Application of solanaceae plant coded PABP protein in prevention and treatment of tobacco mosaic virus

Abstract

The invention relates to the technical field of biotechnology and crop disease prevention and control, in particular to application of PABP protein coded by solanaceae plants to prevention and control of tobacco mosaic virus viruses. The invention finds that the PABP protein (taking NbPABP as an example) encoded by solanaceae plants and the encoding gene thereof play an important role in infection and proliferation of the pepper mild mottle virus PMMoV, are susceptible genes of PMMoV encoded by host plants, can be used for preventing and treating pepper mild mottle virus diseases caused by PMMoV, and can be used for molecular breeding for resisting / resisting PMMoV and the like. Valuable protein is provided for antiviral infection of plants, particularly, the antiviral level of solanaceous plants can be improved, production safety of solanaceous vegetables is guaranteed and promoted, and the protein has wide application prospects in the fields of antiviral breeding of plants and the like.

Description

Technical Field

[0001] The invention relates to the technical field of biotechnology and crop disease prevention and control, and in particular to an application of a PABP protein encoded by a Solanaceae plant in preventing and controlling tobacco mosaic virus. Background Art

[0002] Peppers and tomatoes are important solanaceous vegetable crops. In recent years, the occurrence of pepper mild mottle virus (PMMoV) has been increasing year by year. PMMoV infection causes the leaves of plants to become chlorotic and mottled, and growth to slow down, posing a serious threat to the high and stable yields of peppers and other crops. Since there are no resistant / tolerant varieties and no effective control agents, it is urgent to identify disease-resistant genes and develop resistant materials in order to sustainably and more effectively control PMMoV and its harm, and ensure the production safety of peppers and other solanaceous vegetables.

[0003] As an intracellular obligate parasite, plant viruses must rely on and assist host factors to complete life processes such as virus uncoating, genome replication, protein expression, virus particle assembly, and movement to neighboring cells during infection and proliferation. Therefore, identifying host factors involved in plant virus infection and proliferation can help us gain a deeper understanding of the virus's infection and pathogenicity mechanisms, thereby providing new materials and theoretical basis for the prevention and control of viral diseases. Summary of the invention

[0004] The present invention provides an application of a PABP protein encoded by a Solanaceae plant in preventing and controlling viruses of the genus Tobacco mosaic virus. The present invention finds that the NbPABP protein and its encoding gene play an important role in the infection and proliferation of pepper mild mottle virus PMMoV of the genus Tobacco mosaic virus, are the susceptible genes of PMMoV encoded by the host plant, and can be used to prevent and control pepper mild mottle virus diseases caused by PMMoV.

[0005] In a first aspect, the present invention provides a use of NbPABP protein in any one or more of the following: (1) Application in regulating the expression of tobacco mosaic virus.

[0006] (2) Regulate plant resistance to tobacco mosaic virus diseases.

[0007] (3) Prevent and control tobacco mosaic virus diseases.

[0008] (4) Regulating plant yield.

[0009] (5) Application in plant germplasm improvement.

[0010] (6) Use in the preparation of products for preventing and treating tobacco mosaic virus diseases.

[0011] The amino acid sequence of the NbPABP protein includes any one of the following: (a) As shown in SEQ ID NO.1.

[0012] (b) A protein derived from SEQ ID NO.1 by replacing, deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.1 and retaining the function of the amino acid sequence shown in SEQ ID NO.1.

[0013] (c) an amino acid sequence that has at least 90% homology to the amino acid sequence shown in SEQ ID NO. 1 and has an equivalent functional protein.

[0014] (d) An amino acid sequence obtained by connecting a tag, an enzyme cleavage site and / or a connecting peptide sequence to the N-terminus and / or C-terminus of any one of the amino acid sequences of (a) to (c).

[0015] In a second aspect, the present invention provides a NbPABP The use of genes in any one or more of the following: (1) Application in regulating the expression of tobacco mosaic virus.

[0016] (2) Regulate plant resistance to tobacco mosaic virus diseases.

[0017] (3) Prevent and control tobacco mosaic virus diseases.

[0018] (4) Regulating plant yield.

