Phosphate solubilizing microorganisms isolated from medicinal plants improve growth of mint.

The current research project involves isolation and characterization of PSM (phosphate solubilizing microorganisms) from the rhizospheric soil of certain medicinal plants and to determine their effect on plant growth. Medicinal plants, Aloe vera, Bauhinia variegata, Cannabis sativa, Lantana camara and Mentha viridis were selected for the isolation of PSMs. Soil status of the selected medicinal plants was also checked. Phosphate solubilizing bacteria (PSB) were observed under stereomicroscope for their morphological characteristics and Gram's staining. Phosphate solubilizing fungi (PSF) were also identified microscopically. Colony diameter, halo zone diameter and solubilization index were determined on PVK agar plates. TLC results indicated that citric acid was the most common acid produced by PSM strains. All strains were found to be non-pathogenic in pathogenicity test. A positive plant growth response to PSM inoculation was observed in all studies. In study 1, individual inoculation of PSM showed a significant increased effect on plant growth parameter i.e., fresh and dry weight, plant height and root and shoot length as compared to control. In study2, composite inoculation of PSM along with different P sources revealed that rock phosphate (RP) with PSM increased growth of plants significantly. The present study suggests that PSM inoculation along with RP amendment can be used as biofertilizer.

Scanning electron microscopy for identification of local strain of Aspergillus Parasiticus and its larvicidal efficacy against Aedes Aegypti and non-target toxicity testing on fingerlings of Hypophthalmichthys Molitrix.

Scanning electron microscopy proved to be helpful in identification of Aspergillus parasiticus. A. parasiticus has a worldwide occurrence with high entomopathogenic efficacy against Aedes aegypti. However, assessment of pathogenic effects of entomopathogenic fungi on non-target organisms is required to evaluate its use as bio control agent. Aim of the present research is to study the parricidal activity of the local strain of A. parasiticus (MK371710) against A. aegypti and its toxicity assessment against aquatic model organism Hypophthalmichthys molitrix. A. parasiticus was isolated from the soil of Jallo Park, Lahore. The larvicidal effect of A. parasiticus was evaluated against fourth instar larvae of A. aegypti. Hundred percent of mortality of larvae was observed after exposure to 1 × 107 conidia/ml of fungal suspension after 72 h post treatment. The LC50 value of A. parsiticus in 24 h post treatment and 48 h post treatment was recorded as 1.0 × 107 conidia/ml 2.99 × 105 conidia/ml, respectively. However, an in-depth understanding safety to non-target organisms is necessary, if we are to properly control the action of these entomopathogenic fungi under natural conditions. For the toxicity assessment fingerlings of H. molitrix were exposed to conidial suspensions of A. parasiticus. Eye pop (64%), fin hemorrhage (33%), and scale infection (30%) were the major morphological effects observed during the study. Results reveal that although A. parasiticus is highly pathogenic to dengue vector but also have significant effects on organisms other than insects and its application as biological control agent requires safety considerations. HIGHLIGHTS: Assessment of pathogenic effects of entomopathogenic fungi on non-target organisms is required to evaluate its use as biocontrol agent. SEM was used to identify the microscopic view of Aspergillus parasiticus. Aim of the present research is to study the larvicidal activity of the local strain of A. parasiticus (MK371710) against Aedes aegypti and its toxicity assessment against aquatic model organism Hypophthalmichthys molitrix.

ZnO nanoparticles produced in the culture supernatant of Bacillus thuringiensis ser. israelensis affect the demographic parameters of Musca domestica using the age-stage, two-sex life table.

BACKGROUND: Indiscriminate use of broad-spectrum insecticides can have deleterious effects on insects and the environment. The use of nanoparticles synthesized from microbes has recently gained importance as a safe alternative to conventional insecticides. Recently, zinc oxide (ZnO) nanoparticles synthesized using Bacillus thuringiensis have shown insecticidal potential; however, in addition to its acute toxicity, it is necessary to determine possible sublethal effects at the organismal level to understand the toxicity of a new insecticide. Bt-derived enzymes such as nitrate reductase and other biomolecules play a vital role in the reduction of metal ions to metal nanoparticles. Here, we assessed the acute toxicity and sublethal effects of ZnO nanoparticles produced in the culture supernatant of B. thuringiensis ser. israelensis (Bti) as a reducing agent on the biological traits of Musca domestica. RESULTS: Concentration-response larval bioassays using different concentrations of ZnO-Bti-supernatant nanoparticles revealed LC10 , LC20 , LC50 and LC90 values of 4.17, 6.11, 12.73 and 38.90 µg g-1 of larval diet, respectively. Exposure of M. domestica larvae to two concentrations (LC10 and LC20 ) resulted in a lengthened developmental time (egg to adult) and preoviposition period, and reduced fecundity, survival, longevity and oviposition period. Furthermore, population parameters including net reproductive rate, mean generation time, age-specific survival rate, fecundity, life expectancy and reproductive values, analyzed following age-stage and two-sex life table theory, were significantly decreased after exposure to these concentrations of ZnO-Bti-supernatant nanoparticles compared with the control. CONCLUSION: ZnO-Bti-supernatant nanoparticles were shown to be toxic to M. domestica. Exposure of M. domestica to low concentrations of ZnO-Bti-supernatant nanoparticles resulted in negative transgenerational effects on progeny production in this fly. © 2022 Society of Chemical Industry.

