Application of RNAse Inhibitor 5'-Guanosine Monophosphate in Improving Plant Disease Resistance

Abstract

The present invention belongs to the technical field of agricultural microorganisms, and discloses the application of an RNA enzyme inhibitor 5'-guanosine monophosphate (5'-GMP) in improving plant disease resistance. The present invention discloses the application of an RNA enzyme inhibitor 5'-GMP in preventing and controlling soybean root rot caused by Fusarium graminearum and Rhizoctonia solani, soybean wilt caused by Fusarium oxysporum, and alfalfa root rot caused by Fusarium oxysporum. The present invention proves through experiments that 5'-GMP can significantly improve the disease resistance of soybean to Fusarium graminearum, Fusarium oxysporum, Rhizoctonia solani, and alfalfa to Fusarium oxysporum; in addition, the RNA enzyme inhibitor 5'-GMP of the present application can also significantly promote the growth of soybean, alfalfa, and rice.

Description

Technical Field

[0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to the application of 5'-guanosine monophosphate, an RNA enzyme inhibitor, in improving the disease resistance of plants. Background Art

[0002] In recent years, Fusarium graminearum ( Fusarium graminearum ), Fusarium oxysporum ( Fusarium oxysporum ) and Rhizoctonia solani ( Rhizoctonia solani ) have caused frequent diseases, which have seriously affected the yield and quality of crops. These pathogens have different characteristics. Fusarium graminearum is one of the main pathogens of soybean root rot; Fusarium oxysporum is transmitted through the soil, causing wilt, leading to duct blockage and plant wilting; and Rhizoctonia solani mainly infects the rhizomes during the seedling stage, causing seedling lodging and death. The occurrence of diseases is closely related to environmental factors such as continuous cropping, poor soil management, high temperature and high humidity.

[0003] For these diseases, the current prevention and control strategies include agricultural prevention, chemical prevention, biological prevention and soil disinfection. Agricultural prevention emphasizes crop rotation, improving soil microecology, breeding disease-resistant varieties and optimizing irrigation conditions; chemical prevention reduces pathogenic bacteria by seed dressing and spraying fungicides; biological prevention mainly uses microbial preparations and antagonists to inhibit the growth of pathogens; and soil disinfection reduces the base number of pathogens through lime nitrogen or high temperature treatment. However, these prevention and control measures also have many shortcomings in practice. For example, long-term use of chemical fungicides leads to drug resistance in pathogens and the efficacy gradually decreases; excessive application of chemical agents causes soil and water pollution, threatening ecological security; agricultural prevention and control measures such as crop rotation and the promotion of disease-resistant varieties are difficult to implement in full due to arable land resources and technical limitations; biological prevention is affected by environmental factors and the effect is not stable enough; in addition, some farmers lack comprehensive prevention and control awareness, resulting in limited effect of single prevention and control means and easy recurrence of diseases.

[0004] Therefore, developing and utilizing new drugs to prevent and control diseases will be an important direction for preventing and controlling plant diseases, which will not only help increase crop yield and quality, but also reduce the use of other chemical agents and protect the ecological environment. Summary of the invention

[0005] The purpose of the present invention is to provide an application of 5'-guanosine monophosphate (5'-GMP), an RNase inhibitor, to solve the problems existing in the prior art.

[0006] In one aspect, the present invention provides an application of 5'-guanosine monophosphate (5'-GMP), wherein the application specifically comprises one or more of the following:

[0007] 1) Reduce Fusarium graminearum ( Fusarium graminearum ), Fusarium oxysporum ( Fusarium oxysporum ) and / or Rhizoctonia solani ( Rhizoctonia solani )’s toxicity;

[0008] 2) Prevent and control plant root rot;

[0009] 3) Improving the antibacterial capacity of soybean against Fusarium graminearum, Fusarium oxysporum and / or Rhizoctonia solani, or improving the antibacterial capacity of alfalfa against Fusarium oxysporum;

[0010] 4) preparing a composition having the ability to improve the antibacterial ability of soybean to Fusarium graminearum, Fusarium oxysporum and / or Rhizoctonia solani, or preparing a composition having the ability to improve the antibacterial ability of alfalfa to Fusarium oxysporum;

[0011] 5) Promoting plant growth or preparing a composition having the effect of promoting plant growth.

