Evaluating yeast filamentous growth at the single-cell level

Area of Science:

• Microbiology
• Cell Biology
• Genetics

Background:

• Budding yeast exhibits phenotypic plasticity, including filamentous growth, triggered by nutrient scarcity.
• Filamentous growth is a crucial adaptation for yeast survival in nutrient-poor environments.
• Understanding yeast filamentous growth provides insights into microbial adaptation and multicellularity.

Purpose of the Study:

• To describe and compare two established assays for studying yeast filamentous growth: the single-cell invasive growth assay and the pseudohyphal growth assay.
• To highlight the applicability of both assays across different yeast cell types (haploid and diploid).
• To provide researchers with a clear methodology for investigating yeast filamentous growth.

Main Methods:

• Single-cell invasive growth assay: Examines filamentous growth under glucose-limiting conditions, quantifying changes in budding patterns and cell length.
• Pseudohyphal growth assay: Assesses filamentous growth under nitrogen-limiting conditions, revealing patterns in microcolonies and cell subpopulations.
• Both assays can be adapted for use with either haploid or diploid yeast cells.

Main Results:

• The single-cell assay provides quantitative data on budding patterns and cell elongation, with a high percentage of cells responding.
• The pseudohyphal growth assay visualizes filamentous structures in larger cell clusters and adjacent cell groups.
• Both methods are versatile and can be applied to study filamentous growth in both haploid and diploid yeast strains.

Conclusions:

• The single-cell invasive growth and pseudohyphal growth assays are effective methods for studying nutrient-induced filamentous growth in budding yeast.
• These assays offer complementary approaches, providing quantitative and qualitative insights into yeast adaptation.
• The flexibility of both assays for use with haploid and diploid cells broadens their utility in yeast research.