Preparation method of crape myrtle protoplast

A technology of protoplasts and crape myrtle, applied in the field of preparation of crape myrtle protoplasts, to achieve the effect of high integrity and activity

Active Publication Date: 2021-11-16
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there is no prior art for the successful isolation of protoplasts from Lagerstroemia indica, and the development of efficient preparation methods for Lagerstroemia indica protoplasts is of great significance for the molecular breeding of Lagerstroemia indica

Method used

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  • Preparation method of crape myrtle protoplast
  • Preparation method of crape myrtle protoplast
  • Preparation method of crape myrtle protoplast

Examples

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Embodiment 1

[0045] The present embodiment provides a kind of preparation method of crape myrtle protoplast, and concrete steps are as follows:

[0046] (1) Take the leaves on the young shoots that germinate after pruning the branches of Lagerstroemia in the same year, wash and dry them, and place the leaves of Lagerstroemia in a shading climate box at 25°C for 20-30 minutes to make the leaves slightly dehydrated and wilted. It is standard that the leaves begin to soften and dehydrate but do not appear to be curled at the edge. After the upper epidermis of the treated blade is fixed with adhesive tape, the lower epidermis is torn off, and the upper epidermis is put into the enzymatic hydrolysis solution upwards ( figure 1 );

[0047] The composition of the enzymolysis solution is: 1% cellulase, 0.5% isolated enzyme, 0.4% pectinase, 0.6M mannitol, 20mM KCl, 20mM MES (pH5.7), 10mM CaCl 2 , 0.1% BSA and 5mM DTT;

[0048] (2) Open the lid of the petri dish containing the enzymolysis solution...

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Abstract

The invention relates to the technical field of plant protoplast, in particular to a preparation method of crape myrtle protoplast. The invention provides a preparation method of protoplast for crape myrtle, which comprises the following steps: taking crape myrtle leaves as a material, removing lower epidermis from the crape myrtle leaves, carrying out enzymolysis treatment on the leaves without the lower epidermis in an enzymolysis solution to obtain an enzymolysis solution, separating the enzymolysis solution to obtain a mesophyll enzymolysis product, washing and separating to obtain the mesophyll protoplast. According to the method, the protoplast can be efficiently extracted from the lagerstroemia indica leaves, the prepared protoplast has high integrity and activity, and a basis is provided for molecular breeding of lagerstroemia indica.

Description

technical field [0001] The invention relates to the technical field of plant protoplasts, in particular to a method for preparing crape myrtle protoplasts. Background technique [0002] A protoplast is a living plant cell surrounded by a plasma membrane after the cell wall has been removed. Protoplasts are easy to prepare, and can survive for several days under suitable conditions after separation and purification. They still have all the characteristics of living cells and have certain reproductive capabilities. Since there is no cell wall, protoplasts can ingest genetic material including nuclei, organelle genomes or exogenous DNA fragments. It is a very good transient expression system and an ideal experimental material for basic life sciences and crop breeding and improvement. At present, protoplasts have been isolated from a large number of species, distributed in more than 40 families and more than 160 genera, mainly Solanaceae, Fabaceae, Poaceae, Compositae and Rosac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N9/42C12N9/18C12N9/26C12N9/88
CPCC12N5/04C12N9/2437C12N9/88C12N9/2408C12N9/18C12Y301/01011C12Y302/01015C12Y402/02002
Inventor 潘会堂刘洁茹池秀凤冯露程堂仁王佳张启翔
Owner BEIJING FORESTRY UNIVERSITY
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