[0012] In certain embodiments, the plants are soybean, alfalfa, and rice.

[0013] In certain embodiments, the pathogen of plant root rot is Fusarium graminearum, Fusarium oxysporum and / or Rhizoctonia solani.

[0014] On the other hand, the present invention provides a composition having any of the following effects, wherein the active ingredient in the composition comprises 5'-guanosine monophosphate (5'-GMP), wherein the effect is:

[0015] 1) Reduce the virulence of Fusarium graminearum, Fusarium oxysporum and / or Rhizoctonia solani;

[0016] 2) Prevent and control plant root rot;

[0017] 3) Improving the antibacterial capacity of soybean against Fusarium graminearum, Fusarium oxysporum and / or Rhizoctonia solani, or improving the antibacterial capacity of alfalfa against Fusarium oxysporum;

[0018] 4) Promote plant growth.

[0019] In certain embodiments, the composition is a seed coating agent, preferably a suspension seed coating agent.

[0020] In certain embodiments, the plants are soybean, alfalfa, and rice.

[0021] In another aspect, the present invention also provides a method, wherein the composition is applied to the soil around plant seeds or around the rhizosphere of plant seedlings.

[0022] The method is any of the following:

[0023] 1) Reduce the virulence of Fusarium graminearum, Fusarium oxysporum and / or Rhizoctonia solani;

[0024] 2) Prevent and control plant root rot;

[0025] 3) Improving the antibacterial capacity of soybean against Fusarium graminearum, Fusarium oxysporum and / or Rhizoctonia solani, or improving the antibacterial capacity of alfalfa against Fusarium oxysporum;

[0026] 4) Promote plant growth.

[0027] In certain embodiments, the plants are soybean, alfalfa, and rice.

[0028] Compared with the prior art, the present invention shows through soybean pot experiments and Fusarium graminearum RNase activity detection experiments that 5'-GMP can reduce the toxicity of Fusarium graminearum by inhibiting the RNase activity of Fusarium graminearum, thereby improving the soybean resistance to Fusarium graminearum ( Fusarium graminearum ), Fusarium oxysporum ( Fusarium oxysporum )、Rhizoctonia solani( Rhizoctonia solani ) and alfalfa against Fusarium oxysporum ( Fusarium oxysporum ) and the application of 5'-GMP can significantly promote the growth of soybean, rice and alfalfa under the condition of no pathogen infection. At the same time, 5'-GMP is made into seed coating agent by dry seed dressing method to coat soybean, and the seed coating agent can significantly improve the disease resistance of soybean to Fusarium graminearum, Fusarium oxysporum and Rhizoctonia solani. The present invention provides new drug resources and technical support for chemical control of soybean and alfalfa root rot. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the specific implementation manner of the present invention or the technical solution in the prior art, the drawings required for describing the specific implementation manner are briefly introduced below.

[0030] Figure 1 This is the 5'-monophosphate guanosine (5'-GMP) antibacterial plate test in Example 1; the left picture shows the growth of Fusarium graminearum under different concentrations of 5'-monophosphate, and the right bar graph shows the diameter of the Fusarium graminearum strain.

[0031] Figure 2 This is the detection of 5'-guanosine monophosphate (5'-GMP) inhibiting the activity of Fusarium graminearum RNase in Example 1; in the picture, Fusarium graminearum RNase Fg12 can degrade soybean RNA, 5'-guanosine monophosphate can effectively inhibit the activity of Fusarium graminearum RNase, and the inhibitory effect increases with the increase of 5'-guanosine monophosphate concentration.

