Method of improving soybean genetic transformation efficiency by virtue of synergism of surfactant and ultrasonic waves and application of method
A genetic transformation method and surfactant technology are applied in the field of the synergistic improvement of soybean genetic transformation efficiency by surfactants and ultrasonic waves, and can solve the problems of low yield of clump buds, difficulty in accurate and effective wounding of explants, and low transformation efficiency.
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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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