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Method for screening dwarf apple stocks by aid of reverse genetics

A technology of reverse genetics and dwarfing rootstocks, applied in the field of apple breeding, can solve the problems of Pingyi sweet tea not having dwarfing characteristics, the dwarfing characteristics of mutants cannot be used, and restricting the application and development, and improving the dwarfing ability. , easy to operate, simple steps

Inactive Publication Date: 2018-01-30
SHANDONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, Pingyi sweet tea does not have dwarfing characteristics, which limits its further application and development
[0005] The green revolution in the 20th century originated from the screening of gibberellin (GA)-insensitive dwarf mutants. The acceptance and conduction of GA hormone signals in such mutants were blocked, although the endogenous GA content did not change much, even compared with wild type is still taller, but the dwarf traits of mutants cannot be restored by exogenous GA

Method used

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  • Method for screening dwarf apple stocks by aid of reverse genetics
  • Method for screening dwarf apple stocks by aid of reverse genetics
  • Method for screening dwarf apple stocks by aid of reverse genetics

Examples

Experimental program
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experiment example 1

[0025] Experimental Example 1: Screening of GA-insensitive dwarf mutants in Pingyi sweet tea mutagenesis seedling population by forward genetic method

[0026] The wild-type Pingyi sweet tea (WT) seeds that were mutagenized by Co60 dose of 5000 roentgen radiation for 15 min were stratified at 4 °C for 60 days, grown for 30 days, and then sown into the substrate. Progeny—the dwarf mutant (dwarf1) was harvested for seed treatment, and the dwarf mutant (dwarf1) and wild-type Pingyi sweet tea (WT) were treated with 0 μM, 60 μM, and 150 μM of GA3, respectively. GA3, a homologue of gibberellin, commonly known as 920, has strong physiological activity. The response of dwarf mutant (dwarf1) and wild-type Pingyi sweet tea (WT) to GA was detected, and the results showed that the dwarf mutant (dwarf1) exhibited dwarf characteristics compared with wild-type Pingyi sweet tea (WT) , is not sensitive to GA response. like figure 1 shown.

experiment example 2

[0027] Experimental example 2: Grafting experiments to verify the dwarfing characteristics of dwarf1

[0028] The wild-type Pingyi sweet tea (WT) and the dwarf mutant (dwarf1) were grafted with Gala apple (hereinafter referred to as Gala) respectively, and managed under the same conditions to detect the dwarfing ability of the two rootstocks to scion. The results showed that the dwarf mutant (dwarf1) had dwarfogenicity to the scion. like figure 2 shown.

experiment example 3

[0029] Experimental example 3: Grafting experiment to verify that DELLA protein in dwarf1 is not sensitive to gibberellin

[0030] Gala self-rooted seedlings, Gala / wild-type Pingyi sweet tea (WT), and Gala / dwarf mutant (dwarf1) corresponding to 1-5 years old were treated with GA to detect the stability of MhGA1 protein. The results showed that GA treatment significantly promoted the degradation of MhGAI1 protein in Gala self-rooted seedlings and Gala / wild-type Pingyi sweet tea (WT) stock-spike combination, while Gala / dwarf mutant (dwarf1) significantly inhibited this degradation, indicating that The DELLA protein in the dwarf mutant (dwarf1) is insensitive to gibberellin. like image 3 shown.

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Abstract

The invention discloses a method for screening dwarf apple stocks by the aid of reverse genetics and belongs to the technical field of apple breeding. The method includes: subjecting Pingyi sweet-teaseeds with an apomixes feature to Co60 radiation mutation; sowing mutated seeds; screening offspring with a dwarfing trait in group offspring and subjecting the offspring to exogenous GA treatment; screening the offspring insensitive to GA; through MhGAI1 antibody detection, performing screening to obtain DELLA mutants. Through the radiation mutation method, specific antibodies for Pingyi sweet tea subjected to radiation mutation are detected by the aid of the reverse genetics for the first time, plants insensitive to GA are screened, and genetic resources with the dwarf trait and capable of keeping apomixes capacity at the same time are further screened.

Description

technical field [0001] The invention relates to a method for screening apple dwarf rootstocks by using reverse genetics, and belongs to the technical field of apple breeding. Background technique [0002] Dwarfing cultivation is an important new cultivation mode in today's apple production process. Dwarfing cultivation mode has the characteristics of labor-saving and labor-saving. Therefore, it has become the future development direction of apple industry. At present, dwarf rootstocks imported from the West are generally used in dwarf cultivation. These rootstocks have a disadvantage, that is, the root system is underdeveloped and the main root is too short. [0003] Since apples in my country are generally planted in hilly and mountainous areas and in the west with water-deficient conditions, they are often faced with drought and water shortages during the cultivation process, and dwarf cultivation is not conducive to promotion in these areas. Therefore, it is urgent to re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/06A01H1/04A01G7/06G01N33/53
Inventor 郝玉金王小非沙广利
Owner SHANDONG AGRICULTURAL UNIVERSITY
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