Autofluorescence as a Signal to Sort Developing Glandular Trichomes by Flow Cytometry

Area of Science:

• Plant Biology
• Molecular Biology
• Biochemistry

Background:

• Plant glandular trichomes (GTs) produce valuable metabolites, driving research into their function.
• Existing research primarily focuses on secondary metabolite pathways, with limited understanding of GT development and differentiation.
• Studying GT development requires stage-specific molecular and metabolic data, necessitating efficient isolation of trichome subtypes.

Purpose of the Study:

• To develop a method for isolating distinct developmental stages of glandular trichomes (GTs).
• To enable stage-specific transcriptomic and metabolomic analyses of GTs.
• To provide a broadly applicable technique for isolating specific plant cell types.

Main Methods:

• A novel method was established for separating and isolating intact young and mature type VI trichomes from Solanum habrochaites.
• The key step involves cell sorting using flow cytometry, exploiting distinct autofluorescence signals between young and mature trichomes.
• The purity and viability of sorted trichomes were confirmed for downstream molecular analyses.

Main Results:

• Flow cytometry successfully isolated pure fractions of young and mature glandular trichomes (GTs).
• The sorted trichomes are suitable for subsequent transcript and metabolite analyses.
• The autofluorescence-based sorting method demonstrates high efficiency and purity.

Conclusions:

• A robust method for isolating distinct GT developmental stages using autofluorescence and flow cytometry has been developed.
• This technique facilitates in-depth studies of GT development, differentiation, and the regulation of specialized metabolism.
• The principles of this method are transferable to isolating other plant cell types based on autofluorescence properties.