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Root agrobacterium mediated turf grass-transforming method

A technology of Agrobacterium rhizogenes and turfgrass, applied in the field of Agrobacterium rhizogenes-mediated transformation of turfgrass, can solve problems such as establishment and implementation difficulties

Active Publication Date: 2008-02-27
BEIJING NORTH ELITE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The Agrobacterium rhizogenes-mediated method has its irreplaceable superiority in the genetic transformation technology of plants, but it is limited by factors such as the genotype of the recipient plant and the state of the recipient tissue material, so this technology is in a new Establishment and implementation on recipient plants is very difficult

Method used

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  • Root agrobacterium mediated turf grass-transforming method
  • Root agrobacterium mediated turf grass-transforming method
  • Root agrobacterium mediated turf grass-transforming method

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1, Agrobacterium rhizogenes mediated transformation of tall fescue grass

[0032] 1. Induction culture of tall fescue (Festuca arundinacea Schreb.) callus

[0033] 1. Callus induction

[0034] Select ripe and plump tall fescue grass (Festuca arundinacea Schreb.) seeds, soak in distilled water for 4 hours at room temperature, peel off the seed coat, and then put them in distilled water to continue soaking overnight. The soaked peeled seeds are first washed twice with 70% alcohol, then disinfected with 30% sodium hypochlorite for 30 minutes, and rinsed with sterile water three times. Under sterile conditions, the embryos are completely peeled off from the sterilized seeds, and then the embryos are cut in half transversely and inserted into the callus induction medium respectively. Seeding density: 100 grains / dish, the diameter of the dish is 75mm.

[0035] The composition of each liter of callus induction medium is as follows: on the basis of MS basic medium...

Embodiment 2

[0071] Embodiment 2, Agrobacterium rhizogenes mediated transformation of tall fescue

[0072] 1. Induction culture of tall fescue (Festuca arundinacea Schreb.) callus

[0073] 1. Callus induction

[0074] Select mature and plump tall fescue (Festuca arundinacea Schreb.) seeds, soak them in distilled water for 3.5 hours at room temperature, peel off the seed coat, and then put them into distilled water to continue soaking overnight. The soaked peeled seeds are first washed twice with 70% alcohol, then disinfected with 30% sodium hypochlorite for 30 minutes, and rinsed with sterile water three times. The embryos are completely peeled off from the sterilized seeds, and then the embryos are cross-cut in half and inserted into callus induction medium respectively. Seeding density: 100 grains / dish, the diameter of the dish is 75mm.

[0075] The composition of each liter of callus induction medium is as follows: on the basis of MS basic medium, add 3 mg of auxin substance 2,4-dich...

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Abstract

The method includes the following steps: 1. inducing callus with embryonal explant; 2. setting the embryonal explant onto the callus state regulating culture medium for developing toward embryonal callus; 3. soaking the callus inside root agrobacterium liquid with plant expression vector; 4. setting the soaked callue onto co-culture medium for co-culturing; 5. setting the acceptor tissue onto inducing and screening culture medium for inducing resistive callus; and 6. setting the growing resistive callus onto differentiating culture medium for differentiating regenerated plant. The present invention solves the difficult problem of transgenic breeding of turf grass, provides one reliable and practical platform for transgenic breeding of turf grass, and has important theoretical and practical significance.

Description

technical field [0001] The invention relates to a method for producing transgenic lawn grass, in particular to a method for transforming lawn grass mediated by Agrobacterium rhizogenes. Background technique [0002] Turfgrass plays an important role in improving the ecological environment and improving people's quality of life, and exerts significant ecological benefits. However, turfgrass has some disadvantages such as large water consumption, inability to tolerate over-cold or over-heated climate, and short green period, which limit its wider application. The rapid development of modern molecular biology provides a new technical platform for the cultivation of new varieties of turfgrass. [0003] At present, known transgenic techniques include direct introduction of exogenous genes, such as silicon carbide fiber-mediated method, polyethylene glycol (PEG)-mediated protoplast transformation method, gene gun method, electric shock transformation method, etc.; and Agrobacteri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63A01H4/00
Inventor 陈蕾王蕾陈宛新韩晶
Owner BEIJING NORTH ELITE BIOTECH
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