Truncation of MAT1-2-7 Deregulates Developmental Pathways Associated with Sexual Reproduction in Huntiella omanensis

Abstract

ABSTRACT The MAT1-1-1 and MAT1-2-1 genes are thought to be the master regulators of sexual development in most ascomycete fungi, and they are often essential for this process. In contrast, it has been suggested that the secondary mating-type genes act to calibrate the sexual cycle and can be dispensable. Recent functional characterization of genes such as Aspergillus fumigatus MAT1-2-4, Huntiella omanensis MAT1-2-7, and Botrytis cinerea MAT1-1-5 has, however, shown that these secondary genes may play more central roles in the sexual pathway and are essential for the production of mature fruiting structures. We used a comparative transcriptome sequencing (RNA-seq) experiment to show that the truncation of MAT1-2-7 in the wood inhabiting H. omanensis residing in the Ceratocystidaceae is associated with the differential expression of approximately 25% of all the genes present in the genome, including the transcriptional regulators ste12, wc-2, sub1, VeA, HMG8, and pro1. This suggests that MAT1-2-7 may act as a transcription factor and that ΔMAT1-2-7 mutant sterility is the result of layered deregulation of a variety of signaling and developmental pathways. This study is one of only a few that details the functional characterization of a secondary MAT gene in a nonmodel species. Given that this gene is present in other Ceratocystidaceae species and that there are diverse secondary MAT genes present throughout the Pezizomycotina, further investigation into this gene and others like it will provide a clearer understanding of sexual development in these eukaryotes. IMPORTANCE Secondary mating-type genes are being described almost as quickly as new fungal genomes are being sequenced. Understanding the functions of these genes has lagged behind their description, in part due to limited taxonomic distribution, lack of conserved functional domains, and difficulties with regard to genetic manipulation protocols. This study aimed to address this by investigating a novel mating-type gene, MAT1-2-7, for which two independent mutant strains were generated in a previous study. We characterized the molecular response to the truncation of this gene in a nonmodel, wood-infecting fungus and showed that it resulted in widespread differential expression throughout the transcriptome of this fungus. This suggests that secondary MAT genes may play a more important role than previously thought. This study also emphasizes the need for further research into the life cycles of nonmodel fungi, which often exhibit unique features that are very different from the systems understood from model species.

Keywords

• CRISPR-Cas9
• functional characterization
• fungal genetics
• MAT gene
• MAT locus
• MAT1-2-7