Fast Detection of Auxins by Microplate Technique

Anguiano-Cabello, Julia Cecilia; Flores-Olivas, Alberto; Fuentes, Yisa María Ochoa; Arredondo-Valdés, Roberto; Olalde‐Portugal, Víctor

doi:10.4236/ajps.2017.82013

Cited by 5 publications

(5 citation statements)

References 17 publications

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“…The production of auxins was determined using a microplate method as described by Anguiano-Cabello et al (2017) . For the colorimetric assay, strain 169 was precultivated in R2A growth medium and incubated according to the procedure of Suckstorff and Berg (2003) .…”

Section: Methodsmentioning

confidence: 99%

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Genomic Analysis of the Endophytic Stenotrophomonas Strain 169 Reveals Features Related to Plant-Growth Promotion and Stress Tolerance

et al. 2021

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Plant-associated Stenotrophomonas isolates have great potential for plant growth promotion, especially under stress conditions, due to their ability to promote tolerance to abiotic stresses such as salinity or drought. The endophytic strain Stenotrophomonas sp. 169, isolated from a field-grown poplar, increased the growth of inoculated in vitro plants, with a particular effect on root development, and was able to stimulate the rooting of poplar cuttings in the greenhouse. The strain produced high amounts of the plant growth-stimulating hormone auxin under in vitro conditions. The comparison of the 16S rRNA gene sequences and the phylogenetic analysis of the core genomes showed a close relationship to Stenotrophomonas chelatiphaga and a clear separation from Stenotrophomonas maltophilia. Whole genome sequence analysis revealed functional genes potentially associated with attachment and plant colonization, growth promotion, and stress protection. In detail, an extensive set of genes for twitching motility, chemotaxis, flagella biosynthesis, and the ability to form biofilms, which are connected with host plant colonization, could be identified in the genome of strain 169. The production of indole-3-acetic acid and the presence of genes for auxin biosynthesis pathways and the spermidine pathway could explain the ability to promote plant growth. Furthermore, the genome contained genes encoding for features related to the production of different osmoprotective molecules and enzymes mediating the regulation of stress tolerance and the ability of bacteria to quickly adapt to changing environments. Overall, the results of physiological tests and genome analysis demonstrated the capability of endophytic strain 169 to promote plant growth. In contrast to related species, strain 169 can be considered non-pathogenic and suitable for biotechnology applications.

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Section: Methodsmentioning

confidence: 99%

“…The test is based on the Salkowski’s reagent and is not specific for only IAA, but the indole derivates indole-pyruvate and indole-acetamide could also form complex with the reagent. However, the principal auxin detected is IAA ( Anguiano-Cabello et al, 2017 ).…”

Section: Methodsmentioning

confidence: 99%

Genomic Analysis of the Endophytic Stenotrophomonas Strain 169 Reveals Features Related to Plant-Growth Promotion and Stress Tolerance

et al. 2021

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“…Screening for IAA production was done by using Salkowsky's reagent [32] and concentrations of IAA were determined using a standard curve prepared from pure IAA solutions [33]. Thin layer chromatography [34] was performed with ethanolic extracts of bacterial secondary metabolites.…”

Section: Evaluation Of Pgp Traitsmentioning

confidence: 99%

Drought tolerance induction and growth promotion by indole acetic acid producing Pseudomonas aeruginosa in Vigna radiata

