Agrobacterium-rhizogene-mediated stable transformation method of peach root systems and application of method

A technology of Agrobacterium rhizogenes and root system, which is applied in the field of peach biotechnology and molecular breeding, can solve the problem of difficult access to peach transgenic materials, and achieve the effect of short cycle, simple operation and high conversion rate

Active Publication Date: 2019-04-19
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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Problems solved by technology

By summarizing previous studies, it is found that it is extremely difficult to obtain peach transgenic materials by using Agrobacterium tumefaciens to mediate the transformation of peach explants

Method used

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  • Agrobacterium-rhizogene-mediated stable transformation method of peach root systems and application of method
  • Agrobacterium-rhizogene-mediated stable transformation method of peach root systems and application of method
  • Agrobacterium-rhizogene-mediated stable transformation method of peach root systems and application of method

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Embodiment 1

[0048] This method uses MSU440 Agrobacterium rhizogenes carrying the red fluorescent protein gene 35S::DsRED1 expression cassette to mediate the transformation of jujube nectarine to obtain a transgenic root+wild-type bud peach compound line expressing red fluorescent protein; the size of the DsRED1 gene is 443bp, the expressed red fluorescent protein can excite a strong red fluorescent signal under a stereofluorescence microscope. The specific implementation steps of this system are as follows:

[0049] ① Aseptic seedling culture: see figure 1 a, It can be seen that the seedlings were obtained.

[0050] A. Collect 7 mature fruits of jujube nectarine, remove the pulp, soak with 20% commercial bleach for 30 minutes, remove the bleach, air-dry the peach core, and store at 4°C for 2 months;

[0051] B. Take out the peach core at 4°C, remove the endocarp, put the jujube nectarine seeds in a beaker on an ultra-clean bench, sterilize the seeds with 75% alcohol for 30 seconds and mix...

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Abstract

The invention discloses an agrobacterium-rhizogene-mediated stable transformation method of peach root systems and application of the method, and relates to the field of peach biotechnology and molecular breeding. The method comprises the steps of 1, culture of sterile seedlings; 2, preparation of a transformation receptor material explant; 3, preparation of an infecting bacterial liquid; 4, infection; 5, co-culture; 6, transgenic root induction culture; 7, transgenic root detection; 8, acquisition of compound peach plants. Through the application of the method, the transgenic root+wild bud compound peach plants containing important agronomic trait genes are obtained; research on resistance improvement of peach root-knot nematodes is carried out; research on drought and saline-alkali stress resistance of peach roots is carried out. By means of the method, the transgenic root+wild bud compound peach plants can be obtained, and therefore important technical support is provided for the further implementation of research on diseases and pests and resistance breeding of the peach root systems.

Description

technical field [0001] The invention relates to the fields of peach biotechnology and molecular breeding, in particular to a method for stable transformation of peach roots mediated by Agrobacterium rhizogenes and its application; The research on salt-alkali tolerance, heavy metal poisoning resistance, waterlogging resistance, nematode resistance and strengthening peach resistance breeding has laid a foundation. Background technique [0002] Plant transgenic technology is a method of transferring interested exogenous genes into the genome of plants to obtain plants with target traits. It is mainly used in basic research on molecular mechanisms of plant traits, improvement of plant varieties, and cultivation of new varieties. At present, some important crops such as cotton, soybean, rice and apple have been improved through transgenic technology to obtain new transgenic varieties with excellent traits such as high yield, high resistance, multi-resistance and high quality, lay...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/65A01H4/00A01H5/00A01H6/74
CPCA01H4/00C12N15/65C12N15/8205
Inventor 韩月彭徐胜利郑蓓蓓赖恩惠
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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