Quantitative Monitoring of Mycelial Growth of Aspergillus fumigatus in Liquid Culture by Optical Density

Miyazawa, Ken; Umeyama, Takashi; Hoshino, Yasutaka; Abe, Keietsu; Miyazaki, Yoshitsugu

doi:10.1128/spectrum.00063-21

Cited by 10 publications

(11 citation statements)

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“…During the hyphal growth stage, filamentous fungi secrete an extracellular matrix composed mainly of proteins, lipids, and polysaccharides such as galactosaminogalactan, α-glucan, and galactomannan [ 13 , 14 ]. Surface exposure of adhesive α-glucan and galactosaminogalactan during germination is believed to cause hyphal aggregation that is responsible for the pellet formation usually observed in liquid shake cultures [ 15 , 16 ]. The pellet formation causes the heterogenous growth of filamentous fungal species, as the surface and the interior of pellets differ significantly with respect to supply of nutrients and oxygen [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

“…The pellet formation causes the heterogenous growth of filamentous fungal species, as the surface and the interior of pellets differ significantly with respect to supply of nutrients and oxygen [ 17 ]. Recently, impairment of biosynthesis of α-glucan and galactosaminogalactan was shown to lead to dispersed hyphal growth facilitating biomass determination via optical density [ 16 ] but these genetic modifications severely affect the physiology.…”

Section: Introductionmentioning

confidence: 99%

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Online Biomass Monitoring Enables Characterization of the Growth Pattern of Aspergillus fumigatus in Liquid Shake Conditions

Bauer

Abt

Yap

et al. 2022

JoF

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Numerous filamentous fungal species are extensively studied due to their role as model organisms, workhorses in biotechnology, or as pathogens for plants, animals, and humans. Growth studies are mainly carried out on solid media. However, studies concerning gene expression, biochemistry, or metabolism are carried out usually in liquid shake conditions, which do not correspond to the growth pattern on solid media. The reason for this practice is the problem of on-line growth monitoring of filamentous fungal species, which usually form pellets in liquid shake cultures. Here, we compared the time-consuming and tedious process of dry-weight determination of the mold Aspergillus fumigatus with online monitoring of biomass in liquid shake culture by the parallelizable CGQ (“cell growth quantifier”), which implements dynamic biomass determination by backscattered light measurement. The results revealed a strong correlation of CGQ-mediated growth monitoring and classical biomass measurement of A. fumigatus grown over a time course. Moreover, CGQ-mediated growth monitoring displayed the difference in growth of A. fumigatus in response to the limitation of iron or nitrogen as well as the growth defects of previously reported mutant strains (ΔhapX, ΔsrbA). Furthermore, the frequently used wild-type strain Af293 showed largely decreased and delayed growth in liquid shake cultures compared to other strains (AfS77, A1160p+, AfS35). Taken together, the CGQ allows for robust, automated biomass monitoring of A. fumigatus during liquid shake conditions, which largely facilitates the characterization of the growth pattern of filamentous fungal species.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Online Biomass Monitoring Enables Characterization of the Growth Pattern of Aspergillus fumigatus in Liquid Shake Conditions

Bauer

Abt

Yap

et al. 2022

JoF

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“…Growth differences among strains at different temperatures have been investigated in several fungi, including species of Aspergillus, Penicillium, Paecilomyces , and Metarhizium ( 27 – 31 ). For filamentous fungi, previous studies measured the radial growth of colonies on solid media at different temperatures as indicators of their growths ( 32 – 34 ). Larger colonies represent higher tolerance to the environmental conditions tested.…”

Section: Discussionmentioning

confidence: 99%

Assessing thermal adaptation of a global sample of Aspergillus fumigatus: Implications for climate change effects

