Pesticide-tolerant microbial consortia: Potential candidates for remediation/clean-up of pesticide-contaminated agricultural soil.

Reclamation of pesticide-polluted lands has long been a difficult endeavour. The use of synthetic pesticides could not be restricted due to rising agricultural demand. Pesticide toxicity has become a pressing agronomic problem due to its adverse impact on agroecosystems, agricultural output, and consequently food security and safety. Among different techniques used for the reclamation of pesticide-polluted sites, microbial bioremediation is an eco-friendly approach, which focuses on the application of resilient plant growth promoting rhizobacteria (PGPR) that may transform or degrade chemical pesticides to innocuous forms. Such pesticide-resilient PGPR has demonstrated favourable effects on soil-plant systems, even in pesticide-contaminated environments, by degrading pesticides, providing macro-and micronutrients, and secreting active but variable secondary metabolites like-phytohormones, siderophores, ACC deaminase, etc. This review critically aims to advance mechanistic understanding related to the reduction of phytotoxicity of pesticides via the use of microbe-mediated remediation techniques leading to crop optimization in pesticide-stressed soils. The literature surveyed and data presented herein are extremely useful, offering agronomists-and crop protectionists microbes-assisted remedial strategies for affordably enhancing crop productivity in pesticide-stressed soils.

Bacterial ACC deaminase: Insights into enzymology, biochemistry, genetics, and potential role in amelioration of environmental stress in crop plants.

Growth and productivity of crop plants worldwide are often adversely affected by anthropogenic and natural stresses. Both biotic and abiotic stresses may impact future food security and sustainability; global climate change will only exacerbate the threat. Nearly all stresses induce ethylene production in plants, which is detrimental to their growth and survival when present at higher concentrations. Consequently, management of ethylene production in plants is becoming an attractive option for countering the stress hormone and its effect on crop yield and productivity. In plants, ACC (1-aminocyclopropane-1-carboxylate) serves as a precursor for ethylene production. Soil microorganisms and root-associated plant growth promoting rhizobacteria (PGPR) that possess ACC deaminase activity regulate growth and development of plants under harsh environmental conditions by limiting ethylene levels in plants; this enzyme is, therefore, often designated as a "stress modulator." TheACC deaminase enzyme, encoded by the AcdS gene, is tightly controlled and regulated depending upon environmental conditions. Gene regulatory components of AcdS are made up of the LRP protein-coding regulatory gene and other regulatory components that are activated via distinct mechanisms under aerobic and anaerobic conditions. ACC deaminase-positive PGPR strains can intensively promote growth and development of crops being cultivated under abiotic stresses including salt stress, water deficit, waterlogging, temperature extremes, and presence of heavy metals, pesticides and other organic contaminants. Strategies for combating environmental stresses in plants, and improving growth by introducing the acdS gene into crop plants via bacteria, have been investigated. In the recent past, some rapid methods and cutting-edge technologies based on molecular biotechnology and omics approaches involving proteomics, transcriptomics, metagenomics, and next generation sequencing (NGS) have been proposed to reveal the variety and potential of ACC deaminase-producing PGPR that thrive under external stresses. Multiple stress-tolerant ACC deaminase-producing PGPR strains have demonstrated great promise in providing plant resistance/tolerance to various stressors and, therefore, it could be advantageous over other soil/plant microbiome that can flourish under stressed environments.

Metabolomics-Based Mechanistic Insights into Revealing the Adverse Effects of Pesticides on Plants: An Interactive Review.

In plant biology, metabolomics is often used to quantitatively assess small molecules, metabolites, and their intermediates in plants. Metabolomics has frequently been applied to detect metabolic alterations in plants exposed to various biotic and abiotic stresses, including pesticides. The widespread use of pesticides and agrochemicals in intensive crop production systems is a serious threat to the functionality and sustainability of agroecosystems. Pesticide accumulation in soil may disrupt soil-plant relationships, thereby posing a pollution risk to agricultural output. Application of metabolomic techniques in the assessment of the biological consequences of pesticides at the molecular level has emerged as a crucial technique in exposome investigations. State-of-the-art metabolomic approaches such as GC-MS, LC-MS/MS UHPLC, UPLC-IMS-QToF, GC/EI/MS, MALDI-TOF MS, and 1H-HR-MAS NMR, etc., investigating the harmful effects of agricultural pesticides have been reviewed. This updated review seeks to outline the key uses of metabolomics related to the evaluation of the toxicological impacts of pesticides on agronomically important crops in exposome assays as well as bench-scale studies. Overall, this review describes the potential uses of metabolomics as a method for evaluating the safety of agricultural chemicals for regulatory applications. Additionally, the most recent developments in metabolomic tools applied to pesticide toxicology and also the difficulties in utilizing this approach are discussed.

