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A gene gun-mediated genetic transformation method of flax

A genetic transformation method and gene gun technology, applied in the field of plant molecular breeding, can solve problems such as affecting the hit rate of the gene gun

Active Publication Date: 2022-07-29
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The gene gun method is to use compressed gas as the driving force to hit the gold powder with DNA attached to the cells to complete the gene transfer operation. After the gene gun is fired, a large airflow will be generated. The material is light, and the airflow generated easily blows away the receptors, which affects the hit rate of the gene gun

Method used

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  • A gene gun-mediated genetic transformation method of flax
  • A gene gun-mediated genetic transformation method of flax
  • A gene gun-mediated genetic transformation method of flax

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1. Construction of flax male sterility vector

[0062] Taking the flax male sterility gene MS2-F sequence as the target, two different fragments 172 and 944 of the gene were selected, and two double-stranded RNA expression cassette vectors pHANNIBAL-172 and pHANNIBAL-944 were constructed. The vector pART27, as well as the cloned Arabidopsis tapetum-specific A9 promoter and herbicide resistance gene EPSPS, constructed two herbicide resistance genes and flax male sterility gene-linked flax male sterility expression vector.

Embodiment 2

[0063] Embodiment 2. Hypocotyl genetic transformation method of Linxuan No. 3

[0064] 1. The preparation methods of MS mass, MS trace, MS organic, inositol and iron salt involved in the present embodiment are as follows:

[0065] MS bulk is MS bulk basic salt, MS bulk basic salt is composed of solvent water and solute, said solute is potassium nitrate, ammonium nitrate, magnesium sulfate heptahydrate, potassium dihydrogen phosphate and calcium chloride dihydrate or anhydrous calcium chloride , the addition amount is shown in Table 1:

[0066] Table 1

[0067] Element Use concentration (g / L) Potassium nitrate KNO 3

19 Ammonium nitrate NH 4 NO 3

16.5 Magnesium Sulfate Heptahydrate MgSO 4 ·7H 2 O

3.7 Potassium dihydrogen phosphate KH 2 PO 4

1.7 Calcium chloride dihydrate or calcium chloride anhydrous CaCI 2 ·2H 2 O or CaCI 2

4.4 or 3.32

[0068] MS trace is MS trace base salt, and MS trace base salt i...

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Abstract

The invention discloses a method for genetic transformation of flax mediated by gene gun. ; Inoculate the young embryos after the shot into a callus induction medium for cultivation to obtain embryogenic callus; inoculate the embryogenic callus into a selection medium for cultivation; inoculate the resistant callus The seedling is inoculated into a regeneration medium for cultivation to obtain a seedling; the seedling is cultivated in a rooting medium to obtain a transgenic plant of flax rotation selection No. 3. The invention develops a regeneration system suitable for the tissue culture of flax round selection No. 3 through the optimized combination of macroelements, trace elements, organic matter and hormones, and realizes the stable transformation of the variety by means of gene gun mediation.

Description

technical field [0001] The present invention relates to the technical field of plant molecular breeding. Specifically, it is a gene gun-mediated genetic transformation method of flax. Background technique [0002] Flax (Linum usitatissimum L.) is generally divided into oil flax and fiber flax, mainly distributed in North China and Northwest China. Flax has the characteristics of drought resistance and barren resistance, adapts to the harsh ecological environment, has become an irreplaceable drought-resistant crop in dryland agriculture, and plays an important role in the adjustment of agricultural structure. Flax is an important raw material for the textile and oil industries. Flax is not only a very good edible health oil, but also can be used in chemical paint, high-end car spray paint and other industries. Flax gum extracted from flax seeds has very important application value in food, medicine and daily chemical industries. The oil content of flaxseed is about 40%, p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/89A01H5/00A01H6/58
CPCC12N15/895C12N15/8289C12N15/8274
Inventor 伊六喜史树德张晓明刘坤斯钦巴特尔赵小庆包文泉侯建华王树彦李志伟苏少锋
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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