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A kind of Agrobacterium-mediated transformation method of corn mature embryo stem tip

A technology for Agrobacterium-mediated and mature embryos, applied in biochemical equipment and methods, botany equipment and methods, fermentation, etc., can solve the problems of large sex line variation, restricting the research and development of maize genetic transformation, and long cycle

Inactive Publication Date: 2019-06-28
CHINA AGRI UNIV
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Problems solved by technology

However, although both barley and corn are monocotyledonous plants, barley and corn as two different crops also determine the difference in transformation, and there is currently no technical solution suitable for corn
[0007] At present, the genetic transformation methods of maize mainly use Agrobacterium-mediated method and gene gun method, and the receptors are mainly immature embryos and embryogenic callus, and the shortcomings of callus-formed clones of regenerated plants are large variation and long cycle. The Development of Maize Genetic Transformation Research

Method used

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  • A kind of Agrobacterium-mediated transformation method of corn mature embryo stem tip
  • A kind of Agrobacterium-mediated transformation method of corn mature embryo stem tip
  • A kind of Agrobacterium-mediated transformation method of corn mature embryo stem tip

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Experimental program
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Embodiment 1

[0089] 1. Maize stem tip culture and carrier construction:

[0090] 1. Corn stem tip culture

[0091] 1.1 Select plump and undamaged seeds, put them into a triangular flask and carry out two sterilization treatments:

[0092] The first pass: Sterilize with 70% ethanol for 8 minutes, sterilize with 0.1% mercuric chloride aqueous solution for six minutes, wash with sterile water three times: wash with sterile water for 2 times the seed volume for one minute for the first time, and 3 times for the second time Wash the seed volume with sterile water for 2-3 minutes, wash with sterile water of 2 times the seed volume for one minute for the third time, then add 1.5 times the seed volume of sterile water to soak the seeds for 6 hours (24-26 degrees Celsius, if the temperature is too high, The time can be shortened appropriately), the time of soaking must be well grasped, if the time is too long, the seeds will easily turn black, and the rate of germination will decrease;

[0093] T...

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Abstract

The invention relates to transgenic maize, in particular to an agrobacterium-mediated mature maize embryo shoot apex transformation method. The method, which adopts maize shoot apex as a receptor, is not limited by genotype; and a set of strict culture medium system which is suitable for using the maize shoot apex as the transformation receptor is also developed, so that the efficient genetic transformation of the maize is achieved. The method disclosed by the invention shortens a maize transformation cycle, avoids genotype limitation by using the shoot apex as the receptor and also avoid such links as plant regeneration and the like in a transgenosis process. The method breaks through the limitation of explants transformation and improves the transformation efficiency of the maize. The method is simple and convenient to operate, simple and easy to operate, short in transformation cycle, high in transformation efficiency and reliable in result, and a transgenic maize plant can be rapidly obtained.

Description

technical field [0001] The invention relates to transgenic corn, in particular to an Agrobacterium-mediated method for transforming mature corn embryo stem tips. Background technique [0002] The history of corn cultivation in my country is about 470 years. At present, my country's sown area is about 300 million mu, second only to rice and wheat, ranking third among food crops, and second only to the United States in the world. Corn is mainly used as food, feed, energy, nutrition and raw materials in the pharmaceutical industry. Its industrial value is widely used in papermaking, food, textile, pharmaceutical and other industries. It has the characteristics of high yield, storage resistance and high reproduction value. [0003] As of 2014, the planting area of ​​GM crops increased from 1.7 million hectares in 1996 to 181.5 million hectares in 2014, of which the planting area of ​​herbicide-tolerant GM corn was about 8 million hectares, and the planting area of ​​insect-resi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/46
Inventor 康定明刘永健曹金伶张少蔓赵传森严慧慧
Owner CHINA AGRI UNIV
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