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Kiwi fruit protoplast extracting solution, kiwi fruit protoplast as well as preparation method and application of kiwi fruit protoplast

A protoplast and kiwifruit technology, applied to plant cells, etc., can solve the problems of long enzymatic hydrolysis time and low production efficiency, and achieve the effects of improving extraction effect, shortening enzymatic hydrolysis time, and maintaining vitality

Active Publication Date: 2022-07-29
JIANGXI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main method for preparing kiwifruit protoplasts is enzymatic hydrolysis. However, when using conventional enzymatic hydrolysis to prepare kiwifruit protoplasts, the enzymatic hydrolysis takes a long time and the preparation efficiency is not high.

Method used

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  • Kiwi fruit protoplast extracting solution, kiwi fruit protoplast as well as preparation method and application of kiwi fruit protoplast
  • Kiwi fruit protoplast extracting solution, kiwi fruit protoplast as well as preparation method and application of kiwi fruit protoplast
  • Kiwi fruit protoplast extracting solution, kiwi fruit protoplast as well as preparation method and application of kiwi fruit protoplast

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0037] The present invention also provides a method for preparing the kiwifruit protoplast extract, comprising the following steps:

[0038] Mix the preheated mixed enzyme solution with mannitol, CaCl 2 , DTT and MES were mixed to obtain the kiwifruit protoplast extract.

[0039] In the present invention, the cellulase R-10 and the pectinase Y-23 are mixed to obtain a mixed enzyme solution and then preheated to obtain a preheated mixed enzyme solution. The preheating temperature of the present invention is preferably 52-55°C, more preferably 53-54°C; the preheating time is preferably 10-15 min, more preferably 12-14 min. The preheating treatment of the mixed enzyme solution in the present invention can stimulate the activity of the enzyme, promote the dissolution of the enzyme, realize the degradation of the cell wall in a short time, and avoid the reduction of the extraction effect caused by the shrinkage of the protoplast.

[0040]In the present invention, the preheated en...

Embodiment 1

[0062] Materials and reagents used in the present invention include mannitol, CaCl 2 , DTT, MES, cellulase R-10, pectinase Y-23 and sterile water;

[0063] 1. Preparation of washing solution:

[0064] Sterile water, mannitol, CaCl 2 , DTT and MES to prepare washing solution, in which mannitol, CaCl 2 The molar ratios of , DTT and MES were 800:65:1 and 10.

[0065] Place the washing solution in a 4°C refrigerator for 2-4 hours, take out 20-30 mL of washing solution and place it in a 50-mL sterile centrifuge tube, and place it in the refrigerator for future use; take 26-30 mL of washing solution into a 50-mL beaker for later use.

[0066] 2. Preparation of mixed enzyme solution:

[0067] A mixed enzyme solution was prepared with sterile water, cellulase R-10 and pectinase Y-23, wherein the mass concentration ratio of cellulase R-10 and pectinase Y-23 was 1:0.3.

[0068] 3. Preparation of protoplast extract:

[0069] Put the mixed enzyme solution in step 2 in a water bath at ...

Embodiment 2～6

[0071] Using the materials and reagents of Example 1, protoplast extracts with the same components as those in Example 1 but with different concentrations were prepared.

[0072] Table 1 Concentration of each component in the protoplast extract of Examples 1 to 6

[0073] component Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Mannitol (mM) 800 900 500 300 300 300 CaCl 2 (mM)

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Abstract

The invention belongs to the technical field of protoplast preparation, and particularly relates to a kiwi fruit protoplast extracting solution, a kiwi fruit protoplast and a preparation method and application of the kiwi fruit protoplast. Each liter of the kiwi fruit protoplast extracting solution comprises 1.3%-2% W / V of mixed enzyme solution, 300-900 mM of mannitol, 50-65 mM of CaCl2, 1-2 mM of DTT and 10-20 mM of MES. The mixed enzyme solution is prepared from cellulase R-10 and pectinase Y-23. When the kiwi fruit protoplast extracting solution is used for performing enzymolysis on kiwi fruit pulp, the enzymolysis time can be shortened, so that the prepared protoplast keeps the activity for a long time, and the extraction efficiency is improved.

Description

technical field [0001] The invention belongs to the technical field of protoplast preparation, and in particular relates to a kiwifruit protoplast extract, a kiwifruit protoplast and a preparation method and application thereof. Background technique [0002] Kiwifruit is a deciduous woody vine of the Actinidiaceae (Actinidiaceae) genus (Actinidia), most of which are dioecious and very few are monoecious. Kiwifruit is distributed in the vast area from Malaysia to eastern Siberia. At present, there are 66 species of kiwifruit, of which 62 species are native to my country. my country is the original center of kiwifruit. Wuduhe Town, Yiling District, Yichang City, Hubei Province, is the original source of kiwifruit in the world. Origin. Kiwifruit has high nutritional value, has medicinal value such as anti-inflammatory and anti-tumor, and can also be processed into fruit juice and other foods that can be stored for a long time. [0003] Plant protoplast refers to the general te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
CPCC12N5/04C12N2509/00Y02A50/30
Inventor 匡柳青樊友富刘勇杨莉胡威刘德春贾东峰
Owner JIANGXI AGRICULTURAL UNIVERSITY
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