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Method and culture medium for efficiently inducing soybean cotyledon explants to produce clustered buds

A cotyledon node and explant technology, which is applied in the field of efficiently inducing soybean cotyledon node explants to produce clustered buds, can solve the problems of long time for inducing soybean cotyledon node explants and low germination rate, and achieve clustered buds in a good state, The effect of low pollution rate and shortened germination time

Inactive Publication Date: 2017-09-26
刘寒冬
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the above-mentioned technical problems, the present invention provides a method and medium for efficiently inducing soybean cotyledon explants to produce clustered buds, which solves the problems of long time and low germination rate in the prior art for inducing soybean cotyledon explants , fills the technical gap, applies this method to related genetic transformation research and production work, and provides an important technical platform and support for transgenic research

Method used

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  • Method and culture medium for efficiently inducing soybean cotyledon explants to produce clustered buds
  • Method and culture medium for efficiently inducing soybean cotyledon explants to produce clustered buds
  • Method and culture medium for efficiently inducing soybean cotyledon explants to produce clustered buds

Examples

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Effect test

Embodiment 1

[0024] 1) Configure the medium, dissolve MSB basic medium in 1 liter of distilled water, MES 0.5g, sucrose 25g, glutamine 50mg, asparagine 50mg, 6-benzylaminopurine 1.3mg, agar 6.5g, heat to dissolve completely, Adjust the pH value of the culture medium to 5.6-5.8, autoclave it, pour it on the plate when the temperature drops to 60°C, and cool it for later use. The above operations are all carried out under sterile conditions, and the composition of the MSB basic medium is as follows:

[0025]

[0026]

[0027] 2) Soybean seeds are sterilized, sterilized and germinated. In an ultra-clean workbench, put the selected high-quality soybean seeds in a sterilized triangular flask, add 70-80% ethanol for disinfection for 5-10 minutes, pour After draining the ethanol, add hydrogen peroxide with a volume fraction of 15-20% to the soybean seeds, place in a shaking incubator at 25-28°C for 15-30 minutes, and sterilize by shaking in the dark at a speed of 100-200r / min, and then use ...

Embodiment 2

[0032] The difference from Example 1 is that when configuring the medium in step 1), dissolve the MSB basic medium in 1 liter of distilled water, MES 0.7g, sucrose 28g, glutamine 45mg, asparagine 45mg, 6-benzylamino Purine 1.4 mg, agar 7 g, and other steps are the same as in Example 1.

Embodiment 3

[0034] The difference from Example 1 is that when configuring the medium in step 1), dissolve the MSB basic medium in 1 liter of distilled water, MES 0.9g, sucrose 28g, glutamine 50mg, asparagine 50mg, 6-benzylamino Purine 1.5 mg, agar 7.5 g, and other steps are the same as in Example 1.

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Abstract

The invention belongs to the technical field of plant tissue culture and induction and in particular relates to a method and a culture medium for efficiently inducing soybean cotyledonary node explants to generate cluster buds. The culture medium comprises an MSB culture medium, 0.5g / L-1.5g / L of 2-(N-morpholin)ethanesulfonic acid, 20g / L-30g / L of sucrose, 40mg / L-60mg / L of glutamine, 40mg / L-60mg / L of asparaginate, 0.5mg / L-2mg / L of 6-benzyl aminopurine and 6g / L-8g / L of agar, and the pH value is 5.6 to 5.8. The method comprises the following steps: standing and culturing sterilized soybean seeds at 4 DEG C for 14-18 hours and then removing seed coats; longitudinally cutting epicotyls along middle lines of plumular axes to obtain soybean cotyledonary nodes with 3mm-5mm of plumular axes and top meristems; and then inoculating the soybean cotyledonary nodes into the culture medium and culturing in a greenhouse for 20-25 days, wherein about 20-40 cluster buds can be finally induced from each explant. According to the method provided by the invention, the induction rate of the cluster buds is extremely improved in short time, and important technical platform and support are provided for researches of an efficient expanding propagation system and a genetic transformation system taking the cotyledonary nodes as the basis.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture and induction, and specifically relates to a method and a culture medium for efficiently inducing soybean cotyledon node explants to produce clustered buds. Background technique [0002] Soybean is originally produced in my country, and it is an important oil crop and economic crop in the world at present, and it is also an important source of vegetable protein. Soybeans produced by conventional breeding can no longer meet the needs of society, and genetically modified genetic breeding is needed to produce new soybean varieties with high yield and stress resistance. However, soybean genetic transformation technology has always been a difficult point in the field of plant genetic engineering, and is limited by many factors, the most important of which is the proper treatment of soybean transformed cotyledonary node explants and the rapid induction of a large number of clustered buds. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01H4/00
CPCA01H4/00A01H4/001
Inventor 郑志民
Owner 刘寒冬
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