Test tube screening method of isaria cicadae strain and culture medium

A technology of Isyria cicadae and screening method, which is applied in the field of microorganisms, can solve the problems such as the production cannot meet the market demand, seasonal restrictions, and the scarcity of wild cicadae resources, and achieve the effect of good selective cultivation, convenient operation and short cultivation period

Active Publication Date: 2014-02-12
ZHEJIANG BIOASIA PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Cicada has been widely recognized by the common people for its medicinal value, but due to the scarcity of wild cicada resources and seasonal restrictions, its output is far from meeting market demand

Method used

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  • Test tube screening method of isaria cicadae strain and culture medium
  • Test tube screening method of isaria cicadae strain and culture medium
  • Test tube screening method of isaria cicadae strain and culture medium

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Collect wild cicadae (No. 1) from the wild, bring it back to the laboratory, use the inoculation needle to pick the conidia to the potato glucose plate medium containing antibacterial antibiotics, the composition of the plate medium is: potato 20wt%, glucose 2wt% , agar 2wt%, antibiotic 0.01%. Incubate in the dark at 25°C for 4 days. Pick the colony of I. cicadae from the plate medium to the potato glucose slant medium, the composition of the slant medium is: 15wt% potato, 2.5wt% glucose, 2.5wt% agar, cultivated at 25°C for 10 days, and obtain the number 1 strain. Scrape the mature conidia from the inclined plane with an inoculation loop, add them into sterilized water, shake and mix well, and prepare 10 6 / ml of spore suspension. Soak the wheat overnight, drain it, add 1 / 4 volume of imbibition wheat (4g) to the test tube, add 6ml of water, plug it with a cotton plug, sterilize at 121°C for 50 minutes, and cool down. Transfer the spore suspension to the wheat cultur...

Embodiment 2

[0037] Collect wild cicadae (No. 2) from the field, bring it back to the laboratory, use an inoculation needle to pick conidia into the potato sucrose plate medium containing antibacterial antibiotics, the composition of the plate medium is: 25wt%, glucose 1.5wt %, agar 2.5wt%, antibiotics 0.02%, cultured in the dark at 23°C for 5 days. Pick the colony of I. cicadae from the plate culture medium to the potato sucrose slant medium, the composition of the slant medium is: 20wt% potato, 1.5wt% sucrose, 1.5wt% agar, cultivated at 23°C for 15 days, and obtain No. 2 strain. Scrape the mature conidia from the inclined plane with an inoculation loop, add them into sterilized water, shake and mix well, and prepare 10 7 / ml of spore suspension. Soak the wheat overnight, drain it, add 1 / 7 of the tube volume of imbibition wheat (2.5g) to the test tube, add 3.75ml of water, put on a cotton plug, and sterilize at 121°C for 50 minutes. Inoculate the spore suspension on the wheat medium ac...

Embodiment 3

[0039] Collect wild cicadae (No. 3) from the field, bring it back to the laboratory, use the inoculation needle to pick the conidia to the potato sucrose plate medium containing antibacterial antibiotics, the composition of the plate medium is: 15wt%, sucrose 2.5wt% , 1.5wt% agar, 0.015% antibiotics, cultured in the dark at 24°C for 5 days. Pick the colony of I. cicadae from the plate culture medium to the potato sucrose slant medium, the composition of the slant medium is: 25wt% potato, 2.0wt% sucrose, 2.5wt% agar, cultivated at 24°C for 13 days, and obtain No. 3 strain. Scrape the mature conidia from the inclined plane with an inoculation loop, add to sterilized water, shake and mix well, and prepare 5×10 6 / ml of spore suspension. Soak the wheat overnight, drain it, and add 1 / 3 of the tube volume of imbibition wheat (5g) to the test tube, add 5ml of water, plug it with a cotton plug, and sterilize at 121°C for 50 minutes. The spore suspension was inoculated on the wheat ...

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Abstract

The invention relates to the field of microorganisms, and in particular discloses a test tube screening method of an isaria cicadae strain. The method comprises the following steps: collecting wild cordyceps cicadae or conidium thereof, and separating and purifying through a panel culture medium to obtain isaria cicadae single colony; picking the isaria cicadae single colony from the panel culture medium in the former step to a slope culture medium, and culturing to obtain conidium; collecting the conidium obtained from the former step, preparing spore solution, and inoculating to a test tube equipped with a test tube culture medium made of wheat; culturing and observing sporocarp, spore or hypha production condition. The test tube culture is adopted by the test tube screening method of the isaria cicadae strain, the occupied culture area is small, the operation is convenient, the culture period is short, the seed culture process in a shake flaks is not needed, the labor intensity is greatly reduced, and the energy consumption in the screening process is reduced; the wheat culture medium has good isaria cicadae strain selective culture effect.

Description

technical field [0001] The invention relates to the field of microbes, and specifically discloses a test tube screening method and a culture medium for I. cicadae strains. Background technique [0002] Cicada is a medicinal insect-growing fungus, which is a traditional precious Chinese herbal medicine in my country. Its medicinal use was first recorded in "Lei Gong Pao Shao Lun" in Liu Song Dynasty in the Southern and Northern Dynasties. Traditional medicine believes that cicadae has the effects of evacuating wind-heat, rash, relieving wind and spasm, and improving eyesight. Modern medical research shows that it has various physiological functions such as immune regulation, improving the nutritional status of the body, improving kidney function, anti-tumor, anti-virus, antipyretic and analgesic, sedative and hypnotic, nourishing and strong. Studies have confirmed that cicadae is rich in biologically active substances and nutrients. Cicadae is rich in protein, amino acids, ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12R1/645
Inventor 谭悠久陈祝安樊美珍吕雯婷陈超曾奕婷
Owner ZHEJIANG BIOASIA PHARMA CO LTD
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