[0032] Figure 3 This is the effect of 5'-monophosphate guanosine on soybean root rot caused by Fusarium graminearum PH-1 in Example 2; the left photo shows the survival of potted soybean seedlings under different treatments, and the right bar graph shows the survival rate and fresh weight of potted soybean seedlings under different treatments.

[0033] Figure 4 The figure shows the effect of 5'-monophosphate guanosine on soybean wilt caused by Fusarium oxysporum in Example 2; the left picture shows the survival of potted soybean seedlings under different treatments, and the right bar graph shows the survival rate and fresh weight of potted soybean seedlings under different treatments.

[0034] Figure 5 The figure shows the effect of 5'-monophosphate guanosine on soybean damping-off disease caused by Rhizoctonia solani in Example 2; the picture on the left shows the survival of potted soybean seedlings under different treatments, and the bar graph on the right shows the survival rate and fresh weight of potted soybean seedlings under different treatments.

[0035] Figure 6 The figure shows the effect of 5'-monophosphate guanosine on alfalfa root rot caused by Fusarium oxysporum in Example 2; the picture on the left shows the survival of potted soybean seedlings under different treatments, and the bar graph on the right shows the survival rate and fresh weight of potted soybean seedlings under different treatments.

[0036] Figure 7 The effect of 5'-monophosphate guanosine on soybean growth in Example 3; the left photo shows the growth of potted soybean seedlings under different 5'-monophosphate guanosine concentrations, and the right bar graph shows the fresh weight of potted soybean seedlings under different treatments.

[0037] Figure 8 The effect of 5'-monophosphate guanosine on the growth of alfalfa in Example 3; the left photo is a picture of the growth of potted alfalfa under the treatment of 5'-monophosphate guanosine, and the right bar graph is the fresh weight of potted alfalfa under different treatments.

[0038] Fig. 9 The effect of 5'-monophosphate guanosine on rice growth in Example 3; the left photo is a picture of the growth of potted rice under 5'-monophosphate treatment, and the right bar graph is the fresh weight of potted rice under different treatments.

[0039] Fig.10 This is the effect of 5'-monophosphate guanosine seed coating agent in Example 4 on soybean root rot caused by Fusarium graminearum PH-1; the left photo shows the survival of potted soybean seedlings under different treatments, and the right bar graph shows the survival rate and fresh weight of potted soybean seedlings under different treatments.

[0040] Fig.11 The figure shows the effect of 5'-monophosphate guanosine seed coating agent in Example 4 on soybean wilt caused by Fusarium oxysporum; the picture on the left shows the survival of potted soybean seedlings under different treatments, and the bar graph on the right shows the survival rate and fresh weight of potted soybean seedlings under different treatments.

[0041] Fig.12The figure shows the effect of 5'-monophosphate guanosine seed coating agent in Example 4 on soybean root rot caused by Rhizoctonia solani; the picture on the left shows the survival of potted soybean seedlings under different treatments, and the bar graph on the right shows the survival rate and fresh weight of potted soybean seedlings under different treatments. DETAILED DESCRIPTION

[0042] The following examples are provided to facilitate a better understanding of the present invention, but are not intended to limit the present invention. The test methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples are purchased from conventional biochemical reagent stores unless otherwise specified.

[0043] The PDA medium formula involved in the following examples is as follows: 6 g potato powder, 20 g glucose, dilute to 1 L with deionized water, 20 g agar powder, and sterilize at 121°C for 15 min.

[0044] Example 1 5'-Guanosine monophosphate (5'-GMP) inhibits the RNase activity of Fusarium graminearum

[0045] 1. 5'-Guanosine monophosphate (5'-GMP) does not inhibit the growth of Fusarium graminearum

[0046] 1) Prepare PDA plates containing 0 mM, 0.1 mM, and 1 mM 5'-monophosphate (5'-GMP) concentrations. Pour 15 mL of PDA medium containing different concentrations of 5'-monophosphate (5'-GMP) into each culture dish.

