Efficient cordyceps sinensis ascospore separating method

A technology for spore separation and ascospores, which is applied in the field of ascospore separation of Cordyceps sinensis, which can solve the problems of time-consuming and labor-consuming, fragile ascospores, and easy contamination, and achieve the effects of low cost, high success rate and strong practicability

Inactive Publication Date: 2019-05-31
LANZHOU JIAOTONG UNIV
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One is to obtain Cordyceps sinensis through tissue separation. This method is relatively simple and common. As long as the surface is properly sterilized, there is a high probability of obtaining Cordyceps sinensis. It is widely used, but this method has a disadvantage that it cannot guarantee the obtained Cordyceps sinensis is an absolute purebred
There is also a method for obtaining Cordyceps sinensis by separating ascospores. The disadvantage of this method is that the ascospores are exposed to the external environment during the ejection period, and the ascospores are fragile and cannot be disinfected on the surface. It is easy to be contaminated. Although micromanipulation separation with the help of related advanced instruments can be successful, it is very time-consuming and labor-intensive, and is not suitable for large-scale separation operations

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Efficient cordyceps sinensis ascospore separating method
  • Efficient cordyceps sinensis ascospore separating method
  • Efficient cordyceps sinensis ascospore separating method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, germination culture of Cordyceps sinensis ascospores.

[0029] The Cordyceps sinensis used in the present invention is the fresh Cordyceps sinensis of the ascospore ejection period in Tianzhu area, Gansu Province. Its main steps are: 1. Cover the sterilized cellophane bag on the fruiting body of Cordyceps sinensis during the ascospore ejection period to collect ascospores ( figure 1 a). 2. Dilute the collected ascospores with a small amount of sterile water. After calculating the ascospore density with a hemocytometer, dilute the ascospore suspension to 45 / ml. 3. Pipette gun to quantitatively draw 1ml of ascospore dilution on a 90mm germination medium plate, spread it evenly with a middle coating rod, and then transfer it to a 15°C constant temperature incubator for germination culture. During this period, continuously observe the ascospore germination with a microscope.

Embodiment 2

[0030] Embodiment 2, the isolation and cultivation of Cordyceps sinensis ascospores

[0031] After 3-5 days of germination and culture, the ascospores will begin to germinate and can enter the stage of isolation and culture. The main steps are: 1. Transfer the 90mm germination medium plate into the ultra-clean workbench, and observe the germination of ascospores with an ordinary optical microscope. 2. After finding a single ascospore, confirm it repeatedly with a high-power lens to ensure that the ascospore in the field of view is a germinated single spore ( figure 1 d). 3. Turn off the fluorescent lamp of the ultra-clean workbench and all ambient light sources to create a dark environment. 4. Use a self-made ring sampler ( figure 1 b) Aim at the bright circular spot formed by the light path of the microscope on the surface of the germination medium in a dark environment ( figure 1 c) Hole sampling. 4. Inoculate the sampling blocks containing germinated monospores one ...

Embodiment 3

[0032] Embodiment 3, the vegetative growth cultivation of Cordyceps sinensis

[0033] When isolated and cultured for about 30 days, the Cordyceps sinensis fungus developed from a single germinated ascospore in the petri dish began to produce signs of growth visible to the naked eye ( figure 2 a), the colony morphology was obvious at 60 days ( figure 2 b), the diameter of the colony is about 1cm, and the diameter of the colony further increases after 90 days, and the pigment is obvious ( figure 2 c) At this time, the Cordyceps sinensis fungus developed from a single ascospore has been successfully obtained. If further research and identification is required, the vegetative growth link can be carried out first. The main steps are: 1. The Cordyceps sinensis obtained through isolation and culture Army, use a 5mm puncher to punch a sample, and inoculate it on a 70mm nutrient medium plate. 2. After the sealing film is sealed, transfer them to a constant temperature incubator at...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a cordyceps sinensis ascospore separating method. The method is characterized in that fresh ascospores popped by cordyceps sinensis are collected, after being diluted, sterilewater coats a germination culture medium, a common optical microscope is used for observing the germination condition of ascospores, under the dark environment, bright light spots formed by a light path of a microscope are used for marking the positions where ascospores are germinated in real time, a customized annular sampling device is used for being aligned to the light spot position for punching and sampling, the germinated monospores are transferred to an isolation medium for culture, and purebred ophiocordyceps sinensis obtained by developing single ascopspores is obtained. The method has the advantages of being simple, precise, high in efficiency, low in cost, capable of completing one separation operation every 90 s with the success rate being 70% or above, capable of being used for separation of large batches of cordyceps sinensis ascospores to obtain a large amount of purebred ophiocordyceps sinensis.

Description

Technical field [0001] The invention relates to the field of pure culture of Cordyceps sinensis, and specifically provides a method for isolating Cordyceps ascospores, which is simple, efficient, low in cost and high in success rate, and can be applied to the large-scale separation of Cordyceps ascospores, and the obtained Cordyceps ascospores All are pure species developed from a single ascospore, no secondary purification is required. Background technique [0002] Cordyceps sinensis (Cordyceps sinensis) is a rare and precious wild Chinese medicinal material distributed in the Qinghai-Tibet Plateau and its surrounding areas. It is famous all over the world for its high medicinal value. Its essence is Cordyceps sinensis fungus ( Ophiocordyceps sinensis ) infects the fungus complex formed by the bat moth larvae in the alpine meadow. Cordyceps sinensis has important research and application value. The Bailing Capsules developed by using the fermented bacteria powder of Co...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12R1/645
Inventor 陈晨谢放夏樱霞
Owner LANZHOU JIAOTONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap