Method for inoculating maize yellow mosaic virus based on aphid mediator and application

Abstract

The invention belongs to the technical field of biology, and relates to a method for virus inoculation of maize yellow mosaic virus based on aphid mediators and application of the method, the optimum virus inoculation system of MaYMV is determined by performing MaYMV virus inoculation on maize seedlings through aphids, and research finds that when five maize aphids are inoculated for 5 days in the maize VE or V1 period, the virus inoculation efficiency of the MaYMV on maize is the highest; through virus inoculation, it is clear that after MaYMV infection, the corn can have dwarf and small ear symptoms; high-susceptibility varieties zhu3151 and B73 are used for carrying out MaYMV virus inoculation, red leaf symptoms appear after infection, transcriptome analysis is carried out, gene expression differences of different samples or under different conditions are compared, the change rule of gene expression is disclosed, and deep understanding of expression modes and regulatory mechanisms of genes under different conditions is facilitated; the research on the pathogenic mechanism has important significance on disease prevention and control, and development of a prevention and control method and cultivation of resistant varieties are facilitated.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology and relates to a method for inoculating corn yellow mosaic virus. Background Art

[0002] corn( Zeamays L.) is my country's largest crop, with an annual total output accounting for 23% of the world's total. my country is the world's major corn producer and consumer, and corn has been my country's largest food crop variety for many years. In addition to its edible value, corn is also an important raw material for excellent feed and light industry. Pests and diseases are the main disasters affecting corn production, among which corn virus disease is one of the main diseases threatening corn production safety. Under natural conditions, there are many viruses that can infect corn and cause yield reduction. Maize yellow mosaic virus (MaYMV), as a new type of baculovirus, is widely distributed in Asia, America, Africa and other places. It usually co-infects with other RNA viruses and can be detected in cereal crops such as corn and weeds. Infected corn plants usually show symptoms such as leaf yellowing, mosaic, and dwarfing. In severe cases, it can lead to plant death, affecting corn yield and quality, and seriously endangering corn agricultural production safety.

[0003] MaYMV is mainly transmitted by aphids in a non-persistent manner. Aphids transmit the virus mainly through their piercing-sucking mouthparts when sucking the sap of diseased plants, and spread the virus when sucking the sap of other healthy plants. When aphids move on plant leaves, stems and other parts, their mouthparts will pierce plant tissues, thereby contacting the virus in the plant body. When aphids move from one diseased plant to another healthy plant, the virus will spread. The transmission of viruses by aphids has a significant impact on agricultural production. On the one hand, viral infection can cause plant growth and development to be hindered, yield to decrease, and quality to deteriorate; on the other hand, viral infection can also increase the incidence of diseases and insect pests in crops, further exacerbating agricultural production losses. At present, research on maize yellow mosaic virus is mostly focused on detection methods, epidemic law surveys, genetic diversity, and disease prediction and early warning.

[0004] Patent CN201910302612.9 discloses an RT-LAMP detection kit for maize yellow mosaic virus, which can detect maize yellow mosaic virus simply and quickly, overcoming the limitation of virus detection that is highly dependent on instruments. The kit can be used for maize yellow mosaic virus detection in corn and sorghum. The method can monitor whether the corn sample is infected with maize yellow mosaic virus simply and quickly from corn. Ma Chao et al. investigated and counted the occurrence of maize yellow mosaic virus in Anhui Province, and conducted genetic diversity analysis, laying a certain foundation for monitoring and preventing MaYMV virus in corn and other crops in Anhui Province (Ma Chao, Gao Le, Han Kelei, et al. Occurrence and genetic diversity analysis of maize yellow mosaic virus in Anhui Province [J / OL]. Acta Phytopathologica Sinica, 1-11 [2025-01-16].). Xie Lina et al. identified the occurrence and distribution of MaYMV in wheat and corn in different regions of Henan Province, and found that the evolution of the virus may be related to the geographical distribution, which provides a scientific basis for the monitoring, early warning and prevention and control of the virus disease (Xie Lina, Zhang Yuyang, Yu Lianwei, et al. Distribution of Maize Yellow Mosaic Virus in Wheat and Corn in Henan Province and Its CP Sequence Analysis [J]. Acta Phytopathologica Sinica, 2024, 54(02): 476-480.).

