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An engineered strain with inactivated Cordyceps militaris rhf1 gene and application of the Cordyceps militaris rhf1 gene

A technology of Cordyceps militaris and engineering bacteria, which is applied to the engineering bacteria with inactivated Cordyceps militaris Rhf1 gene, the application field of Cordyceps militaris Rhf1 gene, and can solve the problems of decreased yield of Cordyceps militaris, incapability of artificial cultivation, high price, etc.

Active Publication Date: 2016-02-24
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • An engineered strain with inactivated Cordyceps militaris rhf1 gene and application of the Cordyceps militaris rhf1 gene
  • An engineered strain with inactivated Cordyceps militaris rhf1 gene and application of the Cordyceps militaris rhf1 gene
  • An engineered strain with inactivated Cordyceps militaris rhf1 gene and application of the Cordyceps militaris rhf1 gene

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Experimental program
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Effect test

Embodiment 1

[0061] 1. Experimental materials and methods

[0062] 1. Strain culture

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

[0064] E. coliDH5α culture: LB medium, cultured at 37°C in the dark.

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

[0066] 2. Minimal extraction of genomic DNA (E.Z.N.AHPFungalDNAKit)

[0067] 3. Cordyceps militaris total RNA extraction

[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 wild-type Cordyceps militaris JM4 and mutant g38 Cordyceps militaris conidia: directly pick the mycelium with an inoculating needle and plate it on solid PPDA medium, culture at 23°C in the dark, and after 7-10 days, add 10 mL without The conidia were washed from the plate with ultrapure water and filtered twice with th...

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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 with Rhf1 gene of Cordyceps militaris inactivated and the application of the Rhf1 gene of Cordyceps militaris. Background technique: [0002] Cordyceps sinensis (Cordyceps sinensis) is a precious traditional Chinese medicine with high demand and high price. However, the yield of Cordyceps sinensis has dropped sharply, and so far it cannot be cultivated artificially. Cordyceps militaris (Cordyceps militaris) has similar efficacy to Cordyceps sinensis and has become the preferred substitute for Cordyceps sinensis. In the process of artificial commercial cultivation of Cordyceps militaris, how to solve the degradation of bacteria, 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, etc., are the core technical pr...

Claims

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Application Information

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