Use of biological antimicrobial agents in antagonizing plant pathogenic fungi, preventing plant diseases, preparing plant disease control agents, plant disease control agents

Abstract

The present invention relates to the use of a biological antimicrobial agent in antagonizing plant pathogenic fungi, preventing and controlling plant diseases, and preparing a plant disease control agent, and to the plant disease control agent, belonging to the technical field of microorganisms. The volatile metabolites of the fungus Rhacocystis albicans have a significant inhibitory effect on the growth of tomato gray mold, pepper alternating spot pathogen, and cucumber wilt pathogen, and can be used to antagonize plant pathogenic fungi (tomato gray mold, pepper alternating spot pathogen, and cucumber wilt pathogen), thereby avoiding the serious harm to human and animal health caused by pesticide residues when used.

Description

TECHNICAL FIELD

[0001] The present application relates to the application of biological antibacterial agent in antagonizing plant pathogenic fungi, preventing and treating plant diseases, preparing plant disease prevention and treatment agent, and belongs to the field of microbial technology. BACKGROUND

[0002] At present, chemical control is still the main method for preventing and treating plant pathogenic fungi, but long-term and large-scale use of chemical pesticides without scientific basis can easily cause the generation of pathogenic fungi resistance, resulting in reduced control effect. In order to improve the effect, people often increase the amount of pesticides, which eventually leads to pesticide residues in crops, seriously endangering the health of humans and animals.

[0003] In the past few decades, volatile metabolomics has developed rapidly. In the early stage, people mainly explored the complex mechanisms of interaction between plants and plants, plants and insects, and insects and insects. In recent years, since the volatile metabolites produced by bacteria or fungi can also produce induction effects between plants or insects, people have also begun to study the inhibitory effect of volatile metabolites of biocontrol bacteria on pathogenic bacteria or pests and the induction resistance effect of volatile metabolites of biocontrol bacteria on plants. Irpex lacteus is a white rot fungus with worldwide distribution. However, there is no report on the use of volatile metabolites of Irpex lacteus to inhibit plant pathogenic fungi. SUMMARY

[0004] The purpose of the present application is to provide an application of biological antibacterial agent in antagonizing plant pathogenic fungi, which can solve the problem of pesticide residues caused by using pesticides to antagonize plant pathogenic fungi, which seriously endangers the health of humans and animals.

[0005] The second purpose of the present application is to provide an application of biological antibacterial agent in preventing and treating plant diseases, which can solve the problem of pesticide residues caused by using pesticides to prevent and treat plant diseases, which seriously endangers the health of humans and animals.

[0006] The third purpose of the present application is to provide an application of biological antibacterial agent in preparing plant disease prevention and treatment agent, which can solve the problem of pesticide residues caused by using pesticides to prevent and treat plant diseases, which seriously endangers the health of humans and animals.

[0007] The fourth purpose of the present application is to provide a plant disease prevention and treatment agent, which can solve the problem of pesticide residues caused by using pesticides to prevent and treat plant diseases, which seriously endangers the health of humans and animals.

[0008] In order to achieve the above purposes, the technical scheme adopted by the application of biological antibacterial agent in antagonizing plant pathogenic fungi is as follows:

[0009] The application of biological antibacterial agent in antagonizing plant pathogenic fungi, wherein the biological antibacterial agent is H. albidum, and the plant pathogenic fungi are Alternaria solani and / or Fusarium oxysporum f.sp.cucumeris; or the biological antibacterial agent is volatile metabolite of H. albidum, and the plant pathogenic fungi are Botrytis cinerea, Alternaria solani, Fusarium oxysporum f.sp.cucumeris.

[0010] The experimental results show that H. albidum and / or its volatile metabolite have obvious inhibitory effect on the growth of Botrytis cinerea, Alternaria solani and Fusarium oxysporum f.sp.cucumeris, and can be used to antagonize the plant pathogenic fungi (Botrytis cinerea, Alternaria solani and Fusarium oxysporum f.sp.cucumeris), thereby avoiding the harm to human and livestock caused by pesticide residues.

