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Method for improving soybean resistant cluster bud induction efficiency by using 5-azacytidine

An azacytosine and bud induction technology is applied in the field of improving the induction efficiency of soybean resistant clump buds, and can solve the problems of low soybean transgenic efficiency, limited application of transgenic technology, lack of transformation efficiency of varieties with low transformation efficiency, etc. The effect of improving induction efficiency and transformation efficiency

Pending Publication Date: 2019-07-23
SHENYANG AGRI UNIV
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  • Application Information

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Problems solved by technology

However, previous studies have shown that Agrobacterium-mediated soybean transformation efficiency is closely related to soybean genotype, which significantly limits the application of transgenic technology in some varieties with good agronomic traits
At present, the problem of low transgenic efficiency of soybeans is mainly solved by screening soybean varieties with high transformation efficiency, but there is a lack of effective methods to improve the transformation efficiency of varieties with low transformation efficiency

Method used

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  • Method for improving soybean resistant cluster bud induction efficiency by using 5-azacytidine
  • Method for improving soybean resistant cluster bud induction efficiency by using 5-azacytidine
  • Method for improving soybean resistant cluster bud induction efficiency by using 5-azacytidine

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Experimental program
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Effect test

Embodiment 1

[0023] Utilize 5-azacytidine nucleoside to improve the method for inducing efficiency of soybean resistant clustering buds, comprising the following steps:

[0024] (1) Sterilization of soybean seeds

[0025] Soybean seeds were sterilized by chlorine gas fumigation. Select plump soybean grains without disease spots, place the soybean seeds in a petri dish (diameter 90mm) with a half-open lid, place the petri dish in a seed desiccator, put a 150ml small beaker in the desiccator, and pour it into the beaker Add 100ml of sodium hypochlorite solution and 4ml of concentrated hydrochloric acid, quickly cover the upper cover of the desiccator, and use a parafilm to seal the interface of the desiccator. After 8 hours of disinfection, take out the culture dish and place it in a sterilized ultra-clean workbench. Cover the Petri dish and blow off the chlorine gas for 10 minutes.

[0026] (2) Germination of soybean seeds

[0027] Soybean seeds with hilum down, inoculate in petri dishes...

Embodiment 2

[0041] In this embodiment, only step (5) was added to the resistant cluster bud induction medium in the second week, and 10 μM 5-azacytidine nucleoside was added, and the other steps were the same as in Example 1.

Embodiment 3

[0043] In this embodiment, only step (5) is added to the resistant cluster bud induction medium in the second week, and 50 μM 5-azacytidine nucleoside is added, and other steps are the same as in Example 1.

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Abstract

The present invention relates to a method for improving the soybean resistant cluster bud induction efficiency by using 5-azacytidine, and belongs to the technical field of transgene. The main technical scheme is as follows: sterilization, germination, preparation of agrobacterium liquid, soybean cotyledon node infection by agrobacterium, and resistant cluster bud induction. In an agrobacterium-mediated soybean cotyledon node genetic transformation system, the method uses a methylation inhibitor of the 5-azacytidine for the first time for carrying out demethylation treatment on the soybean cotyledon explants which induce resistant cluster buds, improves the induction efficiency of the resistant cluster buds of the soybean cotyledon explants, and may improve the transformation efficiency ofthe T0 generation.

Description

technical field [0001] The invention relates to the technical field of transgenics, in particular to a method for improving the induction efficiency of soybean resistant clustered buds by using 5-azacytosine nucleoside. Background technique [0002] Soybean is the main edible oil, edible protein and feed crop in my country, and the improvement of its yield and quality is of great significance to the economy and agricultural industry. Soybean quality improvement using conventional breeding methods has disadvantages such as long breeding period, cumbersome operation, and poor interspecific combining ability. However, transgenic technology provides an effective method to break species barriers, which greatly promotes the genetic improvement process of crops. The selection process of high quality / high yielding crop varieties is thereby accelerated. In recent years, transgenic technology has been greatly developed, but compared with other species, the low transformation efficien...

Claims

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Application Information

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IPC IPC(8): C12N15/82
CPCC12N15/8201C12N15/8205
Inventor 赵明哲赵强谢甫绨
Owner SHENYANG AGRI UNIV
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