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CGMCC5.26 is a laboratory equipment used for the storage and preservation of microbiological cultures. It is designed to maintain the viability and purity of microbial samples under controlled temperature and humidity conditions.

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6 protocols using cgmcc5

1

Cultivation of Ganoderma lucidum CGMCC5.26

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The strain G. lucidum CGMCC5.26 was purchased from China General Microbiological Culture Collection Center and maintained on potato dextrose agar slants at 4 °C. The seed medium and preculture details of G. lucidum have been described previously [9 (link),11 (link)]. Briefly, the medium was composed of glucose (20 g/L), yeast nitrogen base without amino acids (5 g/L), tryptone (5 g/L), KH2PO4 (4.5 g/L), and MgSO4·7H2O (2 g/L) at initial pH. The seed was in a 250 mL flask containing 80 mL of medium and was kept at 30 °C on a rotary shaker (150 rpm). In the fermentation medium, the carbon sources were glucose and xylose, and the other components were the same as those in the seed medium. Before inoculating, the seeds were homogenized using an IKA T10 basic homogenizer (IKA, Königswinter, Germany). Briefly, the sterilized dispersing elements of homogenizer inserted the seed medium into a 250 mL flask, and homogenized with the fifth level for 1 min. Then, the homogenized seed medium was centrifuged at 10,000× g for 5 min, and 3.5 g of wet weight cells/L were inoculated into a 500 mL flask with 150 mL of medium. The submerged culture was grown on a rotary shaker at 150 rpm and 30 °C for eight days.
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2

Submerged Fermentation of Ganoderma lucidum

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Ganoderma lucidum CGMCC5.26 was purchased from the China General Microbiological Culture Collection Center (Beijing). It was preserved on potato dextrose agar (PDA) slants at 4°C. The composition of the seed and submerged fermentation medium (g/L) is as follows: glucose (20), tryptone (5), yeast nitrogen base without amino acids (YNB) (5), and MgSO4⋅7H2O (2), KH2PO4 (4.5), with culture conditions of 30°C at 150 rpm (Ma et al., 2018 (link)).
Medium A (g/L) (Tang and Zhong, 2002 (link)) contained glucose (35), tryptone (5), yeast extract (2.5), KH2PO4⋅H2O (1), MgSO4⋅7H2O (0.5), and Vitamin B1 (0.05), with culture conditions of 30°C at 150 rpm. Medium B (g/L) (Hu et al., 2018 (link)) contained glucose (20), tryptone (2), yeast extract 2, KH2PO4 (4.6), and MgSO4⋅7H2O (0.5), with culture conditions of 30°C at 150 rpm.
The seed culture included four mycelium agar squares (3 mm × 3 mm), which were transferred into a 250 mL flask and cultured for 7 days. The fermentation culture was developed as follows. After homogenization with a high-speed tissue homogenate machine (IKA T10 basic ULTRA-TURRAX), a 3 mL of the seed culture solution was inoculated into a 500 mL flask and cultured for 7 days (Peng et al., 2015 (link)).
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3

Cultivation of Ganoderma lucidum Strain CGMCC5.26

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Ganoderma lucidum CGMCC5.26 was purchased from China General Microbiological Culture Collection Center (CGMCC) and maintained on potato dextrose agar (PDA) slants at 4 °C. In order to get consistent metabolites and transcriptome data, a semi‐synthetic media was selected in this study. The medium was composed of Glucose 20 g l−1, yeast nitrogen base without amino acids (YNB) 5 g l−1, tryptone 5 g l−1, KH2PO4 4.5 g l−1 and MgSO4·7H2O 2 g l−1 at initial pH. Glucose, KH2PO4 and MgSO4·7H2O purchased from Sinopharm (Beijing, China), YNB purchased from BioDee Biotechnology Co., Ltd (Beijing, China), and tryptone purchased from OXOID (Hants, UK). The medium has been used many years to study the secondary metabolism of G. lucidium (Peng et al., 2015; Ma et al., 2018, 2019). The seed was in a 250‐ml flask containing 80 ml medium and kept at 30 °C on a rotary shaker (150 rpm) for ten days.
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4

Cultivating Ganoderma lucidum with Selenium

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Ganoderma lucidum (CGMCC 5.26) was purchased from the China General Microbiological Culture Collection Center (CGMCC) and preserved on potato dextrose agar slants at 4 °C. The seed and liquid culture medium was composed of glucose (20 g/L), yeast nitrogen base without amino acids (5 g/L), tryptone (5 g/L), KH2PO4 (4.5 g/L), MgSO4·7H2O (2 g/L), and 0–500 ppm Na2SeO3 at initial pH. Liquid culture was performed in a 500 mL flask with 150 mL medium and kept at 30 °C at 150 rpm on a rotary shaker.
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5

Cultivation of Ganoderma lingzhi Strains

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The G. lingzhi strain CGMCC 5.0026 was purchased from China General Microbiological Culture Collection Center (CGMCC) and the wild-type strain of RWY-0 was donated from the Anhui Science and Technology University. The Ganoderma lingzhi strains were incubated in liquid cultures. The mycelium pellets were grown in 250 mL flasks containing 150 mL of a Potato Dextrose Broth (PDB) culture and placed on a rotary shaker incubator at 150 rpm at 28°C for 14 days before collection for experiments.
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6

Cultivation of Ganoderma lucidum Dikaryotic Strain

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The G. lucidum dikaryotic strain CGMCC5.0026 was obtained from China General Microbiological Culture Collection Center (Beijing, China). Microcrystalline cellulose (MCC), lignin, D-cellobiose, D-galactose, L-arabinose, D-xylose and D-rhamnose were purchased from Sangon Biotech (Shanghai, China). Vegetative mycelia were grown on PDA plate at 28 °C in the dark. Liquid cultures were shaken at 28 °C with 125 revolutions per minute (rpm). Seed cultures were grown in 250 ml flasks containing 100 ml preculture medium (g/L: glucose 35, peptone 5, yeast extract 2.5, KH2PO4 0.883, Vitamin B1 0.05, MgSO4·7H2O 0.5, initial pH 5.5) at 125 rpm and 28 °C for 7 d. The fermentation experiments were performed in 250 ml flasks containing 100 ml fermentation medium (g/L: glucose 35, peptone 5, yeast extract 5, KH2PO4 0.883, Vitamin B1 0.05, MgSO4·7H2O 0.5, pH5.5) at 125 rpm at 28 °C for 7 days. All chemicals were added to the fermentation media at 72 h post of inoculation at indicated concentration. MCC and lignin were sterilized at 115 °C for 30 min. Instead, the rest of saccharide and sugars were sterilized by filtration through a 0.2 μm membrane.
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