Ecotoxicological implications of residual pesticides to beneficial soil bacteria: A review.

Optimization of crop production in recent times has become essential to fulfil food demands of constantly increasing human populations worldwide. To address this formidable challenge, application of agro-chemicals including synthetic pesticides in intensive farm practices has increased alarmingly. The excessive and indiscriminate application of pesticides to foster food production however, leads to its exorbitant deposition in soils. After accumulation in soils beyond threshold limits, pesticides harmfully affect the abundance, diversity and composition and functions of rhizosphere microbiome. Also, the cost of pesticides and emergence of resistance among insect-pests against pesticides are other reasons that require attention. Due to this, loss in soil nutrient pool cause a substantive reduction in agricultural production which warrant the search for newer environmentally friendly technology for sustainable crop production. Rhizosphere microbes, in this context, play vital roles in detoxifying the polluted environment making soil amenable for cultivation through detoxification of pollutants, rhizoremediation, bioremediation, pesticide degradation, and stress alleviation, leading to yield optimization. The response of soil microorganisms to range of chemical pesticides is variable ranging from unfavourable to the death of beneficial microbes. At cellular and biochemical levels, pesticides destruct the morphology, ultrastructure, viability/cellular permeability, and many biochemical reactions including protein profiles of soil bacteria. Several classes of pesticides also disturb the molecular interaction between crops and their symbionts impeding the overall useful biological processes. The harmful impact of pesticides on soil microbes, however, is poorly researched. In this review, the recent findings related with potential effects of synthetic pesticides on a range of soil microbiota is highlighted. Emphasis is given to find and suggest strategies to minimize the chemical pesticides usage in the real field conditions to preserve the viability of soil beneficial bacteria and soil quality for safe and sustainable crop production even in pesticide contaminated soils.

An Insight into Efflux-Mediated Arsenic Resistance and Biotransformation Potential of Enterobacter Cloacae RSC3 from Arsenic Polluted Area.

Indiscriminate discharge of heavy metals/metalloids from different sources into the sustainable agro-ecosystem is a major global concern for food security and human health. Arsenic (As), categorized as group one human carcinogen is a quintessential toxic metalloid that alters the microbial compositions and functions, induce physiological and metabolic changes in plants and contaminate surface/ground water. The management of arsenic toxicity, therefore, becomes imminent. Acknowledging the arsenic threat, the study was aimed at identifying arsenic resistant bacteria and evaluating its arsenic removal/detoxification potential. Of the total 118 bacterial isolates recovered from arsenic rich environment, the bacterial strain RSC3 demonstrating highest As tolerance was identified as Enterobacter cloacae by 16S rRNA gene sequence analysis. Enterobacter cloacae tolerated high concentration (6000 ppm) of As and exhibited 0.55 h-1 of specific growth rate as calculated from growth kinetics data. Strain RSC3 also displayed varying level of resistance to other heavy metals and many antibacterial drugs in plate bioassay. The bacterial strain RSC3 possessed gene (arsC) which causes transformation of arsenate to arsenite. The arsenate uptake and efflux of the bacterial cells was revealed by high throughput techniques such as AAS, SEM/TEM and EDX. The simultaneous As reducing ability, and multi metal/multi-antibiotics resistance potentials of E. cloacae provides a promising option in the microbes based remediation of As contaminated environments.

Understanding the phytotoxic impact of Al3+, nano-size, and bulk Al2O3 on growth and physiology of maize (Zea mays L.) in aqueous and soil media.

The release and accumulation of metal-oxide nanoparticles in soils have threatened terrestrial plants. However, limited knowledge is available on the accumulation of nano-Al2O3 (22 nm), bulk-Al2O3 (167 nm), and Al3+ by maize plants and the subsequent impact on its physiology and growth in agar (0.7% w/v), hydroponic (1X), and soil. Maize plants were cultivated with 0.05-2 mg g-1 or ml-1 of three Al types and their biological attributes, oxidative status, Al bioaccumulation, and translocation were measured. The ICP-MS results revealed a dose-dependent increase (P ≤ 0.05 or ≤0.01) in Al content in maize tissues following nano-Al2O3 and Al3+ exposure, however, plants exposed to bulk-Al2O3 showed no significant uptake of Al. Atomic mapping by EDX during SEM analysis and TEM revealed varied distributions of nano-Al2O3 from roots to aerial parts and intracellular transportation. Al deposition in tissues followed the order: Al3+ > nano-Al2O3 > bulk-Al2O3 and therefore, a similar trend of toxicity was observed for seed germination, the emergence of plant organs, length, biomass accumulation, total chlorophyll, phosphorus content, and total soluble protein. Oxidative stress was profoundly induced dose-dependently and was highest at 2 mg ml-1 or g-1 of Al3+ and nano-Al2O3 when superoxide radical formation, proline induction, activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (GPX), and glutathione reductase (GR) and membrane lipid peroxidation were measured. Aluminum toxicity was found higher in hydroponically grown maize compared to soil-grown maize. Forty days exposure in soil showed greater inhibition of maize growth compared to 20 days exposure. This study is significant in understanding the maize response to different Al types in soil and soil-free media.