[0011] The application of biological antibacterial agent in preventing and treating plant diseases adopts the technical scheme that:

[0012] The application of biological antibacterial agent in preventing and treating plant diseases, wherein the biological antibacterial agent is H. albidum, and the plant diseases are Alternaria solani and / or Fusarium oxysporum f.sp.cucumeris; or the biological antibacterial agent is volatile metabolite of H. albidum, and the plant diseases are Botrytis cinerea, Alternaria solani and / or Fusarium oxysporum f.sp.cucumeris.

[0013] The experimental results show that H. albidum and / or its volatile metabolite have obvious inhibitory effect on the growth of Botrytis cinerea, Alternaria solani and Fusarium oxysporum f.sp.cucumeris, and can be used to prevent and treat Botrytis cinerea, Alternaria solani and Fusarium oxysporum f.sp.cucumeris, thereby avoiding the harm to human and livestock caused by pesticide residues.

[0014] The application of biological antibacterial agent in preparing plant disease control agent adopts the technical scheme that:

[0015] The application of biological antibacterial agent in preparing plant disease control agent, wherein the biological antibacterial agent is H. albidum, and the plant disease control agent is used for preventing and treating Alternaria solani and / or Fusarium oxysporum f.sp.cucumeris; or the biological antibacterial agent is volatile metabolite of H. albidum, and the plant disease control agent is used for preventing and treating Botrytis cinerea, Alternaria solani and / or Fusarium oxysporum f.sp.cucumeris.

[0016] The experimental results show that H. albidum and / or its volatile metabolite have obvious inhibitory effect on the growth of Botrytis cinerea, Alternaria solani and Fusarium oxysporum f.sp.cucumeris, and can be used to prevent and treat Botrytis cinerea, Alternaria solani and Fusarium oxysporum f.sp.cucumeris, thereby avoiding the harm to human and livestock caused by pesticide residues.

[0017] The plant disease control agent adopts the technical scheme that:

[0018] A plant disease control agent comprising a biological antibacterial agent, the biological antibacterial agent being Irpex lacteus, the plant disease control agent being used for preventing and treating pepper alternaria leaf spot, cucumber fusarium wilt; or the biological antibacterial agent being a volatile metabolite of Irpex lacteus, the plant disease control agent being used for preventing and treating tomato botrytis blight, pepper alternaria leaf spot, cucumber fusarium wilt.

[0019] The experimental results show that the Irpex lacteus and / or the volatile metabolite thereof have obvious inhibitory effect on the growth of tomato botrytis blight, pepper alternaria leaf spot and cucumber fusarium wilt, and can be used for preparing a plant disease control agent for preventing and treating tomato botrytis blight, pepper alternaria leaf spot and cucumber fusarium wilt, thereby avoiding the harm to human and livestock caused by pesticide residues.

[0020] In the present application, the classification name of the Irpex lacteus is Irpex lacteus LL210, the preservation number is CGMCC No. 21057; the tomato botrytis blight is Botrytis cinerea Pers; the pepper alternaria leaf spot is Alternaria alternata; and the cucumber fusarium wilt is Fusarium oxysporum. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 It is a picture of the growth of the mycelium in the plate containing only the Irpex lacteus in the experimental example 4 of the present application;

[0022] Figure 2 It is a picture of the growth of the mycelium in the plate containing only the tomato botrytis blight in the experimental example 4 of the present application;

[0023] Figure 3 It is a picture of the growth of the mycelium in the plate containing the Irpex lacteus and the tomato botrytis blight in the experimental example 4 of the present application;

[0024] Figure 4 It is a picture of the growth of the mycelium in the plate containing only the pepper alternaria leaf spot in the experimental example 4 of the present application;

[0025] Figure 5 It is a picture of the growth of the mycelium in the plate containing the Irpex lacteus and the pepper alternaria leaf spot in the experimental example 4 of the present application;

[0026] Figure 6 It is a picture of the growth of the mycelium in the plate containing only the cucumber fusarium wilt in the experimental example 4 of the present application;

[0027] Figure 7 It is a picture of the growth of the mycelium in the plate containing the Irpex lacteus and the cucumber fusarium wilt in the experimental example 4 of the present application;

[0028] Figure 8 Figure 4 is a mycelium radius growth chart of the experimental group and the control group containing tomato gray mold fungus in Example 4 of the present application at different times;

