Method for extracting apoplast antifreeze proteins from winter rape roots

A technology of antifreeze protein and extraction method, which is applied to the preparation method of peptides, chemical instruments and methods, plant peptides, etc., and can solve the problem that there is no mature method for antifreeze protein in rapeseed

Inactive Publication Date: 2013-10-02
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Rapeseed antifreeze prote...

Method used

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  • Method for extracting apoplast antifreeze proteins from winter rape roots
  • Method for extracting apoplast antifreeze proteins from winter rape roots
  • Method for extracting apoplast antifreeze proteins from winter rape roots

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: First time sampling, extraction of apoplast protein from the root of No. 6 winter rapeseed Longyou:

[0034] (1) Sampling: On October 5th, Longyou No. 6 roots were dug out in the field, put in an ice box within 3-5 minutes, brought back to the laboratory, and stored in a 4°C refrigerator;

[0035] (2) 1000ml of extraction buffer is prepared according to the following weight and volume:

[0036] 0.05M / L Tris-Hcl6.057g to adjust the pH to 8.0;

[0037] 0.01M / L EDTA-Na2.2H2O (disodium ethylenediaminetetraacetic acid) 3.72g;

[0038] 0.02M / LVC (vitamin c) 3.52g;

[0039] Dilute to 1000ml with ddH2O, and store the prepared buffer at 4°C;

[0040] (3) Cut into strips: remove the soil carried by the root of rapeseed, remove the dead part of the root, rinse with running water for 10-12 minutes, until the sundries are washed, rinse with ddH2O twice, dry the surface moisture with filter paper, wash the root and dissect it Cut into 1-2cm long strips;

[0041] (4) S...

Embodiment 2

[0052] Embodiment 2: sampling for the second time, extraction of apoplast protein from the root of Longyou No. 6:

[0053] (1) Sampling: On November 15th, Longyou No. 6 roots were dug out in the field, put in an ice box within 3-5 minutes, brought back to the laboratory, and stored in a 4°C refrigerator;

[0054] (2) 1000ml of extraction buffer is prepared according to the following weight and volume:

[0055] 0.05M / L Tris-Hcl6.057g to adjust the pH to 8.0;

[0056] 0.01M / L EDTA-Na2.2H2O (disodium ethylenediaminetetraacetic acid) 3.72g;

[0057] 0.02M / LVC (vitamin c) 3.52g;

[0058] Dilute to 1000ml with ddH2O, and store the prepared buffer at 4°C;

[0059] (3) Cut into strips: remove the soil carried by the root of rapeseed, remove the dead part of the root, rinse with running water for 10-12 minutes, until the sundries are washed, rinse with ddH2O twice, dry the surface moisture with filter paper, wash the root and dissect it Cut into 1-2cm long strips;

[0060] (4) Immer...

Embodiment 3

[0071] Example 3: The third sampling, the extraction of the root and leaf apoplast proteins of Longyou No. 6:

[0072] (1) Sampling: On December 1, dig out rapeseed roots in the field, put them in an ice box within 3-5 minutes, bring them back to the laboratory, and store them in a refrigerator at 4°C;

[0073] (2) 1000ml of extraction buffer is prepared according to the following weight and volume:

[0074] 0.05M / L Tris-Hcl6.057g to adjust the pH to 8.0;

[0075] 0.01M / L EDTA-Na2.2H2O (disodium ethylenediaminetetraacetic acid) 3.72g;

[0076] 0.02M / LVC (vitamin c) 3.52g;

[0077] Dilute to 1000ml with ddH2O, and store the prepared buffer at 4°C;

[0078] (3) Cut into strips: remove the soil carried by the root of rapeseed, remove the dead part of the root, rinse with running water for 10-12 minutes, until the sundries are washed, rinse with ddH2O twice, dry the surface moisture with filter paper, wash the root and dissect it Cut into 1-2cm long strips;

[0079] (4) Immer...

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Abstract

The invention relates to a method for extracting antifreeze proteins, in particular to a method for extracting apoplast antifreeze proteins from winter rape roots. According to the method, apoplast antifreeze proteins are successfully extracted from cold-induced winter rape roots. The method for extracting apoplast antifreeze proteins from winter rape roots is characterized by comprising the following steps: (1) sampling; (2) extracting a buffer solution for preparing; (3) slitting; (4) immersing; (5) vacuumizing; and (6) centrifuging. According to the method, apoplast antifreeze proteins are successfully and rapidly extracted from cold-induced winter rape roots, so that technical support is provided for further research of the cold resistance mechanism of winter rape.

Description

technical field [0001] The invention relates to a method for extracting antifreeze protein, in particular to a method for extracting antifreeze protein from winter rapeseed root apoplast. The method successfully extracts the apoplast antifreeze protein from cold-induced winter rapeseed root. Background technique [0002] Low temperature is one of the main environmental factors affecting the normal growth and development of plants in northern my country. When plants encounter low temperature stress, a series of physiological and biochemical changes will occur in the body to adapt to low temperature stress, and at the same time they will synthesize and secrete a variety of proteins - low temperature induced proteins. Antifreeze proteins (AFPs) are a class of low-temperature-induced proteins, which can be produced in different organs such as roots, stems, and leaves of plants, and can also exist in callus under tissue culture conditions, suspension culture cells and suspension ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K1/30C07K1/14
Inventor 孙万仓杨刚史鹏辉王丽萍刘自刚曾秀存方彦武军艳杨宁宁孔德晶鲁美宏赵艳宁董红业王月
Owner GANSU AGRI UNIV
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