[0019] (5) Application in plant germplasm improvement.

[0020] (6) Use in the preparation of products for preventing and treating tobacco mosaic virus diseases.

[0021] Said NbPABP The nucleotide sequence of the gene is at least one of the following.

[0022] (a) As shown in SEQ ID NO.2.

[0023] (b) A nucleotide sequence in which one or more nucleotides are substituted, deleted and / or added to the nucleotide sequence shown in SEQ ID NO. 2 and the nucleotide sequence expresses a protein with the same function.

[0024] (c) A nucleotide sequence that hybridizes with the sequence shown in SEQ ID NO. 2 under stringent conditions and expresses a protein with the same function, wherein the stringent conditions are hybridization at 65°C in a 0.1×SSPE solution containing 0.1% SDS or a 0.1×SSC solution containing 0.1% SDS, and the membrane is washed with the solution.

[0025] (d) A nucleotide sequence that has more than 90% homology with the nucleotide sequence shown in SEQ ID NO.2 and expresses a protein with the same function.

[0026] Preferably, the above application regulates the plant's disease resistance to tobacco mosaic virus diseases, or controls crop yield, or prevents tobacco mosaic virus diseases, or improves germplasm resources by regulating the content or activity of NbPABP protein.

[0027] Further preferably, the above application is achieved by regulating the content or activity of NbPABP protein through genetic engineering technology.

[0028] Further preferably, the above application inhibits the replication and proliferation of tobacco mosaic virus by reducing the content or activity of NbPABP protein, so as to improve the plant's disease resistance to tobacco mosaic virus disease, or increase crop yield, or improve germplasm resources.

[0029] Preferably, the above application promotes the replication and proliferation of tobacco mosaic virus by increasing the content or activity of NbPABP protein, so as to construct a disease model of tobacco mosaic virus.

[0030] In a third aspect, the present invention provides the use of an inhibitor for reducing the content or activity of NbPABP protein in plants, including any of the following uses: (1) Improve the resistance of plants to tobacco mosaic virus diseases.

[0031] (2) Increase plant yield.

[0032] (3) Prevent and control tobacco mosaic virus diseases.

[0033] (4) Preparation of pesticides, fertilizers or additives for preventing and controlling tobacco mosaic virus diseases.

[0034] The amino acid sequence of the Solanaceae plant NbPABP protein includes any one of the following: (a) As shown in SEQ ID NO.1.

[0035] (b) A protein derived from SEQ ID NO.1 by replacing, deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.1 and retaining the function of the amino acid sequence shown in SEQ ID NO.1.

[0036] (c) an amino acid sequence that has at least 90% homology to the amino acid sequence shown in SEQ ID NO. 1 and has an equivalent functional protein.

[0037] (d) An amino acid sequence obtained by connecting a tag, an enzyme cleavage site and / or a connecting peptide sequence to the N-terminus and / or C-terminus of any one of the amino acid sequences of (a) to (c).

[0038] Preferably, the inhibitor can inhibit the expression or activity of NbPABP protein by inhibiting its transcription, inhibiting its transcriptional processing, or inhibiting its function after transcriptional processing. The inhibitor can be used as a pesticide, fertilizer or fertilizer additive, and its types include but are not limited to: (1) Interfering RNA such as siRNA or shRNA.

[0039] (2) Small molecule inhibitors.

[0040] (3) Enzyme inhibitors.

[0041] (4) Antibody drugs.

[0042] In the specific implementation process, the content or activity of NbPABP protein can be regulated by regulating cDNA, RNA and other substances. For example, it can be RNA encoded by the nucleotide sequence shown in positions 996-1295 of SEQ ID NO.2 (silent NbPABP Gene).

[0043] In the specific implementation process, the content or activity of NbPABP protein in plant cells is regulated by biotechnology methods including but not limited to using Ti plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation, etc.

[0044] In a fourth aspect, the present invention provides a method for inhibiting the replication and proliferation of tobacco mosaic virus in plants, comprising: reducing the content or activity of NbPABP protein by genetic engineering technology.

