Cytological characterization of an Aspergillus nidulans mutant from a strain with chromosomic duplication

A development mutant, named V103, was obtained spontaneously from the A strain of A. nidulans. The A strain contains a duplicated segment of chromosome I that has undergone translocation to chromosome II (I ¨ II). It is mitotically unstable and generates phenotypically deteriorated types, some with enhanced stability. The deteriorated variants of A. nidulans show abnormal development, exhibiting slower colony growth, variations in colony pigmentation and changes in conidiophore structure. The alterations observed in the conidiophore include fewer metulae and phialides, further elongation and ramification of these structures, delayed nuclear migration and the presence of secondary conidiophores.

Mechanism of sterigmatocystin biosynthesis regulation by pH in Aspergillus nidulans

External pH constitutes one of the most important environmental factors that control growth, metabolism and differentiation in microorganisms, including fungi. We have analyzed the effect of external pH on sterigmatocystin biosynthesis in Aspergillus nidulans. It was observed in repeated experiments that alkaline pH, in opposition to acid pH, increased sterigmatocystin production and the transcript levels of aflR, the master gene that regulates expression of the sterigmatocystin cluster in A. nidulans. It is known that pH effects in fungi operate mostly through the Pal/Pac signaling pathway, originally described in Aspergillus nidulans. Accordingly, we studied the role of this signaling pathway in ST biosynthesis. It was observed that aflR transcript levels were increased in the "alkalinity mimicking" mutant pacCc14 and were minimal in the "acidity mimicking" mutant palA1. No sterigmatocystin was produced by palA1 or pacC- mutants at neither acid or alkaline pH of incubation. Finally, fluG and flbA, genes known to regulate both conidiation and sterigmatocystin synthesis upstream in the regulatory cascade, were up-regulated at alkaline pH.

Identification of nutrient-dependent changes in extracellular pH and acid phosphatase secretion in Aspergillus nidulans

The present study was designed to identify nutrient-dependent changes in extracellular pH and acid phosphatase secretion in the biA1 palC4 mutant strain of Aspergillus nidulans. The palC4 mutant was selected as lacking alkaline phosphatase, but having substantially increased acid phosphatase activity when grown on solid minimal medium under phosphate starvation, pH 6.5. Gene palC was identified as a putative member of a conserved signaling cascade involved in ambient alkaline sensing whose sole function is to promote the proteolytic activation of PacC at alkaline pH. We showed that both poor growth and conidiation of the palC4 mutant strain on solid medium, alkaline pH, were relative to its hypersensitivity to Tris (hydroxymethyl) aminomethane buffer. Also, the secretion of acid phosphatase was repressed when both the wild-type and palC4 mutant strains were grown in low-phosphate yeast extract liquid medium, pH 5.0, indicating that the secretion of this enzyme is not necessary to regenerate inorganic phosphate from the organic phosphate pool present in yeast extract.

Parasexuality in asexual development mutants of Aspergillus nidulans

The parasexual cycle with parameiosis has been characterized previously by the occurrence of genetic recombination and haploidization inside heterokaryotic hyphae prior to conidial formation. The aim of current research was to characterize, through genetic and cytological analyses, an asexual development mutant strain of A. nidulans and to use it to obtain parameiotic segregants. Analyses showed the medusa phenotype of the B84 strain, whose mutant allele was mapped in the chromosome I. The heterokaryons B84(med)//G422(med+) and B84(med)//G839(brl) were formed in liquid MM+2% CM and inoculated in the appropriate selective media. Two mitotic segregant groups were obtained: aneuploids and haploid stable recombinants. Mitotic segregants, wild-types, and developmental mutants, which did not produce new visible mitotic sectors in the presence of Benomyl and which showed normal meiotic behavior during the sexual cycle, were classified as parameiotics.