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Method for inducing embryonic callus

A technique for embryogenic callus and callus, which is applied in the field of rapid induction of embryogenic callus, can solve the problems of unstable transgenic and low transformation rate of Chinese cabbage, and achieves good effect.

Inactive Publication Date: 2018-03-30
SOUTH SUBTROPICAL CROPS RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main means of genetic transformation of cabbage heart is to infect cotyledons or flower spikes with Agrobacterium. These methods have the problems of low transformation rate of cabbage heart and unstable transgene
At present, there is no induction technique and operation method for Chinese cabbage callus

Method used

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  • Method for inducing embryonic callus
  • Method for inducing embryonic callus
  • Method for inducing embryonic callus

Examples

Experimental program
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Effect test

Embodiment 1

[0045] The cabbage seeds were sterilized with 70% v / v ethanol for 1 minute, then sterilized with 1.5wt% sodium hypochlorite aqueous solution for 2 minutes, and finally washed twice with sterile deionized water, and the sterilized seeds were used to induce healing. hurt.

[0046] The obtained sterile seeds were inoculated on the light medium for inducing callus, and the light medium was composed of MS+2.0mg / L6-BA+0.5mg / LNAA+4.0mg / LAgNO 3 +0.8% agar powder+3% sucrose composition, and pH=5.8. Carry out light culture for 14 days, induce callus to take place, obtain light yellow, dry callus of texture (such as figure 1 shown).

[0047] Then, cut the callus obtained by light culture into small pieces with a diameter of about 0.5cm, and continue to transfer to the subculture medium MS+2.0mg / L6-BA+0.5mg / LNAA+4.0mg / LAgNO 3 +0.8% agar powder +3% sucrose. Subculture once every 10 days, subculture 3 times in a row, obtain granular, light yellow embryogenic callus. figure 2 a~2c show...

Embodiment 2

[0051] The cabbage seeds were sterilized with 70% v / v ethanol for 1 minute, then sterilized with 1.5wt% sodium hypochlorite aqueous solution for 2 minutes, and finally washed twice with sterile deionized water, and the sterilized seeds were used to induce callus .

[0052] The obtained sterile seeds were inoculated on the light medium for inducing callus, and the light medium was composed of MS+2.0mg / L6-BA+0.5mg / LNAA+4.0mg / LAgNO 3 +0.8% agar powder+3% sucrose composition, and pH=5.7. Light culture was carried out for 14 days to induce callus, and the obtained callus was light yellow and dry in texture.

[0053]Then, cut the callus obtained by light culture into small pieces with a diameter of about 0.5cm, and continue to transfer to the subculture medium MS+2.0mg / L6-BA+0.5mg / LNAA+4.0mg / LAgNO 3 +0.8% agar powder +3% sucrose. Subculture once every 10 days, subculture 3 times in a row, obtain granular, pale yellow embryogenic callus.

[0054] The daily light conditions for li...

Embodiment 3

[0057] The cabbage seeds were sterilized with 70% v / v ethanol for 1 minute, then sterilized with 1.5wt% sodium hypochlorite aqueous solution for 2 minutes, and finally washed twice with sterile deionized water, and the sterilized seeds were used to induce healing. hurt.

[0058] The obtained sterile seeds were inoculated on the light medium for inducing callus, and the light medium was composed of MS+2.0mg / L6-BA+0.5mg / LNAA+4.0mg / LAgNO 3 +0.6% agar powder+2.5% sucrose composition, and pH=5.8. Light culture was carried out for 14 days to induce callus and obtain bright yellow callus with dry texture.

[0059] Then, cut the callus obtained by light culture into small pieces with a diameter of about 0.5cm, and continue to transfer to the subculture medium MS+2.0mg / L6-BA+0.5mg / LNAA+4.0mg / LAgNO 3 +0.8% agar powder +3% sucrose. Subculture once every 10 days, subculture 3 times in a row, obtain granular, light yellow embryogenic callus.

[0060] The daily light conditions of light...

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Abstract

The invention relates to a method for inducing embryonic callus. The method comprises the following steps of sterilizing seeds with a sterilization reagent; using the sterilized seeds as a material for inducing callus, inoculating the seeds to a first culture medium for inducing the callus, and performing illumination cultivation; and transferring the callus undergone illumination cultivation to asecond culture medium for subculturing, so as to obtain the embryonic callus. The method is easy to conduct, and can obtain a large quantity of the embryonic callus efficiently.

Description

technical field [0001] The invention relates to a method for inducing embryogenic callus, in particular to a method for rapidly inducing embryogenic callus for cabbage seeds. Background technique [0002] Caixin (BrassicacampestrisL.ssp.chinensisvar.utilisTsenetLee) is a specialty vegetable of the genus Brassica in the family Brassicaceae. It eats flower moss and is an important part of vegetable crops in South China. It is currently widely planted throughout the country and exported to Hong Kong, Macao and Southeast Asia. countries. Cabbage has a short growth cycle and is easy to plant, and has become an important material for scientific research. At present, the main means of genetic transformation of cabbage heart is to infect cotyledons or flower spikes with Agrobacterium. These methods have the problems of low transformation rate of cabbage heart and unstable transgene. At present, there is no induction technique and operation method about Chinese cabbage callus. The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 张鲁斌孙曼丽宋康华贾志伟
Owner SOUTH SUBTROPICAL CROPS RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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