Korfanty

Heifetz

2023

Front. Public Health

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Aspergillus fumigatus is a common environmental mold and a major cause of opportunistic infections in humans. It's distributed among many ecological niches across the globe. A major virulence factor of A. fumigatus is its ability to grow at high temperature. However, at present, little is known about variations among strains in their growth at different temperatures and how their geographic origins may impact such variations. In this study, we analyzed 89 strains from 12 countries (Cameroon, Canada, China, Costa Rica, France, India, Iceland, Ireland, New Zealand, Peru, Saudi Arabia, and USA) representing diverse geographic locations and temperature environments. Each strain was grown at four temperatures and genotyped at nine microsatellite loci. Our analyses revealed a range of growth profiles, with significant variations among strains within individual geographic populations in their growths across the temperatures. No statistically significant association was observed between strain genotypes and their thermal growth profiles. Similarly geographic separation contributed little to differences in thermal adaptations among strains and populations. The combined analyses among genotypes and growth rates at different temperatures in the global sample suggest that most natural populations of A. fumigatus are capable of rapid adaptation to temperature changes. We discuss the implications of our results to the evolution and epidemiology of A. fumigatus under increasing climate change.

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“…The disruptant of the Aspergillus luchuensis agsE gene, an ortholog to A. nidulans agsB , shows better protoplast formation than the wild-type strain when treated with the cell wall lytic enzyme Yatalase [ 135 ]. The A. fumigatus mutants in which the ags1 gene, an ortholog of A. nidulans agsB , is disrupted, form smaller hyphal pellets than the wild type [ 136 ]. Taken together, α-1,3-glucan is an aggregation factor for hyphae and conidia in Aspergillus fungi.…”

Section: Improvement Of Productivity By Modification Of Macromorpholo...mentioning

confidence: 99%

“…In the background of the defect of α-1,3-glucan biosynthesis (Δ agsA Δ agsB Δ agsC ) in A. oryzae , disruption of the sphZ and ugeZ genes (AGΔ-GAGΔ), which are speculative GAG biosynthetic genes of A. oryzae , leads to dispersion of hyphae under submerged culture conditions, suggesting that GAG also contributes to aggregation in A. oryzae [ 138 ]. A simultaneous defect of α-1,3-glucan and GAG biosynthesis also leads to hyphal dispersion in A. fumigatus [ 136 ]. In B. cinerea and Cochlioborus heterostrophus , GAG also contributes to hyphal aggregation [ 139 ].…”

Section: Improvement Of Productivity By Modification Of Macromorpholo...mentioning

confidence: 99%

Cell Wall Integrity and Its Industrial Applications in Filamentous Fungi

Yoshimi,

Miyazawa,

Kawauchi

et al. 2022

JoF

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Signal transduction pathways regulating cell wall integrity (CWI) in filamentous fungi have been studied taking into account findings in budding yeast, and much knowledge has been accumulated in recent years. Given that the cell wall is essential for viability in fungi, its architecture has been analyzed in relation to virulence, especially in filamentous fungal pathogens of plants and humans. Although research on CWI signaling in individual fungal species has progressed, an integrated understanding of CWI signaling in diverse fungi has not yet been achieved. For example, the variety of sensor proteins and their functional differences among different fungal species have been described, but the understanding of their general and species-specific biological functions is limited. Our long-term research interest is CWI signaling in filamentous fungi. Here, we outline CWI signaling in these fungi, from sensor proteins required for the recognition of environmental changes to the regulation of cell wall polysaccharide synthesis genes. We discuss the similarities and differences between the functions of CWI signaling factors in filamentous fungi and in budding yeast. We also describe the latest findings on industrial applications, including those derived from studies on CWI signaling: the development of antifungal agents and the development of highly productive strains of filamentous fungi with modified cell surface characteristics by controlling cell wall biogenesis.

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Quantitative Monitoring of Mycelial Growth of Aspergillus fumigatus in Liquid Culture by Optical Density

Abstract: Filamentous fungi generally form hyphal pellets in liquid culture. This property prevents filamentous fungi to apply the methods used for unicellular organisms such as yeast and bacteria.

Cited by 10 publications

References 50 publications

Online Biomass Monitoring Enables Characterization of the Growth Pattern of Aspergillus fumigatus in Liquid Shake Conditions

Online Biomass Monitoring Enables Characterization of the Growth Pattern of Aspergillus fumigatus in Liquid Shake Conditions

Assessing thermal adaptation of a global sample of Aspergillus fumigatus: Implications for climate change effects

Cell Wall Integrity and Its Industrial Applications in Filamentous Fungi

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