Bioprospecting Plant Growth Promoting Rhizobacteria for Enhancing the Biological Properties and Phytochemical Composition of Medicinally Important Crops.

Traditionally, medicinal plants have long been used as a natural therapy. Plant-derived extracts or phytochemicals have been exploited as food additives and for curing many health-related ailments. The secondary metabolites produced by many plants have become an integral part of human health and have strengthened the value of plant extracts as herbal medicines. To fulfil the demand of health care systems, food and pharmaceutical industries, interest in the cultivation of precious medicinal plants to harvest bio-active compounds has increased considerably worldwide. To achieve maximum biomass and yield, growers generally apply chemical fertilizers which have detrimental impacts on the growth, development and phytoconstituents of such therapeutically important plants. Application of beneficial rhizosphere microbiota is an alternative strategy to enhance the production of valuable medicinal plants under both conventional and stressed conditions due to its low cost, environmentally friendly behaviour and non-destructive impact on fertility of soil, plants and human health. The microbiological approach improves plant growth by various direct and indirect mechanisms involving the abatement of various abiotic stresses. Given the negative impacts of fertilizers and multiple benefits of microbiological resources, the role of plant growth promoting rhizobacteria (PGPR) in the production of biomass and their impact on the quality of bio-active compounds (phytochemicals) and mitigation of abiotic stress to herbal plants have been described in this review. The PGPR based enhancement in the herbal products has potential for use as a low cost phytomedicine which can be used to improve health care systems.

Tolerance of pesticides and antibiotics among beneficial soil microbes recovered from contaminated rhizosphere of edible crops.

A total of 45 beneficial soil bacterial isolates (15 each of Pseudomonas, Azotobacter and phosphate solubilizing bacteria: PSB) recovered from polluted rhizosphere soils were morphologically and biochemically characterized. Bacterial isolates produced indole-3-acetic acid (IAA), phenolate siderophores; SA (salicylic acid) and 2, 3-dihydroxy benzoic acid (2, 3-DHBA), 1-amino cyclopropane 1-carboxylate (ACC) deaminase, solubilised insoluble phosphate (Pi), secreted exopolysaccharides (EPS) and produced ammonia and cyanogenic compound (HCN). Isolates were tested for their tolerance ability against 12 different agrochemicals (chemical pesticides) and 14 antibiotics. Among Pseudomonas, isolate PS1 showed maximum (2183 µg mL-1) tolerance to all tested agrochemicals. Likewise, among all Azotobacter isolates (n = 15), AZ12 showed maximum (1766 µg mL-1) while AZ7 had lowest (950 µg mL-1) tolerance ability to all tested agrochemicals. Moreover, among phosphate solubilizing bacterial isolates, maximum (1970 µg mL-1) and minimum (1308 µg mL-1) tolerance to agrochemicals was represented by PSB8 and PSB13 isolates, respectively. The antibiotic sensitivity/resistance among isolates varied considerably. As an example, Pseudomonas spp. was susceptible to several antibiotics, and inhibition zone differed between 10 mm (polymyxin B) to 34 mm (nalidixic acid). Also, isolate PS2 showed resistance to erythromycin, ciprofloxacin, methicillin, novobiocin and penicillin. The resistance percentage to multiple antibiotics among Azotobacter isolates varied between 7 and 33%. Among PSB isolates, inhibition zone differed between 10 and 40 mm and maximum and minimum resistance percentage to multiple antibiotics was recorded as 47% and 20%, respectively. The persistence of pesticides in agricultural soil may contribute to an increase in multidrug resistance among soil microorganisms. In conclusion, plant growth promoting (PGP) substances releasing soil microorganisms comprising of inherent/intrinsic properties of pesticides tolerance and antibiotics resistance may provide an attractive, agronomically feasible, and long-term prospective alternative for the augmentation of edible crops. However, in future, more research is needed to uncover the molecular processes behind the development of pesticide tolerance and antibiotic resistance among soil microorganisms.

Sorghum-Phosphate Solubilizers Interactions: Crop Nutrition, Biotic Stress Alleviation, and Yield Optimization.

