A. fumigatus is the most common airborne pathogen causing fatal mycoses in immunocompromised patients. Polarized growth is one of the critical factors for establishing fungal pathogenesis, but little is known about the genes involved in early polar growth and their regulation. To understand the spatial distribution of polarity related mRNA, we performed spatial gene expression analysis of germlings. A. fumigatus Af293 was cultured in complete medium for 8hr which is the time just before septation. Tip, base, and conidium regions were captured by Laser Microdissection Pressure Catapulting (LMPC) and whole germlings were collected as a reference. Total RNA was extracted and a cDNA library was constructed for each region and for whole germlings. The quality of each cDNA library was confirmed by performing qRT-PCR for highly expressed genes. mRNA sequencing of each library was performed using a Next- Generation Sequencer (454 GS FLX). By comparing each region, we found that more than 1000 mRNAs are asymmetrically localized. To confirm mRNA localization, we performed Fluorescence in situ Hybridization (FISH) with some of the highly expressed genes in each region and found that their distribution was consistent with sequence results. These data suggest that many mRNAs are asymmetrically localized in tip, base, and conidium region of germlings.
Full conference title:
26th Fungal Genetics Conference
- Fungal Genetics Conference 26th (2005)