Plant Hormones Analysis Service

Plant Metabolomics, as a research field focusing on plants, primarily investigates the metabolic changes of plants in different growth and developmental stages or in response to various external stimuli. This discipline utilizes metabolomics techniques to comprehensively study the variations in metabolites. It has found widespread applications in the realms of plant biology and related fields.

Plant hormones, also known as phytohormones, are endogenously synthesized compounds in plants. They play crucial roles in regulating responses to external environmental changes, modulating growth states, resisting adverse conditions, and maintaining essential metabolic processes. Phytohormones are of significant importance in plant growth and development, pest and disease resistance, breeding, and protection. Consequently, there is an increasing interest in metabolomics techniques specifically designed for the targeted quantification of plant hormones.

Why Is It Necessary To Detect Plant Hormones?

On one hand, plant hormones play a crucial regulatory role in plant growth, development, and responses to the environment. On the other hand, as essential agricultural and horticultural products, plants can significantly improve crop yield and quality through the precise regulation of plant hormones (such as auxins, gibberellins, and cytokinins), which can influence plant architecture and yield composition. Hormones like jasmonic acid, salicylic acid, and brassinosteroids also play pivotal roles in plant defense against pests and diseases. Therefore, the qualitative and quantitative analysis of plant hormones is essential for understanding hormone action mechanisms, plant life processes, improving crop and horticultural plant quality, and targeted genetic modification.

Challenges in Plant Hormone Detection

Plant hormones are secondary metabolites in plants and exist in extremely low quantities. They are typically present in the range of nanograms (ng) or even picograms (pg) per gram of fresh plant material.

Substantial variations in hormone content can exist among different samples, with fresher samples often yielding better results.

Matrix effects can cause significant interference. Matrix refers to components in the sample other than the analyte, which can disrupt the analysis and affect the accuracy of the results; this interference is known as the matrix effect.

Plant Hormone Detection Methods

Currently, there are three main categories of plant hormone testing methods: Enzyme-Linked Immunosorbent Assay (ELISA), High-Performance Liquid Chromatography (HPLC), and Liquid Chromatography-Mass Spectrometry (LC-MS/MS).

ELISA is the simplest and most cost-effective method, offering a short detection cycle. However, its precision is lower compared to the other two methods. It is suitable for large-scale screening when a limited budget is available. Nevertheless, its acceptance in high-level journals for hormone determination is decreasing, and it is less suitable for low-concentration hormone detection.

HPLC, in combination with various detectors, can directly analyze multiple plant hormones and is one of the widely used methods. HPLC offers higher sensitivity than ELISA and can detect a broader range of hormones, including IAA, ABA, and SA, among others.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) integrates liquid chromatography's exceptional separation capabilities with mass spectrometry's remarkable specificity and sensitivity. LC-MS/MS attains unparalleled accuracy, achieving detection precision at the picogram level. This methodology is particularly well-suited for discerning clients who demand meticulous data accuracy and offers comprehensive coverage for the detection of nearly all known plant hormones.

Workflow

Da Cao et al,. Frontiers in Plant Science 2020

Our Plant Hormone Quantification Service

With state-of-the-art high-specificity, high-resolution mass spectrometry instrumentation and an expert bioinformatics team, our company has effectively developed the proficiency to quantitatively assess numerous plant hormones across diverse plant sample types. We provide a comprehensive research service encompassing sample preparation, analytical testing, and bioinformatics analysis. We have curated a comprehensive library of targeted plant hormones, employing internal standards for precise quantification of nine primary classes and 107 distinct categories of plant hormones. These encompass Auxin, Cytokinin (CK) , Jasmonic Acid (JA), Salicylic Acid (SA), Abscisic Acid (ABA), Gibberellic Acid (GA), 1-Aminocyclopropane-1-carboxylic Acid (ACC), Brassinosteroids (BR), and Strigolactones (SL).

Service Advantages

Comprehensive Service: Our service encompasses the full spectrum of plant sample handling, testing, and rigorous bioinformatics analysis. Leveraging advanced technology and extensive expertise, we provide a holistic solution.

Swift Testing Turnaround: Employing batch-based liquid-liquid extraction for sample pre-processing translates to a remarkably brief testing cycle.

Unwavering Precision: Utilizing UPLC-MS/MS enables us to achieve meticulous quantification, coupled with stringent quality control practices, ensuring scientifically sound outcomes.

Comprehensive Substance Profiling: Our focus extends to nine primary classes of plant hormones, with a particular emphasis on functionally significant ones.

Gold Standard Quantification: We adhere to the gold standard by employing isotope internal standards for absolute quantification.