Sweet sorghum [Sorghum bicolor (L.) Moench] is a highly productive, gluten-free cereal crop plant that can be used as an alternative energy resource, human food, and livestock feed or for biofuel-ethanol production. Phosphate fertilization is a common practice to optimize sorghum yield but because of high cost, environmental hazards, and soil fertility reduction, the use of chemical P fertilizer is discouraged. Due to this, the impetus to search for an inexpensive and eco-friendly microbiome as an alternative to chemical P biofertilizer has been increased. Microbial formulations, especially phosphate solubilizing microbiome (PSM) either alone or in synergism with other rhizobacteria, modify the soil nutrient pool and augment the growth, P nutrition, and yield of sorghum. The use of PSM in sorghum disease management reduces the dependence on pesticides employed to control the phytopathogens damage. The role of PSM in the sorghum cultivation system is, however, relatively unresearched. In this manuscript, the diversity and the strategies adopted by PSM to expedite sorghum yield are reviewed, including the nutritional importance of sorghum in human health and the mechanism of P solubilization by PSM. Also, the impact of solo or composite inoculations of biological enhancers (PSM) with nitrogen fixers or arbuscular mycorrhizal fungi is explained. The approaches employed by PSM to control sorghum phytopathogens are highlighted. The simultaneous bio-enhancing and biocontrol activity of the PS microbiome provides better options for the replacement of chemical P fertilizers and pesticide application in sustainable sorghum production practices.

Psychrophilic Bacterial Phosphate-Biofertilizers: A Novel Extremophile for Sustainable Crop Production under Cold Environment.

Abiotic stresses, including low-temperature environments, adversely affect the structure, composition, and physiological activities of soil microbiomes. Also, low temperatures disturb physiological and metabolic processes, leading to major crop losses worldwide. Extreme cold temperature habitats are, however, an interesting source of psychrophilic and psychrotolerant phosphate solubilizing bacteria (PSB) that can ameliorate the low-temperature conditions while maintaining their physiological activities. The production of antifreeze proteins and expression of stress-induced genes at low temperatures favors the survival of such organisms during cold stress. The ability to facilitate plant growth by supplying a major plant nutrient, phosphorus, in P-deficient soil is one of the novel functional properties of cold-tolerant PSB. By contrast, plants growing under stress conditions require cold-tolerant rhizosphere bacteria to enhance their performance. To this end, the use of psychrophilic PSB formulations has been found effective in yield optimization under temperature-stressed conditions. Most of the research has been done on microbial P biofertilizers impacting plant growth under normal cultivation practices but little attention has been paid to the plant growth-promoting activities of cold-tolerant PSB on crops growing in low-temperature environments. This scientific gap formed the basis of the present manuscript and explains the rationale for the introduction of cold-tolerant PSB in competitive agronomic practices, including the mechanism of solubilization/mineralization, release of biosensor active biomolecules, molecular engineering of PSB for increasing both P solubilizing/mineralizing efficiency, and host range. The impact of extreme cold on the physiological activities of plants and how plants overcome such stresses is discussed briefly. It is time to enlarge the prospects of psychrophilic/psychrotolerant phosphate biofertilizers and take advantage of their precious, fundamental, and economical but enormous plant growth augmenting potential to ameliorate stress and facilitate crop production to satisfy the food demands of frighteningly growing human populations. The production and application of cold-tolerant P-biofertilizers will recuperate sustainable agriculture in cold adaptive agrosystems.

Differential bioaccumulations and ecotoxicological impacts of metal-oxide nanoparticles, bulk materials, and metal-ions in cucumbers grown in sandy clay loam soil.

