Agrolbacterium-mediated method for transforming lily from exogenous gene

An Agrobacterium-mediated and Agrobacterium-transformation technology, which is applied in the fields of biotechnology and agriculture, can solve the problems of genetic transformation that have not yet been found to be successful, and achieve the effects of shortening the breeding cycle, high safety, and high transformation rate

Inactive Publication Date: 2005-01-12
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no reports of documents that are closely related to the subject of the present invention, and no successful examples have been found in the genetic transformation of lily through Agrobacterium-mediated exogenous functional genes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The experimental variety is Oriental lily. Peel the bulb from the outside to the inside, rinse it with clean water, soak it in washing powder solution for 30 minutes, and then sterilize it on a sterile operating table: soak it in 75% alcohol for 2 minutes, rinse it three times with 20% concentration of sterile water The commercially available Antiformin was soaked for 15-20min, rinsed with sterile water for 3 times, and cut into 5mm size as explants for later use.

[0016] The explants were inoculated on MS (pH5.8) fixed medium and cultured for 5 days, and fresh wounds were cut out around uncontaminated scales with a scalpel as materials for Agrobacterium infection.

[0017] Centrifuge the Agrobacterium with pinellia lectin gene pta, resuspend it in liquid MS medium, add 100μM AS at the same time, activate it for 1-2hr, and then infect the explants for 20 minutes (OD 600nm ≈0.8), put it on MS+6-BA 1.0mg / L+NAA 0.5mg / +2,4-D 0.1mg / L (pH5.8) solid medium for 5 days, take it...

Embodiment 2

[0020] The experimental variety is Oriental lily. Peel the bulb from the outside to the inside, rinse it with clean water, soak it in washing powder solution for 30 minutes, and then sterilize it on the aseptic operation table: soak it in 75% alcohol for 2 minutes, rinse it three times with sterile water, 20% concentration The commercially available Antiformin was soaked for 15-20min, rinsed with sterile water for 3 times, and cut into 5mm size as explants for later use.

[0021] The explants were inoculated on MS (pH5.8) fixed medium and cultured for 7 days, and fresh wounds were cut out around uncontaminated scales with a scalpel as materials for Agrobacterium infection.

[0022] The Agrobacterium carrying the rapeseed sodium hydrogen pump gene nhx was centrifuged and resuspended in liquid MS medium, while adding 100 μM AS, activated for 1-2 hours, and then infected the explants for 15 minutes (OD 600nm ≈0.8), put it on MS+6-BA1.0mg / L+NAA 0.5mg / +2, 4-D 0.1mg / L (pH5.8) solid ...

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Abstract

This invention relates to a method for transforming lily with agricillin mediaed ex-source function gene applying a way of transforming lily bulb with agricillins, and applying liquid Ms added by acety1 syringone activation mycelium to get the effect of setting up agricillin mediaed lily gene genetic transformation system and increasing its transformation rate. First of all, the lily bulb is disinfected and cut to small blocks and cultivated on a fix MS culture medium, unpolluted lepises are selected to be cut to form new wounds on the edge, an explant is infected by the resuspended agricillin to be cultivated on a co-culture medium to be selectively cultivated on a screen medium with kantlex to induce resistant adrentitious buds finally to be induced on a rooting medium and to be shift to get regeneration shoots and transformed lily plants.

Description

technical field [0001] The invention relates to the fields of biotechnology and agricultural technology. In particular, it is a method for transforming lily with exogenous functional gene mediated by Agrobacterium. Background technique [0002] For the genetic transformation of plants, it is generally believed that the method mediated by Agrobacterium is an ideal way. In recent years, the use of Agrobacterium to introduce foreign genes into monocotyledonous plants has made breakthroughs in both methods and practical application effects. sexual progress. As an important monocotyledonous flower, lily has little research on its genetic transformation. [0003] In the search and analysis of prior art documents, it was found that Watad et al. applied in "Microprojectile bombardment-mediated transformation of Lilium longiflorum", "microprojectile bombardment-mediated transformation of Lilium longiflorum" (Plant cell report, Plant Cell Bulletin, 1998) The reporter gene and marke...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N5/04C12N5/10C12N15/82
Inventor 唐东芹杨学军钱虹妹黄丹枫
Owner SHANGHAI JIAO TONG UNIV
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