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Transcriptomic of Beauveria bassiana during in vitro blastospore production

ABSTRACT: Culturing the entomopathogenic fungus Beauveria bassiana under high glucose concentrations coupled with high aeration results in a fungal developmental shift from hyphae growth to mostly blastospores (yeast-like cells). The underlying molecular mechanisms involved in this shift remains elusive. Therefore, this has motivated us to realize a systematic analysis of the differential gene expression to uncover the fungal transcriptomic response to osmotic and oxidative stresses associated with the resulting high blastospores yield. Differential gene expression was compared under moderate glucose concentration (10% w/v) and high glucose concentration (20% w/v) daily for the three-day culturing. The RNAseq-based transcriptomic results underpinned a higher proportion of down- than up-regulated genes when the fungus was grown under 20% glucose than at 10%. A follow-up study explored a broader glucose range (4, 8, 12, 16, 20% w/v) with phenotype assessment and qRT-PCR of selected genes. Antioxidant, calcium transport, conidiation, and osmosensor related genes were highly up-regulated in higher glucose titers (18-20%) than at lower titers. These findings improved the knowledge of the molecular mechanisms during blastospore development and may help the large scale blastospores industrial production.

ORGANISM(S): Beauveria bassiana

PROVIDER: GSE163673 | GEO | 2020/12/22

REPOSITORIES: GEO

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Project description:The glucose dual-affinity transport system (low- and high-affinity) is a conserved strategy exerted by microorganisms to cope with the naturally fluctuating availability of nutrients in the environment. The glucose sensing and uptaking process were believed to be tightly involved in cellulases expression regulation in cellulolytic fungi. However, both the identities and functions of the major molecular components of this evolutionarily conserved system in filamentous fungi remain elusive. Here, we conducted a systematic identification and characterization of the glucose dual-affinity transport system in the model fungus Neurospora crassa. Using RNA sequencing coupled with functional transport analyses, we were able to assign GLT-1 (Km = 18.42 ± 3.38 mM) and HGT-1/-2 (Km = 16.13 ± 0.95 µM and 98.97 ± 22.02 µM) to low- and high-affinity glucose transport systems, respectively. The high-affinity transporters hgt-1/-2 were able to complement a moderate growth defect under high glucose when glt-1 was deleted. Simultaneous deletion of hgt-1/-2 led to extensive derepression of genes for plant cell wall deconstruction on cellulose. This suppression by HGT-1/-2 was connected to both carbon catabolite repression (CCR) and the cyclic adenosine monophosphate-protein kinase A pathway. Alteration of a residue conserved across taxa for hexose-transporters was found to result in a loss of glucose-transporting function, whereas CCR signal transduction was retained, indicating a dual function for HGT-1/-2 as “transceptors”. In this study, GLT-1 and HGT-1/-2 are identified as the key components of the glucose dual-affinity transport system, which play diverse roles in glucose transport and carbon metabolism. Given their wide conservation across fungal species, the glucose dual-affinity transport components and their pleiotropic roles revealed in this study would shed extensive new light on the molecular basis of nutrient transport, signaling, and plant cell wall degradation in fungi.

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Transcriptomic of Beauveria bassiana during in vitro blastospore production

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