Expanding applications of metal-oxide nanoparticles (NPs) and increased environmental deposition of NPs followed by their interactions with edible crops threaten yields. This study demonstrates the effects of aging (45 days in soil) of four NPs (ZnO, CuO, Al2O3, TiO2; 3.9-34 nm) and their corresponding metal oxide bulk particles (BPs; 144-586 nm) on cucumbers (Cucumis sativus L.) cultivated in sandy-clay-loam field soil and compares these with the phytotoxic effects of readily soluble metal salts (Zn2+, Cu2+, and Al3+). Data revealed the cell-to-cell translocations of NPs, their attachments to outer and inner cell surfaces, nuclear membranes, and vacuoles, and their upward movements to aerial parts. Metal bioaccumulations in cucumbers were found in the order: (i) ZnO-NPs > ZnO-BPs > Zn2+, (ii) CuO-NPs > CuO-BPs > Cu2+, (iii) Al3+> Al2O3-NPs > Al2O3-BPs and (iv) TiO2-NPs > TiO2-BPs. Aging of NPs in soil for 45 days significantly enhanced metal uptake (P ≤ 0.05), for instance aged ZnO-NPs at 1 g kg-1 increased the uptake by 20.7 % over non-aged ZnO-NPs. Metal uptakes inhibited root (RDW) and shoot (SDW) dry weight accumulations. For Cu species, maximum negative impact (%) was exhibited by Cu2+ (RDW:SDW = 94:65) followed by CuO-NPs (RDW:SDW = 78:34) and CuO-BPs (RDW:SDW = 27:22). Aging of NPs/BPs at 1-4 g kg-1 further enhanced the toxic impact of tested materials on biomass accumulations and chlorophyll formation. NPs also induced membrane damage of root tissues and enhanced levels of antioxidant enzymes. The results of this study suggest that care is required when aged metal-oxide NPs of both essential (Zn and Cu) and non-essential (Al and Ti) metals interact with cucumber plants, especially, when they are used for agricultural purposes.

Differential responses of maize (Zea mays) at the physiological, biomolecular, and nutrient levels when cultivated in the presence of nano or bulk ZnO or CuO or Zn2+ or Cu2+ ions.

Expanding applications of metal-based nanoparticles (NPs) in industry and agriculture have influenced agro-ecosystems. However, relatively little is known about the bioaccumulation, distribution, and phytotoxicity of ZnO-NPs, CuO-NPs, ZnO-bulk, CuO-bulk, Zn2+, or Cu2+ in maize. Plants were exposed to 0.05-2 mg ml-1 or g-1 of six tested materials in agar (7 days) in hydroponic medium (20 days), or sandy-clay-loam soil (20 or 40 days). Seed germination, emergence and lengths of plumules, principal and seminal roots were significantly inhibited by ZnO-NPs, CuO-NPs, Zn2+, and Cu2+. Toxicity was more pronounced in hydroponic culture than in soil, and perceptible alterations in biomolecules were evident. ICP-MS analysis exhibited progressive uptake of metals while morphological, elemental, and surface/deeper scanning showed translocation and distribution of NPs in tissues. Tested materials induced enhanced superoxide radical production, lipid peroxidation, and antioxidant enzymes and proline levels. Exposure significantly reduced P-accumulation, photosynthesis, and protein production. Zn2+ and Cu2+ were found to be more toxic than NPs. Compared to 20 days exposure in soil, toxicity slightly increased after 40 days. ZnO-NPs and CuO-NPs increased apoptotic sub-G1 population by 22.4% and 38%, respectively. These results provide a better understanding of the mechanistic aspects responsible for the nanotoxicities of ZnO- and CuO-NPs in maize.

Mesorhizobium ciceri as biological tool for improving physiological, biochemical and antioxidant state of Cicer aritienum (L.) under fungicide stress.

Fungicides among agrochemicals are consistently used in high throughput agricultural practices to protect plants from damaging impact of phytopathogens and hence to optimize crop production. However, the negative impact of fungicides on composition and functions of soil microbiota, plants and via food chain, on human health is a matter of grave concern. Considering such agrochemical threats, the present study was undertaken to know that how fungicide-tolerant symbiotic bacterium, Mesorhizobium ciceri affects the Cicer arietinum crop while growing in kitazin (KITZ) stressed soils under greenhouse conditions. Both in vitro and soil systems, KITZ imparted deleterious impacts on C. arietinum as a function of dose. The three-time more of normal rate of KITZ dose detrimentally but maximally reduced the germination efficiency, vigor index, dry matter production, symbiotic features, leaf pigments and seed attributes of C. arietinum. KITZ-induced morphological alterations in root tips, oxidative damage and cell death in root cells of C. arietinum were visible under scanning electron microscope (SEM). M. ciceri tolerated up to 2400 µg mL-1 of KITZ, synthesized considerable amounts of bioactive molecules including indole-3-acetic-acid (IAA), 1-aminocyclopropane 1-carboxylate (ACC) deaminase, siderophores, exopolysaccharides (EPS), hydrogen cyanide, ammonia, and solubilised inorganic phosphate even in fungicide-stressed media. Following application to soil, M. ciceri improved performance of C. arietinum and enhanced dry biomass production, yield, symbiosis and leaf pigments even in a fungicide-polluted environment. At 96 µg KITZ kg-1 soil, M. ciceri maximally and significantly (p ≤ 0.05) augmented the length of plants by 41%, total dry matter by 18%, carotenoid content by 9%, LHb content by 21%, root N by 9%, shoot P by 11% and pod yield by 15% over control plants. Additionally, the nodule bacterium M. ciceri efficiently colonized the plant rhizosphere/rhizoplane and considerably decreased the levels of stressor molecules (proline and malondialdehyde) and antioxidant defence enzymes viz. ascorbate peroxidise (APX), guaiacol peroxidise (GPX), catalase (CAT) and peroxidises (POD) of C. arietinum plants when inoculated in soil. The symbiotic strain effectively colonized the plant rhizosphere/rhizoplane. Conclusively, the ability to endure higher fungicide concentrations, capacity to secrete plant growth modulators even under fungicide pressure, and inherent features to lower the level of proline and plant defence enzymes makes this M. ciceri as a superb choice for augmenting the safe production of C. arietinum even under fungicide-contaminated soils.

