Microbial detoxification of chlorpyrifos, profenofos, monocrotophos, and dimethoate by a multifaceted rhizospheric Bacillus cereus strain PM38 and its potential for the growth promotion in cotton.

Bacillus genera, especially among rhizobacteria, are known for their ability to promote plant growth and their effectiveness in alleviating several stress conditions. This study aimed to utilize indigenous Bacillus cereus PM38 to degrade four organophosphate pesticides (OPs) such as chlorpyrifos (CP), profenofos (PF), monocrotophos (MCP), and dimethoate (DMT) to mitigate the adverse effects of these pesticides on cotton crop growth. Strain PM38 exhibited distinct characteristics that set it apart from other Bacillus species. These include the production of extracellular enzymes, hydrogen cyanide, exopolysaccharides, Indol-3-acetic acid (166.8 µg/mL), siderophores (47.3 µg/mL), 1-aminocyclopropane-1-carboxylate deaminase activity (32.4 µg/mL), and phosphorus solubilization (162.9 µg/mL), all observed at higher concentrations. This strain has also shown tolerance to salinity (1200 mM), drought (20% PEG-6000), and copper and cadmium (1200 mg/L). The amplification of multi-stress-responsive genes, such as acdS, ituC, czcD, nifH, sfp, and pqqE, further confirmed the plant growth regulation and abiotic stress tolerance capability in strain PM38. Following the high-performance liquid chromatography (HPLC) analysis, the results showed striking compatibility with the first kinetic model. Strain PM38 efficiently degraded CP (98.4%), PF (99.7%), MCP (100%), and DMT (95.5%) at a concentration of 300 ppm over 48 h at 35 °C under optimum pH conditions, showing high coefficients of determination (R2) of 0.974, 0.967, 0.992, and 0.972, respectively. The Fourier transform infrared spectroscopy (FTIR) analysis and the presence of opd, mpd, and opdA genes in the strain PM38 further supported the potential to degrade OPs. In addition, inoculating cotton seedlings with PM38 improved root length under stressful conditions. Inoculation of strain PM38 reduces stress by minimizing proline, thiobarbituric acid-reactive compounds, and electrolyte leakage. The strain PM38 has the potential to be a good multi-stress-tolerant option for a biological pest control agent capable of improving global food security and managing contaminated sites.

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.

Elucidating molecular characterization of chlorpyrifos and profenofos degrading distinct bacterial strains for enhancing seed germination potential of Gossypium arboreum L.

Chlorpyrifos (CP) and profenofos (PF) are organophosphate pesticides (OPs) widely used in agriculture and are noxious to both fauna and flora. The presented work was designed to attenuate the toxicity of both pesticides in the growth parameters of a cotton crop by applying plant growth-promoting rhizobacteria (PGPR), namely Pseudomonas aeruginosa PM36 and Bacillus sp. PM37. The multifarious biological activities of both strains include plant growth-promoting traits, including phosphate solubilization; indole-3-acetic acid (IAA), siderophore, and HCN production; nitrogen fixation; and enzymatic activity such as cellulase, protease, amylase, and catalase. Furthermore, the molecular profiling of multi-stress-responsive genes, including acdS, ituC, czcD, nifH, and sfp, also confirmed the plant growth regulation and abiotic stress tolerance potential of PM36 and PM37. Both strains (PM36 and PM37) revealed 92% and 89% of CP degradation at 50 ppm and 87% and 81% at 150 ppm within 7 days. Simultaneously 94% and 98% PF degradation was observed at 50 ppm and 90% and 92% at 150 ppm within 7 days at 35 °C and pH 7. Biodegradation was analyzed using HPLC and FTIR. The strains exhibited first-order reaction kinetics, indicating their reliance on CP and PF as energy and carbon sources. The presence of opd, mpd, and opdA genes in both strains also supported the CP and PF degradation potential of both strains. Inoculation of strains under normal and OP stress conditions resulted in a significant increase in seed germination, plant biomass, and chlorophyll contents of the cotton seedling. Our findings indicate that the strains PM36 and PM37 have abilities as biodegraders and plant growth promoters, with potential applications in crop sciences and bioremediation studies. These strains could serve as an environmentally friendly, sustainable, and socially acceptable solution to manage OP-contaminated sites.

Efficacy of organic-based carrier material for plant beneficial rhizobacteria application in okra under normal and salt-affected soil conditions.

