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Microscale magnetic microparticle-based immunopurification of cytokinins from Arabidopsis root apex
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SYSNO ASEP 0476413 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Microscale magnetic microparticle-based immunopurification of cytokinins from Arabidopsis root apex Author(s) Plačková, Lenka (UEB-Q) ORCID, RID
Oklešťková, Jana (UEB-Q) RID, ORCID, SAI
Pospíšková, K. (CZ)
Poláková, K. (CZ)
Buček, J. (CZ)
Stýskala, J. (CZ)
Zatloukal, M. (CZ)
Šafařík, Ivo (BC-A) RID, ORCID
Zbořil, R. (CZ)
Strnad, Miroslav (UEB-Q) RID, ORCID
Doležal, Karel (UEB-Q) RID, ORCID
Novák, Ondřej (UEB-Q) RID, ORCID, SAINumber of authors 12 Source Title Plant Journal. - : Wiley - ISSN 0960-7412
Roč. 89, č. 5 (2017), s. 1065-1075Number of pages 11 s. Language eng - English Country US - United States Keywords solid-phase extraction ; tandem mass-spectrometry ; endogenous cytokinins ; plant-samples ; cis-zeatin ; quantitative-analysis ; aromatic cytokinins ; abscisic-acid ; oryza-sativa ; purification ; cytokinins ; magnetic microparticles ; immunoaffinity separation ; metabolite profiling ; Arabidopsis thaliana ; technical advance Subject RIV EB - Genetics ; Molecular Biology OECD category Plant sciences, botany R&D Projects LO1204 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LK21306 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GA14-34792S GA ČR - Czech Science Foundation (CSF) Institutional support UEB-Q - RVO:61389030 ; BC-A - RVO:60077344 UT WOS 000395810800017 EID SCOPUS 85013471725 DOI https://doi.org/10.1111/tpj.13443 Annotation Cytokinins (CKs) are pivotal plant hormones that have crucial roles in plant growth and development. However, their isolation and quantification are usually challenging because of their extremely low levels in plant tissues (pmol g(-1) fresh weight). We have developed a simple microscale magnetic immunoaffinity-based method for selective one-step isolation of CKs from very small amounts of plant tissue (less than 0.1 mg fresh weight). The capacity of the immunosorbent and the effect of the complex plant matrix on the yield of the rapid one-step purification were tested using a wide range of CK concentrations. The total recovery range of the new microscale isolation procedure was found to be 30-80% depending on individual CKs. Immunoaffinity extraction using group-specific monoclonal CK antibodies immobilized onto magnetic microparticles was combined with a highly sensitive ultrafast mass spectrometry-based method with a detection limit close to one attomole. This combined approach allowed metabolic profiling of a wide range of naturally occurring CKs (bases, ribosides and N-9-glucosides) in 1.0-mm sections of the Arabidopsis thaliana root meristematic zone. The magnetic immunoaffinity separation method was shown to be a simple and extremely fast procedure requiring minimal amounts of plant tissue. Workplace Institute of Experimental Botany Contact David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Year of Publishing 2018
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