Mechanism and kinetics of chlorpyrifos co-metabolism by using environment restoring microbes isolated from rhizosphere of horticultural crops under subtropics.

The indiscriminate use of organophosphate insecticide chlorpyrifos in agricultural crops causes significant soil and water pollution and poses a serious threat to the global community. In this study, a microbial consortium ERM C-1 containing bacterial strains Pseudomonas putida T7, Pseudomonas aeruginosa M2, Klebsiella pneumoniae M6, and a fungal strain Aspergillus terreus TF1 was developed for the effective degradation of chlorpyrifos. Results revealed that microbial strains were not only utilizing chlorpyrifos (500 mg L-1) but also coupled with plant growth-promoting characteristics and laccase production. PGP traits, that is, IAA (35.53, 45.53, 25.19, and 25.53 µg mL-1), HCN (19.85, 17.85, 12.18, and 9.85 µg mL-1), and ammonium (14.73, 16.73, 8.05, and 10.87 µg mL-1) production, and potassium (49.53, 66.72, 46.14, and 52.72 µg mL-1), phosphate (52.37, 63.89, 33.33, and 71.89 µg mL-1), and zinc (29.75, 49.75, 49.12, and 57.75 µg mL-1) solubilization tests were positive for microbial strains T7, M2, M6, and TF1, respectively. The laccase activity by ERM C-1 was estimated as 37.53, 57.16, and 87.57 enzyme U mL-1 after 5, 10, and 15 days of incubation, respectively. Chlorpyrifos degradation was associated with ERM C-1 and laccase activity, and the degree of enzyme activity was higher in the consortium than in individual strains. The biodegradation study with developed consortium ERM C-1 showed a decreased chlorpyrifos concentration from the 7th day of incubation (65.77% degradation) followed by complete disappearance (100% degradation) after the 30th day of incubation in the MS medium. First-order degradation kinetics with a linear model revealed a high k -day value and low t 1/2 value in ERM C-1. The results of HPLC and GC-MS analysis proved that consortium ERM C-1 was capable of completely removing chlorpyrifos by co-metabolism mechanism.

Exploration of Klebsiella pneumoniae M6 for paclobutrazol degradation, plant growth attributes, and biocontrol action under subtropical ecosystem.

In recent times, injudicious use of paclobutrazol (PBZ) in mango orchards deteriorates the soil quality and fertility by persistence nature and causes a serious ecosystem imbalance. In this study, a new Klebsiella pneumoniae strain M6 (MW228061) was isolated from mango rhizosphere and characterized as a potent plant growth promoter, biocontrol, and PBZ degrading agent. The strain M6 efficiently utilizes PBZ as carbon, energy and nitrogen source and degrades up to 98.28% (50 mgL-1 initial conc.) of PBZ at 15th day of incubation in MS medium. In the soil system first order degradation kinetics and linear model suggested 4.5 days was the theoretical half-life (t1/2 value) of PBZ with strain M6. Box Behnken design (BBD) model of Response surface methodology (RSM) showed pH 7.0, 31°C temperature, and 2.0 ml inoculum size (8 x 109 CFU mL-1) was optimized condition for maximum PBZ degradation with strain M6. Plant growth promoting attributes such as Zn, K, PO4 solubilization IAA, HCN and NH3 production of strain M6 showed positive results and were assessed quantitatively. The relation between plant growth promotion and PBZ degradation was analyzed by heat map, principal component analysis (PCA) and, clustal correlation analysis (CCA). Strain M6 was also showing a significant biocontrol activity against pathogenic fungi such as Fusarium oxysporum (MTCC-284), Colletotrichum gloeosporioides (MTCC- 2190), Pythium aphanidermatum (MTCC- 1024), Tropical race 1 (TR -1), and Tropical race 4 (TR -4). Hence, results of the study suggested that strain M6 can be utilized as an effective bio-agent to restore degraded land affected by persistent use of paclobutrazol.

Mechanisms and kinetics for the degradation of paclobutrazol and biocontrol action of a novel Pseudomonas putida strain T7.

The present study explores biodegradation kinetics and process optimization of plant growth retardant from triazole group paclobutrazol (PBZ; C15H20ClN3O mol. wt. 293.79 g mol-1) in a batch experiment. A gram-negative rod-shaped bacterium T7 was isolated from PBZ applied agricultural field by enrichment technique and characterized as Pseudomonas putida strain T7. Strain was tested for PBZ biodegradation and plant growth-promoting characteristics. Results revealed that strain T7 utilizes PBZ as a carbon and energy source and showing degradation up to 98.30% on the 15th day. First-order degradation kinetics and a linear model were well fitted and showing a maximum t1/2 value on 9th day. Biodegradation optimization by Box Behnken design (BBD) of Response surface methodology (RSM) showed maximum degradation at pH 7.0, 31 °C temperature, and 2 mL inoculum size (8 × 109 CFU mL-1). The bacterium was also able to solubilize Zn, K, and PO4 and produced a copious amount of IAA, HCN, and Ammonia. The biocontrol activity against plant pathogens like Fusarium oxysporum (MTCC-284), Colletotrichum gloeosporioides (MTCC 2190), Pythium aphanidermatum (MTCC - 1024), Tropical race-1 (TR -1), and Tropical race - 4 (TR-4) showed the great antagonistic effect. Hence, this strain can be employed as an effective bio-agent for eco-friendly cleanup strategies and pathogen suppressive agents in paclobutrazol contaminated soil.

Influence of imidacloprid on bacterial community diversity of mango orchard soil assessed through 16S rRNA sequencing-based metagenomic analysis.

