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Microscale magnetic microparticle-based immunopurification of cytokinins from Arabidopsis root apex

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, SAI}
Number of authors	12
Source Title	Plant Journal. - : Wiley - ISSN 0960-7412 Roč. 89, č. 5 (2017), s. 1065-1075
Number 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	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

Number of the records: 1