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Method for improving heredity transformation efficiency of agrobacterium tumefaciens mediated rice

A technology of genetic transformation efficiency, Agrobacterium-mediated, applied in the direction of biochemical equipment and methods, botanical equipment and methods, using vectors to introduce foreign genetic material, etc. Shorten the time of germination and germination, simplify the process, and obtain the effect easily

Inactive Publication Date: 2019-03-29
JINGXI XIUMEI BIANCHENG AGRI SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the infection of Agrobacterium is chemotaxis, and generally only infects dicotyledonous plants and gymnosperms, and has no infectivity to monocotyledonous plants
In recent years, although Agrobacterium-mediated transformation has been successfully transformed on some monocots, the transformation rate is still not high
In addition, due to the use of plant explants (such as immature embryos, callus, shoot tips, etc.)

Method used

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  • Method for improving heredity transformation efficiency of agrobacterium tumefaciens mediated rice
  • Method for improving heredity transformation efficiency of agrobacterium tumefaciens mediated rice
  • Method for improving heredity transformation efficiency of agrobacterium tumefaciens mediated rice

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Effect test

Embodiment 1

[0027] A method for improving the rice genetic transformation efficiency mediated by Agrobacterium, said method comprising the following steps:

[0028] (1) Treatment of seeds: After soaking the rice seeds in clear water for 12 hours, remove the seeds, put them into 0.1% potassium permanganate solution for immersion and disinfection for 20 minutes, remove the seeds, wash them twice with sterile distilled water, and then use Blot the water on the surface of the seeds with sterile filter paper, put the seeds into the seed culture solution, treat them with 40kHz ultrasonic wave in the dark at 40°C for 30min, then let them stand for 10min, remove the seeds, soak them in hot water at 40°C for 6h, and get germination seed;

[0029] The above seed culture solution is composed of the following components: 15.7 mg / L acetosyringone, 91.0 g / L mannitol, 0.59 mg / L NaCl, 5 mg / L cellulase and 5 mg / L pectinase.

[0030] (2) Cultivation of Agrobacterium: put the Agrobacterium single bacterium...

Embodiment 2

[0039] A method for improving the rice genetic transformation efficiency mediated by Agrobacterium, said method comprising the following steps:

[0040] (1) Treatment of seeds: After soaking the rice seeds in clear water for 14 hours, remove the seeds, put them into 0.15% potassium permanganate solution for immersion and disinfection for 25 minutes, remove the seeds, wash them with sterile distilled water for 3 times, and then use Blot the water on the surface of the seeds with sterile filter paper, put the seeds into the seed culture solution, and treat them with 45kHz ultrasonic waves at 42.5°C in the dark for 40 minutes, then let them stand for 15 minutes, remove the seeds, soak them in hot water at 42.5°C for 8 hours, and obtain germination. seed;

[0041] The above seed culture solution is composed of the following components: 22.6 mg / L acetosyringone, 100.2 g / L mannitol, 0.89 mg / L NaCl, 15 mg / L cellulase and 10 mg / L pectinase.

[0042] (2) Cultivation of Agrobacterium: ...

Embodiment 3

[0051] A method for improving the rice genetic transformation efficiency mediated by Agrobacterium, said method comprising the following steps:

[0052] (1) Treatment of seeds: After soaking the rice seeds in clear water for 16 hours, remove the seeds, put them into 0.2% potassium permanganate solution for immersion and disinfection for 30 minutes, remove the seeds, wash them with sterile distilled water for 3 times, and then use Blot the water on the surface of the seeds with sterile filter paper, put the seeds into the seed culture solution, and treat them with 50kHz ultrasonic waves at 45°C in the dark for 50 minutes, then let them stand for 20 minutes, remove the seeds, soak them in hot water at 45°C for 10 hours, and obtain germination. seed;

[0053] The above seed culture solution is composed of the following components: 29.5 mg / L acetosyringone, 109.3 g / L mannitol, 1.18 mg / L NaCl, 25 mg / L cellulase and 15 mg / L pectinase.

[0054] (2) Cultivation of Agrobacterium: put ...

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Abstract

The invention relates to the technical field of genetic modification of plants, in particular to a method for improving the heredity transformation efficiency of agrobacterium tumefaciens mediated rice. The method comprises the steps of performing ultrasonic treatment on seeds to obtain germinated seeds, culturing agrobacterium tumefaciens containing target genes, soaking and infecting the germinating seeds, performing co-culture, performing bacterium removal and selective culture, and performing molecular detection on transgenic plants so as to obtain the transgenic plants. Through the adoption of the method disclosed by the invention, the transformation time of genetic modification can be effectively shortened, and besides, the heredity transformation efficiency of the agrobacterium tumefaciens mediated rice can be improved.

Description

【Technical field】 [0001] The invention relates to the technical field of plant transgenesis, in particular to a method for improving the efficiency of rice genetic transformation mediated by Agrobacterium. 【Background technique】 [0002] Rice is an annual monocotyledonous herbaceous plant belonging to the Gramineae family. It is native to China and India. Rice was planted in the Yangtze River Basin in China 7,000 years ago. It is one of the important food crops in China and the world. At present, as the contradiction of more people and less land becomes increasingly prominent, how to make full use of limited land resources to feed more than one billion people has become an important challenge for my country's agricultural scientific research. Secondly, with the further improvement of the quality of life of the people, the requirements for rice quality are getting higher and higher. There is an urgent need to breed a large number of new rice varieties to meet the needs of dif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/46
CPCC12N15/8205
Inventor 谢宗宜
Owner JINGXI XIUMEI BIANCHENG AGRI SCI & TECH CO LTD
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