[0047] 2) Inoculate Fusarium graminearum into a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 4 days.

[0048] 3) Observe and measure the diameter of Fusarium graminearum colonies.

[0049] The results are as follows Figure 1 As shown, 5'-guanosine monophosphate (5'-GMP) did not inhibit the growth of F. graminearum as measured by strain diameter.

[0050] 2. 5'-guanosine monophosphate (5'-GMP) inhibits the RNase activity of Fusarium graminearum

[0051] 1) Soybean RNA was incubated with 1 μmol / L Fg12 protein (with RNase activity) and 0.1 mM, 1 mM, and 10 mM GMP at 25°C for 30 min.

[0052] 2) After incubation, add Loading Buffer to the sample and perform agarose gel electrophoresis at 120V for 15 minutes. RNase A is used as a positive control. Buffer and EV (glutathione S-91 transferase, GST protein, 1 μmol / L) are used as negative controls.

[0053] 3) Place the agarose gel in a UV imager to observe the degradation of RNA bands.

[0054] The results are as follows Figure 2 As shown, the Fusarium graminearum Fg12 protein can effectively degrade soybean RNA. With the addition of different concentrations of 5'-monophosphate guanosine (5'-GMP), 5'-monophosphate guanosine (5'-GMP) can effectively inhibit the RNase activity of Fusarium graminearum Fg12, and the inhibition of 5'-monophosphate guanosine (5'-GMP) on Fg12 RNase activity is dose-related.

[0055] Example 2 5'-Guanosine monophosphate (5'-GMP) can improve soybean resistance to pathogens

[0056] 1. Testing the effect of 5'-guanosine monophosphate (5'-GMP) on improving soybean resistance to Fusarium graminearum

[0057] 1) Inoculate Fusarium graminearum PH-1 in a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 6 days until it fills the culture dish. Fusarium graminearum plate PH-1 is used for soil inoculation. Cut the plate containing Fusarium graminearum into 5×5 mm blocks for later use.

[0058] 2) Fill a cylindrical pot (cylindrical inner diameter: 15 cm, inner height: 13 cm) with sterile soil, pour out the vermiculite, moisten it with sterile water, add 5×5 mm2 Fusarium graminearum clumps (2 plates with a diameter of 90 mm), and then put the mixed soil back into the cylindrical pot.

[0059] 3) Add 10 ml of 5'-monophosphate guanosine (5'-GMP) at concentrations of 0 mM, 0.1 mM, and 1 mM to the potted plants, sow 10 soybeans per pot (the soybean variety is Hefeng 47; the height of vermiculite above the soybeans is about 1 cm), grow in a greenhouse at 25°C, 16 h light / 8 h dark, and 70% relative humidity for 12 days, and water once every 3 days. Each treatment was repeated three times.

[0060] 4) Statistical analysis of the survival rate and fresh weight of soybeans under different treatment conditions.

[0061] The results are as follows Figure 3As shown in the picture of the control group, the leaves turned yellow, the plants fell over, and the plants were in an "inverted eight" shape, which are symptoms of soybean root rot caused by Fusarium graminearum; after treatment with 5'-guanosine monophosphate (5'-GMP), the survival rate of soybean seedlings was significantly higher than that of the control group, which indicates that 5'-guanosine monophosphate (5'-GMP) can effectively improve soybean's resistance to soybean root rot caused by Fusarium graminearum PH-1.

[0062] 2. Testing the effect of 5'-guanosine monophosphate (5'-GMP) on improving soybean resistance to Fusarium oxysporum

[0063] 1) Inoculate Fusarium oxysporum in a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 6 days until it fills the culture dish. Fusarium oxysporum is used for soil inoculation. Cut the plate containing Fusarium oxysporum into 5×5 mm blocks for later use.