[0005] There is currently no effective method for inoculating maize yellow mosaic virus. The transmission characteristics of aphids to maize yellow mosaic virus are not clear. In addition, there are few studies on the molecular mechanism of maize yellow mosaic virus. The study of the pathogenic mechanism is of great significance for disease prevention and control, and is helpful for the development of new prevention and control methods and the cultivation of resistant varieties. Summary of the invention

[0006] In order to solve the above problems, the present invention proposes a method and application of corn yellow mosaic virus infection based on aphid vectors.

[0007] The technical solution of the present invention is achieved in this way: On the one hand, the present application provides a method based on the infection of aphids with MaYMV, the steps of which are: feeding aphids with MaYMV on corn plants carrying MaYMV, and placing the fed aphids on corn seedlings for infection.

[0008] Preferably, the aphid is corn aphid or gramineous aphid.

[0009] Preferably, the above-mentioned feeding time is 3-5 days.

[0010] Preferably, the number of aphids used for the above-mentioned inoculation is 1-7, and the inoculation time is 1-5 days.

[0011] Preferably, the corn seedlings are in the VE stage, V1 stage or V2 stage.

[0012] In a second aspect, the application of the drug-infection system constructed by the above method is selected from any one or more of the following: (1) To study the effects of MaYMV infection on the metabolic pathways of maize plants; (2) Study the effects of MaYMV infection on the cell structure of corn plants; (3) Study the effect of MaYMV infection on corn yield; (4) Study the response of corn plants to MaYMV infection.

[0013] Preferably, the above metabolic pathways include photosynthesis, sugar metabolism, energy metabolism and supply, substance synthesis and transport, and cell function maintenance.

[0014] Preferably, the above-mentioned cell structure includes reconstruction and modification of cell wall, cell growth and division, cell stress and apoptosis, gravity sensing and cell orientation.

[0015] Preferably, the above-mentioned effects on corn yield include stunting and growth retardation of corn plants, short ears and reduced grain number.

[0016] Preferably, the corn plants respond to MaYMV infection by enhancing pathogen defense responses and anti-oxidative stress capabilities.

[0017] The present invention has the following beneficial effects: 1. The present application determined the most suitable infection system of MaYMV by using aphids to inoculate corn seedlings with MaYMV. The study found that the efficiency of corn aphids in transmitting MaYMV on corn was 39.59% on average, which was higher than 35.66% of wheat aphids; the infection efficiency of MaYMV in corn VE or V1 stage was 56.16% and 56.14% on average, respectively, which was significantly higher than 50.86% in V2 stage; the efficiency of infection when 5 aphids were used was 47.75% on average, which was significantly higher than 43.22% when 1 aphid was used and 43.61% when 7 aphids were used; 5 days after the infection of corn with MaYMV, the infection rate was 90%-100% for 7-15 days of infection, which was higher than 30%-80% after 1 day of infection and 60%-100% after 3 days of infection; therefore, the most suitable infection system is that 5 corn aphids are infected for 5 days in corn VE or V1 stage. Using the inoculation system established indoors, corn was inoculated with MaYMV outdoors and in potted conditions, which confirmed that corn would show symptoms of dwarfing and developmental retardation after MaYMV infection. The corn ears showed obvious short ears and reduced number of grains.

[0018] 2. This application used highly susceptible varieties zhu3151 and B73 to inoculate with MaYMV, and determined that after MaYMV infection, highly susceptible varieties zhu3151 and B73 would show red leaf symptoms. Transcriptome analysis was performed on the two groups of varieties, and the differences in gene expression under different samples or different conditions were compared to reveal the changing patterns of gene expression. It was found that after MaYMV infection, the metabolic pathways of corn plants were affected, including the regulation of sugar metabolism, energy metabolism and supply, material synthesis and transport, photosynthesis, and maintenance of cell functions; after MaYMV infection, the cell structure of corn plants was affected, including the reconstruction and modification of cell walls, cell growth and division, cell stress and apoptosis, gravity sensing, and cell orientation; corn plants responded to MaYMV infection by enhancing pathogen defense responses. The study of pathogenic mechanisms is of great significance for disease prevention and control, and is helpful for the development of prevention and control methods and the cultivation of resistant varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only 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.

[0020] Figure 1 The MaYMV infection system was established; A is a comparison of the virus transmission abilities of corn aphids and wheat aphids, B is a comparison of the infection efficiencies in corn VE, V1, and V2 stages, C is a comparison of the infection efficiencies using 1, 5, and 7 aphids, and D is a comparison of the efficiency of corn infection after 1, 3, and 5 days.

