Fabrication method of cover glass for conidiospore of Ophio cordyceps sinensis and dynamic counting method of produced conidiospore

A technology of Cordyceps sinensis and conidia, which is applied in the direction of biochemical equipment and methods, measurement/inspection of microorganisms, etc., can solve the problems of long sporulation cycle, low sporulation amount, and poor culture observation and counting, and achieve the goal of working High efficiency, simplified experimental procedures, and good research and application value

Inactive Publication Date: 2012-09-19
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the conidia of Cordyceps sinensis often fails to meet the counting standard of the blood count method due to the small amount of sporulation and the long cycle of sporulation, so there has been no good solution for the cultivation, observation and counting

Method used

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  • Fabrication method of cover glass for conidiospore of Ophio cordyceps sinensis and dynamic counting method of produced conidiospore

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, Cordyceps sinensis conidia count

[0026] 1. The cultivation of Cordyceps sinensis

[0027] Each 1L of solid medium for Cordyceps sinensis is prepared according to Chinese patent ZL200510137457.8: 50.0 grams of wheat bran, 20.0 grams of glucose, 5.0 grams of peptone, 3.0 grams of yeast extract, 200 grams of potatoes, 18.0 grams of agar, and dilute to 1000 milliliters with water.

[0028] Separation and purification from natural Cordyceps sinensis (see Dong, C.-H. and Yao, Y.-J. (2008). In vitroevaluation of antioxidant activities of aqueous extracts from natural and cultured mycelia of Cordyceps sinensis. LWT-Food Science and Technology 41(4): 669-677.) obtained Ophiocordyceps sinensis strain No. 762, which has been preserved in China General Microorganism Culture Collection Center (CGMCC No.2793), a patent strain.

[0029] Cordyceps sinensis (Ophio cordyceps sinensis) strain 762 was preserved on December 5, 2008 in the General Microbiology Center of Chin...

Embodiment 2

[0038] Embodiment 2, Cordyceps sinensis conidia count

[0039] 1. The cultivation of Cordyceps sinensis

[0040] Each 1L of solid medium for Cordyceps sinensis is prepared according to Chinese patent ZL200510137457.8: 50.0 grams of wheat bran, 20.0 grams of glucose, 5.0 grams of peptone, 3.0 grams of yeast extract, 200 grams of potatoes, 18.0 grams of agar, and dilute to 1000 ml with water.

[0041] Cordyceps fungus strain No. 762 CGMCC No.2793 isolated and purified from natural Cordyceps sinensis.

[0042] The Cordyceps sinensis strain No. 762 obtained by the above isolation and purification was extracted, amplified and sequenced by DNA, and analyzed by using the molecular database dedicated to Cordyceps sinensis, and identified as a true Cordyceps sinensis strain by molecular biology.

[0043] Cordyceps sinensis strain No. 762 CGMCC No.2793 was inoculated in the center of a petri dish containing a solid special medium for Cordyceps sinensis, placed at 18°C, and cultured in th...

Embodiment 3

[0049] Embodiment 3, Cordyceps sinensis conidia count

[0050] 1. The cultivation of Cordyceps sinensis

[0051] Each 1L of solid medium for Cordyceps sinensis is prepared according to Chinese patent ZL200510137457.8: 50.0 grams of wheat bran, 20.0 grams of glucose, 5.0 grams of peptone, 3.0 grams of yeast extract, 200 grams of potatoes, 18.0 grams of agar, and dilute to 1000 ml with water.

[0052] Separation and purification from natural Cordyceps sinensis to obtain Cordyceps sinensis strain No. 762 CGMCC No.2793.

[0053] The Cordyceps sinensis strain No. 762 obtained by the above isolation and purification was extracted, amplified and sequenced by DNA, and analyzed by using the molecular database dedicated to Cordyceps sinensis, and identified as a true Cordyceps sinensis strain by molecular biology.

[0054] Inoculate Cordyceps sinensis strain No. 762 CGMCC No.2793 in the center of a petri dish containing a special solid medium for Cordyceps sinensis, place it at 18°C, a...

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Abstract

The invention discloses a fabrication method of cover glass for conidiospores of Ophio cordyceps sinensis and a dynamic counting method of produced conidiospores. The method comprises inoculating Ophio cordyceps sinensis at each hole on a solid culture medium containing at least one hole to obtain at least one inoculated hole; arranging a piece of cover glass above each inoculated hole to obtain a cover glass plate; and culturing the cover glass plate until a colony grows, taking 3 pieces of cover glass every a period of time, and counting to obtain the number of conidiospores produced by Ophio cordyceps sinensis at different moments. According to the experiment, the invention simplifies experimental operation, can in-situ observe and count at fixed points through an inverted microscope, and objectively reflects the conidiospore production cycle of Ophio cordyceps sinensis strains.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a method for the preparation of conidia coverslips of Cordyceps sinensis and the dynamic counting method for sporulation. Background technique [0002] In microbiological research experiments, microscopic counting is required in many research fields such as cell suspension culture, conidia observation, and growth curve drawing. Microscopic counting of conidia is a basic technique in fungal biology research experiments. It is the basis for understanding the sporulation state of cultured strains, drawing the dynamic curve of sporulation, and measuring the effects of different medium formulations, culture methods, and growth stress on the sporulation of strains. important means. Commonly used counting methods include dilution plate method, turbidimetric counting method, MTT colorimetric method, hemocytometer plate counting method, etc. Dilution plate method and turbidimetric c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/06
Inventor 姚一建任蜀豫王文婧
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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