Method and Application of Surfactant and Ultrasound Synergistically Improving Soybean Genetic Transformation Efficiency

A genetic transformation method and surfactant technology, which is applied in the field of synergistically improving soybean genetic transformation efficiency with surfactants and ultrasonic waves, can solve the problems of accurate and effective wounding of explants, low yield of clustered buds, and low transformation efficiency

Active Publication Date: 2015-08-12
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a method for overcoming the low transformation efficiency existing in the existing Agrobacterium-mediated soybean cotyledonary node genetic transformation method, the difficulty of accurate and effective wounding of explants, the serious defense reaction of explants to Agrobacterium, and the overgrowth. The method and application of surfactant and ultrasonic synergistically improving the genetic transformation efficiency of soybean cotyledonary nodes mediated by Agrobacterium

Method used

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  • Method and Application of Surfactant and Ultrasound Synergistically Improving Soybean Genetic Transformation Efficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Surfactants and ultrasonic waves synergistically improve the transformation efficiency of the Agrobacterium tumefaciens-mediated genetic transformation method of soybean cotyledonary nodes

[0048] This example compares the Agrobacterium tumefaciens mediated soybean cotyledon node genetic transformation method (abbreviated as 2S) of the present application with surfactant and ultrasonic co-treatment, Agrobacterium tumefaciens mediated soybean with surfactant and scratch treatment Cotyledonary node genetic transformation method (abbreviated as SUR), soybean cotyledonary node genetic transformation method mediated by Agrobacterium tumefaciens mediated by ultrasonic treatment alone (abbreviated as SAAT) and soybean cotyledonary node genetic transformation method mediated by Agrobacterium tumefaciens mediated by scratch treatment alone (referred to as CK) conversion efficiency, these four methods were tested at the same time.

[0049] 1. The specific steps of the...

Embodiment 2

[0098] Embodiment 2, cultivating transgenic soybean

[0099] This example compares the Agrobacterium tumefaciens mediated genetic transformation method of soybean cotyledon nodes (abbreviated as 2S) using the surfactant and ultrasonic co-treatment of the application, and the Agrobacterium tumefaciens mediated by the surfactant and scratch treatment. Soybean cotyledon node genetic transformation method (SUR for short), soybean cotyledon node genetic transformation method mediated by A. Methods (CK for short) to cultivate the resistant cluster bud rate and explant necrosis rate of transgenic soybean, these four methods were all experimented at the same time.

[0100] 1. The specific steps of the Agrobacterium tumefaciens-mediated genetic transformation method of soybean cotyledon nodes (2S for short) co-treated with surfactants and ultrasonic waves of the present application are as follows:

[0101] 1.1. Preparation of soybean cotyledon node explants without scratches

[0102]...

Embodiment 3

[0161] Example 3. Transformation efficiency of soybean cotyledonary node genetic transformation method (2S for short) mediated by Agrobacterium tumefaciens co-treated with surfactants and ultrasonic waves of the present application on different soybean varieties

[0162] Using the soybean Tianlong No. 1 (National Examination Dou 2008023) sensitive to the method of genetic transformation of soybean cotyledon nodes mediated by Agrobacterium tumefaciens as a control, the method of 1.1-1.4 and step 5 in step 1 of Example 1 was used to determine The overall staining efficiency of explants of 150 soybean varieties in Table 2 and the target cells in cotyledon nodes by the Agrobacterium tumefaciens mediated genetic transformation method of soybean cotyledon nodes co-treated with surfactants and ultrasonic waves of the present application (abbreviated as 2S) dyeing efficiency. Experiments were repeated three times, and 30-35 explants per variety were tested for each replicate.

[0163...

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Abstract

The invention discloses a method of improving the soybean genetic transformation efficiency by virtue of the synergism of a surfactant and ultrasonic waves and an application of the method. The method is an agrobacterium tumefaciens mediated soybean cotyledonary node genetic transformation method and comprises the following steps: infecting a soybean cotyledonary node explant by virtue of a recombinant agrobacterium tumefaciens containing a target DNA (Deoxyribonucleic Acid) to obtain an infected explant, co-culturing the infected explant to obtain the infected explant after co-culture, and completing the transformation, wherein the infected explant obtained by infecting the soybean cotyledonary node explant by virtue of the recombinant agrobacterium tumefaciens containing the target DNA is the infected explant obtained by arranging the soybean cotyledonary node explant into an infection liquid of the recombinant agrobacterium tumefaciens to carry out ultrasonic treatment, the infection liquid contains the surfactant, and scratches are not made at the cotyledonary node position of the soybean cotyledonary node explant. The method can be used for culturing a novel variety of transgenic soybeans as well as excavating and identifying gene functions.

Description

technical field [0001] The invention relates to a method for synergistically improving soybean genetic transformation efficiency with a surfactant and ultrasonic waves and its application. Background technique [0002] Soybean (Glycine max(L.). Merr) originated in China. It is an important oil crop and economic crop, and it is also the main source of edible vegetable oil and plant protein. It occupies an extremely important position in the diet structure of our people. The establishment of a simple and efficient soybean transgenic technology system can make up for the deficiency and defects of the lack of genetic resources in traditional hybrid breeding methods, help to improve soybean pest resistance and stress resistance, and help improve soybean protein and oil quality. Improve the mining of soybean functional genes. However, soybean genetic transformation has always been one of the difficulties in the field of plant genetic engineering, and it is recognized as one of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 邱丽娟郭兵福张丽娟刘章雄郭勇李脉泉常汝镇
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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