[0021] Figure 2 The effects of MaYMV infection on corn yield; A and B are comparisons of plant heights of outdoor corn pots after MaYMV infection when they grew to the trumpet stage, C is polyacrylamide gel electrophoresis detection of indirect poisoning in the field, D and E are comparisons of plant heights of corn after MaYMV infection in the field when they grew to maturity, and F and G are comparisons of corn ear lengths.

[0022] Figure 3 Transcriptome sequencing analysis of highly susceptible corn varieties after MaYMV infection; A and B are the disease symptoms of zhu3151 and B73 after MaYMV infection, C is the PCA analysis of transcriptome data, D is the statistical table of transcription sequencing data, and E is the statistics of differentially expressed genes between zhu3151 and B73.

[0023] Figure 4Figure 3 KEGG analysis and GO annotation of differential expression of zhu3151 and B73 under different treatments; A is the Venn diagram analysis of BCS / BMS and ZCS / ZMS, B is the KEGG analysis of co-regulated differential genes between BCS / BMS and ZCS / ZMS, C is the GO annotation of co-regulated genes between BCS / BMS and ZCS / ZMS, D is the Venn diagram analysis of BCR / BMR and ZCR / ZMR, E is the KEGG analysis of co-regulated differential genes between BCR / BMR and ZCR / ZMR, F is the GO annotation of co-regulated genes between BCR / BMR and ZCR / ZMR, G is the Venn diagram analysis of BMS / BMR and ZMS / ZMR, H is the KEGG analysis of co-regulated differential genes between BMS / BMR and ZMS / ZMR, and I is the GO annotation of co-regulated genes between BMS / BMR and ZMS / ZMR. DETAILED DESCRIPTION

[0024] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only 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.

[0025] Unless otherwise specified, the experimental methods used in the following experimental examples are all conventional methods; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0026] Example 1: Optimal system for inoculation of maize yellow mosaic virus 1. Comparison of the virus transmission ability of corn aphid and graminearum Non-toxic corn aphids and graminearum aphids were fed on corn plants infected with MaYMV. Aphid RNA was extracted 3 days after feeding, and then reverse transcribed and detected by polyacrylamide gel electrophoresis. The gray value of the electrophoresis bands was analyzed. Figure 1 A. The average virus transmission efficiency of corn aphids is 39.59%, and the average virus transmission efficiency of wheat aphids is 35.66%. The virus transmission ability of corn aphids is significantly higher than that of wheat aphids.

[0027] 2. Comparison of virus inoculation efficiency in corn VE, V1 and V2 stages Through the above experiments, it has been confirmed that corn aphids have a better ability to transmit the virus. In order to explore the effect of aphids inoculated with the virus at different growth stages of corn, the corn was inoculated with the virus at VE, V1, and V2 stages. Figure 1B. The infection efficiency of corn at VE or V1 stage was 56.16% and 56.14% on average, respectively, which was significantly higher than the infection efficiency of 50.86% at V2 stage.

[0028] 3. Comparison of the efficiency of using 1, 5, and 7 aphids to inoculate Through the above experiments, it has been confirmed that the best effect is to select corn aphids for each plant to be infected at the VE or V1 stage of corn. In order to explore the efficiency of different numbers of aphids infected with MaYMV on corn infection, 1, 5, and 7 corn aphids were set up for MaYMV infection. Figure 1 C. When five aphids were used for inoculation, the average inoculation efficiency was 47.75%, which was significantly higher than the efficiency of 43.22% when one aphid was used for inoculation and the efficiency of 43.61% when seven aphids were used for inoculation.

[0029] 4. Comparison of the efficiency of corn infection for 1 day, 3 days and 5 days Through the above experiments, it has been confirmed that selecting 5 corn aphids per plant to inoculate at the VE or V1 stage of corn has the best effect. In order to explore the effect of different days of aphid inoculation on the infection efficiency of MaYMV on corn, aphids were inoculated at the V1 stage of corn for 1, 3, and 5 days, and the test was carried out on the 7th, 9th, 11th, 13th, and 15th days. The results are as follows Figure 1 D. As the time of inoculation increases, the incidence of maize increases. Five days after aphid inoculation, the infection rate of MaYMV on maize within 7-15 days is 90%-100%, which is higher than the MaYMV infection rate of 30%-80% after aphid inoculation for 7-15 days and the MaYMV infection rate of 60%-100% after aphid inoculation for 7-15 days.

