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Method for rapid preparation of leaf protoplasts of Stevia rebaudiana

A technology of protoplasts and stevia, applied to plant cells, etc., to achieve the effect of easy access and simple enzymatic hydrolysis

Inactive Publication Date: 2016-03-09
ANHUI SCI & TECH UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

The stevia research group of our school has conducted systematic research on stevia variety breeding, polyploid mutagenesis, tissue culture rapid propagation, seed quality, high-yield cultivation, etc., but there is no research on the preparation of protoplasts from stevia leaves. According to research reports, the isolation and preparation of stevia protoplasts lay the foundation for the study of stevia protoplast fusion, protoplast culture and regeneration of plants, transient expression of genes and subcellular localization of proteins

Method used

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  • Method for rapid preparation of leaf protoplasts of Stevia rebaudiana
  • Method for rapid preparation of leaf protoplasts of Stevia rebaudiana
  • Method for rapid preparation of leaf protoplasts of Stevia rebaudiana

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Embodiment Construction

[0018] In order to deepen the understanding of the present invention, the present invention will be further described below in conjunction with examples. These examples are only used to explain the present invention, and do not constitute a limitation to the protection scope of the present invention.

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Abstract

The invention relates to a method for rapid preparation of leaf protoplasts of Stevia rebaudiana. According to the invention, optimal conditions for separating the leaf protoplasts of Stevia rebaudiana are screened out with small and tender leaves of tissue culture seedlings of Stevia rebaudiana as a material by controlling factors like plasmolysis time, the ratio and concentration of cellulase Onozuka R-10, hemicellulase and macerozyme R-10, the concentration of mannitol in enzymatic hydrolysis liquid, the pH value of the enzymatic hydrolysis liquid, hydrolysis time, etc. Free protoplasts prepared by using the method are great in quantity and high in activity; and the method is simple and fast in preparation of the leaf protoplasts of Stevia rebaudiana.

Description

technical field [0001] The invention relates to a method for rapidly preparing stevia leaf protoplasts, belonging to the field of biotechnology. Background technique [0002] Stevia ( Steviarebaudiana Bertoni), as an ideal sweetener and functional health product, has become a research and development hotspot in the fields of food and medicine. The stevia research group of our university has conducted systematic research on stevia variety breeding, polyploid mutagenesis, tissue culture rapid propagation, seed quality, high-yield cultivation, etc., but there is no research on the preparation of protoplasts from stevia leaves. According to research reports, the isolation and preparation of stevia protoplasts lay the foundation for the study of stevia protoplast fusion, protoplast culture and regeneration of plants, transient expression of genes and subcellular localization of proteins. Contents of the invention [0003] Purpose of the invention: to provide a method for rapi...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 舒英杰王虹周玉丽胡能兵何克勤崔广荣宋桂成
Owner ANHUI SCI & TECH UNIV
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