A kind of protoplast preparation and regeneration method of Lily mushroom

A technology of lotus leaves and protoplasts, applied in the field of bioengineering, to reduce the loss of protoplasts, reduce the cost of preparation, and improve the production rate

Active Publication Date: 2017-04-05
梁倩倩
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research on the preparation process and method of the protoplasts of the lotus leaf chrysanthemum in China. Therefore, it is of great significance to use an efficient and economical method to prepare the protoplasts of the lotus leaf chrysanthemum

Method used

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  • A kind of protoplast preparation and regeneration method of Lily mushroom
  • A kind of protoplast preparation and regeneration method of Lily mushroom
  • A kind of protoplast preparation and regeneration method of Lily mushroom

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Embodiment Construction

[0025] Bacteria of the present invention

[0026] The lotus-leaved mushroom strain (preservation number CGMCC1518, strain number ZY48-1, patent number ZL200510096405.0) was provided by the Edible Fungi Research Institute of Hexi University.

[0027] Reagents of the present invention

[0028] Lysozyme (Guangdong Provincial Institute of Microbiology); Helicase (Beijing Baitai Biochemical Technology Company); Cellulase (US Bettery Company); other reagents were domestic analytically pure.

[0029] Configuration of enzyme solution

[0030] Dissolve lytic enzyme and helicase with 0.6mol / L sucrose osmotic pressure stabilizer, make lytic enzyme mass concentration be 2%, helicase mass concentration be 1%, with 2% lytic enzyme and 1% helicase by volume ratio Mix 1:1, so that the concentration of lysozyme in the mixed enzyme is 1%, and the concentration of helicase is 0.5%, centrifuge at 10000r / min for 10min, take the supernatant and use 0.22 mu m microporous membrane filtration, sta...

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Abstract

The invention relates to a preparation and regeneration method of a lyophyllum decastes protoplast. The preparation and regeneration method comprises the following steps of treating lyophyllum decastes hyphae having fungus age of 72-96h by a cane sugar osmotic pressure stabilizing agent having content of 0.6mol / L, carrying out enzymolysis on a mixed enzyme liquid containing 1% by mass of lywallzyme and 0.5% by mass helicase at a temperature of 30 DEG C for 2.5-3h, preparing a self-made needle filter, and carrying out separation by the self-made needle filter to obtain the lyophyllum decastes protoplast. The preparation and regeneration method has a protoplast yield of 4.79*10<7> per milliliter (5.76*10<5> per milligram) and a protoplast regeneration rate of 7.81%. Compared with single lywallzyme, the mixed enzyme liquid containing lywallzyme and helicase improves protoplast preparation and regeneration rate and reduces a cost. The preparation and regeneration method utilizes a small-needle and small-area filtration method so that a cost is lower than that of the routine method. The preparation and regeneration method lays a foundation for lyophyllum decastes breeding adopting the protoplast technology.

Description

technical field [0001] The invention belongs to the field of bioengineering, and relates to a method for preparing and regenerating protoplasts of the chrysanthemum chrysanthemum. Background technique [0002] Lotus Leaf Shimegum ( Lyophyllum decays ) belongs to Basidiomycetes, Agaricaceae, Agaricaceae, Tricholomaceae, and Liphyme genus edible fungi. Its mycelium and fruiting bodies have high protein content, complete amino acid types, and multivitamins. It is a wild edible fungus. The totipotency of protoplasts of edible fungi is easier to reflect, so protoplast mutagenesis and fusion breeding technology is one of the most efficient and economical methods in the breeding of high-yielding and new strains of edible fungi. The successful preparation of protoplasts is a necessary condition for protoplast mutagenesis and fusion breeding techniques. At present, there is no research on the preparation process and method of the protoplasts of the chrysanthemum chrysanthemum in Ch...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04
Inventor 梁倩倩席亚丽魏生龙
Owner 梁倩倩
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