Seven basidiomata specimens of G. cf. highlandensis were sampled from Japan in 2018-2020 (Table 1 ). An isotype specimen of Clitocybe highlandensis (= G. highlandensis; TENN-F-16020) loaned from the University of Tennessee and included in this study (Table 1 ). Mycelia were isolated from the inner tissue or basidiospores of three specimens using malt extract agar [MA; 15 g malt extract (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 15 g agar powder (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan), and 1,000 mL distilled water]. The external morphology of the basidiomata specimens was observed, and specimens were air-dried at 45 °C for 1-2 d and deposited in the Tottori University Mycological Herbarium (TUMH) at the Fungus/Mushroom Resource and Research Center (FMRC). Several specimens were also deposited at the Tottori Mycological Institute (TMI). Three established strains (TUFC 101584, TUFC 101840, and TUFC 101893), isolated from TUMH 63383, TUMH 63956, and TUMH 64253, were deposited at the FMRC as Tottori University Fungal Culture (TUFC) strains (Table 1 ). Two strains of G. foliicola (TUFC 100720 and TUFC 101392) stored at FMRC were also included for comparison (Table 1 ).
Malt extract
Manufactured by BD
Sourced in United States, Germany
Malt extract is a concentrated liquid form of malted grain, typically barley. It is a common ingredient used in the production of various food and beverage products, particularly in the brewing and baking industries. Malt extract serves as a source of fermentable sugars and contributes to the flavor and color of the final product.
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Lab products found in correlation
Yeast extract, by BD (17 mentions)
Peptone, by BD (10 mentions)
Agar, by BD (3 mentions)
Dextrose, by BD (2 mentions)
Potato dextrose broth (pdb), by BD (2 mentions)
D glucose, by BD (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Methanol, by Thermo Fisher Scientific (2 mentions)
Agar powder, by Junsei (2 mentions)
Glucose, by Junsei (2 mentions)
Bacto peptone, by BD (2 mentions)
Potato dextrose agar (pda), by BD (1 mentions)
Agar powder, by Fujifilm (1 mentions)
Glucose, by Nacalai Tesque (1 mentions)
Xylan from birchwood, by Merck Group (1 mentions)
Fructose, by Nacalai Tesque (1 mentions)
Agar powder, by Nacalai Tesque (1 mentions)
Kh2po4, by Nacalai Tesque (1 mentions)
Mannose, by Nacalai Tesque (1 mentions)
Mgso4 7h2o, by Nacalai Tesque (1 mentions)
29 protocols using malt extract
1
Morphological and Genetic Analysis of Gymnopus highlandensis
2
Bursaphelenchus okinawaensis Propagation
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Bursaphelenchus okinawaensis (SH1 isolate) was propagated on the fungus Botrytis cinerea Pers. Fr. 1794 (Helotiales: Sclerotiniaceae) at 25°C on malt extract agar (1.5% malt extract [Becton Dickinson] and 4% agar [Rikaken]) containing 100 µg/mL chloramphenicol (Fujifilm) in 90 mm Petri dishes (Shinya et al., 2014 (link)).
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Bursaphelenchus okinawaensis (SH1 isolate) was propagated on the fungus Botrytis cinerea Pers. Fr. 1794 (Helotiales: Sclerotiniaceae) at 25°C on malt extract agar (1.5% malt extract [Becton Dickinson] and 4% agar [Rikaken]) containing 100 µg/mL chloramphenicol (Fujifilm) in 90 mm Petri dishes (Shinya et al., 2014 (link)).