AIMS: Plant beneficial rhizobacteria (PBR) improve salt tolerance and plant yield in vegetable plants by producing 1-aminocyclopropane-1-carboxylate-deaminase, indole-3-acetic acid and phosphate solubilization. Organic-based carrier material is needed to ensure the PBR's uniform application, distribution, survival and functioning in a variety of fields. The PBR also use carbon present in the carrier as food and energy source. The selection of a suitable organic-based carrier material for the application of the PBR in normal and saline soils always has received less attention. The current study compared the PBR suitability of different organic-based carrier materials (biochar, biogas residues [BGRs] and coconut powder) and evaluated their effects on okra productivity under normal and saline soil conditions. METHODS AND RESULTS: In a pot experiment, the PBR strain Bacillus sp. MR-1/2 (accession number, MG548383) was applied with/or without organic-based carrier materials to okra grown in three different soils: S1 (EC 1.0 dS m-1 ), S2 (EC 3.0 dS m-1 ) and S3 (EC 5.0 dS m-1 ). The experiment was set up in a completely randomized design with five replicates in factorial arrangement. Results indicated that in soil S1, PBR + BGR increased the number of pods per plant, plant dry weight and indole compounds by 64%, 68% and 17% while reduced the electrolyte leakage (ELL), malonaldehyde (MDA) contents and stress ethylene level by 17%, 55% and 38%, respectively over the PBR application without any carrier. Similarly, in soil S2, the treatment PBR + BGR increased the number of pods by 81%, plant dry weight by 40% and indole compounds by 13% while reduced the ELL by 17%, MDA contents by 50% and stress ethylene by 30% over the PBR alone treatment. In soil S3, PBR + biochar increased the number of pods by 51%, plant dry weight by 62% and indole compounds by 20%, while reduced the ELL by 21%, MDA by 40% and indole compounds by 54% over the PBR alone treatment. CONCLUSIONS: Results concluded that in soil S1 and S2 (normal soils), BGR as carrier for PBR showed best results, while in soil S3, biochar as carrier for PBR resulted in enhanced potassium (K+ ) and calcium (Ca+2 ) uptake and increased the productivity of okra. SIGNIFICANCE AND IMPACT OF STUDY: Response of different carrier materials in supporting PBR under different soil conditions was variable. This study will help in the selection and use of best suitable carrier material for PBR application under different soil conditions. It is recommended that farmer should use BGR as carrier material for PBR application in normal soils while biochar should be used as carrier for the PBR application in saline soil.

Deciphering distinct biological control and growth promoting potential of multi-stress tolerant Bacillus subtilis PM32 for potato stem canker.

Plant growth-promoting rhizobacteria (PGPR) represent a set of microorganisms that play significant role in improving plant growth and controlling the phytopathogens. Unpredictable performance after the application of PGPR has been observed when these were shifted from in-vitro to in-vivo conditions due to the prevalence of various abiotic stress conditions. During growing period, the potato crop is subjected to a combination of biotic and abiotic stresses. Rhizoctonia solani, a soil-borne plant pathogen, causes reduced vigor and yield of potato crop worldwide. In the current study, multi-stress-tolerant rhizobacterial strain, Bacillus subtilis PM32, was isolated from field-grown potato with various plant growth promoting (PGP) traits including zinc and potassium solubilization, biological nitrogen fixation, ammonia and siderophore, as well as extracellular enzyme productions (cellulase, catalase, amylase, protease, pectinase, and chitinase). The strain PM32 exhibited a distinct potential to support plant growth by demonstrating production of indole-3-acetic acid (102.6 µM/mL), ACC-deaminase activity (1.63 µM of α-ketobutyrate/h/mg protein), and exopolysaccharides (2.27 mg/mL). By retarding mycelial growth of R. solani the strain PM32 drastically reduced pathogenicity of R. solani. The strain PM32 also suppressed the pathogenic activity significantly by impeding mycelial expansion of R. solani with inhibition co-efficient of 49.87. The B. subtilis PM32 also depicted significant tolerance towards salt, heavy metal (Pb), heat and drought stress. PCR based amplification of ituC and acds genes coding for iturin and ACC-deaminase activity respectively indicated potential of strain PM32 for lipopeptides production and ACC deaminase enzyme activity. Results of both in-vitro and pot experiments under greenhouse conditions depicted the efficiency of B. subtilis PM32 as a promising bio-control agent for R. solani infection together with enhanced growth of potato plants as deciphered from biomass accumulation, chlorophyll a, b, and carotenoid contents. Therefore, it was envisioned that application of indigenous multi-stress tolerant PGPR may serve to induce biotic and abiotic stress tolerance in crops/plants for pathogen control and sustainable global food supply. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01067-2.

Characterization of bio-fabricated silver nanoparticles for distinct anti-fungal activity against sugarcane phytopathogens.