[0029] Figure 9 Figure 5 is a mycelium radius growth chart of the experimental group and the control group containing pepper alternaria fungus in Example 4 of the present application at different times;

[0030] Figure 10 Figure 6 is a mycelium radius growth chart of the experimental group and the control group containing cucumber fusarium wilt fungus in Example 4 of the present application at different times;

[0031] Figure 11 Figure 7 is a mycelium radius growth chart of the experimental group and the control group containing Irpex lacteus in Example 4 of the present application at different times. DETAILED DESCRIPTION

[0032] The application of the biological antibacterial agent of the present application in antagonizing plant pathogenic fungi is an opening invention. The experimental results show that the white cavity Irpex lacteus LL210 and its volatile metabolites have obvious inhibitory effect on the growth of tomato gray mold fungus (Botrytis cinerea Pers), and the white cavity Irpex lacteus LL210 and its volatile metabolites can be used for preventing and treating tomato gray mold; the white cavity Irpex lacteus LL210 and its volatile metabolites have obvious inhibitory effect on the growth of pepper alternaria fungus (Alternaria alternata), and the white cavity Irpex lacteus LL210 and its volatile metabolites can be used for preventing and treating pepper alternaria; the white cavity Irpex lacteus LL210 and its volatile metabolites have obvious inhibitory effect on the growth of cucumber fusarium wilt fungus (Fusarium oxysporum), and the white cavity Irpex lacteus LL210 and its volatile metabolites can be used for preventing and treating cucumber fusarium wilt.

[0033] The application of the white cavity Irpex lacteus and / or its volatile metabolites in antagonizing plant pathogenic fungi, the plant pathogenic fungi being one, two or three of tomato gray mold fungus, pepper alternaria fungus, and cucumber fusarium wilt fungus.

[0034] The application of the white cavity Irpex lacteus and / or its volatile metabolites in preventing and treating plant diseases, the plant diseases being one, two or three of tomato gray mold, pepper alternaria, and cucumber fusarium wilt.

[0035] The application of the white-cyst Irpex lacteus and / or its volatile metabolites in the preparation of a plant disease control agent for preventing tomato gray mold, pepper anthracnose, and cucumber fusarium wilt.

[0036] A plant disease control agent comprising white-cyst Irpex lacteus and / or its volatile metabolites for preventing tomato gray mold, pepper anthracnose, and cucumber fusarium wilt.

[0037] The technical solutions of the application are further described below in combination with specific embodiments.

[0038] The application of the biological antibacterial agent (white-cyst Irpex lacteus and / or its volatile metabolites) in antagonizing plant pathogenic fungi, preventing plant diseases, preparing a plant disease control agent, and specific embodiments of the plant disease control agent are described through the following experimental embodiments.

[0039] Experimental Embodiment 1

[0040] This experimental embodiment is used to verify the antagonistic effect of the volatile metabolites of white-cyst Irpex lacteus LL210 on tomato gray mold (Botrytis cinerea Pers), and the specific experimental method is as follows: tomato gray mold (Botrytis cinerea Pers) is inoculated into a culture dish, and the culture dish is placed on the upper part of a test container, then a culture medium containing white-cyst Irpex lacteus LL210 is placed in the lower part of the test container, then a fan inside the test container is turned on, and the test container is in a sealed state, after 3 days of culture, the inhibition rate of white-cyst Irpex lacteus on tomato gray mold (Botrytis cinerea Pers) is determined every day, and the test time is 11 days, and this experiment is named as the experimental group experiment. At the same time, a control group experiment is carried out under the same experimental conditions, in which a culture medium without white-cyst Irpex lacteus LL210 is placed in the lower part of the test container. The calculation method of the inhibition rate is as follows: inhibition rate = (diameter of the colony in the control group experiment - diameter of the colony in the experimental group experiment) / (diameter of the colony in the control group experiment - diameter of the initial bacterial cake) x 100%. The antibacterial rate of the volatile metabolites of white-cyst Irpex lacteus LL210 on tomato gray mold (Botrytis cinerea Pers) at different test times is shown in Table 1.