[0045] Preferably, the plant or crop described in any of the above uses, products or methods is a plant of the Solanaceae family, preferably a plant of the genus Nicotiana or Solanum, and more preferably tobacco, pepper, tomato, potato, eggplant or wolfberry.

[0046] The tobacco mosaic virus described in the present invention includes tobacco mosaic virus, tomato mosaic virus, tomato brown wrinkled fruit virus, cucumber green mottle mosaic virus, tobacco mild green mosaic virus, pepper mild mottle virus, etc., preferably pepper mild mottle virus.

[0047] The tobacco mosaic virus disease described in the present invention refers to a viral disease caused by tobacco mosaic virus and occurring in plants such as pepper, tomato, and tobacco, and is preferably a viral disease caused by PMMoV infection and occurring in plants such as pepper and tobacco.

[0048] Above NbPABP Genes from Nicotiana benthamiana NbPABPThe gene is highly conserved with PABP in economic crops of Solanaceae such as pepper, so the present invention can also be used in dicotyledonous host plants of Solanaceae such as pepper.

[0049] Compared with the prior art, the present invention has the following beneficial effects: The present invention found that NbPABP protein and its encoding gene play an important role in PMMoV infection and proliferation, and is a PMMoV susceptibility gene encoded by the host plant. NbPABP In the silence experiment, NbPABP When the expression level was downregulated by 60%, the accumulation level of PMMoV genomic RNA was downregulated by 70%, and the accumulation level of PMMoV CP protein was downregulated by 60%, indicating that silencing the expression of N. benthamiana NbPABP gene, which can significantly reduce the levels of PMMoV genomic RNA and CP protein and inhibit PMMoV infection.

[0050] The above findings can be used to prevent and treat pepper mild mottle virus disease caused by PMMoV, and can be used for PMMoV resistance / resistance molecular breeding and other work. The present invention can provide a basis for studying the role of plant proteins in virus infection, antiviral breeding and other fields, improve the level of antiviral research on solanaceous vegetables, and ensure and promote the production safety of solanaceous vegetables. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0052] Figure 1 The silence in Embodiment 2 of the present invention NbPABP The results of the inoculated leaves of N. benthamiana were shown in Figure 1. The picture was taken under UV light 5 days after PMMoV-GFP inoculation. The dots are PMMoV-GFP observed under UV light, and their number can indicate the accumulation of PMMoV-GFP in the leaves. TRV: mCherry The control group means no silence NbPABP Nicotiana benthamiana leaves; TRV: NbPABP The experimental group, indicating silence NbPABP Nicotiana benthamiana leaves.

[0053] Figure 2 The silence in Embodiment 2 of the present invention NbPABPThe fluorescence points of the inoculated leaves of N. benthamiana were counted and analyzed using Graphpad Prism software. t -test for statistical analysis of significant differences, **** indicates p <0.0001.

[0054] Figure 3 The silence in Embodiment 2 of the present invention NbPABP After that, the leaves of Nicotiana benthamiana were inoculated with NbPABP Results of gene expression. RT-qPCR detection NbPABP The relative expression of genes was analyzed using Graphpad Prism software. t -test for statistical analysis of significant differences, ** indicates p <0.01.

[0055] Figure 4 The results of the accumulation of PMMoV genomic RNA in the inoculated leaves of Nicotiana benthamiana after silencing NbPABP in Example 2 of the present invention are shown in Figure 2. The relative expression of PMMoV genomic RNA was detected by RT-qPCR and analyzed by Graphpad Prism software. t -test for statistical analysis of significant differences, * indicates p <0.05.

[0056] Figure 5 The silence in Embodiment 2 of the present invention NbPABP Figure 2. PMMoV CP accumulation in inoculated leaves of N. benthamiana. Western blotting was used to detect PMMoV CP accumulation levels. Actin was used as a protein loading control. Coomassie Brilliant Blue staining (CBB) indicated the total protein content. The size of PMMoV CP was 17 kDa and that of Actin was 43 kDa. Quantitative values ​​were calculated using ImageJ software. DETAILED DESCRIPTION

[0057] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0058] Unless otherwise specified, the examples are based on conventional experimental conditions, such as Sambrook J & Russell DW, Molecular Cloning: a Laboratory Manual, 2001, or the conditions recommended by the manufacturer's instructions.

