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Special Cryptococcus laurentii culture medium and application thereof

A Cryptococcus lorenti yeast and culture medium technology, which is applied in the field of fermentation and cultivation of postharvest fruit biocontrol yeast, can solve the problems of composition and concentration differences, composition and content uncertainty, etc., and achieve the effect of clear components

Inactive Publication Date: 2011-09-28
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But also because it is a natural mixed substance, the composition and concentration (content) will have certain differences between different manufacturers and different batches, which will have a certain negative impact on the theoretical research based on this medium
For example, when it is necessary to study the effect of a specific substance on yeast culture and activity, the final experimental results and judgment will be adversely affected due to the uncertainty of the composition and content in the original medium

Method used

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  • Special Cryptococcus laurentii culture medium and application thereof
  • Special Cryptococcus laurentii culture medium and application thereof
  • Special Cryptococcus laurentii culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1. Activation of Cryptococcus lorenti strains and seed liquid culture

[0018] Take out Cryptococcus laurentii CGMCC NO.3590 stored in the refrigerator at 4°C, inoculate it in NYDA medium by streak inoculation, activate it in a biochemical incubator at 28°C for 48 hours, and then activate it once under the same conditions .

[0019] Pick a little bacteria from the activated Cryptococcus laurentis slant and inoculate it into a 250mL Erlenmeyer flask containing 50mL of NYDB medium, and culture it in a shaker incubator at 28°C for 24h with a rotation speed of 200rpm.

[0020] 2. Preparation of fermentation medium

[0021] In a 250mL Erlenmeyer flask according to the orthogonal test method L 16 (2 15 ) into each culture medium component (see Table 1 and Table 2, weight g per L), conventional autoclaving (121°C, sterilizing for 20min) for later use (biotin, L-aspartic acid and vitamin B1 Add after filter sterilization), and dilute to 50mL with water.

[0022] 3. Ferment...

Embodiment 2

[0035] 1. Activation of Cryptococcus lorenti strains and seed liquid culture

[0036] With embodiment 1.

[0037] 2. Preparation of fermentation medium

[0038] In a 250mL Erlenmeyer flask, the glucose in each treatment is 10g / L, the ammonium sulfate is 3g / L, and the rest of the ingredients are according to the orthogonal test method L 18 (2×3 7 ) into each medium component (see Table 3 and Table 4, weight g per L), autoclaved (121°C, sterilized for 20min) for later use (vitamin B1 was added after filter sterilization), and water to volume to 50mL.

[0039] 3. Fermentation culture and biomass determination of Cryptococcus lorenti

[0040] Under sterile conditions, draw 2mL of seed solution and inoculate in 250ml of sterilized fermentation medium, and cultivate in a shaker incubator at 25°C for 24h with a rotation speed of 200rpm. When measuring the biomass, centrifuge at 5000 rpm for 5 minutes, discard the fermentation supernatant, wash with water twice, and finally measu...

Embodiment 3

[0051] 1. Activation of Cryptococcus lorenti strains and seed liquid culture

[0052] With embodiment 1.

[0053] 2. Preparation of fermentation medium

[0054] In a 250mL Erlenmeyer flask, the glucose in each treatment is 10g / L, the ammonium sulfate is 3g / L, and the rest of the ingredients are according to the orthogonal test method L 9 (3 4 ) into each medium component (see Table 5 and Table 6, the weight in g per L), autoclave (121°C, sterilize for 20min) for later use, and dilute to 50mL with water.

[0055] 3. Fermentation culture and biomass determination of Cryptococcus lorenti

[0056] Under aseptic conditions, draw 1 mL of seed solution and inoculate in 250 ml of sterilized fermentation medium, and cultivate in a shaker incubator at 25° C. for 24 h with a rotation speed of 200 rpm.

[0057] When measuring the biomass, centrifuge at 5000 rpm for 5 minutes, discard the fermentation supernatant, wash with water twice, and finally measure the absorbance of the yeast s...

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Abstract

The invention discloses a special Cryptococcus laurentii culture medium. One liter of the special culture medium comprises the following components: 10-20g of glucose, 3-6g of ammonium sulfate, 1.5-3g of dipotassium hydrogen phosphate, 1-6g of magnesium sulfate, 0-0.02g of ferrous sulfate, 0-0.01g of manganese sulfate, 0-0.02g of zinc sulfate, 0-0.5g / L of calcium chloride, 0-1g of sodium chloride, 0.4-0.5mg of biotin, 0.2-0.5mg of L-aspartic acid, 0.2-1mg of vitamin B1 and the balance water. The pH value of the special culture medium is 3-8. The invention also discloses an application of the special Cryptococcus laurentii culture medium. The application is as follows: inoculating Cryptococcus laurentii in the special culture medium according to an inoculation amount of 0.2-4vol.% and culturing at 25-28 DEG C in 200rpm for 24-120 hours.

Description

technical field [0001] The invention relates to the technical field of fermentation and cultivation of fruit postharvest biocontrol yeast, in particular to a special culture medium for Cryptococcus lorenti and its application. Background technique [0002] The biological control technology of using antagonistic yeast to control postharvest fruit diseases is one of the new postharvest disease control methods that are very concerned at home and abroad. Cryptococcus laurentii (Cryptococcus laurentii) is a postharvest biological control yeast strain widely studied at home and abroad. Its resistance mechanism involves nutrition and space competition, adhesion with pathogenic bacteria, secretion of β-1,3-glucanase, Anti-oxidative stress, induction of fruit resistance or resistance-related reactions, degradation of mycotoxins secreted by pathogenic bacteria, and degradation of volatiles related to fruit and stimulation of pathogenic spore germination, etc. Chinese invention patent...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12R1/645
Inventor 余挺郑晓冬杨林卢黄娉蓝天演
Owner ZHEJIANG UNIV
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