Application of ZmNuC gene and encoded protein thereof in resistance to maize rough dwarf disease

A protein-encoded, transgenic corn technology, applied in the application field of ZmNuC gene and its encoded protein in the resistance to corn rough dwarf disease, can solve the problem that corn germplasm resources cannot meet the needs of corn production, and achieve the effect supported by reliable data

Inactive Publication Date: 2021-09-28
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a corn nucleolin gene ZmNuC and its application in resistance to rough dwarf disease, in order to solve the technical problem that the corn germplasm resources in the prior art cannot meet the current corn production needs

Method used

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  • Application of ZmNuC gene and encoded protein thereof in resistance to maize rough dwarf disease
  • Application of ZmNuC gene and encoded protein thereof in resistance to maize rough dwarf disease
  • Application of ZmNuC gene and encoded protein thereof in resistance to maize rough dwarf disease

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1ZmNuC gene analysis

[0038] In this example, the full-length cDNA sequence of the corn ZmNuC gene was cloned, and the full-length sequence of the gene was obtained by sequencing technology, as shown in SEQ ID NO.1, and based on this analysis, the ORF Finder software was used for comparative analysis. The gene structure of the ZmNuC gene was determined, and it was found through gene structure analysis that the gene contained four introns and five exons ( figure 1 ).

Embodiment 2

[0039] The subcellular localization of embodiment 2ZmNuC gene

[0040] Digest the pAN580 empty vector with BamHI, recover the pAN580 vector fragment by gel, and then perform homologous recombination to connect the target gene ZmNuC (as shown in SEQ ID NO.1) to the vector to obtain pAN580-ZmNuC, transform and pick a single spot , sequencing; by transfecting maize protoplasts to clarify its subcellular location, laser confocal microscopy to observe the fluorescent signal, the green fluorescent signal of the control sample transformed with empty vector existed in the maize protoplast, while the green fluorescent signal of the ZmNuC fusion protein The signal appears in the nucleus region of the protoplast, indicating that the ZmNuC gene is localized on the nucleus ( figure 2 ).

Embodiment 3

[0041] Example 3 ZmNuC Gene Overexpression Vector and Editing Vector Construction and Transformation

[0042] The vector used for overexpression is pCUB-eGFP-3×FLAG, which is used on the basis of pCUB vector (Song Pan, 2015, Maize ZmPOT gene TALEN knockout and overexpression vector construction and genetic transformation, page 18, paragraph 3) The BamHI restriction site is constructed by inserting GFP protein and 3×FLAG tag between the promoter and terminator on the pCUB vector.

[0043]The ZmNuC gene (as shown in SEQ ID NO.1) was inserted between the promoter and the GFP tag protein on the pCUB-eGFP-3×FLAG vector. Carry out transformation, pick a single spot, sequence, and use the heat shock method to transform the overexpression vector plasmid pCUB-ZmNuC-eGFP-3×FLAG into Agrobacterium EHA105 competent cells, pick a single colony of recombinant Agrobacterium, and extract the plasmid DNA for PCR Identification. Escherichia coli positive plasmid was used as positive control. ...

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Abstract

The invention relates to the technical field of plant genetic engineering, and particularly discloses application of a ZmNuC gene and an encoded protein thereof in resistance to the maize rough dwarf disease. The invention finds for the first time that the expression level of the corn nucleolin gene ZmNuC is in negative correlation with the resistance of corns to the rough dwarf disease; a new way for exploring and creating a new material for resisting to the corn rough dwarf disease is provided; a genetic material foundation is laid for subsequent research; and a good information platform is provided for the resource reservation of the corn rough dwarf disease resistant gene. The research on the rough dwarf disease resistance genetic mechanism and the disease-resistant molecular breeding of plants is promoted; and reliable material and data support is provided for the research on the molecular biology mechanism for enhancing the rough dwarf disease resistance of the plants such as corns and the like.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the application of a ZmNuC gene and its encoded protein in resistance to maize rough dwarf disease. Background technique [0002] Rice black-streaked dwarf virus (RBSDV), which causes maize rough dwarf disease, is mainly transmitted by the brown planthopper (Laodelphax striatellus Fallen) in a persistent manner, and cannot be transmitted through soil, seeds, sap friction, and grafting. After the SBPH acquires the poison, it can carry and transmit the virus for life. Adults and nymphs of the poisonous SBPH overwintering in the weeds on the edge of the field will transmit the virus in the next spring, which can cause serious damage to maize rough dwarf disease. The virus overwinters on hosts such as wheat and perennial gramineous weeds. In spring, the first generation of SBPH adults feed on the infected hosts and migrate from wheat to corn, forming a migration pe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H6/46
CPCC07K14/415C12N15/8283C12N15/8218C12N15/8205
Inventor 周羽张嘉月邸宏张林王振华卢晴李益晨曾兴董玲刘显君
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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