[0059] The quantitative tests in the following examples were all repeated three times, and the results were averaged.

[0060] The biological materials used in the following examples are as follows: Nicotiana benthamiana: See Goodin MM, Zaitlin D, Naidu RA, Lommel SA. 2008. Nicotiana benthamiana : its history and future as a model for plant-pathogeninteractions. Molecular Plant-Microbe Interactions 21:1015-1026. Vector pJM23, vector pJG1024, vector pLoxP-Cre: see Yin, Y., Hua M., Zhao K., Wan Q., Bu S., Lu Y., Zhang, H., Rao, S., Yan F., Peng J., Chen H., and Chen, J. 2022. Construction of chimeric viruses based on pepper mild mottle virus usinga modified Cre / loxP system. Journal of Integrative Agriculture 21:2456-2463. For vectors pTRV1 and pTRV2, see: Liu, Y., Schiff M and Dinesh-Kumar SP.2002.Virus-induced gene silencing in tomato.The Plant journal 32:777-786. Pepper mild mottle virus (PMMoV) See: Yin, Y., Hua M., Zhao K., Wan Q., Bu S., Lu Y., Zhang, H., Rao, S., Yan F., Peng J., Chen H., and Chen, J. 2022. Construction of chimeric viruses based on pepper mild mottle virus using a modified Cre / loxP system. Journal of Integrative Agriculture 21:2456-2463. The NbPABP protein sequence in the following examples is shown in SEQ ID No. 1, and the gene encoding the NbPABP protein is named NbPABP Gene, the sequence is shown in SEQ ID No.2.

[0061] Example 1 Construction of recombinant vector pTRV2-NbPABP 1. Extraction of Nicotiana benthamiana ( Nicotiana benthamiana ) and reverse transcribed into cDNA.

[0062] 2. Using the cDNA obtained in step 1 as a template, use pTRV2:: NbPABP -F (SEQ ID No. 3) and pTRV2:: NbPABP -R (SEQ ID No. 4) was used for PCR amplification to obtain NbPABP (SEQ ID No. 2) The PCR amplification product of the nucleotide sequence gene fragment shown in positions 996-1295 of the coding region; NbPABP PCR product of the gene fragment shown at positions 996-1295 of the coding region.

[0063] 3. Use restriction endonucleases XbaI and KpnI The vector pTRV2 was digested with enzymes and the vector backbone was recovered.

[0064] 4. Connect the recovered product of step 2 and the vector backbone of step 3 to obtain the recombinant vector pTRV2-NbPABP.

[0065] 6. Based on the sequencing results, the structure of the recombinant vector pTRV2-NbPABP is described as follows: XbaI and KpnIA double-stranded DNA molecule shown in SEQ ID No. 11 was inserted between the restriction sites.

[0066] Example 2 NbPABP Gene silencing experiments 1. Extraction of PMMoV-GFP crude extract (1) Streak the Agrobacterium strains carrying the pJM23, pJG1024 and pLoxP-Cre plasmids stored at -80°C on LB resistance plates (containing 50 μg / ml Kan and 100 μg / ml Rif) and culture at 28°C for about 48 h. Inoculate a single colony on the LB resistance plate into 3-4 ml LB liquid resistance medium (containing 50 μg / ml Kan and 100 μg / ml Rif) and culture at 28°C and 180 rpm for 14-18 h.

[0067] (2) Inoculate 100 μL of bacterial culture into 10 mL of LB liquid resistance culture medium (containing 50 μg / ml Kan and 100 μg / ml Rif) at a ratio of 1:100, and culture at 28°C and 180 rpm for 10-12 h to obtain bacterial cultures of Agrobacterium pJM23, pJG1024, and pLoxP-Cre.

[0068] (3) Collect the bacteria by centrifugation at 4000 rpm for 10 min at room temperature, suspend the precipitate in 3 mL of infiltration buffer (10 mM MES, pH 5.7, 10 mM MgCl2, 200 µM acetosyringone), and measure the concentration of the bacterial suspension using a spectrophotometer.

