Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Composition for preparing populus tomentosa leaf and/or root tissue protoplast and reagent and method

A technology of protoplasts and Populus tomentosa, applied in biochemical equipment and methods, plant cells, enzymes, etc., can solve the problems of low protoplast efficiency and achieve the effect of low price, high activity and large quantity

Active Publication Date: 2020-12-08
BEIJING FORESTRY UNIVERSITY
View PDF1 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a certain research foundation for the isolation and cultivation of poplar leaf tissue protoplasts at home and abroad (WANG, 1991; Song Shaoyu, Zhang Junqi, etc., 2015), but these methods based on the enzymatic hydrolysis of Arabidopsis thaliana The efficiency of protoplasts is low, and there is an urgent need for a method for preparing poplar tissue protoplasts with good separation effect

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Composition for preparing populus tomentosa leaf and/or root tissue protoplast and reagent and method
  • Composition for preparing populus tomentosa leaf and/or root tissue protoplast and reagent and method
  • Composition for preparing populus tomentosa leaf and/or root tissue protoplast and reagent and method

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0032] The present invention has no special requirements on the method of preparing the composition, use of those skilled in the art can be.

[0033] Preferred method of preparing the compositions of the present invention comprises the steps of: sequentially cellulase, Macerozyme, mannitol, KCl, and 2- (N- morphine Lin) ethanesulfonic acid monohydrate, and the solution prepared above in that 55 ℃ water bath for 10min, cooled to room temperature. Were sequentially added to the CaCl solution was cooled 2 And bovine serum albumin, and mix well.

[0034] The present invention provides the use of the above technical solution the composition Populus blades and / or root tissue of the protoplast preparation.

[0035] The present invention also provides a composition for preparing a reagent Populus blades and / or root tissue protoplast, separate dispensing of reagent A, reagent B and reagent C; the agent A is a combination of the above technical solution thereof; B is the agent solution W...

Embodiment 1

[0053] A method for the preparation of blade tomentosa and / or root tissue protoplasts composition, the following concentrations of constituents: 15g / L cellulase and 4g / L Macerozyme; wherein the cellulase is a cellulase R10 , macerating enzyme is Macerozyme R10, both enzymes were purchased from Baier Di biotechnology company.

Embodiment 2

[0055] A process for preparing a composition Populus blades and / or root tissue of protoplasts for the concentration of the components of the following composition: cellulose 14g / L, Macerozyme 4g / L, mannitol 0.6mol / L, KCl20mmol / L, 2- (N- morphine) ethanesulfonic acid 10mmol / L, CaCl 2 10mmol / L bovine serum albumin and 100g / L; the same source of the enzyme of Example 1.

[0056] Preparation method:

[0057] 10ml composition system

[0058] (A) were sequentially added to the cellulase R100.15g 50mL centrifuge tube, Macerozyme R100.04g, 1mol / L mannitol 6ml, 2mol / LKCl0.1ml, 0.2mol / LMES 1ml (pH = 5.7, before use 70 deg.] C water bath for 5min), distilled water was used. The solution was thoroughly vortexed in a water bath at 55 ℃ water bath 10min, cooled to room temperature; water bath Enzyme can be a suitable temperature in a sufficiently dissolved and mixed, does not destroy the enzyme activity, improve the efficiency of the subsequent enzymatic hydrolysis.

[005...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
radiusaaaaaaaaaa
concentrationaaaaaaaaaa
radiusaaaaaaaaaa
Login to View More

Abstract

The invention provides a composition for preparing populus tomentosa leaf and / or root tissue protoplasta as well as a reagent and a method, and belongs to the technical field of plant protoplast preparation. The composition for preparing the populus tomentosa leaf and / or root tissue protoplasts comprises cellulase and a digestion enzyme, the enzyme activity of the cellulase is 250-350U / g, the enzyme activity of the digestion enzyme is 700-900U / g, and the mass ratio of the cellulase to the digestion enzyme is (14.5-15.5): (3.5-4.5). Populus tomentosa leaves and / or root tissues are subjected toenzymolysis through combined use of the cellulase and the digestion enzyme, so that the high activity of the protoplasts is kept, the number of the obtained protoplasts is large, the price of the enzymes is relatively low, the cost is reduced, and the requirements of subsequent experimental research can be met.

Description

Technical field [0001] The present invention belongs to the technical field of plant protoplast apparatus, particularly relates to a composition prepared Populus blades and / or root tissue and protoplasts for reagents and methods. Background technique [0002] Maobai Yang is a native species unique to China, widely distributed, are distributed in Liaoning Province (South), Hebei, Shandong, Shanxi, Shaanxi and other provinces to neutralize the Yellow River, downstream of the distribution center, in the north of the Ecological Environment and forestry production construction occupies an important position, is the northern forest cultivation pioneer species. However, poplar perennial species, in a conventional manner crossbreeding genetic improvement take longer. Using protoplast cell fusion and somatic hybridization, can improve this drawback. Poplar is also a model species of woody plants, to obtain protoplasts can provide good material for a number of scientific studies of impor...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N9/42C12N9/18C12N9/26C12N9/88
CPCC12N5/04C12N9/2437C12N9/88C12N9/2408C12N9/18C12Y301/01011C12Y302/01015C12Y402/02002
Inventor 张德强徐伟杰谢剑波
Owner BEIJING FORESTRY UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com