Advanced research, development, and application of silver nanoparticles is proceeding in recent times due to their incredible utilization in various fields. Present study was focused on the production, characterization, and antifungal activities of silver nanoparticles (AgNPs). An environment friendly extracellular biosynthetic approach was adopted to produce the AgNPs by using bacteria, fungi, and sugarcane husk. Agents used for reduction of silver to nanoparticles were taken from culture filtrate of plant growth promoting bacteria, Fusarium oxysporum and supernatant extract of sugarcane husk. Nanoparticles were also characterized by scanning electron microscopy (SEM). Synthesis of colloidal AgNPs was observed by UV-Visible diffused reflectance spectroscopy (UV-Vis DRS). Primary peak of surface plasmon resonance band was noticed around 339.782, 336.735, and 338.258 nm for bacterial, fungal, and sugarcane husk produced AgNPs. Structure of all biologically produced nanoparticles were crystalline cubic with nano size of 45.41, 49.06, and 42.75 nm for bacterial, fungal, and sugarcane husk-based nanoparticles, respectively as calculated by Debye-Scherrer equation using XRD. Fourier transform infrared spectroscopy (FTIR) analysis revealed the presence of various compounds that aid in the reduction, capping, and stability of AgNPs. The antifungal activity of AgNPs was also investigated for sugarcane fungal pathogens Colletotricum falcatum and Fusarium moniliforme. All nanoparticles exhibit prominent antifungal activities. Maximum zone of fungal inhibition was noticed about 18, 19, and 21 mm for C. falcatum while 21, 20, and 24 mm for F. moniliforme in case of bacterial, fungal, and plant-based nanoparticles (15 ppm), respectively. Best fungal inhibition was observed under application of sugarcane husk based AgNPs. Moreover, biologically produced AgNPs responded better towards the suppression of F. moniliforme in comparison to C. falcatum. Mentioned sources in present study can be ecofriendly nano-factories for biosynthesis of AgNPs and mankind should benefit from their commercial application.

Assisted phytoremediation of chromium spiked soils by Sesbania Sesban in association with Bacillus xiamenensis PM14: A biochemical analysis.

Due to anthropogenic activities, chromium (Cr) contamination is ubiquitous with deleterious effects on plant and soil microbiota. Present study was designed to address beneficial effects of Bacillus xiamenensis PM14 on Sesbania sesban. Its physiological and biochemical attributes along with enhanced antioxidant enzyme activities under different levels of Cr toxicity (50, 100 and 200 mg kg-1) were evaluated. After harvesting at 50 days of sowing, plant growth attributes (root and shoot length, fresh and dry weight), physiological parameters (chlorophyll a, b and carotenoid content), antioxidant activities (superoxide dismutase, peroxidase and catalase), malondialdehyde content, electrolyte leakage, proline, relative water content and total Cr uptake in S. sesban were recorded. Experiment was statistically managed as complete randomized design (CRD). Results revealed that Cr stress reduced plant growth, relative water content at all levels of Cr contamination. However, inoculation of B. xiamenensis PM14 positively influence all parameters of S. sesban both under normal and stressed conditions. Inoculation of B. xiamenensis PM14 promoted plant growth (root length 17.08%, shoot length 28.36%) physiological attributes (chlorophyll a 55.26%, chlorophyll b 59.13%), antioxidant activities (superoxide dismutase 30.09%, peroxidase 6.96% and catalase 0.89%), relative water content 25.79%, enhanced total Cr uptake 47.33% and reduced proline 12.33%, malondialdehyde content 27.53% and electrolyte leakage 2.73% in S. sesban at 200 mg kg-1 Cr stress as compared to uninoculated plants grown under the same level of Cr. Our findings revealed first report of B. xiamenensis as phytoremediator and its inoculation on Sesbania plant. It also exposed dual effects of B. xiamenensis to ameliorate Cr stress along with improved plant growth and induced heavy metal stress tolerance in spiked soils.

Induction of tolerance to salinity in wheat genotypes by plant growth promoting endophytes: Involvement of ACC deaminase and antioxidant enzymes.

Plant growth-promoting endophytes (PGPEs) can colonize the internal tissues of plants and are capable of promoting plant growth. These bacteria can improve plant tolerance against various biotic and abiotic stresses via the expression of antioxidant enzymes and the production of 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Two salt-tolerant PGPEs, Kocuria rhizophila: KF875448 (14ASP) and Cronobacter sakazakii: KM042090 (OF115), with ACC deaminase activity were investigated for their potential to ameliorate plant salinity stress. The wheat varieties Pasban 90 and Khirman were subjected to two levels of salt stress (80 and 160 mM NaCl) under greenhouse conditions by using a completely randomized design. Analyses of plant growth parameters, antioxidant enzyme activities, chlorophyll and plant mineral contents were conducted to investigate the stress tolerance induced by the PGPEs. The ACC utilization by the PGPEs directly relates to the promotion of plant growth due to the lowering of excess ethylene production under salt stress. High levels of NaCl exhibited negative effects in both varieties. However, inoculation with PGPEs increased the morphological traits and antioxidant activities of the plants while decreasing the Na+ contents in all treatments compared to uninoculated treatment. Wheat variety Pasban 90 was more tolerant than Khirman in to salt stress in all the measured morphological and biochemical parameters, while the bacterial strain OF115 performed significantly better in all morphological and biochemical parameters, such as fresh dry weight, root shoot length, proline and chlorophyll contents, compared to strain 14ASP. The K+/Na+ ratio in the tissues of bacterial treated plants was higher than the control, probably in order to maintain the nutrient balance. The results of our study revealed that the inoculation of plants by ACC deaminase-producing PGPEs is a potential tool for the enhancement of plant growth and stress tolerance. Moreover, endophytic bacteria allied with host plants are capable of enduring high saline conditions and can interact with plants in a very efficient way.