2022

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Drought accompanied with reduced precipitation is one of the key manacles to global agricultural throughput and is expected to escalate further hence posing major challenges to future food safety. For a sustainable agricultural environment, drought resistant plant growth promoting rhizobacteria (PGPR) are new encouraging prospect, which are inexpensive and have no side effects, as those of synthetic fertilizers. In the present study, five strains of Pseudomonas aeruginosa, the strain MK513745, strain MK513746, strain MK513747, strain MK513748, and strain MK513749 were used as drought tolerant PGPR with multiple traits of IAA production, N fixation, P solubilization, siderophore producing capabilities. The strain MK513745 and strain MK513749 produced higher quantities of indole acetic acid (116±0.13 and 108±0.26 μg ml-1). MK513749 yielded 12 different indole compounds in GCMS analysis. The strain MK513748 yielded maximum S.I. (3.33mm) for phosphate solubilizing test. Maximum nitrogen concentration was produced (0.18 μg ml-1) by strain MK513746. Percent siderophore units ranged from 2.65% to 2.83% as all five pseudomonas strains were siderophore positive. In all growth experiments of plant microbe interaction two varieties of Vigna radiata (AZRI-06, NM-11) plants inoculated with P. aeruginosa strains under drought stress responded significantly (P<0.05) better than control stressed plants. Maximum shoot length was enhanced up-to 125%, pod/plant 172%, number of grains 65%, 100 seed weight 95%, 100 seed straw weight 124% and total yield 293% were recorded in plants inoculated with drought stress tolerant PGPR in both varieties as compared to respective stressed control plants. Photosynthetic activity, membrane stability (45%), water content (68%) and antioxidant efficacy (19%) were improved with PGPR inoculations. The variety NM-11 (V2) was more tolerant to drought stress with inoculations of Pseudomonas strains than AZRI-06 (V1). Inoculations with these indole acetic acid producing strains would be suitable for plant growth promotion in areas facing water deficiency.

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“…Further, the mixtures of perchlorates with oxidizable substances are often explosive 33 . These drawbacks of perchloric acid prompted us to explore alternatives, and we came across a study by Anguiano‐Cabello et al, 10 who substituted perchloric acid with sulfuric acid in Salkowski reagent and used it to quantify auxins from B. subtilis strains. This reagent composition enabled efficient auxin estimation from plant samples (Figures 3, 5, and 6); however, unlike the earlier report, the absorption maximum was found to be 540 nm (Figure 1C).…”

Section: Resultsmentioning

confidence: 99%

“…Since then, this method has been extensively used for detecting IAA from microorganisms. Anguiano‐Cabello et al 10 modified this method a bit by replacing perchloric acid with sulfuric acid (H 2 SO 4 ) and employed the new method to quantify auxins from Bacillus subtilis strains. These colorimetric methods of auxin estimation find widespread use in microbe derived auxin detection/estimation studies.…”

Section: Introductionmentioning

confidence: 99%

A rapid and robust colorimetric method for measuring relative abundance of auxins in plant tissues

Manna,

Rengasamy,

Sinha

2024

Phytochemical Analysis

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IntroductionAuxin estimation in plant tissues is a crucial component of auxin signaling studies. Despite the availability of various high‐throughput auxin quantification methods like LC‐MS, GC‐MS, HPLC, biosensors, and DR5‐gus/gfp‐based assays, auxin quantification remains troublesome because these techniques are very expensive and technology intensive and they mostly involve elaborate sample preparation or require the development of transgenic plants.ObjectivesTo find a solution to these problems, we made use of an old auxin detection system to quantify microbe derived auxins and modified it to effectively measure auxin levels in rice plants.Materials and methodsAuxins from different tissues of rice plants, including root samples of seedlings exposed to IAA/TIBA or subjected to different abiotic stresses, were extracted in ethanol. The total auxin level was measured by the presently described colorimetric assay and counterchecked by other auxin estimation methods like LC‐MS or gus staining of DR5‐gus overexpressing lines.ResultsThe presented colorimetric method could measure (1) the auxin levels in different tissues of rice plants, thus identifying the regions of higher auxin abundance, (2) the differential accumulation of auxins in rice roots when auxin or its transport inhibitor was supplied exogenously, and (3) the levels of auxin in roots of rice seedlings subjected to various abiotic stresses. The thus obtained auxin levels correlated well with the auxin levels determined by other methods like LC‐MS or gus staining and the expression pattern of auxin biosynthesis pathway genes.ConclusionsThe auxin estimation method described here is simple, rapid, cost‐effective, and sensitive and allows for the efficient detection of relative auxin abundances in plant tissues.

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Fast Detection of Auxins by Microplate Technique

Cited by 5 publications

References 17 publications

Genomic Analysis of the Endophytic Stenotrophomonas Strain 169 Reveals Features Related to Plant-Growth Promotion and Stress Tolerance

Genomic Analysis of the Endophytic Stenotrophomonas Strain 169 Reveals Features Related to Plant-Growth Promotion and Stress Tolerance

Drought tolerance induction and growth promotion by indole acetic acid producing Pseudomonas aeruginosa in Vigna radiata

A rapid and robust colorimetric method for measuring relative abundance of auxins in plant tissues

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