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Development of Metarhizium anisopliae as a Mycoinsecticide: From Isolation to Field Performance
Published on: July 30, 2017
Colony sectorization of Metarhizium anisopliae is a sign of ageing
Chengshu Wang1,2, Tariq M Butt1, Raymond J St Leger2
1School of Biological Sciences, University of Wales Swansea, Swansea SA2 8PP, UK.
Abstract:
Spontaneous phenotypic degeneration resulting in sterile sectors is frequently observed when culturing filamentous fungi on artificial medium. Sterile sectors from two different strains of the insect pathogenic fungus Metarhizium anisopliae were investigated and found to contain reduced levels of cAMP and destruxins (insecticidal peptides). Microarray analysis using slides printed with 1730 clones showed that compared to wild-type, sterile sectors down-regulated 759 genes and upregulated 27 genes during growth in Sabouraud glucose broth or on insect cuticle. The differentially expressed genes are largely involved in cell metabolism (18.8 %), cell structure and function (13.6 %) and protein metabolism (8.8 %). Strong oxidative stress was demonstrated in sectorial cultures using the nitro blue tetrazolium assay and these cultures show other syndromes associated with ageing, including mitochondrial DNA alterations. However, genes involved in deoxidation and self-protection (e.g. heat-shock proteins, HSPs) were also upregulated. Further evidence of physiological adaptation by the degenerative sectorial cultures included cell-structure reorganization and the employment of additional signalling pathways. In spite of their very similar appearance, microarray analysis identified 181 genes differentially expressed between the two sectors, and the addition of exogenous cAMP only restored conidiation in one of them. Most of the differentially expressed genes were involved in catabolic or anabolic pathways, but the latter included genes for sporulation. Compared to the mammalian ageing process, sectorization in M. anisopliae showed many similarities, including similar patterns of cAMP production, oxidative stress responses and the involvement of HSPs. Thus, a common molecular machinery for ageing may exist throughout the eukaryotes.
Insights
Phenotypic degeneration in Metarhion anisopliae leads to sterile sectors with reduced cyclic adenosine monophosphate (cAMP) and insecticidal destruxins. These sectors exhibit ageing-like symptoms and altered gene expression, suggesting common eukaryotic ageing mechanisms.
Area of Science:
- Mycology
- Molecular Biology
- Cell Biology
Background:
- Spontaneous phenotypic degeneration is common in filamentous fungi cultured on artificial media.
- Sterile sectors in Metarhizium anisopliae, an insect pathogenic fungus, exhibit reduced levels of cyclic adenosine monophosphate (cAMP) and insecticidal destruxins.
Purpose of the Study:
- To investigate the molecular mechanisms underlying phenotypic degeneration in M. anisopliae sterile sectors.
- To compare the ageing process in M. anisopliae with mammalian ageing.
Main Methods:
- Microarray analysis of gene expression in sterile sectors versus wild-type.
- Nitro blue tetrazolium assay to assess oxidative stress.
- Analysis of mitochondrial DNA alterations.
Main Results:
- Sterile sectors showed significant down-regulation of 759 genes and up-regulation of 27 genes compared to wild-type.
- Genes involved in cell metabolism, structure, function, and protein metabolism were differentially expressed.
- Strong oxidative stress, mitochondrial DNA alterations, and upregulation of heat-shock proteins (HSPs) were observed in sectorial cultures.
- Microarray analysis identified 181 differentially expressed genes between two distinct sterile sectors.
Conclusions:
- Sectorization in M. anisopliae shares similarities with mammalian ageing, including cAMP production patterns, oxidative stress responses, and HSP involvement.
- A common molecular machinery for ageing may exist across eukaryotes.
- Exogenous cAMP partially restored conidiation in only one of the two investigated sterile sectors.

