Method for culturing anti-soybean-mosaic-virus transgenic plants

A soybean mosaic virus and transgenic plant technology, applied in the field of plant biology, can solve the problems of interfering or affecting the pathogenicity of SMV, affecting the host range of SMV, etc.

Inactive Publication Date: 2016-05-11
JILIN ACAD OF AGRI SCI
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Problems solved by technology

Therefore, inhibiting the transcription and translation of the SMV-P3 gene can interf

Method used

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  • Method for culturing anti-soybean-mosaic-virus transgenic plants
  • Method for culturing anti-soybean-mosaic-virus transgenic plants
  • Method for culturing anti-soybean-mosaic-virus transgenic plants

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Example 1. Discovery of Fragments with the Function of Inhibiting Soybean Mosaic Virus

[0031] Sequence comparison of 26 SMV strains registered in GenBank (http: / / www.ncbi.nlm.nih.gov / genbank / ) revealed that there is a conserved region in the P3 gene of soybean mosaic virus. A 302bp fragment was finally determined, located at the 99th-400th nucleotides from the 5' end of the P3 gene, as shown in SEQ-2. The RNA encoded by this fragment is expected to inhibit soybean mosaic virus. The RNA sequence transcribed from the SEQ-2 fragment is the single-stranded RNA shown in SEQ-1.

Embodiment 2

[0032] Example 2. Acquisition and identification of transgenic plants resistant to soybean mosaic virus

[0033] 1. Construction of RNAi recombinant plasmid

[0034] (1) Synthesize the double-stranded DNA molecule shown in SEQ-2 of the sequence listing.

[0035] (2) Using the double-stranded DNA molecule synthesized in step 1 as a template, using the primer pair composed of SMV-P3-F and SMV-P3-R, perform PCR amplification with KODFX high-fidelity enzyme (TOYOBO, Japan),

[0036] Obtain the PCR amplification product.

[0037] SMV-P3-F: 5'-CCG CTCGAGTCTAGA TCTCTTGATGGGCTTGGTTTC-3'

[0038] SMV-P3-R: 5'-GG GGTACCAAGCTT GGTTGTTGGATGCTTTTCTTTC-3'

[0039] In SMV-P3-F and SMV-P3-R, the restriction endonuclease recognition sequence is underlined, where "CTCGAG" is the restriction endonuclease XhoI restriction endonuclease recognition sequence, "TCTAGA" is the restriction endonuclease XbaI restriction endonuclease recognition sequence Recognition sequence, "GGTACC" is the reco...

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Abstract

The invention relates to a method for culturing anti-soybean-mosaic-virus transgenic plants, and belongs to the technical field of plant biology. According to the method provided by the invention, a sequence of an RNA (ribonucleic acid) fragment shown by SEQ-1 or the sequence complementary with the segment is used. The RNA fragment can be used for interfering the copying and the movement of the soybean mosaic virus so as to inhibit the development of the soybean mosaic virus symptom. The method for culturing the anti-soybean-mosaic-virus transgenic plants provided by the invention comprises the following steps of expressing the RNA molecules in the target plant; obtaining the transgenic plants with the obviously improved resistance on the soybean mosaic virus. The method has an important application value on the culturing of wide-spectrum anti-soybean-mosaic-virus plants.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a method for cultivating soybean mosaic virus-resistant transgenic plants. Background technique [0002] Soybean mosaic virus (SMV) is a worldwide soybean disease caused by soybean mosaic virus, and it is also one of the most important diseases in major soybean producing areas in my country. SMV can generally cause an average 10-30% reduction in soybean production, and even cause large-scale extinction in severe years and regions. SMV also often causes mottled grains, seriously affecting the appearance quality and commodity value of soybean grains. In taxonomy, SMV belongs to the family Potyviridae and belongs to the genus Potyvirus. It is mainly transmitted by infected seeds, aphids and mechanical inoculation. With the global exchange of soybean germplasm resources, the emergence of new virus strains and the change of pathogenicity, the situation of my country's ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00
CPCC12N15/1131C12N15/8283C12N2310/14
Inventor 杨向东郭东全董英山李启云牛陆张伟杨静杜茜邢国杰李海云钱雪燕姚瑶
Owner JILIN ACAD OF AGRI SCI
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