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Modified method for transforming gramineous crop by agrobacterium mediation flower-dipping method

A technology of gramineous crops and Agrobacterium, which is applied in the field of transforming sweet sorghum, millet and corn, can solve the problems that gramineous crops have not been reported, and achieve the effect of great practical application value

Active Publication Date: 2008-05-28
未名三农生物农业技术有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the flower soaking method has been applied to the Brassica crops of the dicotyledonous plants Brassicaceae and has been successfully transformed. There is no report on the grass crops transformed by the soaking flower method.

Method used

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  • Modified method for transforming gramineous crop by agrobacterium mediation flower-dipping method
  • Modified method for transforming gramineous crop by agrobacterium mediation flower-dipping method
  • Modified method for transforming gramineous crop by agrobacterium mediation flower-dipping method

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Experimental program
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Effect test

Embodiment

[0021] Embodiment, flower soaking method transformation to obtain sweet sorghum, millet, corn transgenic plants

[0022] 1. Transformation of sweet sorghum, millet, and corn plants by soaking flowers

[0023] 1. Experimental materials

[0024] Agrobacterium strain: The Agrobacterium strain was LBA4404. Contains the plasmid vector pKAT1-AsRed.

[0025] Plasmid: pKAT1-AsRed, containing red fluorescent protein reporter gene AsRed2 from jellyfish, with hpt selection marker gene, resistant to Hygmycin antibiotics. The vector was constructed in our laboratory. The target gene AsRed2 (purchased from Clontech Laboratories Inc, USA) was inserted into the expression vector pCAMBIA1301, and the seed-specific expression promoter GluA3 cloned from rice was inserted into the upstream of the AsRed2 gene to obtain pKAT-AsRed Expression vector. Its map is shown in Figure 1, in which T-DNA left border is the left border of T-DNA insertion; hpt is a plant selection marker gene with Hygmycin ...

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Abstract

The invention provides an improved process for transforming gramineous crops by floral dip method, including the following steps. Firstly, agrobacterium with target gene is suspended by infiltration culture medium which contains Silwet L-17, Triton X-100 and GA. Secondly, under the condition of vivo, the integral inflorescence or partial pistils of the plants which are ready to be transformed are immerged in the infiltration culture medium, or the inflorescence or the pistils are sprayed with the infiltration culture medium, and the target gene is integrated with the genome of a target plant by the agrobacterium. By employing the genetically modified process, the invention resolves transformation problems of certain gramineous crops which are hard to be transformed by regular transforming method, thereby the invention has relatively high practical application value.

Description

technical field [0001] The invention relates to an improved method and application of transforming gramineous crops by flower soaking method mediated by Agrobacterium, in particular to the method and application of transforming sweet sorghum, millet and corn. Background technique [0002] Grain production can be mainly divided into two types, one is to provide food for food, and the other is to provide raw materials for bio-energy. Sweet sorghum is an important energy plant. The oil crisis in the 1970s triggered human research on biofuels instead of petroleum, and the United States and Brazil succeeded in producing fuel ethanol from corn and sugarcane. Now, due to the gradual depletion of fossil energy and the need for environmental protection, the development of biomass industry has become an important development strategy for many countries. my country's food is not so rich. There are other ways to produce liquid biofuels. Sweet sorghum has a high photosynthetic effici...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/87C12N15/84
Inventor 夏勉张会新凌莉
Owner 未名三农生物农业技术有限公司
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