Three-in-one method for high throughput plant multi-omics.

Area of Science:

• Plant molecular biology
• Biochemistry
• Genomics

Background:

• Multi-omics analysis in plant single-cell research is crucial for understanding cellular molecular networks.
• Non-simultaneous molecular extractions introduce variability, data inaccuracies, and resource waste in multi-omics studies.

Purpose of the Study:

• To develop and optimize a streamlined protocol for simultaneous extraction of RNA, metabolites, and proteins from plant single cell-type samples.
• To overcome limitations of non-simultaneous extraction methods in multi-omics sample preparation.

Main Methods:

• Adapted a commercial RNA extraction kit with modifications for simultaneous multi-omics analysis.
• RNA is column-bound, metabolites are extracted in polar solvent, and proteins are acetone-precipitated.
• Developed an all-in-one workflow compatible with standard RNA kits.

Main Results:

• Achieved high-quality starting materials for downstream sequencing, metabolomics (LC-MS/MS), and proteomics.
• The protocol is simple, requires minimal training, and is adaptable to various plant species and sample amounts.
• Successfully demonstrated simultaneous extraction of three key molecular types from single plant cells.

Conclusions:

• The optimized 3-in-1 protocol offers an efficient and reliable method for plant single-cell multi-omics.
• This workflow minimizes sample variation and resource expenditure, advancing plant single-cell research.
• The protocol's ease of use and adaptability make it valuable for diverse plant science applications.