Pesticides including insecticides are often applied to prevent distortion posed by plant insect pests. However, the application of these chemicals detrimentally affected the non-target organisms including soil biota. Fipronil (FIP), a broad-spectrum insecticide, is extensively used to control pests across the globe. The frequent usage calls for attention regarding risk assessment of undesirable effects on non-target microorganisms. Here, laboratory-based experiments were conducted to assess the effect of FIP on plant-beneficial bacteria (PBB); Rhizobium leguminosarum (Acc. No. PQ578652), Azotobacter salinestris (Acc. No. PQ578649) and Serratia marcescens (Acc. No. PQ578651). PBB synthesized growth regulating substances were negatively affected by increasing fipronil concentrations. For instance, at 100 mu g FIPmL-1, a decrease in indole-3-acetic acid (IAA) synthesis by bacterial strains followed the order: A. salinestris (95.6%) S. marcescens (91.6%) > R. leguminosarum (87%). Also, exposure of bacteria cells to FIP hindered the growth and morphology of PBB observed as distortion, cracking, and aberrant structure under scanning electron microscopy (SEM). Moreover, FIP-treated and propidium iodide (PI)-stained bacterial cells displayed an insecticide dose-dependent increase in cellular permeability as observed under a confocal laser microscope (CLSM). Colony counts (log(10) CFU mL(-1)) and growth of A. salinestris was completely inhibited at 150 mu g FIPmL-1. The surface adhering ability (biofilm formation) of PBB was also disrupted/inhibited in a FIP dose-related manner. The respiration loss due to FIP was coupled with a reduction in population size. Fipronil at 150 mu gmL(-1) decreased cellular respiration in A. salinestris (72%) S. marcescens (53%) and R. leguminosarum (85%). Additionally, biomarker enzymes; lactate dehydrogenase (LDH), lipid peroxidation (LPO), and oxidative stress (catalase; CAT) induced by FIP represented significant (p <= 0.05) toxicity towards PBB strains. Conclusively, fipronil suggests a toxic effect that emphasizes their careful monitoring in soils before application and their optimum addition in the soil-plant system. It is high time to prepare both target-specific and slow-released agrochemical formulation for crop protection with concurrent safeguarding of soils.

Pesticides including insecticides are applied in agricultural practices to control insect pests. However, their excessive usage often poses a severe threat to the growth, physiology, and biochemistry of plants. Here, responses of chickpea and greengram seedlings exposed to three fipronil (FIP) concentrations i. e. 100 (1x), 200 (2x) and 300 (3x) mu g mL- 1 was evaluated under in vitro. Among doses, 3x had a greater negative impact on germination attributes, root-shoot elongation, vigor indices, length ratios, and survival of seedlings. Besides, the morphological distortion in root tips, oxidative stress generation, and cellular death in fipronil-supplemented root seedlings were observed under scanning electron (SEM) and confocal laser scanning (CLSM), respectively. A significant (p <= 0.05) and pronounced upsurge in plant stressor metabolites such as proline, malondialdehyde (MDA), electrolyte leakage (EL), hydrogen peroxide (H2O2) content, and antioxidants enzymes in plant seedlings further confirmed the fipronil toxicity. In addition, a concentration-dependent decrease in respiration efficiency (RE) and ATP content in FIP-treated seedlings was observed. Reduced mitotic index (MI) and numerous chromosomal anomalies (CAs) in root meristematic cells of seedlings are a clear indication of insecticide-induced cytotoxicity. Furthermore, a dose-dependent increase in DNA damage in root meristematic cells of greengram revealed the genotoxic potential of fipronil. Conclusively, fipronil suggested phyto and cyto-genotoxic effects that emphasize their careful monitoring in soils before application and their optimum addition in soil-plant systems. It is high time to prepare both target-specific and slow-released agrochemical formulations for crop protection with concurrent safeguarding of the soil.