Impact of metal-oxide nanoparticles on growth, physiology and yield of tomato (Solanum lycopersicum L.) modulated by Azotobacter salinestris strain ASM.

The current study for the first time demonstrates the interference of a free-living, N2-fixing, and nanoparticle (NP) tolerant Azotobacter salinestris strain ASM recovered from metal-polluted soil with tomato plant-metal oxide NPs (ZnO, CuO, Al2O3, and TiO2) interactions in a sandy clay loam soil system with bulk materials as control. Tomato plants were grown till full maturity in soils amended with 20-2000 mg kg-1 of each metal-oxide NP with and without seed biopriming and root-inoculation of A. salinestris. A. salinestris was found metabolically active, producing considerably high amounts of bioactive indole-3-acetic-acid, morphologically unaffected, and with low alteration of cell membrane permeability under 125-1500 µgml-1 of NPs. However, ZnO-NPs slightly alter bacterial membrane permeability. Besides, A. salinestris secreted significantly higher amounts of extracellular polymeric substance (EPS) even under NP exposure, which could entrap the NPs and form metal-EPS complex as revealed and quantified by SEM-EDX. NPs were also found adsorbed on bacterial biomass. EPS stabilized the NPs and provided negative zeta potential to NPs. Following soil application, A. salinestris improved the plant performance and augmented the yield of tomato fruits and lycopene content even in NPs stressed soils. Interestingly, A. salinestris inoculation enhanced photosynthetic pigment formation, flower attributes, plant and fruit biomass, and reduced proline level. Bacterial inoculation also reduced the NP's uptake and accumulation significantly in vegetative organs and fruits. The organ wise order of NP's internalization was roots > shoots > fruits. Conclusively, A. salinestris inoculation could be an alternative to increase the production of tomato in metal-oxide NPs contaminated soils.

Physiological disruption, structural deformation and low grain yield induced by neonicotinoid insecticides in chickpea: A long term phytotoxicity investigation.

Arbitrary use of insecticides in agricultural practices cause severe environmental hazard that adversely affects the growth and productivity of edible crops. Considering theses, the aim of the present study was to evaluate the toxicological potential of two neonicotinoid insecticides, imidacloprid (IMID) and thiamethoxam (THIA) using chickpea as a test crop. Application of insecticides at three varying doses revealed a gradual decrease in biological performance of chickpea plants which however, varied noticeably among insecticides and their doses. Significant (P ≤ 0.05) decline in germination efficiency, length of plant organs under in vitro condition was observed in a dose related manner. Among insecticides, 300 µgIMIDkg-1 (3X dose) maximally and significantly (P ≤ 0.05) inhibited germination efficiency, vigor index, length, dry matter accumulation, photosynthetic pigment formation, nodule volume and mass, nutrient uptake, grain yield and protein over untreated control. In contrast, 75 µgTHIAkg-1 (3X dose) considerably declined the leghaemoglobin content, shoot phosphorus and root nitrogen. Enhanced expression of stress biomarkers including proline, malondialdehyde (MDA), and antioxidant defence enzymes was noticed in the presence of insecticides. For instance, at 3X IMID, shoot proline, MDA, ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and peroxidase (POD) were increased significantly (P ≤ 0.05) by 66%, 81%, 36% and 35%, respectively. Additionally, electrolyte leakage was maximally (77%) increased at 3X dose of IMID, whereas, H2O2 in foliage was maximally accumulated (0.0156 µ moles min-1 g-1 fw) at 3X dose of THIA which was 58% greater than untreated foliage. A clear distortion/damage in tip and surface of roots and ultrastructural deformation in xylem and phloem of plant tissues as indication of insecticidal phytotoxicity was observed under scanning electron microscope (SEM). For oxidative stress and cytotoxicity assessment, root tips were stained with a combination of acridine orange and propidium iodide, and Evan blue dyes and examined. Confocal laser scanning microscopic (CLSM) images of roots revealed a 10-fold and 13.5-fold increase in red and blue fluorescence when 3X IMID treated roots were assessed quantitatively. Conclusively, the present investigation recommends that a careful and protected approach should be adopted before the application of insecticides in agricultural ecosystems.

