In Enzalutamide purchase contrast to the wild type,
the AfuNce102 deletion mutant showed a low frequency of conidiophores after 16 h of incubation (Fig. 2d). The size of conidiophores and the number of spores per conidiophore were reduced markedly, and instead, a large number of undifferentiated aerial hyphae were produced. However, after 2 days, conidiophores were visible at the colony margin with a low density in the colony center. The mutant was also not able to produce any conidia at room temperature in minimal medium (Fig. 2b). Despite the conidiation abnormalities, the growth of mutant under a range of conditions such as variable carbon and nitrogen sources and differing incubation temperatures (30, 37, and 42 °C) were examined. The results showed no significant difference in growth under these conditions when compared with the wild type, indicating that the AfuNce102 is not involved selleck in the growth of A. fumigatus under tested conditions. Germination studies of wild-type and deletant spores in SAB or MM liquid medium confirmed a
similar pattern of germination time and the frequency of germinated spores (data not shown). Conidiophore development can be triggered by various environmental signals, and the brlA gene acts as a key regulator in this process (Adams et al., 1988). To check if the brlA expression has been affected by AfuNce102 deletion, the transcription level of brlA was measured after 16 and 24 h incubation of both mutant and parent strains in minimal medium using semi-quantitative RT-PCR. The results indicated that the lack of AfuNce102 function did not influence the transcriptional level of brlA (data not shown). It has been proposed that fluG gene as the most upstream component of FluG pathway is responsible for the synthesis of a low molecular weight extracellular factor that can activate the fungal sporulation program (Lee & Adams, 1994; Wieser & Adams, 1995). As the contiguous cultivation of fluG deletant and the wild-type strain have resulted in complementation of the fluG defect in the mutant, we tested the possible suppression of conidiation defect in AfuNce102 deletion mutant by growing the strain next to the wild IMP dehydrogenase type on minimal medium agar. The results demonstrated
that the conidiation abnormality in AfuNce102 deletion mutant was not suppressed when it was grown next to the wild-type strain (data not shown). MIC levels against a range of known antifungal drugs or chemical compounds were determined to test their effect on the AfuNce102 mutant. No difference in MIC between the wild type and the mutant was observed for itraconazole, hygromycin B, nystatin, and calcofluor white; however, the mutant showed an eightfold increase in sensitivity to the sphingolipid synthesis blocker, Myriocin t, compared with the parental strain (MIC values: 25 μg mL−1 for mutant and 200 μg mL−1 for parent strain). The AfuNce102 deletion mutant was transformed with a 3.5-kb PCR product containing AfuNce102 and 5′ and 3′ flanking regions.