[0069] (4) Add 3 mL of Agrobacterium pJM23 suspension (OD 600nm concentration of 0.5), 3 mL Agrobacterium pJG1024 bacterial solution (OD 600nm concentration of 0.5) and 3 mL of Agrobacterium pLoxP-Cre bacterial solution (OD 600nm The mixture was placed at room temperature for 2-3 h to obtain a mixed solution. The mixed solution was then injected into the lower epidermis of Nicotiana benthamiana leaves using a syringe without a needle (specification: 1 mL) and cultured at 22-24°C for 6 days.

[0070] (5) Take the Nicotiana benthamiana leaves, add 0.1 M phosphate buffer and grind them. Centrifuge at 4500 g for 5 min, and collect the viral supernatant containing the PMMoV-GFP crude extract.

[0071] 2. Silencing Nicotiana benthamiana using TRV VIGS NbPABP Gene (1) The recombinant plasmid pTRV2-NbPABP obtained in Example 1 was transformed into Agrobacterium GV3101 to obtain recombinant Agrobacterium TRV2-NbPABP, which was then stored in a -80°C refrigerator.

[0072] (2) Streak the Agrobacterium carrying the pTRV1, pTRV2-mCherry and pTRV2-NbPABP plasmids stored at -80°C on LB resistance plates (containing 50 μg / ml Kan and 100 μg / ml Rif) and culture at 28°C for about 48 h. Inoculate a single colony on the LB resistance plate into 3-4 ml LB liquid resistance culture medium (containing 50 μg / ml Kan and 100 μg / ml Rif) and culture at 28°C and 180 rpm for 14-18 h.

[0073] (3) Inoculate 100 μL of bacterial culture into 10 mL of LB liquid resistance culture medium (containing 50 μg / ml Kan and 100 μg / ml Rif) at a ratio of 1:100, and culture at 28°C and 180 rpm for 10-12 h to obtain Agrobacterium culture carrying pTRV1, pTRV2-mCherry, and pTRV2-NbPABP plasmids.

[0074] (4) Collect the bacteria by centrifugation at 4000 rpm for 10 min at room temperature, suspend the precipitate in 3 mL of infiltration buffer (10 mM MES, pH 5.7, 10 mM MgCl2, 200 µM acetosyringone), and measure the concentration of the bacterial suspension using a spectrophotometer.

[0075] (5) Add 3 mL of Agrobacterium culture medium (OD 600nm 1.0) and 3 mL of the bacterial suspension of the recombinant Agrobacterium pTRV2-NbPABP plasmid (OD 600nm concentration of 1.0) and 3 mL of Agrobacterium carrying pTRV2-mCherry (OD 600nm The mixture was then injected into the lower epidermis of the 3rd to 4th true leaves of Nicotiana benthamiana using a syringe without a needle (specification: 1 mL) and cultured at 22-24°C for 6 days.

[0076] 3. PMMoV-GFP challenge inoculation of N. benthamiana plants (1) Complete the silence in step 2 (5) NbPABP Bunsen smoke or silence mCherryThe N. benthamiana plants (as a control) were cultured in alternating light and dark conditions (16 h / 20℃ light, 8 h / 18℃ dark) for 6 days, and a little corundum was sprinkled on the 5th and 6th true leaves of N. benthamiana, and the PMMoV-GFP crude extract obtained in step 1 was rubbed and inoculated (inoculation volume of 50 μL, concentration of 0.3 g / mL). The inoculated N. benthamiana plants were then cultured in alternating light and dark conditions (16 h / 20℃ light, 8 h / 18℃ dark).

