Analysis of indole-3-acetic acid (IAA) production in Klebsiella by LC-MS/MS and the Salkowski method

Gang, Shraddha; Sharma, Sheetal; Saraf, Meenu; Buck, Martin; Schumacher, Julia

doi:10.21769/bioprotoc.3230

Cited by 100 publications

(61 citation statements)

References 11 publications

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“…A laminar airflow, an autoclave, an Erlenmeyer flask, an oven, a hot plate, a bunsen burner, a spatula, an incubator, a shaker, a spectrophotometer, a petri dish, a micropipette, a microtube, a microscope, a centrifuge, a 1 mL syringe, and a digital camera were the tools used in this study. Materials used were soybean rhizospheric soil samples, Nicotiana tobaccum L., alcohol 70%, primer 63F and 1387R, nutrient broth (NB) medium, nutrient agar (NA) medium, Pikovskaya medium containing 10 g glucose; Ca3 (PO4)2, 5 g; (NH4)2SO4, 0,5 g; NaCl 0,2 g; MgSO4.7H2O, 0,1 g; KCl, 0,2 g; yeast extract 0,5 g; MnSO4.H2O, 0,002 g; FeSO4.7H2O, 0,002 g (Nautiyal, 1999); Salkowski reagent (150 ml H2SO4, 7,5 ml FeCl3.6H2O 0,5 M, and 250 ml sterile aqua dest), and L-Tryptophan (Gang et al 2019).…”

Section: Methodsmentioning

confidence: 99%

Pseudomonas plecoglossicida as a novel bacterium for phosphate solubilizing and indole-3-acetic acid-producing from soybean rhizospheric soils of East Java, Indonesia

et al. 2020

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Abstract. Astriani M, Zubaidah S, Abadi AL, Suarsini E. 2020. Pseudomonas plecoglossicida as a novel bacterium for phosphate solubilizing and indole-3-acetic acid-producing from soybean rhizospheric soils of East Java, Indonesia. Biodiversitas 21: 578-586. The use of synthetic fertilizers to grow soybean (Glycine max (L.) Merrill) in a long time, can increase the risk of environmental damage. Therefore, the current study aimed to find phosphate solubilizing bacteria that can produce indole-3-acetic acid (IAA) hormone to minimize the use of chemical fertilizers. Superior isolates selected through characterization of phosphate solubilization activity on Pikovskaya medium, screening of IAA producing bacteria, quantitative estimation of phosphate solubilizing activity using a spectrophotometer, hypersensitivity assay, antagonist within isolate assay, and molecular identification of selected bacterial isolates using 16S rRNA sequencing with primer forward 63f and primer reverse 1387r. Among the isolated bacteria, isolate Arj8 showed the highest phosphate solubilizing activity and IAA production. Molecular identification indicated that isolate Arj8 shared 100% similarity with Pseudomonas plecoglossicida. The highest phosphate solubilizing activity (75.39 mg/L) and IAA production (38.89 ppm) recorded on day-3. Multiple potentialities of P. plecoglossicida as phosphate solubilizing and IAA producing bacterium are a novel finding in the development of bioinoculants as bio-fertilizers that can reduce dependency on synthetic chemical fertilizers.

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Section: Methodsmentioning

confidence: 99%

Pseudomonas plecoglossicida as a novel bacterium for phosphate solubilizing and indole-3-acetic acid-producing from soybean rhizospheric soils of East Java, Indonesia

et al. 2020

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“…Indole-3-acetic acid (IAA) production from L-tryptophan by Methylobacterium strains was detected through use of the Salkowski reagent and quanti ed using a common spectrophotometric technique for measuring IAA production that is of microbial origin (Ivanova et al 2001;Kim et al 2017;Gang et al 2019). Without L-tryptophan supplementation no endogenous IAA was detected; however, with Ltryptophan, the total IAA concentration ranged from 0 to 67.47 µmol mL −1 .…”

Section: Discussionmentioning

confidence: 99%

A Survey of Methylobacterium Species and Strains Reveals Widespread Production and Varying Profiles of Cytokinin Phytohormones

