The pcz1 Gene, which Encodes a Zn(II)2Cys6 Protein, Is Involved in the Control of Growth, Conidiation, and Conidial Germination in the Filamentous Fungus Penicillium roqueforti

Abstract

Proteins containing Zn(II)2Cys6 domains are exclusively found in fungi and yeasts. Genes encoding this class of proteins are broadly distributed in fungi, but few of them have been functionally characterized. In this work, we have characterized a gene from the filamentous fungus Penicillium roqueforti that encodes a Zn(II)2Cys6 protein, whose function to date remains unknown. We have named this gene pcz1. We showed that the expression of pcz1 is negatively regulated in a P. roqueforti strain containing a dominant active Gαi protein, suggesting that pcz1 encodes a downstream effector that is negatively controlled by Gαi. More interestingly, the silencing of pcz1 in P. roqueforti using RNAi-silencing technology resulted in decreased apical growth, the promotion of conidial germination (even in the absence of a carbon source), and the strong repression of conidiation, concomitant with the downregulation of the genes of the central conidiation pathway brlA, abaA and wetA. A model for the participation of pcz1 in these physiological processes in P. roqueforti is proposed.

Changes in the Sclerotinia sclerotiorum transcriptome during infection of Brassica napus

“…The Penicillium roqueforti Pcz1 gene encodes Zn2 Cys6 TF which contributes to the regulation of growth, conidiation, and conidial germination [130]. BcYOH1 from B. cinerea encodes a Cys2 His2 TF which is involved in the regulation of secondary metabolite synthesis [131].…”

Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium roqueforti

“…The protocol described by Gil-Duran et al [ 16 ] was used for DNA isolation. For total RNA purification, the strains were grown on YES agar (Bacto Yeast Extract (Difco, USA) 20 g/l, sucrose (Merck, Germany, biochemical grade) 150 g/l and Bacto Agar (Difco, USA) 20 g/l) for 7 days at 28°C.…”

“…For total RNA purification, the strains were grown on YES agar (Bacto Yeast Extract (Difco, USA) 20 g/l, sucrose (Merck, Germany, biochemical grade) 150 g/l and Bacto Agar (Difco, USA) 20 g/l) for 7 days at 28°C. Total RNA was isolated exactly as described previously [ 16 ]. RT-PCR experiments were performed as described by Ravanal et al [ 17 ].…”

“…RNA-mediated gene silencing technology has been successfully used to silence genes in P . roqueforti [ 16 , 19 ]. Here we used the same experimental approach.…”

“…roqueforti transformants were obtained by introducing the genetic constructs described above into strain CECT 2905 by protoplast transformation. Protoplast obtainment, transformation, selection of transformants on Czapek-sorbitol medium containing phleomycin, and obtainment of homokaryotic strains were carried out exactly as described before [ 16 ].…”