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Transformation method of super-sweet corn

A corn and formula technology, applied in the field of genetic engineering and transgenics, can solve problems such as super sweet corn obstacles, and achieve the effect of efficient genetic transformation

Pending Publication Date: 2022-04-12
CROP RES INST GUANGDONG ACAD OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no mature and efficient Agrobacterium-mediated transformation method for this inbred variety, which brings obstacles to the further improvement of specific traits of super sweet corn through transgenic technology using this inbred variety as a recipient

Method used

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  • Transformation method of super-sweet corn
  • Transformation method of super-sweet corn
  • Transformation method of super-sweet corn

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preliminary test of conversion efficiency of Qun01X01

[0036] Qun01X01 is a super-sweet corn inbred line material. The present invention first tests the transformation efficiency of the Qun01X01 inbred line according to the method disclosed in patent CN 104745622B. The carrier used carries the glufosinate-resistant bar gene (shown in SEQ ID NO.1 ) and GFP reporter gene (shown in SEQ ID NO.2), the vector diagram is shown in figure 1 .

[0037] The results showed that no positive plants were obtained in several transformation batches, and no transformation was successful. It is necessary to optimize the parameters of the transformation system.

Embodiment 2

[0038] Embodiment 2 to the optimization of heat shock time

[0039] During the infection stage, heat shock to young embryos can greatly increase the transformation efficiency. It is also mentioned in CN103114106B patent that heat-shocking immature corn embryos at 45°C for 5 minutes can improve the infection efficiency of Agrobacterium. The present invention tries to use the method of heat shock to treat immature embryos, the heat shock temperature is set at 46° C., and the treatment is performed for 3 minutes and 5 minutes respectively. Other operations are consistent with those disclosed in CN 104745622 B. The results showed that heat shock treatment for 3 minutes had the best effect, and the conversion efficiency could reach 3.3% (Table 1).

[0040] The influence of table 1 heat shock treatment on conversion efficiency (%)

[0041]

[0042] Since Qun01X01 cannot be transformed without heat shock treatment, it will affect the accuracy of evaluating the effects of other pa...

Embodiment 3

[0043] Embodiment 3 is to the optimization of infection medium

[0044] Cysteine ​​is an antioxidant that can make immature maize embryo cells less likely to undergo programmed cell death after Agrobacterium infection. Adding 150-300mg / L cysteine ​​to the co-culture medium can increase Agrobacterium (Geng Wanying. Genetic transformation of ZmSec14p gene in maize and genetic research on somatic embryogenesis [D]. Jilin University, 2020; CN103114106B). Low-concentration cysteine ​​has a limited effect, but high-concentration cysteine ​​will affect the growth of immature embryos. The present invention sets no addition, addition of 50mg / L, addition of 200mg / L, addition of 400mg / L cysteine ​​4 The effects of this treatment on the transformation efficiency of Qun01X01 were tested. After testing, it was found that adding 200 mg / L cysteine ​​to the infection medium instead of the co-culture medium was most beneficial to improve the transformation efficiency of Qun01X01 (Table 2).

...

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Abstract

The invention belongs to the technical field of gene engineering and transgenosis, and particularly relates to a transformation method of super-sweet corn. By optimizing parameters such as heat shock immature embryo, infection culture medium and rest culture medium formula, the transformation efficiency of the super-sweet corn inbred line Qun01X01 is improved to 10.77% from non-transformation. By utilizing the method provided by the invention, efficient genetic transformation with the super-sweet corn inbred line as a receptor can be realized, so that transgenic super-sweet corn materials and varieties with specific characters can be directly cultivated.

Description

technical field [0001] The invention belongs to the field of genetic engineering and transgenic technology, and in particular relates to a method for transforming super-sweet corn. Background technique [0002] Super sweet corn belongs to sweet corn (Zea mays L. Sacchara sturt). In addition to the food properties of corn, it is also known as vegetable and fruit corn. It has become an important cultivated crop in the world today. Fresh buds and their products are beauty, One of the important consumer foods in Japan, Western Europe and some countries and regions in Southeast Asia. my country's super-sweet corn planting area and output both rank first in the world. South China is the main production area of ​​super-sweet corn in my country, and it is a characteristic and dominant crop in Guangdong Province. [0003] Genetic transformation is the basic technical system for breeding transgenic maize varieties. Although super sweet corn has disclosed related genetic transformati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C07K14/415C12N15/82
Inventor 于永涛严建兵胡建广许洁婷李高科韩宝柱
Owner CROP RES INST GUANGDONG ACAD OF AGRI SCI
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