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Method of screening leymus chinensis salt-tolerant somatic mutation

A technology of somatic cell mutation and screening methods, applied in plant cells, horticultural methods, botanical equipment and methods, etc., can solve the problems that no one has conducted research on Leymus chinensis callus

Inactive Publication Date: 2007-08-08
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the studies on the salt-alkali resistance of Leymus chinensis are at the seedling stage, and no one has conducted research on the salt-alkali resistance of Leymus chinensis callus and the screening technology for mutants.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Callus regeneration induced by mature embryos of Leymus chinensis

[0029] 1. Treatment of Leymus chinensis seeds

[0030] Take fully mature and structurally complete Leymus chinensis seeds for testing, peel off the seed coat, and use gibberellin 50ppm GA 3 Treat for 48 hours to break dormancy, and then disinfect with 70% ethanol and 0.1% mercuric chloride.

[0031] 2. Callus Induction and Subculture

[0032] The induction medium uses MS medium, specifically agar 8g / L, sucrose 40g / L, hydrolyzed casein 200mg / L, pH 6.0, hormone 2,4-D, that is, 2,4-2 naoxyacetic acid, concentration 2mg / L, inoculate the sterilized plant seeds on the induction medium, cultivate for 15 days, and obtain callus tissue;

[0033] The callus was inoculated on the subculture medium for culture, subcultured once every 10-20 days, and subcultured three times. The basic subculture medium used MS medium, specifically agar 8g / L, sucrose 40g / L, hydrolyzed casein Protein 200mg / LpH6.0, hormone 2,4-D...

Embodiment 2

[0047] 1. Callus regeneration induced by mature embryos of Leymus chinensis

[0048] 1. Treatment of Leymus chinensis seeds

[0049] Take fully mature and structurally complete Leymus chinensis seeds, peel off the seed coat, and use gibberellin 100ppm GA 3 Treat for 48 hours to break dormancy, and then disinfect with 70% ethanol and 0.1% mercuric chloride.

[0050] 2. Callus Induction and Subculture

[0051] The induction medium uses MS medium, specifically agar 8g / L, sucrose 40g / L, hydrolyzed casein 200mg / L, pH 6.0, hormone 2,4-D, that is, 2,4-2 naoxyacetic acid, concentration 3mg / L, inoculate the sterilized plant seeds on the induction medium, culture for 10 days, and obtain callus tissue;

[0052] Inoculate the callus on the subculture medium for culture, subculture once every 10 to 20 days, subculture 4 times, and use MS medium as the basic subculture medium, specifically agar 8g / L, sucrose 40g / L, hydrolyzed casein Protein 200mg / LpH6.0, hormone 2,4-D, concentration 2mg...

Embodiment 3

[0066] 1. Callus regeneration induced by mature embryos of Leymus chinensis

[0067] 1. Treatment of Leymus chinensis seeds

[0068] Take fully mature and structurally complete Leymus chinensis seeds for testing, peel off the seed coat, and use gibberellin 150ppm GA 3 Treat for 48 hours to break dormancy, and then disinfect with 70% ethanol and 0.1% mercuric chloride.

[0069] 2. Callus Induction and Subculture

[0070] The induction medium uses MS medium, specifically agar 8g / L, sucrose 40g / L, hydrolyzed casein 200mg / L, pH 6.0, hormone 2,4-D, that is, 2,4-2 naoxyacetic acid, concentration 4mg / L, inoculate the sterilized plant seeds on the induction medium, culture for 7 days, and obtain callus tissue;

[0071] Inoculate the callus on the subculture medium for culture, subculture once every 10 to 20 days, subculture 5 times, and use MS medium as the basic subculture medium, specifically agar 8g / L, sucrose 40g / L, hydrolyzed casein Protein 200mg / LpH6.0, hormone 2,4-D, concen...

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Abstract

The invention discloses a sieving method of salt tolerant cellar mutant of goat-weed, which is characterized by the following: regenerating callus induced by matured embryo; building sieving system of cellar mutant; obtaining tube vaccine; obtaining the mature goat-weed seed as explant; improving regenerating rate of matured embryo effectively.

Description

technical field [0001] The invention belongs to the field of Leymus chinensis genetic engineering and genetic breeding. Background technique [0002] Leymus chinensis (Leymus chinensis) is an excellent gramineous forage grass with high nutritional value that grows widely in the western part of our province. Social Significance. [0003] At present, the breeding research of Leymus chinensis is mainly based on selective breeding. The breeding method itself limits the speed and efficiency of the improvement of Leymus chinensis varieties, and the cultivated Leymus chinensis varieties cannot meet the needs of production. Using traditional breeding methods, the selection of salt-tolerant varieties is extremely slow. The development of plant tissue and cell culture technology provides a new way for crop improvement. During the process of tissue culture, various mutations will occur in the somatic cells of plants. Utilizing the totipotency of plant cells, mutant cells can be cult...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04A01H4/00
Inventor 王丕武曲同宝关淑艳曲静
Owner JILIN AGRICULTURAL UNIV
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