Gene for improving cyst nematode resistance of soybeans and application of gene

A cyst nematode and gene technology, which is applied in the field of genes for improving soybean resistance to cyst nematode disease, can solve the problem that the number of cyst nematodes in the roots of susceptible soybean plants is large, the disease resistance operation is difficult, and the technical cost is high. problems, to achieve the effect of improving breeding efficiency, saving manpower and material resources, and reducing the number of females

Active Publication Date: 2020-07-31
NORTHEAST AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0003] In view of the large number of female cyst nematodes in the roots of susceptible soybean plants, and the difficulty of conventional disease resistance operations and high technical costs, the present invention provides a A gene that improves the resistance of soybean to cyst nematode disease, the nucleotide sequence of the gene is shown in SEQ ID NO: 1

Method used

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  • Gene for improving cyst nematode resistance of soybeans and application of gene
  • Gene for improving cyst nematode resistance of soybeans and application of gene
  • Gene for improving cyst nematode resistance of soybeans and application of gene

Examples

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Effect test

Embodiment 1

[0029] Example 1. Acquisition of genes that increase soybean resistance to cyst nematodes.

[0030] The nucleotide sequence of the gene for improving the resistance of soybean to cyst nematode described in this example is shown in SEQ ID NO:1. Obtained by cloning as follows.

[0031] 1) RNA extraction from the root of Dongnong L-10:

[0032] Extract RNA from the root tissue of Dongnong L-10 grown for 10 days, and extract by Trizol method:

[0033] (1) Add a little liquid nitrogen to the 1.5ml centrifuge tube containing the sample, and quickly and fully grind it to powder;

[0034] (2) Add 1 mL of Trizol extract to the tube, vortex for 30 s, oscillate to mix, and stand at room temperature for 5 min;

[0035] (3) Add 200 μL of chloroform to the tube, vibrate vigorously, vortex for 30 seconds, fully transform, and let stand at room temperature for 5 minutes;

[0036] (4) Centrifuge at 12000rpm for 15min at 4°C;

[0037] (5) Take the supernatant into a new 1.5 centrifuge tube...

Embodiment 2

[0060] Example 2. Application of GmRSCN16-1 in soybean breeding.

[0061] 1. Construct the plant overexpression vector pCSXN1250-GmRSCN16-1.

[0062] (1) Obtaining the full-length sequence of the resistance gene, the resistance gene GmRSCN16-1 was obtained through PCR amplification, product recovery and purification, ligation, transformation, identification and sequencing of positive clones, and the specific method was described in Example 1.

[0063] (2) Extraction of plasmid DNA

[0064] Use the E.Z.N.A. plasmid mini-prep kit to extract the plasmid.

[0065] (3) Linearization of the pCSXN1250 empty plasmid vector and gel recovery of the linearized plasmid.

[0066] The pCSXN1250 empty vector was digested with XcmΙ enzyme, 37°C for 1h. The digested products were subjected to 1% agarose gel electrophoresis, and the linearized plasmid vector DNA was recovered from the gel. This vector is an expression vector for TA cloning. The recombinant plasmid can be obtained by directl...

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Abstract

The invention discloses a gene for improving cyst nematode resistance of soybeans and application of the gene, and belongs to the technical field of genetic breeding. Aiming at the problems that: thenumber of cyst nematode female insects in infected soybean plants is large, conventional disease-resistant operation difficulty is high at present, and the technical cost is high, the invention provides the gene for improving cyst nematode resistance of soybeans. The nucleotide sequence of the gene is as shown in SEQ ID NO: 1. An Agrobacterium rhizogenes K599 mediated genetic transformation methodis adopted, pCSXN1250-GmRSCN16-1 recombinant plasmid is transferred into an infected Dongnong 50 plant, rooting is induced, physiological races of soybean cyst nematode No.1, No.3 and No.4 are inoculated in a mixed manner, and the invention proves that the GmRSCN16-1 gene has obvious resistance to the soybean cyst nematode. The gene can be used for breeding soybean varieties with the cyst nematode resistance.

Description

technical field [0001] The invention belongs to the technical field of genetic breeding, in particular to a gene for improving the resistance of soybean to cyst nematode disease and its application. Background technique [0002] The phenomenon of continuous soybean cropping exists widely in the world, and the number of cysts in soil and roots will increase with the increase of planting years in soybean continuous cropping areas. Although the control effect of applying pesticides in the soil to treat seeds is obvious, the amount of pesticides used is very large. Long-term use will easily cause nematodes to develop pesticide resistance and cause great environmental pollution. Regarding the control of this disease, the most effective way is to plant and breed soybean varieties that are not sensitive to it. When breeding disease-resistant varieties, conventional breeding methods are time-consuming, about 7 to 10 years, and require a large area of ​​breeding fields, consuming a ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/54
CPCC07K14/415C12N15/8285Y02A40/146
Inventor 韩英鹏赵雪王伟朱治佳吴德鹏李文滨
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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