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Effect of Glyphosate Herbicide on Nitrogen Fixing Bacteria- Azotobacter Species

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Effect of Glyphosate Herbicide on Nitrogen Fixing Bacteria- Azotobacter Species RESEARCH ARTICLE Effect of Glyphosate Herbicide on Nitrogen Fixing Bacteria- Azotobacter Species Adero VO*, Raju NS, Supreeth M Adero VO, Raju NS, Supreeth M. Effect of Glyphosate Herbicide on glucose agar then these were cultured using Ashby’s mannitol agar. The Nitrogen Fixing Bacteria-Azotobacter Species. J Environ Chem Toxicol bacteria were identified by morphological characterization and biochemical 2020;4(2):1-7. characterization (catalase test) thereafter mass culturing was dine using Luria Bertani hi-veg broth and incubated for 24 hours. Assessment of effect of BACKGROUND: Glyphosate, a broad-spectrum, nonselective, systemic glyphosate on the bacteria was done using agar well method and soil herbicide, exists in the form of acid or salt and the most widely used amendments method. herbicide in the world to control weeds especially grasses. It kills plants by RESULTS: All 5 colonies isolated were gram negative rods to oval or interfering with the synthesis of the aromatic amino acids phenylalanine, coccoid in shape, catalase positive and appeared whitish slimy, raised and tyrosine, and tryptophan in the shikimate pathway by inhibiting the enzyme glistering, orange to red and grey to dark brown in colour. At higher 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), which catalyzes the concentrations, glyphosate 41%, (without dilution) had an inhibitory effect reaction of shikimate-3-phosphate (S3P) and phosphoenolpyruvate to form on the growth of Azotobacter but at field dose it does not inhibit their 5-enolpyruvyl-shikimate-3-phosphate (EPSP). Azotobacter, a gram negative, growth. At field dose or higher doses, (double dose or triple dose) genus of motile, oval or spherical shaped aerobic, free-living soil microbes, glyphosate enhance the growth of Azotobacter. play important role in nitrogen fixation. The use of glyphosate to control CONCLUSION: The use of glyphosate to control weeds at field dose or weeds has been thought to have effects on the growth and survivability of slightly higher doses will have a growth enhancing effect on Azotobacter Azotobacter and hence this study was carried out to assess these effects. bacteria while at higher elevated concentrations it will have growth METHODS: Soil samples were taken from the top layer (0-15 cm) of a inhibiting effect. sugarcane and rice field and Azotobacter species isolated using Ashby’s Key Words: Nitrogen fixing bacteria; Azotobacter; Herbicide; Glyphosate INTRODUCTION ever increasing ranging from 10-90%, with an average of 35 to 40%, for all potential food and fiber crops [5]. ith the rise in population after the preindustrial period, there was Pesticides are classified into different groups based on their chemical W composition including organochlorines, organophosphates, carbamates, need to increase food production to meet the ever-increasing demand for formamidines, thiocyanates, organotins, denitrophenols, synthetic food by the human population. This in turn led to agrarian revolution pyrethroids and antibiotics [6]. Commonly used organophosphate pesticides where farming was taken to the next level through extensive mechanization, are parathion, malathion, methyl parathion, chlorpyrifos, glyphosate, tillage of large tracts of land and increased crop yield in turn [1]. However, diazinon, dichlorvos, phosmet, fenitrothion, tetrachlorvinphos, as this intensification increased, soil potential and fertility drastically azamethiphos, and azinphos-methyl. Among the organophosphate, reduced with the rise in infestation of crops by pests and diseases. glyphosate herbicide is the most widely used in agricultural fields for the Therefore, there was need to control the pests and diseases and also to control of weeds especially the grasses since its inception in the 1970 with improve on soil fertility which gave birth to the emergence of synthetic the main producers being USA followed by China, Japan and the fourth is fertilizers and pesticides [2]. As the global demand for food production India [4]. increased with the ever-rising population, the demand for usage of chemical pesticides also continued to rise. Due to increased pest attacks on crops, Glyphosate about 45% of annual food production was lost and the global consumption of pesticides per year stood at about two million tons and among this, The herbicide N-(phosphonomethyl) glycine, commonly known as Europe is leading in consumption with 45%, followed by USA at 24% and glyphosate, is a broad-spectrum, nonselective, systemic herbicide [7]. It is an the remaining 25% is consumed by the rest of the world. In the Asian organophosphorus compound, specifically a phosphonate. It is used to kill continent countries, China is leading in pesticides consumption closely weeds, especially annual broadleaf weeds and grasses that compete with followed by Korea, Japan then India [3,4]. Indian farmers are using wide crops. It was discovered to be an herbicide by