A rapid method for measuring in vitro growth in entomopathogenic fungi

Slowik, Anna R. ORCID: https://orcid.org/0000-0003-3079-3868; Hesketh, Helen ORCID: https://orcid.org/0000-0003-1794-7658; Sait, Steven M.; de Fine Licht, Henrik H.. 2023 A rapid method for measuring in vitro growth in entomopathogenic fungi [in special issue: Insect–pathogen interactions in mass-reared insects] Insects, 14 (8), 703. 14, pp. https://doi.org/10.3390/insects14080703

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Official URL: http://dx.doi.org/10.3390/insects14080703

Abstract/Summary

Quantifying the growth of entomopathogenic fungi is crucial for understanding their virulence and pathogenic potential. Traditional methods for determining growth, such as biomass determination or colony growth area, are time-consuming and quantitatively and spatially limited in scope. In this study, we introduce a high-throughput method for rapidly measuring fungal growth using spectrophotometry in small-volume, liquid media cultures in 96-well microplates. Optical density (OD) changes were directly correlated with dry weight of samples for six isolates from three species of the genus Metarhizium to validate spectrophotometric growth measurements, and investigate species- and isolate-specific effects. We quantified fungal biomass from the microcultures by extracting, drying, and weighing mycelial mats. From the relationship established between OD and biomass, we generated standard curves for predicting biomass based on the OD values. The OD measurements clearly distinguished growth patterns among six isolates from three Metarhizium species. The logistic growth phase, as captured by the OD measurements, could be accurately assessed within a span of 80 h. Using isolates of M. acridum, M. brunneum, and M. guizhouense, this technique was demonstrated to be an effective, reproducible, and simple method for rapidly measuring filamentous fungal growth with high precision. This technique offers a valuable tool for studying the growth dynamics of entomopathogenic fungi and investigating the factors that influence their growth.

Item Type:	Publication - Article
Digital Object Identifier (DOI):	https://doi.org/10.3390/insects14080703
UKCEH and CEH Sections/Science Areas:	Biodiversity (Science Area 2017-)
ISSN:	2075-4450
Additional Information. Not used in RCUK Gateway to Research.:	Open Access paper - full text available via Official URL link.
Additional Keywords:	Metarhizium, fungal growth, filamentous fungi, bioassay technique, spectrophotometry, microspectrophotometry, microplate reader, biomass quantification
NORA Subject Terms:	Biology and Microbiology
Date made live:	09 Nov 2023 11:51 +0 (UTC)
URI:	https://nora.nerc.ac.uk/id/eprint/536027

Item Type:

Publication - Article

Digital Object Identifier (DOI):

https://doi.org/10.3390/insects14080703

UKCEH and CEH Sections/Science Areas:

Biodiversity (Science Area 2017-)

ISSN:

2075-4450

Additional Information. Not used in RCUK Gateway to Research.:

Open Access paper - full text available via Official URL link.

Additional Keywords:

Metarhizium, fungal growth, filamentous fungi, bioassay technique, spectrophotometry, microspectrophotometry, microplate reader, biomass quantification

NORA Subject Terms:

Biology and Microbiology

Date made live:

09 Nov 2023 11:51 +0 (UTC)

URI:

https://nora.nerc.ac.uk/id/eprint/536027