuous Arrhenius plot of lentil amine oxidase showed two distinct phases with a jump between them. Thermal denaturation of the enzyme, using differential scanning calorimetry under the same experimental conditions, showed a transition at the same temperature ranges in the absence of substrate, indicating the occurrence of conformational changes, with an enthalpy change of about 175.9 kJ/mole. The temperature-induced changes of the activity of lentil amine oxidase are compared with those of bovine serum amine oxidase (taken from the literature).

Diabetes mellitus is a multisystemic metabolic disorder that may affect the eyes, kidneys, vessels, and heart. Chronic hyperglycemia causes non-enzymatic glycation of proteins and elevation of the polyol pathway resulting in oxidative stress that damages organs. The current study aimed to investigate the dose-dependent effects of orally consumed Rosa damascena Mill. hydrosol on hematology, clinical biochemistry, lens enzymatic activity, and lens pathology in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced into male Sprague–Dawley rats by intraperitoneal administration of STZ (40 mg/kg body weight). Rose hydrosols containing 1515 mg/L and 500 mg/L total volatiles (expressed as citronellol) were introduced to rats orally for 45 days. Consumption of 1515 mg/L volatile containing rose hydrosol successfully ameliorated hematologic, hepatic, and renal functions. Hydrosols also attenuated hyperglycemia and decreased the advanced glycation end-product formation in a dose-dependent manner. Rose hydrosol components significantly increased the lens enzymatic activities of glutathione peroxidase and decreased the activity of aldose reductase to prevent cataractogenesis. Histopathological examinations of rat lenses also indicated that increasing the dose of rose hydrosol had a protective effect on lenses in diabetic conditions. Additionally, in silico modeling of aldose reductase inhibition with rose hydrosol volatiles was carried out for extrapolating the current study to humans. The present results suggest that rose hydrosol exerts significant protective properties in diabetes mellitus and has no toxic effect on all studied systems in healthy test groups.

Diabetes mellitus is recognized as a leading cause of new cases of blindness. The prevalence of diabetic eye disease is expected to continue to increase worldwide as a result of the dramatic increase in the number of people with diabetes. At present, there is no medical treatment to delay or prevent the onset and progression of cataract or retinopathy, the most common causes of vision loss in diabetics. The plant Emblica officinalis (gooseberry) has been used for thousands of years as a traditional Indian Ayurvedic preparation for the treatment of diabetes in humans. Extracts from this plant have been shown to be efficacious against the progression of cataract in a diabetic rat model. Aldose reductase (ALR2) is implicated in the development of secondary complications of diabetes including cataract and, therefore, has been a major drug target for the development of therapies to treat diabetic disease. Herein, we present the bioassay-guided isolation and structure elucidation of 1-O-galloyl-β-D-glucose (β-glucogallin), a major component from the fruit of the gooseberry that displays selective as well as relatively potent inhibition (IC(50) = 17 µM) of AKR1B1 in vitro. Molecular modeling demonstrates that this inhibitor is able to favorably bind in the active site. Further, we show that β-glucogallin effectively inhibits sorbitol accumulation by 73% at 30 µM under hyperglycemic conditions in an ex-vivo organ culture model of lenses excised from transgenic mice overexpressing human ALR2 in the lens. This study supports the continued development of natural products such as β-glucogallin as therapeutic leads in the development of novel therapies to treat diabetic complications such as cataract.

Stauntonia hexaphylla (Thunb.) Decne. (Lardizabalaceae) leaves (SHL) have been used traditionally as analgesics, sedatives, diuretics, and so on, in China. To date, no data have been reported on the inhibitory effect of SHL and its constituents on rat lens aldose reductase (RLAR) and advanced glycation end products (AGEs). Therefore, the inhibitory effect of compounds isolated from SHL extract on RLAR and AGEs was investigated to evaluate potential treatments of diabetic complications. The ethyl acetate (EtOAC) fraction of SHL extract showed strong inhibitory activity on RLAR and AGEs; therefore, EtOAc fraction (3.0 g) was subjected to Sephadex LH-20 column chromatography, for further fractionation, with 100% MeOH solvent system to investigate its effect on RLAR and AGEs. Phytochemical investigation of SHL led to the isolation of seven compounds. Among the isolated compounds, chlorogenic acid, calceolarioside B, luteolin-3′-O-β-D-glucopyranoside, quercetin-3-O-β-D-glucopyranoside, and luteolin-7-O-β-D-glucopyranoside exhibited significant inhibitory activity against RLAR with IC50 in the range of 7.34–23.99 μM. In addition, 3-(3,4-dihydroxyphenyl) propionic acid, neochlorogenic acid, and luteolin-3′-O-β-D-glucopyranoside exhibited the most potent inhibitory activity against formation of AGEs, with an IC50 value of 115.07–184.06 μM, compared to the positive control aminoguanidine (820.44 μM). Based on these findings, SHL dietary supplements could be considered for the prevention and/or treatment of diabetes complication.

Background. Increased activity of aldose reductase (AR) is one of the mechanisms involved in the development of diabetic complications. Inhibiting AR can be a target to prevent diabetes complications. This study is aimed at evaluating the effect of cyclohexane (CH) and ethanol extracts (ET) of walnut leaves on AR activity in the lens and testis of diabetic rats. Methods. Fifty-six male rats classified into seven groups as control and treatment groups and treated for 30 days. The treatment groups were treated with different concentrations of ET and CH. The diabetic control (DC) group was exposed to streptozotocin. AR activity was measured in the lens and testis. The expression of AR in the testis was evaluated by the immunohistochemistry method. Results. Both extracts significantly reduced the AR activity (ng/mg of tissue protein) in the testis (0.034±0.004, 0.038±0.010, and 0.040±0.007 in the treatment groups vs. 0.075±0.007 in the DC group) and lens (1.66±0.09, 2.70±0.47, and 1.77±0.20 in the treatment groups vs. 6.29±0.48 in the DC group) of the treatment group compared to those of the DC group (P<0.05). AR expression in the testes of the treatment groups was decreased compared with that of the DC group (P<0.0001). Conclusion. Walnut leaf extracts can reduce the activity and localization of AR in the testes and its activity in the lens of diabetic rats.