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Enzymatic hydrolysate for preparing lonicera caerulea protoplast as well as preparation method and application of enzymatic hydrolysate

A technology of protoplasts and enzymatic hydrolysis, applied in the direction of biochemical equipment and methods, enzymes, plant cells, etc., can solve the problems of high protoplast quantity, protoplast integrity and poor activity, etc., to ensure integrity and activity , large application value, and the effect of increasing the quantity

Active Publication Date: 2022-03-22
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the large differences in the cell structure of different plants, it is difficult to obtain a higher number of protoplasts by using the protoplast isolation method of other plants to process the leaves of Lonicera japonica, and the integrity and activity of the protoplasts obtained are not good. good

Method used

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  • Enzymatic hydrolysate for preparing lonicera caerulea protoplast as well as preparation method and application of enzymatic hydrolysate
  • Enzymatic hydrolysate for preparing lonicera caerulea protoplast as well as preparation method and application of enzymatic hydrolysate
  • Enzymatic hydrolysate for preparing lonicera caerulea protoplast as well as preparation method and application of enzymatic hydrolysate

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preparation example Construction

[0042]The present invention preferably also provides a method for preparing the above-mentioned enzymolysis liquid, comprising the following steps: mixing cellulase, isolated enzyme, MES, mannitol and KCl, and bathing in 55°C water bath for 10 minutes to obtain a mixed liquid; cooling the mixed liquid to 20-25°C After that, the mixture and CaCl 2 and BSA were mixed, and the volume was constant to obtain the enzymolysis solution. In the present invention, the enzyme can be fully dissolved and mixed at a suitable temperature through a water bath without destroying the activity of the enzyme and improving the subsequent enzymatic hydrolysis efficiency.

[0043] The present invention also provides the application of the above-mentioned enzymatic hydrolysis solution in preparing protoplasts of Lonicera japonica. The enzymatic hydrolysis solution provided by the invention can remarkably increase the quantity of the prepared protoplasts of the blue honeysuckle, and at the same time ...

Embodiment 1

[0058] An enzymolysis solution for preparing protoplasts of Lonicera japonica plant, which consists of the following components: Cellulase R-10 20g / L, Isolase R-10 5g / L, MES 20mmol / L, Mannitol 0.7mol / L , KCl 20mmol / L, CaCl 2 20mmol / L and BSA1g / L; the enzyme activity of the cellulase R-10 is >10000U / g; the enzyme activity of the isolated enzyme R-10 is >3000U / g.

[0059] The preparation of the enzymolysis solution consists of the following steps:

[0060] Mix cellulase R-10, isolated enzyme R-10, MES, mannitol and KCl, and bathe in water at 55°C for 10 minutes to obtain a mixed solution; after the mixed solution is cooled to room temperature (20-25°C), mix the mixed solution with CaCl 2 and BSA were mixed, and the volume was constant to obtain the enzymolysis solution.

Embodiment 2

[0062] A kind of enzymatic hydrolysis solution similar to Example 1, the only difference is that the concentration of mannitol in the enzymolysis solution is 0.8mol / L.

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Abstract

The invention relates to the technical field of plant protoplast, in particular to enzymatic hydrolysate for preparing lonicera caerulea protoplast as well as a preparation method and application of the enzymatic hydrolysate. The lonicera caerulea leaf is used as a material to prepare the protoplast, and the enzymatic hydrolysate suitable for lonicera caerulea is obtained by compounding proper components according to the species characteristics of lonicera caerulea, the structure characteristics of the leaf and the like. According to the enzymatic hydrolysate provided by the invention, the number of the protoplast of the prepared lonicera caerulea can be remarkably increased, and meanwhile, the integrity and activity of the protoplast can be guaranteed. Besides, the enzymatic hydrolysate provided by the invention is beneficial to implementation of lonicera caerulea protoplast fusion breeding, provides an important material for lonicera caerulea cell engineering breeding and molecular biology research based on cell fusion and gene transformation, and has a relatively high application value.

Description

technical field [0001] The invention relates to the technical field of plant protoplasts, in particular to an enzymolyzed liquid for preparing protoplasts of Lonicera japonica and its preparation method and application. Background technique [0002] Blueberry honeysuckle (Lonicera caerulea L.) is a perennial deciduous shrub of Lonicera genus Lonicera. It grows in deciduous forests or shrubs in the shade of forest edges, and is widely distributed throughout my country. As the third generation of emerging small berry tree species, blueberry honeysuckle is rich in amino acids, minerals, sugars, vitamins and various active substances. It can be eaten raw, rich in nutritional value, and has extremely high medicinal value and economic value. A pure natural health product. [0003] Plant protoplasts refer to living cells with cell walls removed, which are totipotent and can regenerate cell walls and regenerate new plants under certain conditions. Due to its characteristics of no ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N5/04
CPCC12N9/2437C12N5/04Y02A50/30C12N9/00
Inventor 臧丹丹孙燕赵恒田刘淑华
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S