Fungicide-Tolerant Plant Growth-Promoting Rhizobacteria Mitigate Physiological Disruption of White Radish Caused by Fungicides Used in the Field Cultivation.

Excessive use of fungicides in agriculture may result in substantial accumulation of active residues in soil, which affect crop health and yield. We investigated the response of Raphanus sativus (white radish) to fungicides in soil and potential beneficial interactions of radish plants with fungicide-tolerant plant growth-promoting rhizobacteria (PGPR). The PGPR were isolated from cabbage and mustard rhizospheres. Morphological and biochemical characteristics measured using standard methods, together with analysis of partial 16S rRNA gene sequences, revealed that fungicide-tolerant PGPR, isolates PS3 and AZ2, were closely related to Pseudomonas spp. These PGPR survived in the presence of high fungicide concentrations i.e., up to 2400 µg mL-1 carbendazim (CBZM) and 3200 µg mL-1 hexaconazole (HEXA). Bacterial isolates produced plant growth stimulants even under fungicide stress, though fungicides induced surface morphological distortion and alteration in membrane permeability of these bacteria, which was proved by a set of microscopic observations. Fungicides considerably affected the germination efficiency, growth, and physiological development of R. sativus, but these effects were relieved when inoculated with PGPR isolates. For instance, CBZM at 1500 mg kg-1 decreased whole dry biomass by 71%, whole plant length by 54%, total chlorophyll by 50%, protein content by 61%, and carotenoid production by 29%. After applying isolate AZ2 for white radish grown in CBZM (10 mg kg-1)-amended soil, it could improve plant growth and development with increased whole plant dry weight (10%), entire plant length (13%) and total chlorophyll content (18%). Similarly, isolate PS3 enhanced plant survival by relieving plant stress with declined biomarkers, i.e., proline (12%), malondialdehyde (3%), ascorbate peroxidase (6.5%), catalase (18%), and glutathione reductase (4%). Application of isolates AZ2 and PS3 could be effective for remediation of fungicide-contaminated soil and for improving the cultivation of radish plants while minimizing inputs of fungicides.

Destruction of Cell Topography, Morphology, Membrane, Inhibition of Respiration, Biofilm Formation, and Bioactive Molecule Production by Nanoparticles of Ag, ZnO, CuO, TiO2, and Al2O3 toward Beneficial Soil Bacteria.

The unregulated discharge of nanoparticles (NPs) from various nanotechnology industries into the environment is expected to alter the composition and physiological functions of soil microbiota. Considering this knowledge gap, the impact of five NPs (Ag, ZnO, CuO, Al2O3, and TiO2) differing in size and morphology on growth behavior and physiological activity of Azotobacter chroococcum, Bacillus thuringiensis, Pseudomonas mosselii, and Sinorhizobium meliloti were investigated. Various biochemical and microscopic approaches were adopted. Interestingly, all bacterial strains were found sensitive to Ag-NPs and ZnO-NPs but showed tolerance toward CuO, Al2O3, and TiO2-NPs. The loss of cellular respiration due to NPs was coupled with a reduction in population size. ZnO-NPs at 387.5 µg mL-1 had a maximum inhibitory impact on A. chroococcum and reduced its population by 72%. Under Ag-NP stress, the reduction in IAA secretion by bacterial strains followed the order S. meliloti (74%) > P. mosselii (63%) > A. chroococcum (49%). The surface of bacterial cells had small- or large-sized aggregates of NPs. Also, numerous gaps, pits, fragmented, and disorganized cell envelopes were visible. Additionally, a treated cell surface appeared corrugated with depressions and alteration in cell length and a strong heterogeneity was noticed under atomic force microscopy (AFM). For instance, NPs induced cell roughness for P. mosselii followed the order 12.6 nm (control) > 58 nm (Ag-NPs) > 41 nm (ZnO-NPs). TEM analysis showed aberrant morphology, cracking, and disruption of the cell envelope with extracellular electron-dense materials. Increased permeability of the inner cell membrane caused cell death and lowered EPS production. Ag-NPs and ZnO-NPs also disrupted the surface adhering ability of bacteria, which varied with time and concentration of NPs. Conclusively, a plausible mechanism of NP toxicity to bacteria has been proposed to understand the mechanistic basis of ecological interaction between NPs and resourceful bacteria. These results also emphasize to develop strategies for the safe disposal of NPs.

Effective Inhibition of Phytopathogenic Microbes by Eco-Friendly Leaf Extract Mediated Silver Nanoparticles (AgNPs).