[0064] 2) Fill a cylindrical pot (cylindrical inner diameter: 15 cm, inner height: 13 cm) with sterile soil, pour out the vermiculite, moisten the vermiculite with sterile water, add 5×5 mm2 Fusarium oxysporum fungus blocks (2 plates with a diameter of 90 mm), and then put the mixed soil back into the cylindrical pot.

[0065] 3) 10 ml of 5'-monophosphate guanosine (5'-GMP) with concentrations of 0 mM, 0.1 mM, and 1 mM were added to the pots, 10 soybeans were sown in each pot (the soybean variety was Hefeng 47; the height of vermiculite above the soybeans was about 1 cm), and the plants were grown in a greenhouse at 25°C, 16 h light / 8 h dark, and 70% relative humidity for 12 days, and watered every 3 days. Each treatment was repeated three times.

[0066] 4) Statistical analysis of the survival rate and fresh weight of soybeans under different treatment conditions.

[0067] The results are as follows Figure 4 As shown in the picture, the soybeans in the control group grew short, had stunted root growth, and had a low survival rate; after treatment with 5'-guanosine monophosphate (5'-GMP), the survival rate of the soybean seedlings was significantly higher than that of the control group, indicating that 5'-guanosine monophosphate (5'-GMP) can effectively improve soybean's resistance to Fusarium oxysporum.

[0068] 3. Test 5'-guanosine monophosphate (5'-GMP) to improve soybean resistance to Rhizoctonia solani

[0069] 1) Inoculate Rhizoctonia solani in a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 7 days until it fills the culture dish. Rhizoctonia solani is used for soil inoculation. Cut the plate containing Rhizoctonia solani into 5×5 mm blocks for later use.

[0070] 2) Fill a cylindrical pot (cylindrical inner diameter: 15 cm, inner height: 13 cm) with sterile soil, pour out the vermiculite, moisten the vermiculite with sterile water, add 5×5 mm2 Rhizoctonia solani fungus blocks (2 plates with a diameter of 90 mm), and then put the mixed soil back into the cylindrical pot.

[0071] 3) 10 mL of 0 mM, 0.1 mM, and 1 mM 5'-monophosphate guanosine (5'-GMP) was added to the pots, 10 soybeans were sown in each pot (the soybean variety was Hefeng 47; the height of the vermiculite above the soybeans was about 1 cm), and the plants were grown in a greenhouse at 25°C, 16 h light / 8 h dark, and 70% relative humidity for 12 days, and watered every 3 days. Each treatment was repeated three times.

[0072] 4) Statistical analysis of the survival rate and fresh weight of soybeans under different treatment conditions.

[0073] The results are as follows Figure 5 As shown in the picture, the soybeans in the control group grew short, had stunted root growth, and had a low survival rate. After treatment with 5'-guanosine monophosphate (5'-GMP), the survival rate of the soybean seedlings was significantly higher than that of the control group, indicating that 5'-guanosine monophosphate (5'-GMP) can effectively improve the disease resistance of soybeans to Rhizoctonia solani.

[0074] 4. Testing the effect of 5'-guanosine monophosphate (5'-GMP) on the resistance of alfalfa to Fusarium oxysporum

[0075] 1) Inoculate Fusarium oxysporum in a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 6 days until it fills the culture dish. Fusarium oxysporum is used for soil inoculation. Cut the plate containing Fusarium oxysporum into 5×5 mm blocks for later use.

[0076] 2) Fill a cylindrical pot (cylindrical inner diameter: 15 cm, inner height: 13 cm) with sterile soil, pour out the vermiculite, moisten it with sterile water, add 5×5 mm2 Fusarium oxysporum fungus blocks (2 plates with a diameter of 90 mm), and then put the mixed soil back into the cylindrical pot.

