title: Simultaneous detection and quantification of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) produced by rhizobacteria from l-tryptophan (Trp) using HPTLC.
abstract: A simple, quick and reliable method is proposed for the detection and quantitation of indole-3-acetate (IAA) and indole-3-butyrate (IBA), an auxin phytohormone produced by rhizobacteria from l-tryptophan (Trp) metabolism using high performance thin-layer chromatography (HPTLC). Microbial auxin biosynthesis routes involve Trp as a precursor where other than IAA and IBA, products such as indole-3-pyruvate (IPA), indole-3-acetamide (IAM), tryptamine, indole-3-acetonitrile (IAN), indole-3-lactic acid (ILA) and indole-3-acetaldehyde (IAAld) are also produced. In traditional spectrophotometric method, Salkowski reagent develops color by reacting with indolic compounds. The color development is non-specific contributed by several Trp derivatives produced by rhizobacteria rather than IAA only. To overcome this limitation, HPTLC based protocol is developed to precisely detect and quantify IAA and IBA in the range of 100 to 1000ng per spot. This protocol is applicable to detect and quantify IAA and IBA from microbial samples ignoring other Trp derivatives. For microbial samples, the spectrophotometric method gives larger values as compared to HPTLC derived values which may be attributed by total indolic compounds reacting with Salkowski reagent rather than only IAA and/or IBA.
authors.affiliation: Department of Biotechnology, P.D. Patel Institute of Applied Sciences, Charotar University of Science and Technology, CHARUSAT Campus, Changa-388421, Anand, Gujarat, India.
authors.forename: Dweipayan
authors.initials: D
authors.lastname: Goswami
authors.affiliation: Department of Biotechnology, P.D. Patel Institute of Applied Sciences, Charotar University of Science and Technology, CHARUSAT Campus, Changa-388421, Anand, Gujarat, India.
authors.forename: Janki N
authors.initials: JN
authors.lastname: Thakker
authors.affiliation: Ashok and Rita Patel Institute of Integrated Study and Research in Biotechnology and Allied Sciences, ADIT Campus, New Vidyangar, 388121 Anand, Gujarat, India. Electronic address: pinakindhandhukia@aribas.edu.in.
authors.forename: Pinakin C
authors.initials: PC
authors.lastname: Dhandhukia
mesh_terms: D001407:Bacillus / Q000378:metabolism*; D002855:Chromatography, Thin Layer / Q000379:methods*; D007210:Indoleacetic Acids / Q000032:analysis / Q000378:metabolism; D007211:Indoles / Q000032:analysis* / Q000378:metabolism; D057230:Limit of Detection; D008835:Micrococcaceae / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D012231:Rhizobium / Q000378:metabolism*; D013053:Spectrophotometry; D014363:Tryptamines / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism*
chemical_list: D007210:Indoleacetic Acids; D007211:Indoles; D014363:Tryptamines; C014612:indolebutyric acid; C001655:indole-3-acetaldehyde; C030820:tryptamine; C024139:indole-3-lactic acid; C015950:indoleacetamide; D014364:Tryptophan; C016516:indole-3-acetonitrile
keywords: High performance thin layer chromatography (HPTLC); Indole-3-acetate (IAA); Indole-3-butyrate (IBA); Plant growth promoting rhizobacteria (PGPR); Tryptophan (Trp) derivatives