Method for separating halimasch from endophytic fungi of gastrodia elata

A technology of endophytic fungi and armillaria, applied in the direction of microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve the problems of strain degradation, difficult operation, easy contamination of miscellaneous bacteria, etc., and achieve the goal of reducing mold Pollution, avoidance of miscellaneous bacteria contamination, less stimulation by drugs

Pending Publication Date: 2020-04-17
汉中市汉台区汉麓生物科技有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Separation and purification of Armillaria strains is of great significance for obtaining germplasm resources and carrying out breeding work. Among the currently commonly used isolation methods, the fruiting body isolation method is difficult to obtain wild materials due to the difficulty in artificially cultivating Armillaria fruiting bodies; The separation method of fungus cord and fungus wood is relatively complicated, easy to infect miscellaneous bacteria, difficult to operate, and low success rate; in 2006, Xiao Bo et al. published an academic paper "A New Method for Separation of Armillaria Armillaria - Gastrodia elata tissue separation method, using Armillaria fungus cords to invade white hemp cortex tissue, Armillaria mycelium interspersed between white hemp cortex and cells, so the separation material used the epidermis tissue of seed hemp, but there are separation materials in this method Limited (only white hemp cortex), the problem of isolation and culture of susceptible bacteria, and the strains are highly unstable, often the strains in the year of isolation have good activity, and after transfer to the second and third generations, the strains are prone to degeneration

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  • Method for separating halimasch from endophytic fungi of gastrodia elata
  • Method for separating halimasch from endophytic fungi of gastrodia elata
  • Method for separating halimasch from endophytic fungi of gastrodia elata

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment one: from the experiment of armillaria mellea from the endophytic fungus of arrowroot

[0032] 1 Material:

[0033] Arrow hemp is collected from the main growth point of Gastrodia elata in Qinba mountainous area.

[0034] Improved PDA medium: potato (peeled) 200.0g, corn flour 30.0g, glucose 20.0g, agar 12.0g, KH2PO4 5.0g, MgSO4 3.0g, peptone 5.0g, VB1 10.0mg, pH natural, add water to 1000mL ; Dispense into test tubes, the liquid volume is 1 / 3 of the test tubes, put them upright after autoclaving, and wait for use.

[0035] Antibiotic medium: add chloramphenicol (25 μg / mL) and streptomycin (10 μg / mL) to PDA medium, after the medium is sterilized by high temperature and high pressure, cool to about 40-50°C and add chloramphenicol and streptomycin . Wherein the stock solution concentration of chloramphenicol is 34 mg / mL, and the stock solution concentration of streptomycin is 10 mg / mL.

[0036] 2 methods

[0037] 2.1 Arrow gastrodia pretreatment: select the ...

Embodiment 2

[0054] Embodiment two Gastrodia elata and armillaria cultivation test

[0055] 1 material

[0056] A total of 18 Armillaria strains from ZY-1 to ZY-18 were selected for the test Armillaria, and the germinating bacteria were Dendrobium small mushrooms. Gastrodia elata capsules used in the experiment: including Gastrodia elata red and Gastrodia green stalks, all collected from the natural distribution area of ​​Gastrodia elata in Qinba Mountains. Cultivation fungus material: the fungus material used for cultivation selects the green gang wood material, large wood (length 30cm, diameter 8cm), small wood (length 10cm, diameter 2cm), section wood (length 3cm, diameter 2cm). Cut the fish scales with a depth of about 2cm before using large and small materials.

[0057] 2 methods

[0058] 2.1 Strain preparation: Armillaria tertiary species preparation: select pre-separated and screened Armillaria strains to prepare Armillaria cultivars. The cultivars use 8×17cm plastic cultivation ...

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Abstract

The invention discloses a method for separating halimasch from endophytic fungi of gastrodia elata, thereby solving problems that the existing halimasch separation method is limited in separation material, prone to infectious microbe infection, prone to strain degradation and the like. The invention relates to a method for separating halimasch from endophytic fungi of gastrodia elata creatively; the method comprises the steps of pretreatment of gastrodia elata, tissue separation, tissue culture, separation of endophytic fungi, purification and preservation. The halimasch strain subjected to separation, purification and subculture need to be subjected to cultivation tests with gastrodia elata before application to production; the cultivation tests include asexual cultivation of hemp and halimasch and sexual cultivation of gastrodia elata seeds, germinating bacteria and halimasch. The method is high in practicability and convenient to operate; with the method, effects that the separationpollution rate is low, and the separated halimasch strains are high in infectivity and not prone to degradation in gastrodia elata cultivation can be realized.

Description

technical field [0001] The invention belongs to the technical field of Armillaria armillaria isolation, and in particular relates to a method for isolating Armillaria armillaria from endophytic fungi of arrowroot. Background technique [0002] Armillaria fungus, commonly known as hazel mushroom and corn fungus, belongs to the kingdom of fungi, basidiomycotina, agaricaceae, agaricaceae, and white mushroom family. There are about 40 species of this genus of fungi in the world, and they are widely distributed in China. They are distributed in central China, southwest and northeast regions. Armillaria is an important symbiotic fungus of famous and precious traditional Chinese medicine Gastrodia elata and Polyporus chinensis. 95% of the growth period of Gastrodia elata requires Armillaria armillaria to provide nutrients in the form of rhizome cords. Studies have shown that with different sources of Armillaria strains with Gastrodia elata, the yield and quality of Gastrodia elata...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N1/02C12R1/645
CPCC12N1/14C12N1/02
Inventor 谢海彬赵赟鑫叶彦慧解修超彭浩张晓程李庆勇
Owner 汉中市汉台区汉麓生物科技有限公司
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