3
Fungal Strain Characterization on Diverse Media
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Strains were grown at room temperature on two types of malt extract agar: 1) MEA: 40 g/L malt extract (Becton Dickinson, Heidelberg, Germany), yeast extract 4 g/L (Ohly, Hamburg, Germany), 1.5 % agar (Europäischer Agar, Otto Nordwald, Hamburg, Germany) and 2) MEX: 15 g/L malt extract (Becton Dickinson, containing 6 g malt extract base, 1.8 g maltose, 6 g Bacto glucose, 1.2 g Bacto yeast extract) and 1.5 % agar (Europäischer Agar, Otto Nordwald), as well as on potato dextrose agar (PDA: 39 g/L, Roth, Karlsruhe, Germany) and synthetic Mucor agar (Benny 2008 ) (SMA: 40 g/L dextrose (Roth), 2 g/L asparagine (Reanal Laborvegyszer Kereskedelmi Kft., Budapest, Hungary), 0.5 g/L KH2PO4 (Merck, Darmstadt, Germany), 0.25 g/L MgSO4×7H2O (Roth), 0.5 mg/L thiamine hydrochloride (Roth), 1.5 % agar (Europäischer Agar, Otto Nordwald). Macroscopic and microscopic features were studied after 3 and 7 d using a Zeiss Stemi 1000 (Carl Zeiss, Jena, Germany) and a Nikon Eclipse Ni Microscope (Nikon, Düsseldorf, Germany) with differential interference contrast and the NIS Elements software v. 4.30 (Nikon). Colours of the mycelia were described using the colour charts of Munsell (Anonymous 1990 ).
4
Cultivation Protocol for Streptomyces palmae
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S. palmae CMU-AB204T was grown in the International Streptomyces Project medium 2 (ISP2) agar [32 (link)] at 28 °C. For seed culture, 100 mL of ISP2 medium, consisting of 0.4% yeast extract (Becton, Dickinson and Company, Sparks, MD, USA), 1.0% malt extract (Becton, Dickinson and Company, Sparks, MD, USA), and 0.4% glucose, was prepared in an Erlenmeyer flask and the pH was adjusted to 7.0 before sterilization. The slant culture of S. palmae was scraped by an inoculating loop and inoculated into ISP2 medium. The inoculated flask was incubated at 30 °C for three days on a rotary shaker at 150 rpm. Two mL portions of this seed culture were transferred into 500 mL Erlenmeyer flasks containing 150 mL of ISP2 medium, which was followed by fermentation using a rotary shaker at 150 rpm, 30 °C for seven days.
5
Cultivation of Nigrospora aurantiaca from Melaleuca
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The fungal strain Nigrospora aurantiaca was isolated from a healthy leaf of Melaleuca leucadendra linn collected in the yard of National Taiwan University and was identified by sequencing the internal transcribed spacer regions of the rDNA (ITS). A BLAST search of the sequence led to the best match of Nigrospora aurantiaca. Mycelium Nigrospora aurantiaca#TMU062 was inoculated into two different media—liquid medium and solid medium. Inoculation in liquid medium was done in 5 L serum bottles, each containing 50 g of malt extract (Becton, Dickinson and Company, Sparks, USA) and 3.5 L of deionized water. The fermentation was conducted with aeration at 25–30 °C for 14 days. As for solid medium, 250 mL flasks were used—each containing 20 g of barley and 0.2 g of potato dextrose agar (Becton, Dickinson and Company, Sparks, USA). After adding 15 mL of deionized water, they were fermented for 30 days at 27–30 °C.
6
Diverse Culture Media Preparation
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Agar, peptone, malt extract, and yeast extract (Becton Dickinson, BD and Company East Rutherford, Franklin Lakes, NJ, USA) were used. To prepare saline malt extract agar (20 g malt extract, 10 g NaCl, 17 g agar, 200 ug/L chloramphenicol and 1000 mL bidistilled water) were taken. For the PDA potato dextrose agar were (200 g of potato, 20 g of glucose, 15 g of agar and 1000 mL of bidistilled water). YPD broth was prepared using (10 g yeast extract, 20 g peptone, 20 g dextrose, 0.5 g chloramphenicol, and 1000 mL bidistilled water). To obtain solid media, 15 g/L of agar were added to the broths. The lactose broth was prepared with (meat extract 3 g, peptone 5 g, lactose 5 g, distilled water 1000 mL). The minimal salt medium, MSM was obtained with (NH4NO3 12 g, KH2PO4 8 g, Na2SO4 2 g, KCl 4 g, MgSO4.7H2O 1 g, CaCl2 0.5 g, ZnSO4.7H2O 0.26 g, NaCl 20 g, chloramphenicol 200 ug/L and bidistilled water 1000 mL).
7
Microbial Media Composition Analysis
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All chemicals used were the highest grade available. Glucose, fructose, mannose, agar powder, KH2PO4, (NH4)2SO4 and MgSO4 · 7H2O were purchased from Nacalai Tesque, Inc. (Kyoto, Japan). Malt extract and yeast extract were purchased from Becton, Dickinson and Company (BD, Franklin Lakes, NJ). Starch from corn, crystalline cellulose Sigmacell type 20 and xylan from birchwood were purchased from Sigma-Aldrich (St. Louis, MO).
