Brassinosteroid (BR) Analysis Service

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Overview

Brassinosteroids (BRs) are plant-specific steroid hormones that play essential roles in a wide range of developmental and physiological processes in plant life, including vascular differentiation, photomorphogenesis, male fertility, flowering senescence, and plant architecture. BRs are also known to regulate abiotic and biotic stress responses. BRs have been detected in almost every part of the plant, like in roots, shoots, buds, leaves, pollen, flower, seeds, fruits. More than 70 BRs have been identified in the plant. The extremely low concentrations of BRs and big differences in the polarity of individual members within the brassinosteroid hormone family can be a problem when detecting and quantifying their levels in plants. To address this problem, liquid chromatography-mass spectrometry (LC-MS) is employed to determine BRs and has been become the most effective BR quantification method. Many efforts have been made to improve this analytical approach, such as optimizing the BR isolation and purification procedure.

Figure 1. Brassinosteroid (BR) interaction with the innate immunity pathways in plants (Yu et al., 2018).

Applications of BR Analysis

Elucidate the regulation of BR biosynthesis and BR perception
Clarify the metabolism of BR hormone family
Understand BR signal transduction
Reveal mechanisms of receptor kinase activation
Enhance crop quality for human utility

Advantages of Our BR Analysis Service

Improved sample preprocessing ensures high recovery rates and effectiveness

Accurate detection and quantification of BL, CS, 6-DeoxoCS

Customizable protocols and data analyses

End-to-end SA analysis with fast turnaround time and comparable price

Service Workflow

Our team of experts has been dedicated to overcoming the major problems involved in the determination of BRs in complex plant matrices. Our newly developed sample preprocessing method ensures safety, environmental protection, effectiveness, and recovery rates consistently exceeding 85%, as well as maximized homogeneity between samples. LC-MS/MS in multiple-reaction monitoring (MRM) is employed for data acquisition.

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 quantification of BL, CS, 6-DeoxoCS
Differential metabolites screening
Optimal analyses such as KEGG pathway analysis and hierarchical clustering

List of Detectable BRs at Creative Proteomics

Detectable BRs	CAS	Quantification Methods
Brassinolide (BL)	72962-43-7	External reference method / isotope-labeled internal standard method
Castasterone (CS)	80736-41-0	External reference method / isotope-labeled internal standard method
6-deoxocastasterone (6-DeoxoCS)	87833-54-3	External reference method / isotope-labeled internal standard method

Sample Requirements

1. Fresh plant tissues from leaf, flower, stem, root, or fruit: >2 g. 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: >2 g.

At least 3 biological replicates.

Deliverables

Experimental procedure
Parameters of liquid chromatography and MS
MS raw data files and MS data quality checks
BL, CS and 6-DeoxoCS quantification data
Custom data analysis report

Creative Proteomics provides comprehensive BR analyses for precise determination of hormones within the brassinosteroid hormone family, including brassinolide (BL), castasterone (CS), 6-deoxocastasterone (6-DeoxoCS), powered by state-of-the-art mass spectrometry coupled with liquid chromatography (LC). We are committed to clarifying the roles and functions of BRs in plant developmental and physiological processes.

References

Oklestkova J, Tarkowská D, Eyer L, et al. Immunoaffinity chromatography combined with tandem mass spectrometry: A new tool for the selective capture and analysis of brassinosteroid plant hormones. Talanta, 2017, 170: 432-440.
Tang W, Deng Z, Wang Z Y. Proteomics shed light on the brassinosteroid signaling mechanisms. Current opinion in plant biology, 2010, 13(1): 27-33.
Deng Z, Zhang X, Tang W, et al. A proteomics study of brassinosteroid response in Arabidopsis. Molecular & Cellular Proteomics, 2007, 6(12): 2058-2071.
Yu M H, Zhao Z Z, He J X. Brassinosteroid signaling in plant–microbe interactions. International Journal of Molecular Sciences, 2018, 19(12): 4091.