Unparalleled Specificity: Our meticulously optimized pre-processing techniques ensure an exceptional level of specificity for plant hormones. We pair this with suitable chromatographic and mass spectrometry detection conditions.

Remarkable Sensitivity: Our high-sensitivity mass spectrometry system empowers us to detect hormones at levels as low as picograms and even femtograms, guaranteeing the precise quantification of exceedingly minute hormone concentrations.

Platform

UHPLC-QQQ-MS (Exion LC -Sciex QTRAP® 6500+)

Typical Chromatogram for Simultaneous Detection of Multiple Plant Hormones

Customer Case: Lina Duan, Juan Manuel Pérez-Ruiz, Francisco Javier Cejudo, José R Dinneny, "Characterization of CYCLOPHILLIN38 shows that a photosynthesis-derived systemic signal controls lateral root emergence

Journal: Plant Physiology
Published: 2021

Method: Targeted Quantification of Plant Hormones

Abstract

The research illuminates a critical aspect of plant physiology, shedding light on the intricate relationship between photosynthesis and root architecture, specifically in the context of sucrose and auxin metabolism.

By meticulously conducting mutant screens and delving into the Arabidopsis thaliana CYCLOPHILIN 38 (CYP38) gene, the study uncovers the profound impact of photosynthesis on lateral root emergence. The findings, highlighting the role of CYP38 in stabilizing photosystem II (PSII) and its non-cell-autonomous influence on root development, significantly contribute to our understanding of plant growth mechanisms.

Moreover, the observation that low-light conditions and perturbations in photosynthetic activity affect lateral root emergence independently of shoot size underscores the importance of these insights. The elucidation of the interplay between sucrose, auxin biosynthesis, and CYP38-dependent photosynthetic activity in root architecture refinement under limited light conditions represents a noteworthy advancement in plant science.

Results

CYP38 Mutation Alters Root Development

A recessive mutant (presto) with an 85.4% reduction in lateral root (LR) density and a 51% reduction in primary root (PR) growth was identified. This mutant carried a CYCLOPHILIN 38 (CYP38) gene mutation, confirmed through mapping and complementation. Pre-emergence stage LRs were significantly enriched, indicating LR initiation disruption. Despite larger shoot sizes in mutants, LR emergence defects persisted, suggesting specific root development alterations. These findings highlight the critical role of CYP38 in root architecture, independent of overall plant growth.

CYP38 mutation reduces LR density, alters LR stages, and affects PR length in Arabidopsis.

CYP38 Primarily Localized to Chloroplasts

CYP38, known for its role in PSII assembly and stability, exhibits predominant expression in shoot tissues, with limited expression in roots. Subcellular localization studies confirm its exclusive presence in chloroplasts, emphasizing its chloroplast-specific function in Arabidopsis.

CYP38 is highly expressed in shoots and localized in plastids.

CYP38 Regulates LR Emergence via Shoot Signals

Grafting experiments reveal that LR growth defects are primarily determined by the shoot genotype. CYP38 mRNA and protein show limited movement across the shoot–root junction. Despite low transcript levels in the rootstock, LR development is rescued, highlighting CYP38's non-autonomous role in LR emergence from photosynthetic shoot tissues.

The activity of CYP38 in shoots is essential for LR emergence.

Auxin Downregulation Mediates cyp38 Root Phenotype

Shoot-derived auxin plays a crucial role in LR emergence. The cyp38 mutant and low-light conditions lead to reduced IAA levels in roots, primarily affecting distal mature tissues. Transcriptional reporter assays show decreased auxin response in cyp38 mutants and under low light, particularly in cortex cells surrounding LRP. Exogenous IAA application rescues LR emergence in both cyp38 mutants and low-light-treated WT seedlings, suggesting that limited photosynthetic output may hinder auxin biosynthesis from IPA conversion. These findings highlight the role of auxin biosynthesis and signaling in mediating the cyp38 root phenotype under altered light conditions.

Auxin is involved in CYP38-dependent regulation of LRs.

Conclusions

In summary, the study elucidates the role of photosynthesis in shaping root system architecture, focusing on CYP38 function. The author discovered that shoot-derived systemic signals influence a distinct LR development stage, operating independently of known root-regulating signals like sucrose, HY5, or β-cyclocitral. Instead, their findings propose a connection between auxin biosynthesis and cellular redox status, serving as a long-distance signal to control LR emergence.

Reference

1. Lina Duan, Juan Manuel Pérez-Ruiz, Francisco Javier Cejudo, José R Dinneny, Characterization of CYCLOPHILLIN38 shows that a photosynthesis-derived systemic signal controls lateral root emergence, Plant Physiology, Volume 185, Issue 2, February 2021, Pages 503–518.