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Method for breeding cordyceps militaris by hybridizing with ascospore on basis of molecular biological technology

A molecular biology and spore hybridization technology, applied in the biological field, can solve the problems of low fruiting body yield and active substance content, and achieve the effects of improving economic benefits and medicinal value, shortening the hybrid breeding cycle, and increasing yield and active substances.

Inactive Publication Date: 2017-12-22
CHANGDE YANDI BIOTECH LTD CO
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The invention relates to a method for hybrid breeding of ascospores of Cordyceps militaris based on molecular biology technology, which solves the problem that the yield of fruit bodies and the content of active substances are low due to the genetic characteristics of the species of Cordyceps militaris

Method used

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  • Method for breeding cordyceps militaris by hybridizing with ascospore on basis of molecular biological technology
  • Method for breeding cordyceps militaris by hybridizing with ascospore on basis of molecular biological technology
  • Method for breeding cordyceps militaris by hybridizing with ascospore on basis of molecular biological technology

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Experimental program
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Effect test

Embodiment 1

[0028] (1) Under sterile conditions, suspend the fresh and mature fruiting body of Cordyceps militaris (the fruiting body comes from the megaspore head variety cultivated by the strain bank of Changde Yandi Biotechnology Co., Ltd.) on the water agar medium to avoid The fruiting body is in contact with the culture medium;

[0029] (2) Transfer it to a biochemical incubator, and cultivate it in the dark at 21°C for 4-5 days, so that the ascospores are naturally ejected onto the PDA medium;

[0030] (3) Pick the bacterium colony that has only a single ascospore under a stereomicroscope, then add 2ml of sterile water, stir evenly, prepare a spore suspension, get 200 microliters of the spore suspension, and apply it on the PDA plate medium, Placed in a biochemical incubator at 21°C and cultured in the dark;

[0031] (4) After cultivating for 3-4 days, select a single colony of ascospores visible to the naked eye on the ultra-clean workbench, and transfer it to a new PDA plate medi...

Embodiment 2

[0039] (1) Under sterile conditions, suspend the fresh and mature fruiting body of Cordyceps militaris (the fruiting body comes from the microspore head variety cultivated by the strain bank of Changde Yandi Biotechnology Co., Ltd.) above the water agar medium to avoid The fruiting body is in contact with the culture medium;

[0040] (2) Transfer it to a biochemical incubator, and cultivate it in the dark at 21°C for 4-5 days, so that the ascospores are naturally ejected onto the PDA medium;

[0041](3) Pick a colony with only a single ascospore under a stereomicroscope, then add 2ml of sterile water, stir evenly, and prepare a spore suspension, get 200 microliters of the spore suspension, and apply it on the PDA plate medium, Placed in a biochemical incubator at 21°C and cultured in the dark;

[0042] (4) After cultivating for 3-4 days, select a single colony of ascospores visible to the naked eye on the ultra-clean workbench, and transfer it to a new PDA plate medium, where...

Embodiment 3

[0050] (1) Under sterile conditions, suspend fresh, mature fruiting bodies of Cordyceps militaris (the fruiting bodies come from the acerospore powder variety cultivated in the strain bank of Changde Yandi Biotechnology Co., Ltd.) above the water agar medium, Avoid fruiting bodies in contact with the culture medium;

[0051] (2) Transfer it to a biochemical incubator, and cultivate it in the dark at 21°C for 4-5 days, so that the ascospores are naturally ejected onto the PDA medium;

[0052] (3) Pick a colony with only a single ascospore under a stereomicroscope, then add 2ml of sterile water, stir evenly, and prepare a spore suspension, get 200 microliters of the spore suspension, and apply it on the PDA plate medium, Placed in a biochemical incubator at 21°C and cultured in the dark;

[0053] (4) After cultivating for 3-4 days, select a single colony of ascospores visible to the naked eye on the ultra-clean workbench, and transfer it to a new PDA plate medium, where only a ...

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Abstract

The invention relates to a method for breeding cordyceps militaris by hybridizing with ascospore on the basis of a molecular biological technology. The method specifically comprises the following steps: inducing and isolating cordyceps militaris monospores in a sterile environment and culturing to form colonies; using a PCR technology for detecting mating type genes MAT1-1-1, MAT1-1-2 and MAT1-2-1 of the colonies formed by the isolated monospores; selecting the colonies formed by the monospores containing the mating type genes MAT1-1-1 and MAT1-1-2 but not containing MAT1-2-1 and the colonies formed by the monospores containing the mating type gene MAT1-2-1 but not containing MAT1-1-1 and MAT1-1-2; respectively taking mycelia appropriate in size to be placed in a culture solution for dark culturing; inoculating an appropriate concentration of combination-cultured bacteria solution into an artificial culture medium of cordyceps militaris; culturing for a period of time to grow a new variety of cordyceps militaris sporophore from the artificial culture medium. The new variety of cordyceps militaris cultured by the method is easy to obtain relatively good yield and content of active substances of sporophores; a combination capable of stably producing high-quality cordyceps militaris sporophores can be obtained by counting the yield of sporophores and active substances cultured by different combinations; the method is relatively short in cycle and relatively small in workload, and is an efficient method for breeding cordyceps militaris.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for hybrid breeding of Cordyceps militaris ascospores based on molecular biology technology. Background technique [0002] In the sexual reproduction process of fungi, the mating type can explain the genetic control of fungi and the process of sexual reproduction development. Therefore, the mating type gene plays a decisive role in the sex control and genetic evolution of fungi, and it is also a sex compatible phenotype genetic determinants. The mating systems of known fungi are divided into two categories: homothalmic and heterothallic. Homogenous mating is a form of sexual reproduction that is self-fertile, and heterothallic mating is a form of sexual reproduction that is autogenously infertile. Heterothalism is divided into two types according to whether the incompatibility factors controlling the mating type are one pair or two pairs: bipolar heterothalism with one pai...

Claims

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Application Information

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IPC IPC(8): A01G1/04C12N3/00C12Q1/68C12R1/645
CPCA01G18/00C12N3/00C12Q1/6895
Inventor 阳武雄易思富杨子遥王卫周丰杨胜辉
Owner CHANGDE YANDI BIOTECH LTD CO
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