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. •Imidacloprid and thiamethoxam significantly inhibi