[0077] 4. Detection of silence NbPABP PMMoV replication and proliferation levels in N. benthamiana plants Continue to observe the disease of the leaves of Nicotiana benthamiana after PMMoV inoculation. On the 5th day after PMMoV inoculation, take the inoculated leaves and observe the PMMoV-GFP fluorescence points under ultraviolet light. The results are shown in Figure 1 ; Graphpad Prism software was used to analyze the fluorescence images, and Student's t -test was used for statistical analysis of significant differences. The results are shown in Figure 2 Total RNA was extracted and reverse transcribed into cDNA. The cDNA was used as a template and the fluorescent quantitative PCR kit of Beijing Adlai Biotechnology was used. Actin Gene as internal reference gene, RT-qPCR identification NbPABP The relative expression of genes and the content of PMMoV genomic RNA are shown in Figure 3 , Figure 4 ; Total protein was extracted from Nicotiana benthamiana leaves, TMV CP antiserum was used as primary antibody, HRP-labeled goat anti-rabbit antibody was used as secondary antibody, and the expression level of Actin was used as internal reference for western blot analysis. Western blotting was used to detect the accumulation of PMMoV CP protein, and quantitative calculation was performed using ImageJ software. The results are shown in Figure 5 .

[0078] For identification NbPABP The primer pairs for the genes are as follows: NbPABP-qRT-F: 5'-GCCACAGGGTGGTGTGAATA-3' (SEQ ID No. 5) NbPABP-qRT-R: 5'-TCAGCACTTCCAATGCCCTT-3' (SEQ ID No. 6) For identification Actin The primer pairs for the genes are as follows: NbActin-qRT-F: 5'-TGCCATTCTCCGTCTTGACT-3' (SEQ ID No. 7) NbActin-qRT-R: 5'-TGCAGTCTCGAGTTCCTGTT-3' (SEQ ID No. 8) The primer pairs used to identify PMMoV CP are as follows: PMMoV CP-qRT-F: 5'-TGGAAGACTATTCCGACCGC-3' (SEQ ID No. 9) PMMoV CP-qRT-R: 5'-CCGTGCCACGAACTAACTCA-3' (SEQ ID No. 10) The data obtained is 2 -ΔΔCT The analysis steps are as follows: First, the C of the reference gene (ref) was used to identify the experimental group samples (test) and the control group samples (calibrator). T The C value of the target gene (target) T value: ΔC T (test) = C T (target, test) - C T (ref, test) ΔC T (calibrator) = C T (target, calibrator) - C T (ref, calibrator) Secondly, using the △C of the control group sample T The △C of the normalized experimental group samples T value: ΔΔC T = ΔC T (test) - ΔC T (calibrator) Finally, the expression level ratio is calculated: 2 -ΔΔCT = expression ratio.

[0079] Result analysis: Figure 1 , Figure 2 It can be seen that on the 5th day after PMMoV inoculation, compared with the control group (TRV-mCherry), the silenced NbPABP The number of green fluorescent spots on the infected leaves of Nicotiana benthamiana in the experimental group with the gene (TRV-NbPABP) was even smaller.

[0080] Depend on Figure 3 Compared with the control group, the silent NbPABP Genetic Nicotiana benthamiana plants, NbPABP The expression levels were downregulated by about 60%; Figure 4 It can be seen that compared with the control group, the accumulation level of PMMoV genomic RNA was downregulated by about 70%.

[0081] Depend on Figure 5Compared with the control group, the silent NbPABP In N. benthamiana plants with the gene, the accumulation of PMMoV CP protein was downregulated by about 60%.

[0082] The above research results show that by silencing the NbPABP The results showed that NbPABP protein and its encoding gene played an important role in PMMoV replication and proliferation, and were host susceptibility genes that promoted PMMoV infection. They can be used to prevent and control pepper mild mottle virus disease caused by PMMoV, as well as for molecular breeding of PMMoV resistance.

[0083] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of a NbPABP protein in any one or more of the following: (1) Application in regulating the expression of tobacco mosaic virus; (2) Regulating plant resistance to tobacco mosaic virus diseases; (3) Prevent and control tobacco mosaic virus diseases; (4) Regulating plant yield; (5) Application in plant germplasm improvement; (6) Application in the preparation of products for preventing and treating tobacco mosaic virus diseases; It is characterized in that The amino acid sequence of the NbPABP protein includes any one of the following: (a) as shown in SEQ ID NO.1; (b) a protein derived from SEQ ID NO.1 and retaining the function of the amino acid sequence shown in SEQ ID NO.1 by replacing, deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.1; (c) an amino acid sequence having at least 90% homology to the amino acid sequence shown in SEQ ID NO.1 and having an equivalent functional protein; (d) An amino acid sequence obtained by connecting a tag, an enzyme cleavage site and / or a connecting peptide sequence to the N-terminus and / or C-terminus of any one of the amino acid sequences of (a) to (c).

