Shenzhen No.5 flameray gerbera transgene technology

A transgenic, gerbera technology, applied in the direction of recombinant DNA technology, cells modified by introducing foreign genetic material, and introduction of foreign genetic material using vectors, etc., can solve the limitation, low frequency of plant regeneration, low heat resistance, cold resistance, etc. question

Inactive Publication Date: 2006-05-10
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The plant regeneration frequency of this variety is very low at present
In addition, the invasion of various viruses, microorganisms and pests, as well as its

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Step 1, the plasmid is introduced into Agrobacterium tumefaciens: according to the method of direct introduction of the intermediate vector plasmid DNA into Agrobacterium tumefaciens, pCAMBIA1301 is introduced into Agrobacterium tumefaciens LBA4404; plate culture, the medium adopts the formula of YEB+50mg / L kana Solid medium A of mycin + 50mg / L streptomycin, cultured at 28±1°C for 46 hours;

[0024] Step 2, preparation and preservation of LBA4404 seed liquid: pick the single colony of LBA4404 obtained from the above cultivation containing the pCAMBIA1301 plasmid, and inoculate it into medium B: the liquid of YEB+50mg / L kanamycin+50mg / L streptomycin In the culture medium, shake culture at 28±1°C and 180r / min for 24 hours; mix the prepared glycerol with a volume percentage concentration of 80% with the cultured bacterial liquid to make the final glycerol concentration in the glycerol bacterial liquid mixture up to 20%. Dispense the glycerin bacteria liquid mixture into 1...

Embodiment 2

[0032] Embodiment 2: other is with embodiment 1, and difference is:

[0033] Step 1: medium A is a solid medium of YEB+100mg / L kanamycin+100mg / L streptomycin, and the culture time is 50h;

[0034] Step 2: Medium B is a liquid medium of YEB+100mg / L kanamycin+100mg / L streptomycin, the culture conditions are 28±1°C, 220r / min, and the culture time is 36h; LBA4404 seed liquid is divided into 1.5 ml eppendof tubes and stored at -80°C for future use.

[0035] Step three:

[0036] (1) Activation of Agrobacterium: Take 8 μl of the LBA4404 seed liquid obtained in step 2 and inoculate it into 15 ml of medium B in step 2, and cultivate it for 25 hours according to the temperature and shaking culture conditions of step 2; take 1 ml of the bacterial liquid and transfer it to 50 ml of fresh medium In B, continue to cultivate for 14 hours according to the temperature and shaking culture conditions of step 2; transfer the cultured bacterial liquid into a 10ml eppendof tube, centrifuge at 500...

Embodiment 3

[0042] Embodiment 3: other is with embodiment 1, and difference is:

[0043] Step 1: medium A is a solid medium of YEB+100mg / L kanamycin+100mg / L streptomycin;

[0044] Step 2: medium B is a liquid medium of YEB+100mg / L kanamycin+100mg / L streptomycin, and the culture time is 36h;

[0045] Step 3: (1) After completion, OD 600 The value is 0.5; (2) the standby bacterial solution shaking culture time is 2h; (3) the explants are sterilized and cultured for 3 days under the condition of the sterilized medium pH is 5.8; (4) the medium pH is 5.8; (5) The pH value of the adventitious bud rejuvenation and proliferation medium in ) is 5.6; the time of culturing on the rooting medium is 30 days.

[0046] Test results: The transient expression rate of GUS in single-bud explants was 20.3%, the occurrence frequency of resistant adventitious buds was 20.3%, and the frequency of transgenic adventitious buds finally obtained was 7.35%.

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Abstract

This invention relates to a transfer gene technique of the SHENZHEN 5 Africa chrysanthemum, which characterizes in that it comprises the plasmid embeding of the carcinoma bacillus, the preparing and conserving of the LBA4404 germ solution, the transforming of the bacillus, wherein the third step includes the bacillus activating, the outer plant and the bacterial culturing, degerming, the antibiotics filtering, transfer gene plant rejuvenation and proliferation. And this invention transforms the SHENZHEN 5 Africa chrysanthemum bud with a gene containing the embedded CaMV35S.HPT and embedded CaMV35S.GUS gened to get the transfer gene plant, which provdes technique for transforming other object gene and obtaining the new transfer gene Africa chrysanthemum. The advantage in this invention is that the stability is well and the regeneration and the transform efficiency are high.

Description

(1) Technical field [0001] The invention relates to the transgenic technology of Gerbera hybrida Shenzhen No. 5 (S5), that is, a method for obtaining transgenic plants through the transgenic technology. (2) Background technology [0002] Gerbera, also known as Gerbera, is one of the six major cut flowers in the world and is also an important potted flower. It has many horticultural varieties and high ornamental value. Breeders have been working hard to obtain new varieties with various flower colors, flower types, flower fragrances, long vase life, resistance, heat resistance and cold resistance. The traditional breeding technology has disadvantages such as long time for hybrid breeding, difficulty in introducing good traits of distantly related species through hybridization, and introduction of one or some good traits is often accompanied by the introduction of some bad traits. The use of genetic engineering technology is expected to improve the flower...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H4/00C12N5/04C12N5/10
Inventor 王小菁张妙彬
Owner SOUTH CHINA NORMAL UNIVERSITY
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