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Method for improving agribacterium-mediated transformation survival rate of wheat

A technology for Agrobacterium and wheat, applied in biochemical equipment and methods, botanical equipment and methods, applications, etc., can solve the problems of affecting transformation efficiency and the death of transformed wheat seedlings, and achieve improved survival efficiency, great application value and market. Promotion prospect, low price effect

Active Publication Date: 2013-09-04
BEIJING WEIMING KAITUO CROP DESIGN CENT COMPANYLIMITED
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, when using longitudinally cut wheat seedling leaf bases for Agrobacterium transformation, the massive growth of mold during the co-cultivation process led to the death of most of the transformed wheat seedlings. This phenomenon is very common, which seriously affects the transformation efficiency.

Method used

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  • Method for improving agribacterium-mediated transformation survival rate of wheat
  • Method for improving agribacterium-mediated transformation survival rate of wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Experimental materials

[0023] Plasmid: Containing paraquat resistance gene, pV1 vector with antibiotic selection marker gene, has paraquat resistance. The gene was amplified by PCR and connected to pcambia1300 (purchased from Invitrogen) vector to obtain pV1 vector.

[0024] Agrobacterium strain: AGL0

[0025] Wheat variety for transformation: Jingmai 9158

[0026] 2. Cultivation of Agrobacterium

[0027] Pick the Agrobacterium containing the target gene frozen in glycerol, draw a line on the YEB plate medium, and culture it in a dark incubator at 28°C for 2 days. Pick a single colony and inoculate it in 40ml of resistant YEB liquid medium, shake overnight at 200rpm in a dark shaker at 28°C until OD 600 ≈1. Take 400 μl of the above-mentioned bacterial solution and spread it on the YEB solid medium with a plate diameter of 15 cm. After culturing for 2 days in a dark incubator at 28°C, it is ready for use. Do not keep it in a dark incubator at 28°C or transfer i...

Embodiment 2

[0066] 1. Experimental materials

[0067] Plasmid: pV1 vector containing paraquat resistance gene and antibiotic selection marker gene. The gene was amplified by PCR and connected to pcambia1300 (purchased from Invitrogen) vector to obtain pV1 vector.

[0068] Agrobacterium strain: AGL0

[0069] Wheat variety for transformation: Jingmai 9158

[0070] 2. Cultivation of Agrobacterium

[0071] Pick the Agrobacterium containing the target gene frozen in glycerol, draw a line on the YEB plate medium, and culture it in a dark incubator at 28°C for 2 days. Pick a single colony and inoculate it in 40ml of resistant YEB liquid medium, shake overnight at 200rpm in a dark shaker at 28°C until OD 600 ≈1. Take 400 μl of the above-mentioned bacterial solution and spread it on the YEB solid medium with a plate diameter of 15 cm. After culturing for 2 days in a dark incubator at 28°C, it is ready for use. Do not keep it in a dark incubator at 28°C or transfer it to a refrigerator at 4°C....

Embodiment 3

[0111] 1. Experimental materials

[0112] Plasmid: pV1 vector containing paraquat resistance gene and antibiotic selection marker gene. The gene was amplified by PCR and connected to pcambia1300 (purchased from Invitrogen) vector to obtain pV1 vector.

[0113] Agrobacterium strain: AGL0

[0114] Wheat variety for transformation: Jingmai 9158

[0115] 2. Cultivation of Agrobacterium

[0116] Pick the Agrobacterium containing the target gene frozen in glycerol, draw a line on the YEB plate medium, and culture it in a dark incubator at 28°C for 2 days. Pick a single colony and inoculate it in 40ml of resistant YEB liquid medium, shake overnight at 200rpm in a dark shaker at 28°C until OD 600 ≈1. Take 400 μl of the above-mentioned bacterial solution and spread it on the YEB solid medium with a plate diameter of 15 cm. After culturing for 2 days in a dark incubator at 28°C, it is ready for use. Do not keep it in a dark incubator at 28°C or transfer it to a refrigerator at 4°C....

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Abstract

The invention discloses a method for improving the agribacterium-mediated transformation survival rate of wheat. When length cutting sprout leaf-base cushion is used for agrobacterium-mediated transformation, a preservative agent is added into a co-culture medium, so as to restrain the growth of mucedine and improve the survival rate of transformation plants.

Description

Technical field [0001] The present invention belongs to the field of wheat genetically modified, involving the use of preservatives to improve the efficiency of wheat survival efficiency of longitudinal leaf -based leaf -based leaf bases, thereby improving the transformation efficiency. Background technique [0002] Wheat is an important grain crop with total output. It is widely planted around the world.With the growth of the world's population and the improvement of people's living standards, the demand for wheat is increasing, and the requirements for wheat quality are getting higher and higher. Conventional breeding can not meet people's requirements for wheat varieties and quality.The development of genetically modified technology has broken the racial restrictions of conventional breeding. Many crops such as corn and rapeseed using genetically modified technology have successfully cultivated new high -quality and efficient transgenic varieties.Compared with other crops, the...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 冉冬夏勉张斌何峰郑辰
Owner BEIJING WEIMING KAITUO CROP DESIGN CENT COMPANYLIMITED
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