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. 2019 Nov 19;9:17016. doi: 10.1038/s41598-019-53171-9

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Natural occurrence of isopentenyl adenine and kinetin in lysates of Tohama I and knock-out Tohama I Δ1253 strain. Cytokinins from bacterial lysates, prepared as described in Methods, were purified, dried in SpeedVac and dissolved in 100 µl of 5% methanol in water and centrifuged as described in Methods. The supernatant was then analyzed in LC-MS. (A) Extracted ion chromatogram for isopentenyl adenine in sample Tohama I and knock-out Tohama I Δ1253 strain. The chromatographic peak at retention time of 16.4 min. analyzed in mass spectrometry in a positive ion mode with electrospray, resulted in the experimental elemental composition of C₁₀H₁₄N₅ (−2.21 ppm) (lower panel), with a mass of 204.24 Da (arrow) that corresponded to protonated isopentenyl adenine (203.24). (B) Extracted ion chromatogram for kinetin in sample Tohama I and knock-out Tohama I Δ1253 strain. The chromatographic peak at retention time 8.8 min. analyzed as described above gave the experimental elemental composition of C₁₀H₁₀ON₅ (−8.45 ppm) (lower panel), with a mass of 216.08 (arrow) that corresponded to protonated kinetin (215.16). Experiments were performed in triplicate with identical results. The collision energy used for the fragmentation was 26 a.u.