Cytokinin (CTK) Analysis Service

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Overview

Cytokinins are N⁶-substituted adenine derivatives that have been implicated in numerous aspects of plant growth and development, including cell seed germination, de-etiolation, apical dominance, chloroplast differentiation, sink/source relationships, nutrient uptake, flower and fruit development, leaf senescence, and response to biotic and abiotic factors. The most common cytokinins have isoprenoid side chains, but there are also adenine derivatives with aromatic substituents, as well as synthetic cytokinins derived from diphenylurea (DPU). The structure and conformation of side chains are vital to the activity of cytokinins. For example, trans-zeatin is one of the most abundant cytokinins present in higher plants that displays a high activity in bioassays, but the cis-isomer displays a significantly lower activity. Determination of the distribution and content of the various cytokinins in a given plant tissue provides valuable insights into the molecular mechanisms underlying the action of cytokinins.

Figure 1. Structures of cytokinins (Kieber et al., 2014).

Applications of Cytokinin Analysis

Reveal the involvement of CTKs in plant growth and development
Elucidate the mechanism of action and molecular biological effects of CTKs
Ensure food safety
Crop improvement and management

Advantages of Our Cytokinin Analysis Service

Effective sample preprocessing offers high recovery rates

Enable measurement of IP, IPA, 6-BA, KT, tZR, tZ.

Accurate detection and quantification of CTKs with LC-MS platforms

Customizable bioinformatics analysis

End-to-end CTK analysis with comparable prices

Service Workflow

Creative Proteomics has optimized sample preprocessing methods to ensure efficiency and ultrahigh recovery rates consistently exceeding 85%. Mass spectrometry combined with our optimized sample preprocessing methods and other instrumentation offers precise detection down to ultra-trace levels.

Figure 2. Cytokinins analysis service workflow.

Quantification methods: external reference method or isotope-labeled internal standard method

Mode: MRM, capable of simultaneously detecting more than 1000 MRM ion pairs

Precision: ≤10^-9 g

Positive/Negative polarity switching time: ≤20 ms, allowing for the acquisition of Q1/Q3 MRM transition mass spectra in both ionization modes from a single LC-MS/MS run.

Analysis content:

Standard curve creation
Raw data preprocessing
Absolute quantitation of cytokinins
Differential metabolites screening
Optimal analyses such as KEGG pathway analysis and hierarchical clustering

List of Detectable Cytokinins at Creative Proteomics

Detectable CTKs	CAS	Quantification Methods
N6-(Δ2-isopentenyl)-adenine (IP)	2365-40-4	External reference method / isotope-labeled internal standard method
IPA (isopentenyladenosine)	7724-76-7	External reference method / isotope-labeled internal standard method
Benzylaminopurine (6-BA)	1214-39-7	External reference method / isotope-labeled internal standard method
Kinetin (KT)	525-79-1	External reference method / isotope-labeled internal standard method
trans-zeatin ribotide (tZR)	6025-53-2	External reference method / isotope-labeled internal standard method
(trans-zeatin) tZ	1637-39-4	External reference method / isotope-labeled internal standard method

Sample Requirements

1. Fresh plant tissues: >1 g or > 0.2 mL. Provide young plant tissues for best results. Samples should be frozen in liquid nitrogen immediately after collection, and then transferred to -80°C for storage.

2. Plant seeds.

At least 3 biological replicates.

Deliverables

Experimental procedure
Parameters of HPLC and MS
MS raw data files and MS data quality checks
CTK quantification data
Custom analysis report

Creative Proteomics provides high-throughput detection and quantitation of cytokinins based on state-of-the-art LC-MS platforms in a fast and sensitive manner. Our service offers maximized recovery rates and accurate profiling of cytokinins, as well as tailored statistical analysis and bioinformatics analysis to achieve reliable cytokinins analysis results.

References

Kieber J J, Schaller G E. Cytokinins. The Arabidopsis Book/American Society of Plant Biologists, 2014, 12.
Haberer G, Kieber J J. Cytokinins. New insights into a classic phytohormone. Plant Physiology, 2002, 128(2): 354-362.
Ma Q H. Genetic engineering of cytokinins and their application to agriculture. Critical reviews in biotechnology, 2008, 28(3): 213-232.