Palberg¹,

Kisiała²,

Jorge³

et al. 2021

Preprint

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BackgroundSymbiotic Methylobacterium strains comprise a significant part of plant microbiomes. Their presence enhances plant productivity and stress resistance, prompting classification of these strains as plant growth-promoting bacteria (PGPB). Methylobacteria can synthesize unusually high levels of plant hormones, called cytokinins (CKs), including the most active form, trans-Zeatin (tZ). ResultsThis study provides a comprehensive inventory of 46 representatives of Methylobacterium genus with respect to phytohormone production in vitro, including 16 CK forms, abscisic acid (ABA) and indole-3-acetic acid (IAA). High performance-liquid chromatography - tandem mass spectrometry (HPLC-MS/MS) analyses revealed varying abilities of Methylobacterium strains to secrete phytohormones that ranged from 5.09 to 191.47 pmol mL-1 for total CKs, and 0.46 to 82.16 pmol mL-1 for tZ. Results indicate that reduced methanol availability, the sole carbon source for bacteria in the medium, stimulates CK secretion by Methylobacterium. Additionally, select strains were able to transform L-tryptophan into IAA while no ABA production was detected.ConclusionsTo better understand features of CKs in plants, this study uncovers CK profiles of Methylobacterium that are instrumental in microbe selection for effective biofertilizer formulations.

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“…IAA production and estimation was carried out following the method given by Gang et al [ 46 ]. IAA biosynthesis in the cultures was induced by supplementing nutrient broth with 0.1% ( w / v ) L-tryptophan, and cultures were incubated in the dark at 30 °C on an orbital shaker at 200 rpm.…”

Section: Methodsmentioning

confidence: 99%

Bacterial Indole-3-Acetic Acid Influences Soil Nitrogen Acquisition in Barley and Chickpea

Gang

Saraf

Buck

et al. 2021

Plants

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Farming of barley and chickpea is nitrogen (N) fertilizer dependent. Using strategies that increase the nitrogen use efficiency (NUE) and its components, nitrogen uptake efficiency (NUpE) and nitrogen utilization efficiency (NUtE) would reduce the N fertilizer application in the soil and its adverse environmental effects. We evaluated the effects of three different strains of diazotroph Klebsiella (K.p. SSN1, K.q. SGM81, and K.o. M5a1) to understand the role of biological nitrogen fixation (BNF) and bacterial indole-3-acetic acid (IAA) on NUE of the plants. A field study revealed that K.p. SSN1 results in profound increment of root surface area by eightfold and threefold compared to uninoculated (control) in barley and chickpea, respectively. We measured significant increase in the plant tissue nitrogen, chlorophyll content, protein content, nitrate reductase activity, and nitrate concentration in the inoculated plants (p ≤ 0.05). Treated barley and chickpea exhibited higher NUE and the components compared to the control plants (K.p. SSN1 ≥ K.q. SGM81> K.o. M5a1). Specifically, K.q. SGM81 treatment in barley increased NUpE by 72%, while in chickpea, K.p. SSN1 increased it by 187%. The substantial improvement in the NUpE and NUE by the auxin producers K.p. SSN1 and K.q. SGM81 compared with non-auxin producer K.o. M5a1 was accompanied by an augmented root architecture suggesting larger contribution of IAA over marginal contribution of BNF in nitrogen acquisition from the soil.

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Analysis of indole-3-acetic acid (IAA) production in Klebsiella by LC-MS/MS and the Salkowski method

Cited by 100 publications

References 11 publications

Pseudomonas plecoglossicida as a novel bacterium for phosphate solubilizing and indole-3-acetic acid-producing from soybean rhizospheric soils of East Java, Indonesia

Pseudomonas plecoglossicida as a novel bacterium for phosphate solubilizing and indole-3-acetic acid-producing from soybean rhizospheric soils of East Java, Indonesia

A Survey of Methylobacterium Species and Strains Reveals Widespread Production and Varying Profiles of Cytokinin Phytohormones

Bacterial Indole-3-Acetic Acid Influences Soil Nitrogen Acquisition in Barley and Chickpea

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