[0030] Example 2: Effect of MaYMV infection on corn yield 1. Outdoor potted corn after MaYM infection After the indoor inoculation experiment, the MaYMV inoculation system has been preliminarily established, that is, 5 corn aphids were selected to be inoculated for 5 days in the VE or V1 stage of corn. In order to clarify the outdoor inoculation situation of this system, we first chose to conduct a potted experiment outdoors. Since the inoculation was carried out outdoors, the corn leaves in the V1 stage had already unfolded compared to the VE stage, which was convenient for observation and positioning of the inoculation position, so we chose to inoculate in the V1 stage. First, Zhengdan 958, which is widely used in production, was selected for inoculation, and two treatment groups were set up: Mock group and MaYMV group. The results are as follows: Figure 2 As shown in A and B, after infection with MaYMV, corn grown to the trumpet stage showed obvious dwarfing symptoms.

[0031] 2. Field corn after MaYM infection In order to clarify the infection status of the system in the field and the impact of MaYMV infection on the late growth of corn and scion, a Mock group and a MaYMV group were set up in the field. After MaYMV infection, gel electrophoresis was performed on the field corn. The results are as follows: Figure 2 As shown in C, the MaYMV infection rate in the field was 70%. The plant height was measured when the corn grew to maturity. Figure 2 As shown in D and E, the corn infected with MaYMV showed obvious dwarfing and stunted growth. Figure 2 As shown in F and G, the corn ears infected with MaYMV showed obvious shorter ears and fewer grains than those not infected.

[0032] Example 3: Transcriptome sequencing analysis of highly susceptible maize varieties after MaYMV infection 1. MaYMV infects highly susceptible corn varieties In order to clarify the infection status of this system on highly susceptible corn varieties, the corn variety B73, which is widely used in the experiment, and the MaYMV highly susceptible variety zhu3151 were selected for inoculation. Figure 3 As shown in A and B, after being infected by MaYMV, zhu3151 and B73 showed red leaf symptoms when they grew to the trumpet stage.

[0033] 2. Transcriptome measurement data statistics and quality assessment High-throughput sequencing was used to sequence the two treatments of zhu3151 and B73, namely healthy and infected, and the two treatments of systemic leaves and red leaves. Among them, BCR, BCS, BMR, BMS, ZCR, ZCS, ZMR, and ZMS are B73-non-toxic-red leaves (the position corresponding to the infected red leaves), B73-non-toxic-systemic leaves, B73-virus-red leaves, B73-virus-systemic leaves, zhu3151-non-toxic-red leaves (the position corresponding to the infected red leaves), zhu3151-non-toxic-red systemic leaves, zhu3151-virus-red leaves, and zhu3151-virus-systemic leaves. Figure 3 C shows that principal component 1 (PC1) and principal component 2 (PC2) contribute 20.61% and 15.52%, respectively, and samples treated with different treatments of zhu3151 and B73 are separated from each other with good distinction, showing specificity between groups, while the three replicates of zhu3151 and B73 treated in the same period are mostly close to each other, showing good repeatability within the group.

[0034] like Figure 3As shown in D, a total of 24 samples of zhu3151 and B73 were sequenced by high-throughput sequencing to obtain a total of 600,347,261 cleanreads. The Q30 base percentage was above 93.22%, and the base GC content was between 52.07% and 54.71%, with an average of 53.51. The results of the principal component analysis of the above samples showed that the intra-group duplication, inter-group specificity and data quality of the transcriptome sequencing were good and could be used for the next step of data analysis.

[0035] 3. Statistics of differentially expressed genes in different treatments of zhu3151 and B73 The transcriptome data of zhu3151 and B73 were analyzed, and the differentially expressed genes of zhu3151 and B73 under different treatments were screened and divided into 8 groups: BCS / BMS, BCR / BMR, BMS / BMR, BMS / ZMS, BMR / ZMR, ZCS / ZMS, ZCR / ZMR, ZMS / ZMR, and P The value is less than 0.01 and the relative difference between the 24 samples |log 2 The differentially expressed genes were screened based on FoldChange|>1.5. Figure 3 As shown in E, BCS / BMS, BCR / BMR, BMS / BMR, BMS / ZMS, BMR / ZMR, ZCS / ZMS, ZCR / ZMR, and ZMS / ZMR have 1066, 3919, 6948, 6503, 6020, 443, 1789, and 2804 differentially expressed genes, respectively. Among them, 607 genes were up-regulated and 459 genes were down-regulated in BCS / BMS; 2043 genes were up-regulated and 1876 genes were down-regulated in BCR / BMR; 3296 genes were up-regulated and 3625 genes were down-regulated in BMS / BMR; 3233 genes were up-regulated and 3720 genes were down-regulated in BMS / ZMS; 2899 genes were up-regulated and 3121 genes were down-regulated in BMR / ZMR; 356 genes were up-regulated and 87 genes were down-regulated in ZCS / ZMS; 835 genes were up-regulated and 954 genes were down-regulated in ZCR / ZMR; 1413 genes were up-regulated and 1391 genes were down-regulated in ZMS / ZMR.