ABSTRACT: This study was aimed at producing the eco-friendly, safe, and inexpensive silver (Ag) nanoparticles (NPs) and assessing its antimicrobial activity. Fungal pathogens isolated from diseased leaves and fruits of brinjal and bacterial pathogen obtained from a culture collection were used in this study. Green synthesis of AgNPs was performed and optimized using Azadirachta indica leaf extract. The newly synthesized AgNPs (λmax = 437 nm) showed isotropism in size (crystal size/diameter: 21/29 ± 5 nm) and morphology under transmission and scanning electron microscopy and energy dispersive X-ray analysis. The fourier transform infrared spectroscopy data suggested the role of various aliphatic/aromatic moieties and proteins in AgNPs stabilization. The AgNPs reduced the growth of Penicillium sp. maximally by 92% after 6 days. The sensitivity of test fungi towards AgNPs followed the order: Penicillium sp. (92%) > Fusarium sp. (89%) > Aspergillus sp. (69%). Exposure of Ralstonia solanacearum to AgNPs (MIC/MBC 200/400 µg ml-1) displayed damaged cellular envelopes, bulging of cells, and pit formation. The nucleic acid discharge showed a progressive increase from 8 to 34% (r2 = 0.97). The cellular metabolic activity and surface adhering ability of R. solanacearum were completely lost at 400 µgAgNPs ml-1. Results suggested that the AgNPs synthesized in this study had enough anti-pathogenic potential and could inexpensively and safely be used as a promising alternative to agrochemicals. Moreover, the findings observed in this study is likely to serve as an important indicator for the development of effective nano-control agents which in effect would help to manage some deadly phyto-pathogens capable of causing heavy losses to agricultural production systems. GRAPHICAL ABSTRACT: Effective inhibition of phytopathogenic microbes by eco-friendly neem leaf extract mediated silver nanoparticles (AgNPs).

In vitro investigation to explore the toxicity of different groups of pesticides for an agronomically important rhizosphere isolate Azotobacter vinelandii.

In this work, an attempt was made to evaluate the effect of pesticides on growth pattern, surface morphology, cell viability and growth regulators of nitrogen fixing soil bacterium. Pesticide tolerant Azotobacter vinelandii strain AZ6 (Accession no. MG028654) was found to tolerate maximum level of pesticide and displayed multifarious PGP activities. At higher concentrations, pesticides triggered cellular/structural damage and reduced the cell viability as clearly shown under SEM and CLSM. With increase in concentration, pesticides exhibited a significant (p < 0.05) decrease in PGP traits of strain AZ6. Among all three groups of pesticides, herbicides glyphosate and atrazine were most toxic. Kitazin, hexaconazole, metalaxyl, glyphosate, quizalofop, atrazine, fipronil, monocrotophos and imidacloprid at 2400, 1800, 1500, 900, 1200, 900, 1800, 2100 and 2700 µg mL-1, respectively, decreased the production of IAA by 19.5 ±â€¯1.9 (61%), 18.1 ±â€¯1.2 (64%), 36.4 ±â€¯3.4 (28%), 13.1 ±â€¯0.8 (74%), 15.6 ±â€¯1.0 (69%), 7.6 ±â€¯0.5 (83%), 11.9 ±â€¯0.8 (76%), 24.7 ±â€¯1.7 (51%) and 32 ±â€¯2.3 (37%) µg mL-1, respectively, over control (50.7 ±â€¯3.6 µg mL-1). A maximum reduction of 8.4 ±â€¯1.2 (46%), 5.8 ±â€¯0.6 (62%) and 4 ±â€¯0.2 (74%) µg mL-1 in 2, 3-DHBA at 300 (1×), 600 (2×) and 900 (3×) µg mL-1 glyphosate, respectively, While, 32.8 ±â€¯2.7 (19%), 27.2 ±â€¯2 (33%) and 21.5 ±â€¯1.3 (47%) µg mL-1, respectively in the production of SA was observed at 300 (1×), 600 (2×) and 900 (3×) µg mL-1 atrazine, respectively. Likewise, with increase in concentration of pesticides, decrease in P solubilization ability and change in pH of broth was detected. The order of pesticide toxicity to PSE (percent decline over control) at highest concentration was: atrazine (45) > kitazin (44) > metalaxyl (43) > monocrotophos (43) > glyphosate (41) > hexaconazole (39) > quizalofop (33) > imidacloprid (31) > fipronil (25). The present study undoubtedly suggests that even at higher doses of pesticides, A. vinelandii maintained secreting plant growth regulators and this property makes this strain agronomically important microbe for enhancing the growth of plants.