CN-121991966-A - Virus replication essential protein gene NbelF G and application thereof in inhibiting plant virus infection

Abstract

The virus replication essential protein gene NbelF G and the application thereof in inhibiting plant virus infection belong to the technical fields of plant molecular biology and antivirus. The nucleic acid sequence of the gene is shown as SEQ ID NO. 3, and the encoded amino acid sequence is shown as SEQ ID NO. 4. According to the invention, through the virus-induced gene silencing technology, the endogenous NbelF G gene of tobacco is specifically silenced, so that infection and accumulation of tomato spotted wilt virus and rice stripe virus in Nicotiana benthamiana can be obviously inhibited, and the symptoms of disease are reduced and the expression level of viral proteins is reduced. This finding suggests that NbelF G is a key host factor upon which viral replication depends, and that its use as a silencing target can be effective in conferring antiviral ability to plants. The invention provides important gene resources and theoretical basis for creating new antiviral plant materials and developing broad-spectrum virus prevention and control strategies of targeted host factors.

Inventors

• LI JIA
• ZHUO CEN
• ZHAO YAQIAN
• YU QIUYUE
• CAO YANG
• ZHU MIN
• FENG MINGFENG
• TAO XIAORONG

Assignees

• 南京农业大学

Dates

Publication Date

20260508

Application Date

20251218

Claims (10)

1. 1. Viral replication essential protein gene NbelF G, characterized in that silencing the expression of this gene enhances the resistance of plants to tomato spotted wilt virus and/or rice stripe virus, the nucleotide sequence of said gene being shown in SEQ ID NO. 3.
2. 2. The gene of claim 1, wherein the encoded protein has the amino acid sequence shown in SEQ ID NO. 4.
3. 3. Use of NbelF G gene according to claim 1 or 2 in the cultivation of an antiviral plant.
4. 4. Use according to claim 3, characterized in that the virus is tomato spotted wilt virus and/or rice stripe virus.
5. 5. A method for improving the virus resistance of tobacco to plants is characterized by inhibiting the expression of NbelF G gene in tobacco plants by using a virus-induced gene silencing technology, wherein the nucleotide sequence of the NbelF G gene is shown as SEQ ID NO. 3.
6. 6. The method of claim 5, wherein the vector used in the gene silencing technique is a tobacco embrittlement virus vector.
7. 7. The method of claim 6, comprising introducing a tobacco brittle virus recombinant vector comprising a NbelF G gene portion sequence into a tobacco plant by agrobacterium infiltration.
8. 8. The method of any one of claims 5-7, wherein the plant virus is tomato spotted wilt virus and/or rice stripe virus.
9. 9. A recombinant tobacco brittle virus vector for silencing tobacco NbelF G gene, characterized by comprising a fragment of the nucleotide sequence shown in SEQ ID No. 3.
10. 10. An agrobacterium strain comprising the recombinant vector of claim 9.

Description

Virus replication essential protein gene NbelF G and application thereof in inhibiting plant virus infection Technical Field The invention belongs to the technical field of plant molecular biology and antivirus, and particularly relates to a virus replication essential protein gene NbelF G and application thereof in inhibiting plant virus infection. Background Tomato spotted wilt virus (tomato spotted wilt virus, TSWV) is an important member of the genus Orthotospovirus of the genus positive tomato spotted wilt, mainly by means of thrips persistent transmission and sap friction infection, can infect more than 360 plants including Solanaceae, fabaceae, asteraceae and Cucurbitaceae Cucurbitaceae, and seriously endangers crops such as tomatoes, tobacco, peppers, potatoes, peanuts, lettuce and impatiens. TSWV ranks 2 nd among the ten major plant viruses in the world, being a most potentially threatening virus recognized by major tobacco producing countries as being compromised in many tobacco producing countries to varying degrees. TSWV was first reported on Sichuan sun-cured tobacco in 1992 in China, and after that, the hazard range of TSWV is expanded year by year, and the TSWV occurs on cured tobacco planted in regions such as Yunnan, guizhou, heilongjiang, shandong and the like. Rice stripe virus (RICE STRIPE virus, RSV) is the causative agent of rice stripe disease and is a member of the genus Leptovirus (Tenuiviruses). The shape of the virus particles is often a filament, and irregular shapes such as particles and blocks are also known. Laodelphax striatellus is a transmission medium of RSV, and the Laodelphax striatellus transmits viruses through eggs in a circulating propagation manner. Causing serious loss to crop production. When rice stripe viruses infect rice, the leaf and leaf sheath of the rice are mainly damaged, and obvious chlorosis or mottle symptoms can be displayed on the rice leaf. Once a plant virus disease is exploded, effective restriction control is often difficult, so that the method has become a key and difficult problem for research on the plant disease and virology at present. Starting from the occurrence characteristics of plant virus diseases, the detoxified seeds and seedlings, medium insect control, weak virus inoculation technology, screening and popularization of disease-resistant varieties and the like are all the main technical fields of the current control of plant virus diseases. In the agricultural production link, the plant virus diseases are effectively prevented and treated by adopting a method of comprehensive treatment mainly including prevention. Disclosure of Invention Aiming at the technical problems that the existing plant virus control technology depends on means such as detoxification seedling, medium insect control or traditional disease-resistant variety breeding, the methods often have the problems of limited control range, easy influence of environmental factors or easy generation of adaptability variation of viruses, and the like, especially for viruses with serious harm and wide host range such as Tomato Spotted Wilt Virus (TSWV) and Rice Stripe Virus (RSV), high-efficiency and broad-spectrum control targets based on virus-host interaction key links are still lacking at present. The invention provides a virus replication essential protein gene NbelF G and application thereof in inhibiting plant virus infection. According to the technical scheme, the gene NbelF G of the virus replication essential protein is silenced, the expression of the gene can enhance the resistance of plants to tomato spotted wilt virus and/or rice stripe virus, and the nucleotide sequence of the gene is shown as SEQ ID NO. 3. The amino acid sequence of the protein coded by the gene is shown as SEQ ID NO. 4. The use of NbelF G gene in cultivating antiviral tobacco plants. The virus is tomato spotted wilt virus and/or rice stripe virus. A method for improving the resistance of tobacco to plant virus includes such steps as suppressing the expression of NbelF G gene in tobacco plant by virus-induced gene silencing technique, and the nucleotide sequence of NbelF G gene is shown in SEQ ID NO. 3. The vector used in the gene silencing technology is tobacco brittle fracture virus vector. The method comprises the step of introducing a tobacco brittle fracture virus recombinant vector containing NbelF G gene partial sequence into a tobacco plant by an agrobacterium infiltration method. The plant virus is tomato spotted wilt virus and/or rice stripe virus. A recombinant tobacco brittle virus vector for silencing tobacco NbelF G gene, comprising a fragment of the nucleotide sequence shown in SEQ ID No. 3. An agrobacterium strain containing the recombinant vector. First, the control of Tomato Spotted Wilt Virus (TSWV) and Rice Stripe Virus (RSV) in the prior art has focused on external isolation or vector insect control, and the present invention has discovered and confirmed that tobacco endogeno