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A method for cultivating transgenic plants resistant to soybean mosaic virus

A technology of soybean mosaic virus and transgenic plants, which is applied in the field of plant biology and can solve problems that have not been seen

Inactive Publication Date: 2019-06-04
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing research is mainly through RNAi-mediated SMV-CP or SMV-HC-Pro Gene silencing improves the level of soybean resistance to SMV, but there is no relevant report on the SMV resistance mediated by RNAi silencing of other key genes involved in SMV infection, replication, movement and other processes

Method used

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  • A method for cultivating transgenic plants resistant to soybean mosaic virus
  • A method for cultivating transgenic plants resistant to soybean mosaic virus
  • A method for cultivating transgenic plants resistant to soybean mosaic virus

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 alignment of 26 SMV strains registered in GenBank (http: / / www.ncbi.nlm.nih.gov / genbank / ) revealed that the soybean mosaic virus P3 There is a conserved region in the gene, and a 302bp fragment is finally determined in the conserved region, located in P3 The 99th-400th nucleotides from the 5' end of the gene are 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 KOD FX 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 enzyme recognition sequence is underlined, where "CTCGAG" is a restriction enzyme xho Ⅰ enzyme recognition sequence, "TCTAGA" is a restriction endonuclease Xba Ⅰ enzyme recognition sequence, "GGTACC" is a restriction endonuclease Kpn Ⅰ restriction enzyme recognition sequence, " AAGCTT ...

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Abstract

A method for cultivating transgenic plants resistant to soybean mosaic virus belongs to the field of plant biotechnology. The method provided by the invention comprises the RNA fragment shown in SEQ-1 or the sequence complementary to the fragment. The RNA fragment can interfere with the replication and movement of the soybean mosaic virus, thereby inhibiting the development of symptoms of the soybean mosaic virus. The present invention also provides a method for cultivating transgenic plants resistant to soybean mosaic virus, comprising the following steps: expressing the RNA molecule in target plants to obtain transgenic plants with significantly improved resistance to soybean mosaic virus. The invention has important application value for cultivating broad-spectrum soybean mosaic virus-resistant 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 disease ( soybean mosaic virus , SMV) is a worldwide soybean disease caused by soybean mosaic virus, and it is also one of the most important soybean 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. Taxonomically, SMV belongs to the Potatoviridae ( Potyviridae ) Potatovirus ( Potyvirus ), mainly by infested seeds, aphids and mechanical inoculation. With the global exchange of soybean germplasm resources, the emergence of new virus strains and the change of pathogenicity,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00A01H6/54
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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