Molecular Diversity of Chlorpyrifos Degrading Bacteria Isolated from Apple Orchard Soils of Himachal Pradesh

Anish Kumar Sharma,Khyati Harkhani,Jyotsana Pandit

doi:10.22207/jpam.15.2.31

Abstract

A total of seventy-two bacterial isolates were obtained employing enrichment culture technique from apple orchard soils contaminated with chlorpyrifos. Pure cultures of bacterial isolates were obtained using streak plate method on mineral salt medium. Bacterial isolates were characterized on the basis of morphology, culture and biochemical properties. Six bacterial isolates exhibited high extracellular organophosphorus hydrolase activity along with high tolerance towards high concentrations of chlorpyrifos. Genomic DNA extraction from bacterial isolates was done with phenol/chloroform method. Molecular Diversity of six chlorpyrifos degrading bacterial isolates was done employing RAPD-PCR technique by using 25 decamer primers, where amplification was showed by only 20 primers. A total of 337 amplified bands and 64 unique bands ranging in size from 100 to 4900bps were produced after RAPD analysis. The similarity coefficient estimated by Jaccard’s coefficient for these bacterial isolates was found to range between 31 to 64 percent.

Highlights

Organophosphate pesticides (OPs) are used excessively in agriculture and nearly constitute 38 percent of total pesticides usage worldwide[1].OPs are reported to be biodegradable in nature but at the same time they are toxic to mammals as well as non-target vertebrates &
Chlorpyrifos is an example of organophosphate pesticide which is found to be very effective against broad spectrum of insect pests because chlorpyrifos is capable of irreversibly inhibiting acetylcholinesterase (AChE) enzyme in these pests[2]
In the present study soil samples were collected in triplicates from top soil of apple orchards of Himachal Pradesh

Summary

Introduction

Organophosphate pesticides (OPs) are used excessively in agriculture and nearly constitute 38 percent of total pesticides usage worldwide[1].OPs are reported to be biodegradable in nature but at the same time they are toxic to mammals as well as non-target vertebrates &non-vertebrates. Organophosphate pesticides (OPs) are used excessively in agriculture and nearly constitute 38 percent of total pesticides usage worldwide[1].OPs are reported to be biodegradable in nature but at the same time they are toxic to mammals as well as non-target vertebrates &. Farmer’s uses back-pack reservoirs for pesticide application andhave low safety knowledge due to which they are at high risk of exposure to pesticides. Agrochemicals and their degradation products remains in soil and it has been reported that only a small percentage of total quantity of agrochemical used is involved in the control of pests and diseases

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Methods

Results