Abstract
Cocultivation is an emerging and potential way to investigate microbial interaction in the laboratory. Extensive researches has been carried out over the years, but some microorganism cocultivation are not easy to implement in the laboratory, especially the fungus-fungus (FF) cocultivation, owing to the obstacles such as fungal different growth rate, limited growing space, hyphae intertwining, and difficulty of sample separation, etc. In this research, a double-sided petri dish (DSPD) was designed and carried out as a tool to study FF cocultivation in the laboratory. A natural FF cocultivation of Monascus spp. and Aspergillus niger inspired from black-skin-red-koji (BSRK), were studied. By using DSPD, the aforementioned obstacles in the FF cocultivation study were overcome through co-culturing Monascus spp. and A. niger on each side of DSPD. The characteristics of monocultured and co-cultured Monascus spp. and A. niger were compared and analyzed, including colonial and microscopic morphologies, and main secondary metabolites (SMs) of Monascus spp. analyzed by high performance liquid chromatography. And a novel SM was found to be produced by Monascus ruber M7 when co-cultured with A. niger CBS 513.88. Since the above mentioned obstacles, were overcome, we obtained good quality of transcriptome data for further analysis. These results indicate that DSPD might be an efficient tool for investigation of microbial interaction, in particular, for FF interaction.
Highlights
Black-skin-red-koji (BSRK), known as Wuyihongquۂin Chinese, is a special traditional Chinese mixed starter which has been utilized for brewing rice wine and cereal vinegar for more than 1,000 years mainly in Fujian and Zhejiang provinces (Fang et al, 2011; Huang et al, 2018)
On Czapek yeast extract agar (CYA) media, the colony color of M. ruber M7 changed from red to orange when co-cultured with A. niger CBS 513.88
Monascus spp. and A. niger can grow together, A. niger, especially A. niger CBS 113.46 can inhibit the growth of Monascus spp. strains
Summary
Black-skin-red-koji (BSRK), known as Wuyihongqu (ӊҥߚ )ۂin Chinese, is a special traditional Chinese mixed starter which has been utilized for brewing rice wine and cereal vinegar for more than 1,000 years mainly in Fujian and Zhejiang provinces (Fang et al, 2011; Huang et al, 2018). As a representative model of fungus-fungus (FF) cocultivation, the main filamentous fungi in BSRK are Monascus spp. and Aspergillus niger (Huang et al, 2019). Monascus spp. have been used to produce the food and health products due to their functional secondary metabolites (SMs) such as Monascus pigments (MPs), monacolin K (MK), dimerumic acid, and γ-aminobutyric acid (Chen et al, 2015; Feng et al, 2016). Nephrotoxin citrinin (CIT) can be produced by a few of Monascus spp. strains, which can lead to the food safety concerns of Monascus related products (He and Cox, 2016). To date, as a natural FF cocultivation model, the reason why both of Monascus spp. and A. niger can harmoniously grow in BSRK and the mechanisms relating to how they affect each other are still undiscovered
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