Silicon-mediated growth promotion in maize (Zea mays L.) occurs via a mechanism that does not involve activation of the plasma membrane H+-ATPase.

Silicon (Si)-mediated growth promotion of various grasses is well documented. In the present study, Si-induced changes in maize shoot growth and its underlying mechanisms were studied. Maize plants were grown with various concentrations of Si (0-3 mM) in the nutrient solution. Silicon nutrition improved plant expansion growth. Silicon-supplied maize plants (0.8 and 1.2 mM) showed higher plant height and leaf area compared to no-Si amended plants. It was assumed that Si-induced expansion growth was due to positive Si effects on plasma membrane (PM) H+-ATPase. In this context, western blot analysis revealed an increase in PM H+-ATPase abundance by 77% under Si nutrition. However, in vitro measurements of enzyme activities showed no significant effect on apoplast pH, proton pumping, passive H+ efflux and enzyme kinetics such as Km, Vmax, and activation energy. Further, these results were confirmed by in vivo ratiometric analysis of apoplastic pH, which showed non-significant changes upon Si supply. In contrast, 1 mM Si altered the relative transcripts of specific PM H+-ATPase isoforms. Silicon application resulted in a significant decrease of MHA3, and this decrease in transcription seems to be compensated by an increased concentration of H+-ATPase protein. From these results, it can be concluded that changes in cell wall composition and PM H+-ATPase may be responsible for Si-mediated growth improvement in maize.

Occupational health surveillance: pulmonary function testing and oxidative stress among photocopying workers in Pakistan.

BACKGROUND: The photocopying occupation contributes to various health issues in workers. Elevated levels of oxidative stress have been found to be associated with respiratory problems in occupational workers. The intention of the current research is to evaluate the association between the pulmonary function parameters, a biomarker of oxidative stress, and the photocopying occupation in Lahore, Pakistan. MATERIAL AND METHODS: The study was performed on a group of photocopying operators (N = 100) and a control group (N = 100). A structured questionnaire was developed to gather the demographic attributes. Lung function testing was done with the spirometer. The blood lipid peroxides level, as thiobarbituric acid reactive substances (TBARS), was assessed spectrophotometrically. RESULTS: A statistically significant reduction was found in the pulmonary function parameters as compared to the control group. In the study group, the prevalence of respiratory symptoms, including cough (31%), sneezing (18%), wheezing (9%) and breathing issues (19%), was high as compared to the control group. A significantly higher plasma TBARS concentration was observed in the photocopying workers. A negative correlation prevailed between FVC (-0.081), FVC% predicted (-0.038) and FEV1 (-0.043), and the duration of exposure. CONCLUSIONS: The study showed that the occupational exposure could have a significant effect on the normal pulmonary function and the oxidative potential of well-being. The profound emissions in the vicinity of photocopying centers may be responsible for the variations in the pulmonary function parameters and oxidative stress. Med Pr. 2019;70(4):403-10.

Resistance to pyrethroid insecticides in house flies, Musca domestica L., (Diptera: Muscidae) collected from urban areas in Punjab, Pakistan.

House flies are one of the major public health pests in urban settings. People usually use insecticides containing pyrethroids for the management of house flies; however, there is a lack of information on pyrethroid resistance in house flies from urban areas. In the present study, resistance to four pyrethroids (beta-cyfluthrin, deltamethrin, permethrin, transfluthrin) was assessed in house flies collected from urban areas of Punjab, Pakistan. Significant levels of resistance to all the pyrethroids were found in different strains of house flies. The resistance ratios (RRs) at the median lethal dose (LD50) level were in the range of 5.25- to 11.02-fold for beta-cyfluthrin, 7.22- to 19.31-fold for deltamethrin, 5.36- to 16.04-fold for permethrin, and 9.05- to 35.50-fold for transfluthrin. Pairwise comparison of the log LD50s revealed a highly significant correlation (p < 0.01) between deltamethrin and permethrin, suggesting the possibility of a cross-resistance mechanism. The results revealed the occurrence of pyrethroid resistance in house flies from urban areas of Punjab. Regular resistance monitoring surveys and integrated approaches for the management of house flies are needed to retain the efficacy of these insecticides for a longer period of time.

