Engineering bacteria with inactivated cordyceps militaris Rhf1 gene and application of cordyceps militaris Rhf1 gene

A technology for Cordyceps militaris and engineering bacteria, which is applied in the field of biological genetic engineering and can solve the problems of large demand, inability to artificially cultivate, and decline in the yield of Cordyceps sinensis.

Active Publication Date: 2014-09-10
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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  • Description
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  • Application Information

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Problems solved by technology

[0002] Cordyceps sinensis is a precious traditional Chinese medicine with high demand and high price
However, the production of Cordyceps sinensis has declined sharply, and it is still not artificially cultivated so far

Method used

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  • Engineering bacteria with inactivated cordyceps militaris Rhf1 gene and application of cordyceps militaris Rhf1 gene
  • Engineering bacteria with inactivated cordyceps militaris Rhf1 gene and application of cordyceps militaris Rhf1 gene
  • Engineering bacteria with inactivated cordyceps militaris Rhf1 gene and application of cordyceps militaris Rhf1 gene

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Embodiment 1

[0061] 1. Experimental materials and methods

[0062] 1. Strain culture

[0063] Agrobacterium AGL-1 culture: LB medium, cultivated in the dark at 28°C.

[0064] E.coli DH5α culture: LB medium, cultivated in the dark at 37°C.

[0065] Cordyceps militaris culture: PPDA medium, cultured at 22-23°C.

[0066] 2. Minimized extraction of genomic DNA (E.Z.N.A HP Fungal DNA Kit)

[0067] 3. Extraction of total RNA from Cordyceps militaris

[0068] Total RNA was extracted by TRizol method,

[0069] 4. Agrobacterium-mediated (ATMT) transformation of Cordyceps militaris

[0070] 5. Collection of conidia of Cordyceps militaris JM4 and Cordyceps militaris mutant g38

[0071] Collection of conidia of wild-type Cordyceps militaris JM4 and mutant g38 Cordyceps militaris: directly pick the mycelium with an inoculation needle and spread it on a solid PPDA medium, cultivate it in the dark at 23°C, and after 7-10 days, use 10 mL of The conidia were washed from the plate with ultrapure wate...

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Abstract

The invention discloses engineering bacteria with an inactivated cordyceps militaris Rhf1 gene and an application of the cordyceps militaris Rhf1 gene. A cordyceps militaris muton g38 with varied hyphae branches, increased fungus pellet biomass, advanced primordium differentiation, sturdy sporocarp, shortened sporocarp harvest time and increased sporocarp dry weight is obtained in a screening manner. Experiments find that the cordyceps militaris Rhf1 gene is inactivated due to insertion of a split hypha protein gene. Furthermore, the condition that the split hypha protein gene is related to hyphae branch, conidiophore differentiation, primordium differentiation rate, thickness of the sporocarp or sporocarp yield is confirmed by the invention. Therefore, a solution channel is provided for shortening the cultivation time, improving the sporocarp yield and the like in the cultivation process of the cordyceps militaris.

Description

Technical field: [0001] The invention belongs to the field of biological genetic engineering, and in particular relates to an engineering bacterium for inactivating the Rhf1 gene of Cordyceps militaris and the application of the Rhf1 gene of Cordyceps militaris. Background technique: [0002] Cordyceps sinensis is a precious traditional Chinese medicine with high demand and high price. However, the output of Cordyceps sinensis has declined sharply, and it is still not artificially cultivated so far. Cordyceps militaris has similar effects to Cordyceps sinensis, and has become the preferred substitute of Cordyceps sinensis. In the process of artificial commercial cultivation of Cordyceps militaris, how to solve the degradation of strains, shorten the cultivation time, increase the yield of fruiting bodies and the content of active ingredients, and enhance the ability of Cordyceps militaris to infect insects are core technical issues. [0003] As a traditional Chinese herbal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/15C12N15/09C12R1/645
Inventor 江克清韩日畴
Owner INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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