[0077] 3) Add 10 mL of 1 mM 5'-monophosphate guanosine (5'-GMP) to the potted plants, sow 10 alfalfa seeds per pot (the alfalfa variety is Zhongmu No. 1; the height of the vermiculite above the alfalfa is about 1 cm), grow in a greenhouse at 25°C, 16 h light / 8 h dark, and 70% relative humidity for 12 days, and water every 3 days. Each treatment was repeated three times.

[0078] 4) Statistical analysis of the survival rate and fresh weight of alfalfa under different treatment conditions.

[0079] The results are as follows Figure 6 As shown in the picture, the leaves of the control group turned yellow and the growth of root tissue was inhibited, which were symptoms of alfalfa root rot caused by Fusarium oxysporum. After treatment with 5'-guanosine monophosphate (5'-GMP), the survival rate of soybean seedlings was significantly higher than that of the control group, indicating that 5'-guanosine monophosphate (5'-GMP) can effectively improve the disease resistance of alfalfa to alfalfa root rot caused by Fusarium oxysporum.

[0080] Example 3 5'-Guanosine monophosphate (5'-GMP) promotes the growth of soybean, alfalfa and rice

[0081] 1. Testing 5'-guanosine monophosphate (5'-GMP) to promote soybean growth

[0082] 1) Fill a cylindrical pot (cylindrical inner diameter of 15 cm, inner height of 13 cm) with sterile vermiculite, pour sterile water to moisten the vermiculite, add 10 mL of 5'-monophosphate guanosine (5'-GMP) at concentrations of 0 mM, 0.1 mM, and 1 mM to the pot, sow 10 soybeans per pot (soybean variety is Hefeng 47; the height of vermiculite above the soybeans is about 1 cm), grow in a greenhouse at 25℃, 16 h light / 8 h dark, and 70% relative humidity for 14 days, and water once every 3 days. Each treatment was repeated three times.

[0083] 2) Count the fresh weight of soybeans under different treatment conditions.

[0084] The results are as follows Figure 7 As shown in the figure, after treatment with 5'-guanosine monophosphate (5'-GMP), the fresh weight of soybean seedlings was significantly higher than that of the control group, which indicates that 5'-guanosine monophosphate (5'-GMP) can promote the growth of soybean.

[0085] 2. Test 5'-guanosine monophosphate (5'-GMP) to promote alfalfa growth

[0086] 1) Fill a cylindrical pot (cylindrical inner diameter of 15 cm, inner height of 13 cm) with sterile vermiculite, pour sterile water to moisten the vermiculite, add 10 mL of 1 mM 5'-monophosphate guanosine (5'-GMP) to the pot, sow 10 alfalfa seeds per pot (alfalfa variety is Zhongmu No. 1; the height of vermiculite above the alfalfa is about 1 cm), grow in a greenhouse at 25℃, 16 h light / 8 h dark, and 70% relative humidity for 14 days, and water once every 3 days. Each treatment was repeated three times.

[0087] 2) Count the fresh weight of alfalfa under different treatment conditions.

[0088] The results are as follows Figure 8 As shown in the figure, after treatment with 5'-guanosine monophosphate (5'-GMP), the fresh weight of alfalfa was significantly higher than that of the control group, which indicates that 5'-guanosine monophosphate (5'-GMP) can promote the growth of alfalfa.

[0089] 3. Test whether 5'-guanosine monophosphate (5'-GMP) promotes rice growth

[0090] 1) Fill a cylindrical pot (13 cm inner diameter, 13 cm inner height) with 9 cm high black soil, water with sterile water to moisten the soil, add 10 mL of 1 mM 5'-monophosphate guanosine (5'-GMP) to the pot, sow 30 germinated rice seeds per pot (rice variety CO39, germinated to about 1.2 mm long), grow in a greenhouse at 25°C, 16 h light / 8 h dark, and 70% relative humidity for 14 d, and water once every 3 d. Each treatment was repeated three times.