8
Cultivating Cordyceps militaris Mycelia
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C. militaris mycelia (BCRC 32219) were purchased from the Bioresource Collection and Research Center at the Food Industry Research and Development Institute (Hsinchu, Taiwan). Glucose was purchased from J. T. Baker (EU). Malt extract, peptone, and yeast extract were purchased from Becton Dickinson (Franklin Lakes, NJ, USA). Vitamin A was purchased from Sigma-Aldrich (St. Louis, MO, USA) and agar was purchased from High Standard Enterprise Co., Ltd. (Taiwan), respectively. Biochemical assay kits for kidney function detection were obtained from Arkray (Kyoto, Japan).
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C. militaris mycelia (BCRC 32219) were purchased from the Bioresource Collection and Research Center at the Food Industry Research and Development Institute (Hsinchu, Taiwan). Glucose was purchased from J. T. Baker (EU). Malt extract, peptone, and yeast extract were purchased from Becton Dickinson (Franklin Lakes, NJ, USA). Vitamin A was purchased from Sigma-Aldrich (St. Louis, MO, USA) and agar was purchased from High Standard Enterprise Co., Ltd. (Taiwan), respectively. Biochemical assay kits for kidney function detection were obtained from Arkray (Kyoto, Japan).
9
Microbial Culture Protocols for Diverse Fungi
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All the microorganisms were obtained from the Korean Collection for Type Cultures (KCTC, Jeongeup, Korea) and Korean Culture Center of Microorganisms (KCCM, Seoul, Korea). Eight cultures were used for the preliminary screening process as listed below: Absidia coerulea KCTC 6936, Aspergillus fumigatus 6145, Cunninghamella elegans var. elegans 6992, Mortierella ramanniana var. angulispora 6137, Mucor hiemalis 26779, Penicillium chrysogenum 6933, Trichoderma koningii 6042, and Aspergillus niger KCCM 60332.
All the ingredients for microbial media, including dextrose, peptone, malt extract, yeast extract, and potato dextrose broth were purchased from Becton, Dickinson and Co. (Sparks, MD, USA). Two types of media were used in the fermentation experiments and are listed below: A. coerulea, A. fumigatus, A. niger, M. hiemalis, P. chrysogenum, and T. koningii were cultured on malt medium (malt extract 20 g/L, dextrose 20 g/L, peptone 1 g/L); C. elegans var. elegans and M. ramanniana var. angulispora were cultured on potato dextrose medium (24 g/L).
All the ingredients for microbial media, including dextrose, peptone, malt extract, yeast extract, and potato dextrose broth were purchased from Becton, Dickinson and Co. (Sparks, MD, USA). Two types of media were used in the fermentation experiments and are listed below: A. coerulea, A. fumigatus, A. niger, M. hiemalis, P. chrysogenum, and T. koningii were cultured on malt medium (malt extract 20 g/L, dextrose 20 g/L, peptone 1 g/L); C. elegans var. elegans and M. ramanniana var. angulispora were cultured on potato dextrose medium (24 g/L).
10
Synthesis and Microbial Transformation of Isoxanthohumol
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Isoxanthohumol was prepared by chemical cyclization in aqueous NaOH solution at 0 °C as described by Stevens et al. [4 (link)], and also by a microbial transformation method using the fungus R. oryzae KCTC 6946 as previously reported by Kim and Lee [19 (link)]. Isoxanthohumol prepared by both methods was extracted with EtOAc and then purified by chromatographic methods including silica gel and reversed-phase C18 MPLC. The spectroscopic data of isoxanthohumol (1 ) were in good agreement with data in the literature [1 (link)] and its structure was also confirmed by 2D NMR experiments. Optical rotation and CD measurements revealed that the substrate isoxanthohumol was a racemic mixture of (2S)- and (2R)-isoxanthohumol. Ingredients for media including D-glucose, peptone, malt extract, yeast extract, and potato dextrose medium were purchased from Becton, Dickinson and Co. (Sparks, MD, USA), and sucrose was purchased from Sigma-Aldrich Co. (St Louis, MO, USA).
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