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Preparation method of duckweed protoplast

A technology of protoplast and duckweed, applied in the direction of plant cells and the like, can solve the problem that duckweed protoplast cannot be obtained, and achieve the effects of short time-consuming, simple and convenient operation, and few materials.

Inactive Publication Date: 2019-08-27
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Carrying out experiments with current technology, duckweed protoplasts cannot be obtained, but duckweed cell fragments

Method used

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  • Preparation method of duckweed protoplast
  • Preparation method of duckweed protoplast
  • Preparation method of duckweed protoplast

Examples

Experimental program
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Embodiment Construction

[0034] In a specific embodiment of the present invention, duckweed (Duckweed family L. minor NK 1) is collected from lake water in Xiqing District, Tianjin. The compounds and medicines used in the examples of the present invention were all purchased from Takola Experimental Supplies Sales Department, Xiqing District, Tianjin, with a purity of 99.5%; the enzymes used in the examples of the present invention were all purchased from Tianjin Weike Biology Technology Co., Ltd.

[0035] To prepare the Datko medium mother liquor required for propagating intact duckweed plants, mix the Datko medium mother liquor with water at a volume ratio of 3:1000 to obtain the Datko medium, which is stored for later use (stored at 4°C).

[0036] Datko medium stock solution consists of water and compounds at the following concentrations:

[0037]

[0038] Datko medium consisted of water and compounds at the following concentrations:

[0039]

[0040]

[0041] The method for preparing duc...

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Abstract

The invention discloses a method for preparing duckweed protoplast. The method comprises the following steps: 1, adjusting the pH value of a Datko medium to 5-6, and autoclaving the Datko medium; 2, collecting duckweeds, culturing the duckweeds on the Datko medium in step 1, and subculturing the duckweeds 1-2 times every week to obtain duckweed plants; 3, clamping the duckweed plants obtained in step 2, cleaning the duckweed plants with a solution A, placing the duckweed plants in a centrifuge tube, adding a tissue fixing solution, and fixing the duckweed plants for at least 15 min; and 4, carrying out enzymatic hydrolysis on the duckweed plants obtained in step 3 under a dark condition at an enzymatic hydrolysis temperature of 23-27 DEG C by using a tissue enzymatic hydrolysis solution for 25-30 min, and carrying out blow beating and uniform mixing once every 10-15 min to obtain the suspending duckweed protoplast. The method for preparing the duckweed protoplast has the advantages ofless material drawing, short time, and simplicity and convenience in operation, the obtained duckweed protoplast has a high concentration, and data analysis is representative.

Description

technical field [0001] The invention belongs to the field of duckweed protoplast preparation, and in particular relates to a method for preparing duckweed protoplast. Background technique [0002] Duckweed is a monocotyledon water surface floating plant, which belongs to the Duckweed family and is widely distributed all over the world. It also has certain advantages in the development and utilization of bioenergy, and is an aquatic plant with environmental advantages. In recent years, the application of duckweed in water purification has received more and more attention, but the physiological and biochemical mechanism of duckweed in water purification has not yet been elucidated. The preparation of protoplasts is an indispensable key operation for the biological analysis of duckweed cells. Most of the preparation methods of plant protoplasts focus on terrestrial plants, while duckweed is a floating plant on the water surface, and its cell wall strength, cell osmotic pressu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04
Inventor 杨琳韩玉洁吴迪王苏桐魏莹李红雅张小兰刘文鑫陆美伊王家乐
Owner TIANJIN NORMAL UNIVERSITY
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