Application of R1R6 genotype apple as receptor material in preparation of red pulp transgenic apple

A receptor material, 1.R1R6 technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as difficult to obtain, low nutritional and health ingredients, and low transformation efficiency

Active Publication Date: 2021-05-25
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the extremely difficult acquisition of genetically modified apple materials, the functions of many genes cannot be fully verified
At present, the only apple material used for transgenic function verification at home and abroad is the tissue culture seedling 'GL3' of 'Gala', which has a low transformation efficiency.
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Method used

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  • Application of R1R6 genotype apple as receptor material in preparation of red pulp transgenic apple
  • Application of R1R6 genotype apple as receptor material in preparation of red pulp transgenic apple
  • Application of R1R6 genotype apple as receptor material in preparation of red pulp transgenic apple

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, basic research work

[0063] In the early stage, the inventor cooperated with Professor Fei Zhangjun of Cornell University in the United States to conduct genome resequencing and bioinformatics analysis on 117 apple resources worldwide, including Xinjiang wild apple and European forest apple. Studies have shown that Xinjiang red-flesh apples (Malus sieversii f.niedzwetzkyana) native to my country have preserved high homologous consistency due to the geographical isolation of the Tianshan Mountains, and have not been infiltrated by foreign genes. Therefore, the Xinjiang red-flesh apple and apple variety hybrid first generation and backcross first and second generation segregation populations were constructed, and the genetic structure and genetic diversity characteristics of Xinjiang wild apple populations, technical parameters for core germplasm construction, flavonoid content, etc. were clarified. The genetic variation characteristics and development mech...

Embodiment 2

[0064] Embodiment 2, preparation tissue culture seedling

[0065] 1. Obtain the F1 generation group

[0066] The "Zihong No. 1" apple plant (as the male parent) and the "Gala" apple plant (as the female parent) were crossed, and the seeds of mature fruit were harvested, which were the F1 generation seeds. The F1 generation seeds are sown, and the grown plants are the F1 generation plants, and the population formed by the F1 generation plants is the F1 generation population.

[0067] The "Zihong No. 1" apple plants are of R6R6 genotype. "Gala" apple plants were of R1R1 genotype. Each plant of the F1 population was of the R1R6 genotype. Sequence 3 of the sequence table shows the sequencing result of the PCR amplification product of a certain "Zihong No. 1" apple plant when the above-mentioned genotype is identified. Sequence 4 of the sequence table shows the sequencing result of the PCR amplification product of a "Gala" apple plant when the above-mentioned genotype is identi...

Embodiment 3

[0099] Example 3. Taking the MdMYB10 gene as an example to verify whether it can be used as an effective transgenic acceptor material

[0100] The MdMYB10 gene is shown in sequence 5 of the sequence listing.

[0101] 1. Preparation of recombinant Agrobacterium

[0102] Insert the double-stranded DNA molecule shown in the 1st-729th nucleotides in Sequence 5 of the sequence listing between the SalI and BamHI restriction sites of the pRI101 vector to obtain a recombinant plasmid. The recombinant plasmid was introduced into Agrobacterium LBA4404 to obtain recombinant Agrobacterium.

[0103] 2. Carry out Agrobacterium infection and transformation on tissue cultured seedlings of red meat apple

[0104] Test seedlings: 10 strains (H1 strain, H2 strain, H3 strain, G1 strain, G2 strain, G3 strain, G4 strain, G5 strain) that can be normally differentiated and subcultured in embodiment 1 line, G6 strain or G7 strain), and cultured seedlings for 25 days in the proliferation medium.

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Abstract

The invention discloses an application of an R1R6 genotype apple as a receptor material in preparation of a red pulp transgenic apple, provides application of an R1R6 genotype apple as a receptor material in preparation of a transgenic apple, provides an application of the apple (Malus sp.) BCF2 as an acceptor material in preparation of a transgenic apple, and also provides a method for preparing the transgenic apple, which comprises the following step: performing genetic transformation on the Malus sp. BCF2 serving as a receptor material to obtain the transgenic apple. The inventor of the invention finds that in an F2 generation obtained by taking the apples of "purplish red No.1" and "Gala" varieties parents, and it is found that a strain with a genotype of R1R6 is easier to obtain a tissue culture seedling and has higher transformation efficiency as a genetic transformation receptor, the transformation rate of a G2 strain being the highest. The method disclosed by the invention has an important application value for the creation of the transgenic red-pulp apples.

Description

technical field [0001] The invention relates to the application of R1R6 genotype apples as acceptor materials in the preparation of red meat transgenic apples. Background technique [0002] Apple is the largest tree species of deciduous fruit trees in my country. According to statistics in 2016, my country's apple cultivation area and output reached 2.4669 million hm2 and 43.8823 million tons respectively, ranking first in the world and being the world's largest apple resource, production and consumption country . Apple has good storability and long supply cycle. Its fruit contains a high proportion of free polyphenols that are easily absorbed by the human body. It has good anti-oxidation, anti-tumor, prevention of cardiovascular and cerebrovascular diseases, and liver protection. It is nutritious and healthy high value. [0003] In recent years, with the completion of apple genome sequencing, the basic research on apples at home and abroad has made great progress, and more...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H6/74A01H5/12A01H5/08A01H1/02
CPCC12N15/8205C12N15/825C12N15/8243A01H1/02A01H5/08A01H6/7418Y02E50/10
Inventor 刘文军王楠陈学森房鸿成于蕾李志强毛志泉
Owner SHANDONG AGRICULTURAL UNIVERSITY
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