Method for improving agrobacterium-mediated genetic transformation efficiency of maize

A genetic transformation efficiency, Agrobacterium-mediated technology, applied in biochemical equipment and methods, botanical equipment and methods, introduction of foreign genetic material using vectors, etc., can solve the problem of unpredictable phenotype of hybrid offspring and low genetic transformation efficiency. , it is difficult to develop to meet the needs and other problems, to achieve the effect of shortening the time of germination and germination, high genetic transformation rate and easy to obtain

Inactive Publication Date: 2019-02-22
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

[0003] Simple traditional breeding methods have exposed shortcomings: long breeding cycle, low breeding efficiency, and difficult to predict the phenotype of hybrid offspring, which has been difficult to develop to meet the demand. The combination of molecular breeding technology and traditional breeding technology can improve the quality of maize. The output and quality have become the main development trend in the future
Although this method shortens the time for genetic transformation of maize to a certain extent, due to the hard seed coat protection on the outside of the seeds, the infection efficiency is not high, so the genetic transformation efficiency obtained is not high

Method used

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  • Method for improving agrobacterium-mediated genetic transformation efficiency of maize
  • Method for improving agrobacterium-mediated genetic transformation efficiency of maize

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Effect test

Embodiment 1

[0027] A method for improving Agrobacterium-mediated genetic transformation efficiency of maize, said method comprising the following steps:

[0028] (1) Treatment of seeds: corn seeds are soaked and sterilized in 45% sodium hypochlorite solution for 5 minutes, washed with sterile distilled water for 2 times, dry the surface moisture of the seeds with sterile filter paper, put the seeds into the seed culture solution, Ultrasonic treatment at 40kHz in the dark at 40°C for 20 minutes, let stand for 10 minutes, remove the seeds, soak in hot water at 40°C for 30 minutes, and obtain germinated seeds;

[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 containing the target gene into the Agrobacterium culture solution, shake and cultivate it at 25°C and 220rmp, and wait unti...

Embodiment 2

[0039] A method for improving Agrobacterium-mediated genetic transformation efficiency of maize, said method comprising the following steps:

[0040] (1) Treatment of seeds: corn seeds are soaked and sterilized in 50% sodium hypochlorite solution for 10 minutes, washed with sterile distilled water for 3 times, dry the surface moisture of the seeds with sterile filter paper, put the seeds into the seed culture solution, Ultrasonic treatment at 45kHz in the dark at 47.5°C for 25 minutes, let stand for 15 minutes, remove the seeds, soak in hot water at 47.5°C for 40 minutes, and obtain germinated seeds;

[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: put the Agrobacterium single bacterium containing the target gene into the Agrobacterium culture medium, shake and cultivate it at 26.5°C at 250rmp, and wa...

Embodiment 3

[0051] A method for improving Agrobacterium-mediated genetic transformation efficiency of maize, said method comprising the following steps:

[0052] (1) Treatment of seeds: corn seeds are soaked and sterilized in 55% sodium hypochlorite solution for 15 minutes, cleaned 3 times with sterile distilled water, dry the surface moisture of the seeds with sterile filter paper, put the seeds into the seed culture solution, Ultrasonic treatment at 50kHz in the dark at 55°C for 30min, let stand for 20min, remove the seeds, soak in hot water at 55°C for 50min, and obtain germinated seeds;

[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 the Agrobacterium single bacterium containing the target gene into the Agrobacterium culture solution, shake and cultivate it at 28°C and 280rmp, and wait for the OD 600 At...

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Abstract

The invention relates to the field of plant transgenic technology, in particular to a method for improving the agrobacterium-mediated genetic transformation efficiency of maize. Transgenic plants areobtained by the steps of ultrasonic treatment of seeds to obtain germinated seeds, culture of agrobacteria containing target genes, seed infection and co-culture, agrobacterium removal, selective culture, and molecular detection of transgenic plants. The method can effectively shorten the time of transgene transformation and improve the agrobacterium-mediated genetic transformation efficiency of maize.

Description

【Technical field】 [0001] The invention relates to the technical field of plant transgenesis, in particular to a method for improving the efficiency of genetic transformation of maize mediated by Agrobacterium. 【Background technique】 [0002] Maize is an annual monocotyledonous herbaceous plant of the Poaceae Zea genus and one of the most important food crops in the world. With the growth of the world population and the continuous improvement of people's living standards, higher requirements are placed on the variety and quality of corn. At the same time, with the continuous influx of foreign seed industries into the market, the adjustment of the country's planting industry structure, and the influence of various biological and abiotic factors, the corn area has been greatly reduced. In order to improve the quality of corn to ensure national food security, meet people's higher quality needs, improve market competitiveness, and provide support for the development of new energ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/46A01C1/00
CPCA01C1/00C12N15/8205
Inventor 谢宗宜
Owner JINGXI XIUMEI BIANCHENG AGRI SCI & TECH CO LTD
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