[0091] 2) Count the fresh weight of rice under different treatment conditions.

[0092] The results are as follows Fig. 9 As shown, the fresh weight of rice treated with 5'-guanosine monophosphate (5'-GMP) was significantly higher than that of the control group, which indicates that 5'-guanosine monophosphate (5'-GMP) can promote the growth of rice.

[0093] Example 4: Preparation of 5'-guanosine monophosphate (5'-GMP) as seed coating agent to improve soybean disease resistance

[0094] 1. Testing the effect of 5'-guanosine monophosphate (5'-GMP) seed coating on improving soybean resistance to Fusarium graminearum

[0095] 1) Inoculate Fusarium graminearum PH-1 in a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 6 days until it fills the culture dish. Fusarium graminearum plate PH-1 is used for soil inoculation. Cut the plate containing Fusarium graminearum into 5×5 mm blocks for later use.

[0096] 2) The seed coating agent was prepared according to the following method: 5'-guanosine monophosphate (5'-GMP): 1%, xanthan gum: 2.2%, dispersant FS3000: 6%, calcium lignin sulfonate: 5%, ethylene glycol: 3%, magnesium aluminum silicate: 0.3%, bentonite: 0.5%, Elite anhydrous fast T: 1.5%, silicone defoamer: 2%, film-forming agent: 5%, dye: 2%, and water as the balance. The above components (except film-forming agent and dye) were weighed in proportion, added to the ball mill for 60 minutes, filtered, pumped into a dispersion tank for high-speed shearing for 3 minutes, and then fully ground in a sand mill to control the particle diameter of the solid component within 1μm. After grinding, stir evenly, and add film-forming agent and dye to obtain a suspended seed coating agent. Soybean seeds (Hefeng 47) were placed in a 10 cm long ziplock bag, and the seed coating agent was added. The ziplock bag was shaken to fully mix the soybean seeds and the seed coating agent. The blank seed coating agent control was without GMP.

[0097] 3) Fill a cylindrical pot (cylindrical inner diameter: 15 cm, inner height: 13 cm) with sterile soil, pour out the vermiculite, moisten it with sterile water, add 5×5 mm2 Fusarium graminearum blocks (2 plates with a diameter of 90 mm), and then put the mixed soil back into the cylindrical pot.

[0098] 4) 10 soybeans were sown in each pot (the soybean variety was Hefeng 47; the height of the vermiculite above the soybeans was about 1 cm), and the pots were grown in a greenhouse at 25°C, 16h light / 8h dark, and 70% relative humidity for 12 days, and watered every 3 days. Each treatment was repeated three times.

[0099] 5) Statistical analysis of the survival rate and fresh weight of soybeans under different treatment conditions.

[0100] The results are as follows Fig.10 As shown in the results, after treatment with 5'-guanosine monophosphate (5'-GMP) seed coating agent, the survival rate of soybean seedlings and the fresh weight of soybeans were significantly higher than those of the control group, indicating that 5'-guanosine monophosphate (5'-GMP) seed coating agent can effectively improve the disease resistance of soybean to soybean root rot caused by Fusarium graminearum PH-1.

[0101] 2. Testing the effect of 5'-guanosine monophosphate (5'-GMP) seed coating on improving soybean resistance to Fusarium oxysporum

[0102] 1) Inoculate Fusarium oxysporum in a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 6 days until it fills the culture dish. The Fusarium oxysporum plate is used for soil inoculation. Cut the plate containing Fusarium oxysporum into 5×5 mm blocks for later use.

[0103] 2) Mix the soybean seeds and seed coating agent thoroughly according to the above method.

[0104] 3) Fill a cylindrical pot (cylindrical inner diameter: 15 cm, inner height: 13 cm) with sterile soil, pour out the vermiculite, moisten the vermiculite with sterile water, add 5×5 mm2 Fusarium oxysporum fungus blocks (2 plates with a diameter of 90 mm), and then put the mixed soil back into the cylindrical pot.