2. A Nbsp The use of genes in any one or more of the following: (1) Application in regulating the expression of tobacco mosaic virus; (2) Regulating plant resistance to tobacco mosaic virus diseases; (3) Prevent and control tobacco mosaic virus diseases; (4) Regulating plant yield; (5) Application in plant germplasm improvement; (6) Application in the preparation of products for preventing and treating tobacco mosaic virus diseases; It is characterized in that Said Nbsp The nucleotide sequence of the gene is at least one of the following: (a) as shown in SEQ ID NO.2; (b) a nucleotide sequence in which one or more nucleotides are substituted, deleted and / or added to the nucleotide sequence shown in SEQ ID NO.2 and the nucleotide sequence expresses a protein with the same function; (c) a nucleotide sequence that hybridizes with the sequence shown in SEQ ID NO. 2 under stringent conditions and expresses a protein with the same function, wherein the stringent conditions are hybridization at 65°C in a 0.1×SSPE solution containing 0.1% SDS or a 0.1×SSC solution containing 0.1% SDS, and the membrane is washed with the solution; (d) A nucleotide sequence that has more than 90% homology with the nucleotide sequence shown in SEQ ID NO.2 and expresses a protein with the same function.

3. The use according to any one of claims 1 or 2, characterized in that: By regulating the content or activity of NbPABP protein, the plant's disease resistance to tobacco mosaic virus diseases, or the crop yield, or tobacco mosaic virus diseases, or germplasm resources can be improved.

4. The use according to any one of claims 1 to 3, characterized in that: The application is achieved by regulating the content or activity of NbPABP protein through genetic engineering technology.

5. The use according to any one of claims 1 to 4, characterized in that: By reducing the content or activity of NbPABP protein, the replication and proliferation of tobacco mosaic virus can be inhibited, so as to improve the disease resistance of plants to tobacco mosaic virus diseases, increase crop yields, or improve germplasm resources.

6. A method for constructing a disease model of a tobacco mosaic virus, characterized in that: By introducing or overexpressing the NbPABP protein, the replication and proliferation of tobacco mosaic virus in plants can be promoted or plants that do not originally contain the protein can be infected with tobacco mosaic virus.

7. Use of an inhibitor for reducing the content or activity of NbPABP protein in plants, characterized in that: Applications including any of the following: (1) Improve the resistance of plants to tobacco mosaic virus diseases; (2) Increase plant yield; (3) Prevent and control tobacco mosaic virus diseases; (4) Preparation of pesticides, fertilizers or additives for the prevention and treatment of tobacco mosaic virus diseases; The amino acid sequence of the Solanaceae plant NbPABP protein includes any one of the following: (a) as shown in SEQ ID NO.1; (b) a protein derived from SEQ ID NO.1 and retaining the function of the amino acid sequence shown in SEQ ID NO.1 by replacing, deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.1; (c) an amino acid sequence having at least 90% homology to the amino acid sequence shown in SEQ ID NO.1 and having an equivalent functional protein; (d) An amino acid sequence obtained by connecting a tag, an enzyme cleavage site and / or a connecting peptide sequence to the N-terminus and / or C-terminus of any one of the amino acid sequences of (a) to (c).

8. The use according to claim 7, characterized in that: The inhibitor can inhibit the expression or activity of NbPABP protein by inhibiting its transcription, inhibiting its transcription processing, and inhibiting its function after transcription processing. The inhibitor can be used as a pesticide, fertilizer or fertilizer additive.

9. A method for inhibiting the replication and proliferation of tobacco mosaic virus in plants, characterized in that: include: The content or activity of NbPABP protein is reduced through genetic engineering technology.

10. The use, product or method according to any one of claims 1 to 9, characterized in that: The plant or crop is a plant of the Solanaceae family, preferably a plant of the genus Nicotiana or Solanum, and more preferably tobacco, pepper, tomato, potato, eggplant or wolfberry.

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