Improved method for effective screening of ACC (1-aminocyclopropane-1-carboxylate) deaminase producing microorganisms

Aminocyclopropane-1-carboxylate deaminase (ACCD) producing microorganisms support plant growth under a variety of biotic and abiotic stress conditions such as drought, soil salinity, flooding, heavy metal pollution and phyto-pathogen attack. Available screening methods for ACCD give idea only about...

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Veröffentlicht in:Journal of microbiological methods 2016-12, Vol.131, p.102-104
Hauptverfasser: Patil, Chandrashekhar, Suryawanshi, Rahul, Koli, Sunil, Patil, Satish
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Suryawanshi, Rahul
Koli, Sunil
Patil, Satish
description Aminocyclopropane-1-carboxylate deaminase (ACCD) producing microorganisms support plant growth under a variety of biotic and abiotic stress conditions such as drought, soil salinity, flooding, heavy metal pollution and phyto-pathogen attack. Available screening methods for ACCD give idea only about its primary microbial ACCD activity than the actual potential. In the present investigation, we have simply improved screening method by incorporating pH indicator dyes (phenol red and bromothymol blue) in ACC containing medium. This modification is based on the basic principle that ACCD action releases ammonia which can be detected by color change and zone around the bacterial colony. High color intensity and zone around the colony indicates most potent producer, colony showing only a color change indicates moderate potential and no change in colony color indicates least efficiency. Enzymatic bioassays as well as root elongation studies revealed that ACC-deaminase activity of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Bacillus subtilis clearly corresponds to their growth on dye incorporated ACC medium. This method could be used to complement the existing screening methods and to speed up the targeted isolation of agriculturally important microorganisms. [Display omitted] •Effectively screens ACCD producing microorganisms by incorporating pH indicator dye in ACC deaminase Agar medium•Methods give the visible ACCD production potential on the basis of color change around colonies.•Quantitative, ACC deaminase production by spectrophotometric method are in accordance with plate assay results.•Plant root elongation assays also suggested efficacy of plate based screening method.
doi_str_mv 10.1016/j.mimet.2016.10.009
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Available screening methods for ACCD give idea only about its primary microbial ACCD activity than the actual potential. In the present investigation, we have simply improved screening method by incorporating pH indicator dyes (phenol red and bromothymol blue) in ACC containing medium. This modification is based on the basic principle that ACCD action releases ammonia which can be detected by color change and zone around the bacterial colony. High color intensity and zone around the colony indicates most potent producer, colony showing only a color change indicates moderate potential and no change in colony color indicates least efficiency. Enzymatic bioassays as well as root elongation studies revealed that ACC-deaminase activity of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Bacillus subtilis clearly corresponds to their growth on dye incorporated ACC medium. This method could be used to complement the existing screening methods and to speed up the targeted isolation of agriculturally important microorganisms. [Display omitted] •Effectively screens ACCD producing microorganisms by incorporating pH indicator dye in ACC deaminase Agar medium•Methods give the visible ACCD production potential on the basis of color change around colonies.•Quantitative, ACC deaminase production by spectrophotometric method are in accordance with plate assay results.•Plant root elongation assays also suggested efficacy of plate based screening method.</description><identifier>ISSN: 0167-7012</identifier><identifier>EISSN: 1872-8359</identifier><identifier>DOI: 10.1016/j.mimet.2016.10.009</identifier><identifier>PMID: 27769905</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>ACC deaminase ; Agriculture ; Arachis - growth &amp; development ; Arachis - microbiology ; Bacillus subtilis ; Bacteria - classification ; Bacteria - enzymology ; Bacteria - isolation &amp; purification ; Biological Assay ; Bromthymol Blue ; Carbon-Carbon Lyases - analysis ; Carbon-Carbon Lyases - metabolism ; Carbon-Carbon Lyases - pharmacology ; Colony Count, Microbial - methods ; Color ; Environmental Monitoring - methods ; Environmental Pollution ; Enzyme Assays ; Hydrogen-Ion Concentration ; India ; Lycopersicon esculentum - growth &amp; development ; Lycopersicon esculentum - microbiology ; Mass Screening - methods ; PGPR ; pH dye ; Phenol red ; Phenolsulfonphthalein ; Plant Development - drug effects ; Plant Roots - growth &amp; development ; Plant Roots - microbiology ; Pseudomonas aeruginosa ; Rhizosphere ; Screening ; Soil ; Soil Microbiology ; Stenotrophomonas maltophilia ; Stress, Physiological ; Triticum - growth &amp; development ; Triticum - microbiology</subject><ispartof>Journal of microbiological methods, 2016-12, Vol.131, p.102-104</ispartof><rights>2016 Elsevier B.V.</rights><rights>Copyright © 2016 Elsevier B.V. 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Available screening methods for ACCD give idea only about its primary microbial ACCD activity than the actual potential. In the present investigation, we have simply improved screening method by incorporating pH indicator dyes (phenol red and bromothymol blue) in ACC containing medium. This modification is based on the basic principle that ACCD action releases ammonia which can be detected by color change and zone around the bacterial colony. High color intensity and zone around the colony indicates most potent producer, colony showing only a color change indicates moderate potential and no change in colony color indicates least efficiency. Enzymatic bioassays as well as root elongation studies revealed that ACC-deaminase activity of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Bacillus subtilis clearly corresponds to their growth on dye incorporated ACC medium. This method could be used to complement the existing screening methods and to speed up the targeted isolation of agriculturally important microorganisms. 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subjects ACC deaminase
Agriculture
Arachis - growth & development
Arachis - microbiology
Bacillus subtilis
Bacteria - classification
Bacteria - enzymology
Bacteria - isolation & purification
Biological Assay
Bromthymol Blue
Carbon-Carbon Lyases - analysis
Carbon-Carbon Lyases - metabolism
Carbon-Carbon Lyases - pharmacology
Colony Count, Microbial - methods
Color
Environmental Monitoring - methods
Environmental Pollution
Enzyme Assays
Hydrogen-Ion Concentration
India
Lycopersicon esculentum - growth & development
Lycopersicon esculentum - microbiology
Mass Screening - methods
PGPR
pH dye
Phenol red
Phenolsulfonphthalein
Plant Development - drug effects
Plant Roots - growth & development
Plant Roots - microbiology
Pseudomonas aeruginosa
Rhizosphere
Screening
Soil
Soil Microbiology
Stenotrophomonas maltophilia
Stress, Physiological
Triticum - growth & development
Triticum - microbiology
title Improved method for effective screening of ACC (1-aminocyclopropane-1-carboxylate) deaminase producing microorganisms
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