[0105] 4) 10 soybeans were sown in each pot (the soybean variety was Hefeng 47; the height of the vermiculite above the soybeans was about 1 cm), and the pots were grown in a greenhouse at 25°C, 16h light / 8h dark, and 70% relative humidity for 12 days, and watered every 3 days. Each treatment was repeated three times.

[0106] 5) Statistical analysis of the survival rate and fresh weight of soybeans under different treatment conditions.

[0107] The results are as follows Fig.11 As shown in the results, after treatment with 5'-guanosine monophosphate (5'-GMP) seed coating agent, the survival rate of soybean seedlings and the fresh weight of soybeans were significantly higher than those of the control group, indicating that 5'-guanosine monophosphate (5'-GMP) seed coating agent can effectively improve the disease resistance of soybean to soybean wilt caused by Fusarium oxysporum.

[0108] 3. Testing the effect of 5'-guanosine monophosphate (5'-GMP) seed coating on improving soybean resistance to Rhizoctonia solani

[0109] 1) Inoculate Rhizoctonia solani in a culture dish containing 15 mL of PDA medium and culture at 25°C in the dark for 7 days until it fills the culture dish. The Rhizoctonia solani plate is used for soil inoculation. Cut the plate containing Rhizoctonia solani into 5×5 mm blocks for later use.

[0110] 2) Mix the soybean seeds and seed coating agent thoroughly according to the above method.

[0111] 3) Fill a cylindrical pot (cylindrical inner diameter: 15 cm, inner height: 13 cm) with sterile soil, pour out the vermiculite, moisten the vermiculite with sterile water, add Rhizoctonia solani fungus blocks with an area of ​​5 × 5 mm (2 plates with a diameter of 90 mm), and then put the mixed soil back into the cylindrical pot.

[0112] 4) 10 soybeans were sown in each pot (the soybean variety was Hefeng 47; the height of the vermiculite above the soybeans was about 1 cm), and the pots were grown in a greenhouse at 25°C, 16h light / 8h dark, and 70% relative humidity for 12 days, and watered every 3 days. Each treatment was repeated three times.

[0113] 5) Statistical analysis of the survival rate and fresh weight of soybeans under different treatment conditions.

[0114] The results are as follows Fig.12As shown, after treatment with 5'-GMP seed coating agent, the survival rate of soybean seedlings and the fresh weight of soybeans were significantly higher than those of the control group, which indicates that 5'-GMP seed coating agent can effectively improve the disease resistance of soybean to soybean root rot caused by Rhizoctonia solani.

[0115] The soybean pot experiment and the RNase activity detection experiment of Fusarium graminearum showed that 5'-GMP can reduce the virulence of Fusarium graminearum by inhibiting the RNase activity of Fusarium graminearum, thereby improving the disease resistance of soybean to Fusarium graminearum, Fusarium oxysporum, Rhizoctonia solani and alfalfa to Fusarium oxysporum, and the application of 5'-GMP can significantly promote the growth of soybean, rice and alfalfa under the condition of no pathogen infection. At the same time, 5'-GMP is made into seed coating agent for soybean coating by dry seed dressing method, and this seed coating agent can significantly improve the disease resistance of soybean to Fusarium graminearum, Fusarium oxysporum and Rhizoctonia solani.

[0116] Unless otherwise specifically stated, the numerical value set forth in these embodiments does not limit the scope of the present invention. In all examples shown and described here, unless otherwise specified, any specific value should be interpreted as merely exemplary, rather than as restriction, and therefore, other examples of exemplary embodiments may have different values.

Claims

The application of 1.5'-monophosphate guanosine is characterized in that: The applications specifically include: 1) Reduce Fusarium graminearum Fusarium graminearum the toxicity of 2) Prevention and control of Fusarium graminearum Fusarium graminearum Caused by soybean root rot.

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