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Fusion method for protoplast of Cordyceps sinensis and Cordyceps militaris

A technology of protoplast fusion and Cordyceps sinensis, which is applied in biochemical equipment and methods, botanical equipment and methods, fungal products, etc., can solve problems such as poor results, achieve strong repeatability, be beneficial to food quality and safety, and improve food quality and safety. The effect of probability of success

Inactive Publication Date: 2013-08-28
TIANJIN NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the breeding of edible fungi is mostly artificial mutagenesis. Due to the non-directional characteristics of mutagenesis, the effect of solving the above problems is not good. There is no report on the fusion breeding of Cordyceps sinensis and Cordyceps militaris at home and abroad.

Method used

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  • Fusion method for protoplast of Cordyceps sinensis and Cordyceps militaris
  • Fusion method for protoplast of Cordyceps sinensis and Cordyceps militaris
  • Fusion method for protoplast of Cordyceps sinensis and Cordyceps militaris

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Embodiment 1

[0034] (1) Take the front end of the vigorously growing mycelium on the PDA comprehensive medium, inoculate it into a 100mL triangular flask filled with Cordyceps sinensis liquid medium, the filling volume is 50mL, and each bottle is connected with 3 to 4 pieces of 0.5cm bacteria block, at 25°C Leave it to culture in the dark for 5-11 days; pick the mycelium under aseptic conditions, absorb the water with sterile filter paper, and obtain the fresh mycelium of Cordyceps sinensis.

[0035] (2) Take fresh Cordyceps sinensis and Cordyceps militaris mycelium, and carry out enzymatic hydrolysis at the ratio of 0.1g wet weight to 1ml enzyme solution. The enzymatic hydrolysis temperature of silk protoplasts is 30°C, and the enzymatic hydrolysis time is 2 hours. The preparation of Cordyceps sinensis protoplasts according to the present invention adopts the mycelia of liquid static culture for 3 days, and uses 0.6 mol / L KCL as an osmotic pressure stabilizer. % Lyticase (mass volume rati...

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Abstract

The invention provides a fusion method for protoplast of Cordyceps sinensis and Cordyceps militaris. The method is characterized by performing enzymatic hydrolysis for mycelia cells of the Cordyceps sinensis and the Cordyceps militaris to prepare the protoplast, performing thermal inactivation for the protoplast of the Cordyceps sinensis, performing UV-inactivation for the protoplast of the Cordyceps militaris, using a PEG-mediated chemical fusion method, fusing the protoplast of the Cordyceps sinensis and the Cordyceps militaris, and obtaining the fusion strain through colony morphology, an antagonism test and an RAPD (random amplified polymorphic DNA) integrated verification. The method can effectively fuse the protoplast of the Cordyceps sinensis and the Cordyceps militaris to obtain the new strain as an artificial substitute for the Cordyceps sinensis, thereby providing an effective method for breeding objective strains by protoplast fusion.

Description

technical field [0001] The invention belongs to the field of edible fungus biotechnology breeding, and provides a method for merging the biomass of Cordyceps sinensis and Cordyceps militaris, and obtains a fusion strain with both directors, which can be used as an artificial substitute for Cordyceps sinensis. The construction of new target strains by interspecific fusion within the genus of edible fungi provides an effective method. Background technique [0002] In recent years, due to many factors, the resources of wild Cordyceps sinensis are scarce, the output has dropped sharply, and its market price is very expensive. Due to the constraints of its unique habitat and other conditions, artificial cultivation is difficult, and no breakthrough has been made yet. In view of the fact that Cordyceps sinensis and Cordyceps militaris have similar genesis mechanism and nutritional requirements and both belong to the genus Cordyceps, the breeding of Cordyceps sinensis and Cordyceps...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/05A01H15/00
Inventor 郭成金刘西周
Owner TIANJIN NORMAL UNIVERSITY
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