[0036] The results showed that BMS / BMR had the most differentially expressed genes, while ZCS / ZMS had the least differentially expressed genes. In the two varieties of zhu3151 and B73, a large number of up-regulated or down-regulated genes appeared in different treatments. These large number of differentially expressed genes were generated by complex transcriptional regulatory mechanisms.

[0037] 4. KEGG analysis and GO annotation of differential expression of zhu3151 and B73 under different treatments (1) KEGG analysis and GO annotation of genes co-regulated by BCS / BMS and ZCS / ZMS A Venn diagram was drawn for the significantly differentially expressed genes in BCS / BMS and ZCS / ZMS, as shown in Figure 4 A, there are 46 genes with significant differences in both comparison groups. KEGG metabolic pathway analysis was performed on these 46 differentially expressed genes. The results are as follows: Figure 4 B, significantly enriched in Starch and sucrose metabolism, Circadian rhythm–plant, Glyoxylate and dicarboxylate metabolism, Carbon fixation in photosynthetic organisms, ABC transporters, Arginine and proline metabolism, alpha-linolenic acid metabolism, Other types of O-glycan biosynthesis, Linoleic acid metabolism, and also enriched in Plant hormone signal transduction. These biological processes play an important role in energy metabolism and material transport, stress response, and growth and development regulation.

[0038] GO annotation analysis was performed on the 46 co-regulated differentially expressed genes, including biological process, cellular component, and molecular function, such as Figure 4C. In the biological process category, it was significantly enriched in protein autophosphorylation. In addition, it was also significantly enriched in CAAX-box protein processing, microgametogenesis, cold acclimation, megagametogenesis, phosphate transport, reductive pentose-phosphate cycle, cell cycle arrest, L-proline biosynthetic process, amylopectin biosynthetic process and SNARE complex assembly. These processes play an important role in plant response to stress and plant reproductive development. In the cell composition category, it was significantly enriched in endosome, amyloplast, lytic vacuole, and in nucleus and integral component of It is enriched in membrane (membrane integration components), and these cell components play an important role in material transport and metabolism, cell growth and division, cell stress and apoptosis, gravity sensing and cell orientation; in the molecular function category, it is significantly enriched in glucan exo-1,3-beta-glucosidase activity, starch synthase activity, glutamate-5-semialdehyde dehydrogenase activity, and is also enriched in transferase activity, transferring glycosyl groups, transferase activity, and kinase activity. In terms of plant growth and development and stress resistance, these enzyme activities are involved in the reconstruction and modification of cell walls, the regulation of energy metabolism, and the regulation of signal transduction. In addition, in terms of pathogen defense, these enzyme activities may also work synergistically to enhance plant disease resistance.

[0039] (2) KEGG analysis and GO annotation of genes co-regulated by BCR / BMR and ZCR / ZMR A Venn diagram was drawn for the significantly differentially expressed genes in BCR / BMR and ZCR / ZMR, such as Figure 4 D, there are 363 genes with significant differences in both comparison groups. KEGG metabolic pathway analysis was performed on these 363 differentially expressed genes. The results are as follows: Figure 4 E is significantly enriched in Starch and sucrose metabolism, and is greatly enriched in Proteinprocessing in endoplasmic reticulum and Carbonmetabolism. These vital biochemical processes in organisms play a key role in energy supply, material synthesis and transport, and maintenance of cell function.

[0040] GO annotation analysis was performed on the 363 co-regulated differentially expressed genes, such as Figure 4F. In biological process, it was significantly and abundantly enriched in response to heat, protein complex oligomerization, response to hydrogen peroxide, response to reactive oxygen species, cellular response to unfolded protein, response to unfolded protein, and chaperone cofactor-dependent protein refolding. These physiological processes play an important role in maintaining cell homeostasis, enhancing anti-oxidative stress ability, regulating signal transduction and gene expression, and improving cell adaptability and survival. In cell composition, it was significantly and abundantly enriched in cell-cell contact zone, axon initial segment, costamere, node of Ranvier, T-tubule, intercalated disc, nuclear MIS12 / MIND complex, chaperone complex, mitochondrial intermembrane space protein transporter complex, apical plasma membrane, plasma membrane, plasma membrane region anchored component of plasma membrane, Atg1 / ULK1 kinase complex and cell surface are significantly enriched. These structures and complexes involve a wide range of functions, including intercellular communication, nerve conduction, muscle contraction, cell division, protein folding and transport, energy metabolism and many other aspects.In the molecular function category, it is significantly enriched in protein self-association, unfolded protein binding, misfolded protein binding, protein folding chaperone, pyridoxalphosphate binding, and also in ATPase activity. These functions or enzyme activities play an important role in the body and participate in the regulation of protein folding, quality control, and energy metabolism. ;

