Photomorphogenesis in Phycomyces: differential display of gene expression by PCR with arbitrary primers

Abstract

The zygomycete fungus Phycomyces blakesleeanus develops two types of fruiting bodies of very different size, macrophores and microphores. Blue light stimulates macrophorogenesis and inhibits microphorogenesis. I have adapted a method based on the polymerase chain reaction with arbitrary primers to investigate the role of differential gene expression during photophorogenesis in Phycomyces. Several cDNAs for genes induced in vegetative mycelium have been observed, but only one gene induced by blue light has been detected. I have demonstrated the feasibility of this approach by the isolation of a cDNA segment for the heat-shock protein HSP100 that is induced by blue light at the onset of sporangiophore development. The heat-shock protein HSP100 is an ATP-binding protein that has the ability to disassemble protein complexes. In plants, the gene for HSP100 is induced by light. The cDNA segment for HSP100 obtained from Phycomyces is 686 bp long and the predicted amino acid sequence contains one of the ATP-binding sites.

Involvement of G Proteins in the Mycelial Photoresponses of Phycomyces¶

“…This fragment is part of the third chsl (chitin synthase 1) exon(37). 32 Plabeled carB and chsl probes were prepared by random priming(38).…”

“…Pharmacological and biochemical results have suggested various protein kinase‐phosphatases as blue‐light signaling components in both responses or a nitric oxide synthase in sporangiophore development (30,31). It was only recently that a pure molecular approach enabled the partial cloning of another blue‐light‐induced putative component, a heat shock protein, expressed at the onset of sporangiophore development(32).…”

Photoregulated tyrosinase gene in <i>Polyporus arcularius</i>

“…The light governs many developmental transitions and accompanying expression of genes in the life cycles of fungi (Kumagai 1988;Bell-Pedersen et al 1996;Linden et al 1997;Kues et al 1998;Kues 2000;Corrochano 2002;Froehlich et al 2002;He et al 2002). The basidiomycete Polyporus arcularius is a polypore mushroom that needs light for two stages of formation of the fruiting body.…”

Regulation by blue light and heat shock of gene transcription in the fungus Phycomyces: proteins required for photoinduction and mechanism for adaptation to light

“…The development of sporangiophores in Phycomyces should bring about differential gene expression that could be modulated by blue‐light. In order to investigate the role of differential gene expression during sporangiophore development, we adapted a method based on the polymerase chain reaction (PCR) with arbitrary primers that allowed the isolation of hspA , the Phycomyces gene for the heat‐shock protein HSP100 (Corrochano, 2002; Rodríguez‐Romero and Corrochano, 2004). HSP100s are ATP‐binding proteins with the ability to disassemble protein complexes and are involved in the tolerance to high temperatures, proteolysis, and the regulation of gene transcription (Schirmer et al ., 1996; Lee et al ., 2004; Maurizi and Xia, 2004).…”