Draft Genome Sequence of Bacillus pumilus SCAL1, an Endophytic Heat-Tolerant Plant Growth-Promoting Bacterium.

Bacillus pumilus strain SCAL1 is an endophytic, thermophilic plant that was isolated from the leaf of a plant, Solanum lycopersicum L., in Sindh, Pakistan. B. pumilus strain SCAL1 has usually exhibited high resistance to environmental stresses, with a growth temperature ranging from 30 to 60°C. An approximately 3.75-Mb draft genome was assembled into 68 contigs.

Combined application of compost and Bacillus sp. CIK-512 ameliorated the lead toxicity in radish by regulating the homeostasis of antioxidants and lead.

Lead (Pb) contamination is ubiquitous and usually causes toxicity to plants. Nevertheless, application of compost and plant growth promoting rhizobacteria synergistically may ameliorate the Pb toxicity in radish. The present study assessed the effects of compost and Bacillus sp. CIK-512 on growth, physiology, antioxidants and uptake of Pb in contaminated soil and explored the possible mechanism for Pb phytotoxicity amelioration. Treatments comprised of un-inoculated control, compost, CIK-512, and compost + CIK-512; plants were grown in soil contaminated with Pb (500mgkg-1) and without Pb in pot culture. Lead caused reduction in shoot dry biomass, photosynthetic rate, stomatal conductance, relative water contents, whereas enhanced root dry biomass, ascorbate peroxidase, catalase, malondialdehyde and electrolyte leakage in comparison with non-contaminated control. Plants inoculated with strain CIK-512 and compost produced significantly higher dry biomass, photosynthetic rate and stomatal conductance in normal and contaminated soils. Bacterial strain CIK-512 and compost synergy improved growth and physiology of radish in contaminated soil possibly through homeostasis of antioxidant activities, reduced membrane leakage and Pb accumulation in shoot. Possibly, Pb-induced production of reactive oxygen species resulted in increased electrolyte leakage and malondialdehyde contents (r = 0.88-0.92), which led to reduction in growth (r = -0.97) and physiology (r = -0.38 to -0.80), however, such negative effects were ameliorated by the regulation of antioxidants (r = 0.78-0.87). The decreased activity of antioxidants coupled with Pb accumulation in aerial part of the radish indicates the Pb-phytotoxicity amelioration through synergistic application of compost and Bacillus sp. CIK-512.

Study of the anatomical position of the femoral nerve by magnetic resonance imaging in patients with fractured neck of femur: relevance to femoral nerve block.

STUDY OBJECTIVE: To determine the anatomical location of the femoral nerve in patients who have sustained fracture of the neck of femur, and its relevance to femoral nerve block technique. DESIGN: Prospective, observational clinical study. SETTING: Orthopedic and Radiology departments of a regional hospital. SUBJECTS: 10 consecutive adult ASA physical status II and III patients (mean age, 78.5 yrs) and 4 adult healthy volunteers. INTERVENTIONS: A T1 magnetic resonance imaging scan was performed of both upper thighs in patients and healthy volunteers successfully. MEASUREMENTS: The distance (mm) between the midpoint of the femoral artery and the midpoint of the femoral nerve, and the distance of the femoral nerve from the skin was measured at the mid-inguinal ligament, the pubic tubercle, and at the mid-inguinal crease. Data are shown as means (SD). Differences between both sides were compared using paired Student's t-tests. P < 0.05 was significant. MAIN RESULTS: In patients the mean distance (mm) between the midpoint of the femoral nerve from the midpoint of femoral artery at the mid-inguinal crease on the fractured and non-fractured sides was 10.7 and 11.0, respectively (P = 0.87). The mean distance (mm) between the midpoint of the femoral nerve from the midpoint of the femoral artery at the mid-inguinal ligament on the fractured and non-fractured sides was 9.64 and 12.5, respectively (P = 0.03). The mean distance (mm) between the midpoint of the femoral nerve from the midpoint of the femoral artery at the pubic tubercle on the fractured and non-fractured sides was 8.74 and 10.49, respectively (P = 0.18). CONCLUSIONS: Blockade of the femoral nerve may be easier to perform at the mid-inguinal crease in patients with fractured neck of femur.