Efficient genetic transformation method of brassica juncea based on agrobacterium-mediated method

A genetic transformation method and Agrobacterium-mediated technology are applied in the field of high-efficiency genetic transformation of Cardamom mustard based on the Agrobacterium-mediated method, which can solve problems such as insufficient perfection, and achieve the effect of obvious transformation and perfecting of the genetic transformation system.

CN114058638APending Publication Date: 2022-02-18NINGBO ACAD OF AGRI SCI
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Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2022-02-18

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Abstract

The invention discloses an efficient genetic transformation method of brassica juncea based on an agrobacterium-mediated method, the method comprises the following steps of: performing sowing culture, co-culture, pre-culture, differentiation screening and rooting culture on seeds of brassica juncea, and then performing seedling hardening. An acceptor material is brassica juncea, on the basis of previous research, cotyledons of brassica juncea are selected as explants, genetic transformation research is carried out on no-load of a plant genetic transformation vector PBI101 through tissue culture and an agrobacterium-mediated method, buds which are not successfully transferred into genes are whitened and then dead due to resistance screening of kana, survival buds areexplants successfully transferred into genes, the redundant parts of the explants with budded points are cut off, and are transferred into a rooting culture medium for continuous culture so as to screen out the transgenic brassica juncea bud system containing the PBI101 no-load fragment, so that an agrobacterium-mediated brassica juncea genetic transformation system is established.
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Description

technical field

[0001] The invention relates to the technical field of high-efficiency genetic transformation of mustard mustard, in particular to a method for high-efficiency genetic transformation of mustard mustard based on an Agrobacterium-mediated method. Background technique

[0002] Baoxin mustard is heading mustard, also known as cabbage gall and Chaozhou mustard. It belongs to Brassica vegetables of the Brassicaceae family and is a variety of mustard. Mustard mustard has a short growth period, strong growth adaptability, and high yield. Its bulbs and leaves are edible and can be processed into sauerkraut, which has high nutritional value and industrial value. Germplasm resources are the source of scientific and technological innovation in the seed industry, and the innovation and application of breeding technology, especially biotechnology, is an important means for improving the quality and efficiency of variety improvement. At present, biotechnology represented b...

Examples

Embodiment 1

[0038] see Figure 1-Figure 10 , the present invention provides a method for highly efficient genetic transformation of Mustard mustard based on the Agrobacterium-mediated method, comprising: S1. Sowing and culturing: the seeds of Mustard mustard are preferred for use, after aseptic treatment, they are sown on the sowing medium, and cultivated under light ;

[0039] Seeding medium: MS 519 4.43g / L sucrose 20g / L, agar 9.3g / L, and adjust the pH to 5.7-5.9 with 2 mol / L NaOH solution;

[0040] S2. Co-cultivation: select the mustard seedlings, cut off the cotyledons in MS solution as explants, inoculate Agrobacterium into the liquid co-culture medium, shake overnight, resuspend using the liquid seeding medium, and select the cotyledons to inoculate into Cultivate in the dark for 2 days in a co-culture medium covered with sterilized filter paper;

[0041] Co-culture medium: MS0+2mg / L 6-BA+0.2mg / L NAA;

[0042] S3. Pre-cultivation: transfer the cotyledons that have been co-cultured f...