Monsanto chemist John E. ranges of chemical pesticides to limit the losses from pests and diseases, in Franz in 1970. Monsanto brought it to market in 1974 under the trade which insecticides account 73%, herbicide 14%, fungicides 11% and others name roundup. Glyphosate is the most widely used herbicide in the world 2%. Out of these, 40% of the pesticides used are organochlorine and 30% and its demand continues to grow [8]. Farmers quickly adopted it, especially is of organophosphate category with the remaining percentage other forms after Monsanto introduced glyphosate-resistant roundup ready crops, of pesticides [3]. This was due to increased insect pest attack caused mainly enabling farmers to kill weeds without killing their crops [4]. by the prevailing warm humid climatic condition. From the changing Glyphosate has the following properties; chemical formula C H NO P; conditions, it is inevitable that there would be a rise in attack of crops by 3 8 5 molar mass 169.07 g•mol−1; appearance white crystalline powder; density pests and diseases hence reduced. The advances in agricultural sciences to 1.704 g/cm3 (at 20°C); melting point 184.5°C (364.1°F; 457.6 K); boiling date could not reduce the losses due to pests and diseases which have been point, decomposes at 187°C (369°F; 460 K); solubility in water 1.01 g/100 mL (at 20°C) and its acidity (pKa) <2, 2.6, 5.6, 10.6 [9] (Figure 1). Department of Studies in Environmental Sciences, Manasagangothri University of Mysore, Mysuru, India *Correspondence: Victor Onyango Adero, Department of Studies in Environmental Sciences, Manasagangothri University of Mysore, Mysuru 570 006, India, Tel: 0821-2419819/2419885; E-mail: [email protected] Received date: May 22, 2020; Accepted date: June 05, 2020; Published date: June 12, 2020 Copyright: © 2020 Adero VO, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. This open-access article is distributed under the terms of the Creative Commons Attribution Non-Commercial License (CC BY-NC) (http:// creativecommons.org/licenses/by-nc/4.0/), which permits reuse, distribution and reproduction of the article, provided that the original work is properly cited and the reuse is restricted to noncommercial purposes. For commercial reuse, contact [email protected] J Environ Chem Toxicol Vol.4 No.2 2020 1 Adero VO, et al. the soil (nitrogen fixation) [21]. In addition to being a model organism for studying diazotrophs, it is used by humans for the production of biofertilizers, food additives, and some biopolymers. Azotobacter genus is known to fix on an average 10 mg of N/g of carbohydrate under in vitro. A. chroococcum happens to be the dominant inhabitant in arable soils capable of fixing N2 (2-15 mg N2 fixed/g of carbon source) in culture medium. Most efficient strains of Azotobacter would need to oxidize about 1000 kg of organic matter for fixing 30 kg of N/ha. Besides, soil is inhabited by a large variety of other microbes, all of Figure 1) Glyphosate structure. which compete for the active carbon. Plant needs nitrogen for its growth and Azotobacter fixes atmospheric nitrogen non-symbiotically. Therefore, plants get benefited especially cereals, vegetables, fruits, etc. are known to The half-life of glyphosate in soil ranges between 2 to 197 days; with the get additional nitrogen requirements from Azotobacter [22-25]. Owing to typical field half-life of 47 days. Its persistence in the soil is governed by their ability to fix molecular nitrogen and therefore increase the soil fertility several factors but majorly soil type and characteristics and the prevailing and stimulate plant growth, Azotobacter species are widely used in climatic conditions. The median half-life of glyphosate in water varies from agriculture, particularly as nitrogen biofertilizers [26]. a few to 91 days [10]. The objective of the study was to isolate Azotobacter strains from the Glyphosate kills plants by interfering with the synthesis of the aromatic rhizosphere soil, identify and characterize the Azotobacter strains isolated amino acids’ phenylalanine, tyrosine, and tryptophan in the shikimate and to assess the effects of application of glyphosate herbicide on the pathway. It does this by inhibiting the enzyme 5-enolpyruvylshikimate-3- growth and number of free-living nitrogen fixing bacteria; Azotobacter phosphate synthase (EPSPS), which catalyzes the reaction of shikimate-3- species. phosphate (S3P) and phosphoenolpyruvate to form 5-enolpyruvyl- shikimate-3-phosphate (EPSP) [11,12]. It’s absorbed through foliage and MATERIAL AND METHODS minimally through roots, thereby only effective on actively growing plants and cannot prevent seeds from germinating [10]. After application, Isolation and characterization of Azotobacter bacteria from the soil glyphosate is readily transported around the plant to growing roots and leaves and this systemic activity is important for its effectiveness [11,12]. The Soil sample collection inhibition of the enzyme causes shikimate to accumulate in plant tissues Soil samples were taken from the top layer (0-15 cm) of a sugarcane and rice and diverts energy and resources away from other processes.
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