Imidacloprid, used against mango hopper, is a persistent insecticide in soil. Microbes have the ability to remove toxic pesticides from soil surface. Metagenomic is an approach for understanding the diversity and related metabolic activities in any environmental sample without culturing the microbes. Metagenomic analysis of mango orchard soil was carried out using 16S rRNA gene sequencing to understand the impact of imidacloprid on soil microbial population. In control and imidacloprid applied soil samples, representative sequences clustered were 0.142930 and 0.082320 million, respectively. At the kingdom level, 85 and 88 percent represented to bacteria, 2 and 1 percent to archaea, and 13 and 11 percent to unassigned for control and treated metagenomes, respectively. At phylum level, 16 and 17 percent of OTUs (operational taxonomic units) were assigned with Proteobacteria, while 13 and 11 percent of OTUs were unassigned in control and imidacloprid-treated samples, respectively. The other abundant phyla in both the samples were Planctomycetes, Bacteroidetes, and Actinobacteria. At class level, 9 and 11 percent of OTUs were assigned with Planctomycetia in control as well as imidacloprid-treated samples, respectively. A number of OTUs present in control and imidacloprid applied samples are 31,173 and 21,909, respectively, with 18,018 number of OTUs shared between the two samples. The genus Gemmata totally disappeared in imidacloprid applied soil, while those belonging to class Phycisphaerae, genus Prevotella and species copri were identified in imidacloprid treatment. Bacterial community transformation was evident from this study indicating possible microbial bioremediation of imidacloprid in mango orchard soil.

Dissipation kinetics and risk assessment of thiamethoxam and dimethoate in mango.

Thiamethoxam and dimethoate are two insecticides used to control hoppers and inflorescence midges in mango. Thiamethoxam (0.008 and 0.016%) and dimethoate (0.06 and 0.12%) were sprayed on Dashehari mango trees during the pre-mature stage of fruit (first week of May) to study their dissipation kinetics and risk assessment in mango fruit. Thiamethoxam dissipated in fruit from 1.93 and 3.71 mg kg(-1) after 2 h of spraying to 0.08 and 0.13 mg kg(-1) after 20 days of spraying at single and double doses, respectively. Its residue did not persist beyond 20 days in fruit. Dimethoate dissipated in fruit from 2.81 and 5.34 mg kg(-1) after 2 h of application to 0.12 and 0.19 mg kg(-1) after 10 days of application at single and double doses, respectively. No residue was detected in fruit beyond 10 days after its application. Both ready-to-harvest mature mango fruit and pulp (after 40 days of spraying) were free from any residues of these insecticides at both the concentration levels. The rate of dissipation of these insecticides followed first-order kinetics in fruit with residual half-lives of 4.0 to 4.5 days for thiamethoxam and 2 days for dimethoate. Based on their MRL values of 0.5 and 2.0 mg kg(-1) in mango, pre-harvest intervals of 7 and 11 days, and 6 and 7 days were suggested for thiamethoxam and dimethoate, respectively, after spraying at single and double doses. The theoretical maximum residue contribution (TMRC) values for both the insecticides, calculated for residues corresponding to each sampling date, were found to be below the maximum permissible intake (MPI) values on mango fruit (except for dimethoate double dose up to 3 days); hence, both thiamethoxam and dimethoate could be considered non-hazardous to consumers at the above doses and time intervals.

Persistence behavior of imidacloprid and carbosulfan in mango (Mangifera indica L.).

Imidacloprid was sprayed on mango cv. Dashehari at 0.3 mL L(-1) of water during pre-bloom stage with 6-8 cm panicle size (first week of March) to control hopper and carbosulfan was sprayed at 2.0 mL L(-1) of water in the trees of mango hybrid (H-1000) during fruit development stage (first week of May) to control leaf webber. Residues of both the insecticides were analysed in peel, pulp and fruit at different stages of fruit development and maturity. The initial residues of imidacloprid, after 30 days of spraying, were 1.21, 0.56 and 1.77 mg kg(-1) in peel, pulp and whole fruit, respectively. The residues persisted in peel for 60 days and in pulp for 50 days and dissipated with a half-life of 38 days. Mature Dashehari fruits at harvest (after 85 days of spraying) were free from imidacloprid residues. Carbosulfan in mango peel dissipated from 5.30 mg kg(-1) (after 1 h of spraying) to 0.05 mg kg(-1) at the time of harvest (after 45 days of spraying). Carbosulfan residue in pulp was very low (0.08 mg kg(-1)) after 1 h of spraying, which increased gradually to 0.90 mg kg(-1) after 10 days and finally came down to 0.04 mg kg(-1) after 26 days of spraying. The insecticide residue was not detected in the pulp at the time of harvest. The residues persisted in pulp for 26 days and in peel for 45 days and degraded with a half-life of 7 days. The dissipation of both imidacloprid and carbosulfan followed first order rate kinetics in whole fruit (peel + pulp). Therefore, the safe pre-harvest intervals were suggested to be 55 days for imidacloprid and 46 days for carbosulfan before consumption of mango fruits after spraying of these insecticides.

Simultaneous estimation of tribenuron-methyl and its major metabolites by high-performance liquid chromatography.

A rapid reversed phase HPLC method was developed for the separation of tribenuron-methyl and its three major metabolites (methyl-2-aminosulphonyl benzoate, 4-methoxy-6-methyl-2-methylamino-1,3,5-triazine and saccharin) using RP-18 column and methanol-water (60:40, v/v) as mobile phase. The limit of detection of all the four compounds was 0.01 ppm.