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Updated: May 26, 2026
Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants
Published on: August 5, 2020
The application of laser microdissection to profiling fungal pathogen gene expression in planta
Wei-Hua Tang1, Yan Zhang, Jon Duvick
1National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Abstract:
Laser microdissection (LM) of plant tissues infected with a fluorescent protein-tagged fungus is a useful method for obtaining samples highly enriched in fungal RNA for downstream analysis such as hybridization to a microarray. This paper outlines the requirements for successful LM of infected tissues and details a set of protocols for (1) preparing and sectioning infected tissue samples under conditions that preserve both RNA integrity and cytological features; (2) capturing fungal structures via LM; and (3) extraction and amplification of transcripts for further analysis.
Insights
Laser microdissection (LM) effectively isolates fungal RNA from infected plant tissues. This method ensures high-quality RNA for downstream analyses like microarrays, preserving cellular details.
Area of Science:
- Plant pathology
- Molecular biology
- Biotechnology
Background:
- Studying fungal infections in plants requires isolating pathogen-specific molecules.
- Traditional methods may not efficiently separate fungal RNA from host plant RNA.
- Fluorescent protein tagging aids in visualizing and isolating fungal components.
Purpose of the Study:
- To establish protocols for laser microdissection (LM) of fungal-infected plant tissues.
- To optimize sample preparation for preserving RNA integrity and cellular morphology.
- To enable efficient isolation and amplification of fungal RNA for molecular analysis.
Main Methods:
- Developing protocols for preparing and sectioning infected plant tissues.
- Utilizing laser microdissection to capture fluorescently tagged fungal structures.
- Implementing RNA extraction and amplification techniques for isolated fungal transcripts.
Main Results:
- Successful preparation and sectioning of infected tissues maintaining RNA integrity and cytological features.
- Efficient capture of fungal structures using laser microdissection.
- Obtained amplified fungal RNA suitable for downstream applications like microarrays.
Conclusions:
- Laser microdissection is a valuable technique for obtaining highly purified fungal RNA from infected plant tissues.
- The outlined protocols facilitate RNA isolation for gene expression studies in plant-fungal interactions.
- This method enhances the study of fungal pathogens by providing enriched RNA samples.