Impact of copper toxicity on stone-head cabbage (Brassica oleracea var. capitata) in hydroponics.

Arable soils are frequently subjected to contamination with copper as the consequence of imbalanced fertilization with manure and organic fertilizers and/or extensive use of copper-containing fungicides. In the present study, the exposure of stone-head cabbage (Brassica oleracea var. capitata) to elevated Cu(2+) levels resulted in leaf chlorosis and lesser biomass yield at ≥2 µ M. Root nitrate content was not statistically affected by Cu(2+) levels, although it was substantially decreased at ≥5 µ M Cu(2+) in the shoot. The decrease in nitrate contents can be related to lower nitrate uptake rates because of growth inhibition by Cu-toxicity. Shoot sulfate content increased strongly at ≥2 µ M Cu(2+) indicating an increase in demand for sulfur under Cu stress. Furthermore, at ≥2 µM concentration, concentration of water-soluble non-protein thiol increased markedly in the roots and to a smaller level in the shoot. When exposed to elevated concentrations of Cu(2+) the improved sulfate and water-soluble non-protein thiols need further studies for the evaluation of their direct relation with the synthesis of metal-chelating compounds (i.e., phytochelatins).

Modulatory ATP binding to the E2 state of maize plasma membrane H+-ATPase indicated by the kinetics of vanadate inhibition.

P-type ATPases, as major consumers of cellular ATP in eukaryotic cells, are characterized by the formation of a phosphorylated enzyme intermediate (E2P), a process that is allosterically coupled to translocation of cations against an electrochemical gradient. The catalytic cycle comprises binding of Mg-ATP at the nucleotide-binding domain, phosphorylation of the E1 state (E1), conformational transition to the E2P state, and dephosphorylation through the actuator domain and re-establishment of the E1 state. Recently, it has been suggested that, for several P-type ATPases, Mg-ATP binds to the phosphorylated enzyme, thereby accelerating the transition to the E1 state, before then becoming the enzyme's catalytic substrate. Here, we provide evidence supporting this viewpoint. We employed kinetic models based on steady-state kinetics in the presence and absence of the reversible inhibitor orthovanadate. Vanadate is generally considered to be a conformational probe that specifically binds to the E2 state, arresting the enzyme in a state analogous to the E2P state. Hydrolytic H(+) -ATPase activities were measured in inside-out plasma membrane vesicles isolated from roots and shoots of maize plants. For root enzymes, kinetic models of vanadate inhibition that allow simultaneous binding of Mg-ATP and vanadate to the same enzyme state were most plausible. For shoot enzymes, application of the competitive inhibitor Mg-free ATP attenuated vanadate inhibition, which is consistent with a model in which either Mg-free ATP or Mg-ATP is bound to the enzyme when vanadate binds. Therefore, data from roots and shoots indicate that binding of ATP species before transition to the E1 state plays an important role in the catalytic cycle of plant plasma membrane H(+) -ATPase.

Changes in cytosolic Mg2+ levels can regulate the activity of the plasma membrane H+-ATPase in maize.

Plant PM (plasma membrane) H+-ATPase, a major consumer of cellular ATP, is driven by the MgATP complex which may dissociate at low cytosolic Mg2+ activity. We investigated whether hydrolytic activity of PM H+-ATPase is inhibited at ATP concentrations exceeding the Mg2+ concentration. Activity in isolated maize PMs was measured at pH 6.5 in the presence of 5 mM Mg2+ (high) or 2 mM Mg2+ (low), whereas K+ was applied at concentrations of 155 mM (high) or 55 mM (low). In all experiments, with membrane vesicles either from roots or leaves, the enzyme activity decreased in the presence of Mg2+-free ATP. At inhibitory ATP concentrations, the activity was not influenced by the K+ concentration. The activity was restored after increasing the Mg2+ concentration. ATP inhibition also occurred at pH 7.5. Kinetic modelling shows that Mg2+-free ATP acted as a competitive inhibitor with a Ki in the range of the Km. Ki decreased by 75% at low K+ concentration. Ki was one order of magnitude lower at pH 7.5 compared with pH 6.5. The observed inhibition is consistent with a concept in which down-regulation of the cytosolic Mg2+ activity is involved in (phyto)hormonal stress responses.