Trichoderma sporulation medium and sporulation method

A culture medium, Trichoderma technology, applied in the methods of using spores, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of restricting in-depth research on anti-cancer substances, and achieve easy control, stable material properties, and production. Mild effect of spore conditions

Inactive Publication Date: 2013-11-20
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

) is the unique strain of this laboratory protected by patent, so there is no report about the sporulation culture method of this kind of Trichoderma at home and abroad. The land restricts further in-depth research on the endophytic fungus of this species of Juglans catalpa and the anticancer substances produced by its fermentation

Method used

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  • Trichoderma sporulation medium and sporulation method
  • Trichoderma sporulation medium and sporulation method
  • Trichoderma sporulation medium and sporulation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A kind of Trichoderma sporulation culture medium each raw material content relation is as follows:

[0028]

[0029]

[0030] When preparing the sporulation medium, the above-mentioned substances are mixed and dissolved according to the weight percentage content, and then sterilized at 120° C. for 40 minutes under high pressure, and stored at 4° C. for later use;

[0031] Sporulation conditions: 200 microliters of spore suspension or bacteria balls with a diameter of about 5 mm are inoculated into a conical flask containing 20 milliliters of sporulation medium, at room temperature, with 10 hours of light per day, and cultivated for 5 days. The curve graph of the relationship between the number of spores of this Trichoderma and the culture time is as follows: figure 1 shown by figure 1 It can be known that the amount of sporulation produced after five days of cultivation can reach 7.4×10 7 spores / ml medium.

[0032] From the comparison of Example 1 and Comparati...

Embodiment 2

[0035] The content relationship of each raw material in the sporulation medium is as follows:

[0036]

[0037]

[0038] When preparing the sporulation medium, the above-mentioned substances are mixed and dissolved according to the weight percentage content, and then sterilized at 100°C for 60 minutes under high pressure, and stored at 4°C for later use;

[0039] Sporulation conditions: 200 microliters of spore suspension or bacteria balls with a diameter of about 5 mm are inoculated into a Erlenmeyer flask containing 20 milliliters of solid medium, cultured at 22°C for 5 days. The spore yield can reach 1.2×10 after five days of cultivation 7 spores / ml medium.

Embodiment 3

[0041] The content relationship of each raw material in the sporulation medium is as follows:

[0042]

[0043] When preparing the spore-forming medium, mix and dissolve the above-mentioned substances according to the weight percentage content, then sterilize at 140°C for 20 minutes under high pressure, and store at 4°C for later use;

[0044] Sporulation conditions: Inoculate 200 microliters of spore suspension or Trichoderma spheroids with a diameter of about 5mm into a conical flask containing 20ml of solid medium, at 28°C, with 15 hours of light per day, and cultivate for 5 days. The spore yield can reach 5.2×10 after five days of cultivation 6 spores / ml medium.

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PUM

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Abstract

The invention discloses a trichoderma sporulation medium and a sporulation method. The trichoderma sporulation medium is composed of fructose, xylose and a basal medium (a yeast extract product, ammonium tartrate, ammonium nitrate, potassium dihydrogen phosphate, bitter salt, sodium chloride, calcium dichloride dehydrate, green vitriol, disodium ethylene diamine tetraacetate and water). Fructose and xylose are respectively used as carbon sources, and ammonium tartrate and the yeast extract product are used as nitrogen sources. Added inorganic salt has a component for forming microbial cells and has functions of adjusting enzyme activity, cell osmotic pressure, hydrogen ion concentration, oxido reduction potential and the like. Hydrophosphate adjusts the acid-base value of the medium and makes the strain maintain the most appropriate pH value for its growth. The medium provided by the invention satisfies the growth need of the strain. The sporulation method provided by the invention is carried out at 22-28 DEG C by lighting culture. If the Juglans mandshurica endophytic fungi trichoderma sporulation medium is used for culture for 5 days, the sporulation amount can reach 7.4*107 spore / milliliter medium.

Description

technical field [0001] The invention relates to an efficient spore-producing method for endophytic fungi of Juglans catalpa, in particular to a spore-producing medium and a spore-producing method for Trichoderma. Background technique [0002] The endophytic fungus is a bacterial strain isolated from the bark of Juglans catalpa, and its fermentation product has anti-cancer activity. The fungus is identified as Deuteromycotina, Hyphomycetes, Polygonales, and Trichoderma. The anticancer activity of the fermentation product of the bacteria was tested, and the fermentation product could significantly inhibit the growth of liver cancer cells, and the anticancer ability of the fermentation broth was basically equivalent to that of 1 mg / L paclitaxel solution. Therefore, the study of the conidia of the Trichoderma plays an important role in the anticancer activity of the fermentation product and its mechanism and the preservation of the strain. At present, this Trichoderma (Trichode...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N3/00C12R1/885
Inventor 苗志奇张星星陈榕华
Owner SHANGHAI JIAO TONG UNIV
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