The Zn(II)2Cys6-Type Transcription Factor ADA-6 Regulates Conidiation, Sexual Development, and Oxidative Stress Response in Neurospora crassa

Abstract

Conidiation and sexual development are critical for reproduction, dispersal and better-adapted survival in many filamentous fungi. The Neurospora crassa gene ada-6 encodes a Zn(II)2Cys6-type transcription factor, whose deletion resulted in reduced conidial production and female sterility. In this study, we confirmed the positive contribution of ada-6 to conidiation and sexual development by detailed phenotypic characterization of its deletion mutant and the complemented mutant. To understand the regulatory mechanisms of ADA-6 in conidiation and sexual development, transcriptomic profiles generated by RNA-seq from the Δ ada-6 mutant and wild type during conidiation and sexual development were compared. During conidial development, differential expressed genes (DEGs) between the Δ ada-6 mutant and wild type are mainly involved in oxidation-reduction process and single-organism metabolic process. Several conidiation related genes are positively regulated by ADA-6, including genes that positively regulate conidiation ( fluffy and acon-3 ), and genes preferentially expressed during conidial development ( eas , con-6 , con-8 , con-10 , con-13 , pcp-1 , and NCU9357), as the expression of these genes were lower in the Δ ada-6 mutant compared to wild type during conidial development. Phenotypic observation of deletion mutants for other genes with unknown function down-regulated by ada-6 deletion revealed that deletion mutants for four genes (NCU00929, NCU05260, NCU00116, and NCU04813) produced less conidia than wild type. Deletion of ada-6 resulted in female sterility, which might be due to that ADA-6 affects oxidation-reduction process and transmembrane transport process, and positively regulates the transcription of pre-2 , poi-2 , and NCU05832, three key genes participating in sexual development. In both conidiation and the sexual development process, ADA-6 regulates the transcription of cat-3 and other genes participating in reactive oxygen species production according to RNA-seq data, indicating a role of ADA-6 in oxidative stress response. This was further confirmed by the results that deletion of ada-6 led to hypersensitivity to oxidants H 2 O 2 and menadione. Together, these results proved that ADA-6, as a global regulator, plays a crucial role in conidiation, sexual development, and oxidative stress response of N. crassa .

Glucose-lactose mixture feeds in industry-like conditions: a gene regulatory network analysis on the hyperproducing Trichoderma reesei strain Rut-C30

“…We also note that in [63], authors show that Tr -MedA is repressed by CRE-1 in QM9414 ∆cre1, thus suggesting a role of the partial carbon catabolite derepression regarding the observed down-regulation of ID 123713. In N. crassa, acon-3 is positively regulated by the transcription factor ADA-6 involved in conidiation, sexual developement, and oxidative stress response [60]. Interestingly, ypr1 (ID 102499), the yellow pigment regulator, DE in our data, displayed 35 % identity with ada-6 .…”

Glucose-lactose mixture feeds in industry-like conditions: a gene regulatory network analysis on the hyperproducingTrichoderma reeseistrain Rut-C30

“…We determined that the N. crassa ortholog of MedA is acon-3 , a gene required for early conidiophore development and female fertility. In N. crassa , acon-3 is positively regulated by the transcription factor ADA-6 involved in conidiation, sexual developement, and oxidative stress response [58]. Interestingly, ypr1 (ID 102499), the yellow pigment regulator, DE in our data, displayed 35% identity with ada-6 .…”

“…Cluster , whose median profile exhibits a slight repression across lactose concentrations encompasses transcription factors whose ortholog are involved in the development: Tr –WET-1 (ID 4430, [55]), Tr –PRO1 (ID 76590, [56, 57]) and Tr –ACON-3 (ID 123713, [58]). We recall that the Tr –XXX notation refers to the gene in T. reesei for which the ortholog in an other specie is XXX (see the Functional analysis section in Materials and Methods).…”

A WD40 Protein Encoding Gene Fvcpc2 Positively Regulates Mushroom Development and Yield in Flammulina velutipes

“…We detected the expression of more than 70 genes which were reported in fruiting body development in N. crassa by qRT-PCR (Kim and Nelson, 2005;Sun et al, 2012Sun et al, , 2019Garud, 2013;Chinnici et al, 2014;Lehr et al, 2014;, and found that poi-2, an essential gene for female fertility in N. crassa, had a dramatic response to cpc-2 deletion (more than 100-fold change) after fruiting induction in N. crassa (Figure 5), indicating that CPC-2 regulates transcription of poi-2. To see whether FvCPC2 can functionally replace the role of CPC-2 in transcriptional regulation, transcript levels of poi-2 were analyzed by qPCR in wild type, the cpc-2 strain, the cpc-2; cpc-2 strain and the Fvcpc2; cpc-2 strain.…”