[0041] (3) KEGG analysis and GO annotation of genes co-regulated by BMS / BMR and ZMS / ZMR A Venn diagram was drawn for the significantly differentially expressed genes in BMS / BMR and ZMS / ZMR, as shown in Figure 4 G, there are 1697 genes with significant differences in both comparison groups. KEGG metabolic pathway analysis was performed on these 1697 differentially expressed genes. The results are as follows Figure 4 H is significantly enriched in Photosynthesis, Carbon fixation in photosyntheticorganisms, Photosynthesis - antenna proteins, Carbon metabolism, Pentose phosphate pathway, Biosynthesis ofamino acids, Glycolysis / Gluconeogenesis, and Fatty acid degradation. These processes are interrelated in organisms and jointly regulate energy metabolism, material metabolism, growth and development, and maintain the normal physiological functions of organisms.

[0042] GO annotation analysis was performed on the 1697 co-regulated differentially expressed genes, such as Figure 4I. In biological process, it is significantly enriched in photosynthesis, photosynthesis, light harvesting in photosystem l, fructose 1,6-bisphosphate metabolic process, photosystem ll assembly, photosynthetic electron transport chain, and is also greatly enriched in carbohydrate metabolic process. These processes regulate photosynthesis and its related processes, and are of great significance for maintaining the growth and development of plants, the balance of ecosystems, and life activities on Earth. In cell composition, it is significantly enriched in chloroplastthylakoid membrane, chloroplast, photosystemll, photosystem II, photosystem l, chloroplast envelope, plastoglobule, chloroplast thylakoid, chloroplast stroma, photosystem lreaction center, plastid It was significantly and abundantly enriched in thylakoid membrane, photosystem II oxygen evolving complex, thylakoid, plastid thylakoid, photosynthetic membrane, and obsolete thylakoid part.In the molecular function category, it is significantly enriched in chlorophyllbinding, beta-glucosidase activity, fructose-bisphosphate aldolase activity, oxidoreductase activity, acting on NAD(P)H, quinone or similar compound as acceptor, glucose-1-phosphate adenylyltransferase activity, and also in carbohydratebinding. These enzyme activities play an important role in photosynthesis and sugar metabolism in plants. ;

[0043] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method based on the inoculation of aphid vectors with MaYMV, characterized in that: The steps are as follows: aphids are fed with poison on corn plants carrying MaYMV, and the aphids after feeding with poison are placed on corn seedlings to be inoculated with the poison.

2. The method according to claim 1, characterized in that: The aphid is corn aphid or gramineous aphid.

3. The method according to claim 2, characterized in that: The poison feeding time is 3-5 d.

4. The method according to claim 3, characterized in that: The number of aphids used for inoculation is 1-7, and the inoculation time is 1-5 days.

5. The method according to claim 4, characterized in that: The corn seedlings are in the corn VE stage, V1 stage or V2 stage.

6. Application of the drug-inoculation system constructed by the method described in any one of claims 1 to 5, characterized in that: The application is selected from any one or more of the following: (1) To study the effects of MaYMV infection on the metabolic pathways of maize plants; (2) Study the effects of MaYMV infection on the cell structure of corn plants; (3) Study the effect of MaYMV infection on corn yield; (4) Study the response of corn plants to MaYMV infection.

7. The use according to claim 6, characterized in that: The metabolic pathways include photosynthesis, sugar metabolism, energy metabolism and supply, material synthesis and transport, and cell function maintenance.

8. The use according to claim 6, characterized in that: The cell architecture includes cell wall remodeling and modification, cell growth and division, cell stress and apoptosis, gravity sensing and cell orientation.

9. The use according to claim 6, characterized in that: The effects on corn yield include stunted and stunted corn plants, short ears, and reduced kernel number.

10. The use according to claim 6, characterized in that: The maize plants responded to MaYMV infection by enhancing pathogen defense responses and anti-oxidative stress capabilities.

Images