[0041] Table 1 Antibacterial rate of the volatile metabolites of white-cyst Irpex lacteus LL210 on tomato gray mold (Botrytis cinerea Pers) at different test times

[0042] Time 4th d 5th d 6th d 7th d 8th d 9th d 10th d 11th d Bacteriostatic rate 3.05％ 5.37％ 16.36％ 26.94％ 32.04％ 35.51％ 40.84％ 43.78％

[0043] As shown in Table 1, the volatile metabolites of Irpex lacteus LL210 have obvious inhibitory effect on the growth of Botrytis cinerea Pers, and the volatile metabolites of Irpex lacteus LL210 can be used for preventing and treating tomato gray mold, and can also be used for preparing a plant disease control agent for preventing and treating tomato gray mold.

[0044] Experimental Example 2

[0045] This experimental example is used to verify the antagonistic effect of the volatile metabolites of Irpex lacteus LL210 on Alternaria alternata. The specific experimental method is as follows: Alternaria alternata is inoculated into a culture dish, and the culture dish is placed on the upper part of a test container, then a culture medium containing Irpex lacteus LL210 is placed in the lower part of the test container, then a fan inside the test container is turned on, and the test container is in a sealed state. After 3 days of culture, the inhibition rate of Irpex lacteus on Alternaria alternata is determined every day, and the test time is 11 days. This experiment is named as experimental group experiment. At the same time, a control group experiment is carried out under the same experimental conditions, in which a culture medium without Irpex lacteus LL210 is placed in the lower part of the test container. The calculation method of the inhibition rate is as follows: inhibition rate = (diameter of the colony in the control group experiment - diameter of the colony in the experimental group experiment) / (diameter of the colony in the control group experiment - diameter of the initial bacterial cake) x 100%. The antibacterial rate of the volatile metabolites of Irpex lacteus LL210 on Alternaria alternata at different test times is shown in Table 2.

[0046] Table 2 Antibacterial rate of volatile metabolites of Irpex lacteus LL210 on Alternaria alternata at different test times

[0047] Time 4th d 5th d 6th d 7th d 8th d 9th d 10th d 11th d Bacteriostatic rate 8.00％ 9.38％ 11.31％ 21.71％ 27.68％ 31.89％ 35.37％ 38.36％

[0048] As shown in Table 2, the volatile metabolites of Irpex lacteus LL210 have obvious inhibitory effect on the growth of Alternaria alternata, and the volatile metabolites of Irpex lacteus LL210 can be used for preventing and treating pepper alternaria leaf spot, and can also be used for preparing a plant disease control agent for preventing and treating pepper alternaria leaf spot.

[0049] Experimental Example 3

[0050] The present experimental example is used for verifying the antagonistic effect of the volatile metabolites of Irpex lacteus LL210 on Fusarium oxysporum, and the specific experimental method is as follows: Fusarium oxysporum is inoculated into a culture dish, and the culture dish is placed on the upper part of a test container, then a culture medium containing Irpex lacteus LL210 is placed in the lower part of the test container, then a fan inside the test container is started, and the test container is in a sealed state, after 3 days of culture, the inhibition rate of Irpex lacteus on Fusarium oxysporum is determined every day, and the test time is 11 days, and the experiment is named as experimental group experiment. At the same time, a control group experiment in which a culture medium not containing Irpex lacteus LL210 is placed in the lower part of the test container is carried out under the same experimental conditions. The calculation method of the inhibition rate is as follows: inhibition rate = (diameter of the colony in the control group experiment - diameter of the colony in the experimental group experiment) / (diameter of the colony in the control group experiment - diameter of the initial bacterial cake) x 100%. The antibacterial rate of the volatile metabolites of Irpex lacteus LL210 on Fusarium oxysporum at different test times is shown in Table 3.

[0051] Table 3 Antibacterial rate of volatile metabolites of Irpex lacteus LL210 on Fusarium oxysporum at different test times

[0052] Time 4th d 5th d 6th d 7th d 8th d 9th d 10th d 11th d Bacteriostatic rate 5.11％ 6.96％ 7.64％ 12.96％ 12.96％ 12.96％ 12.96％ 12.96％

[0053] As shown in Table 3, the volatile metabolites of Irpex lacteus LL210 have obvious inhibitory effect on the growth of Fusarium oxysporum, and the volatile metabolites of Irpex lacteus LL210 can be used for preventing and treating cucumber fusarium wilt, and can also be used for preparing a plant disease control agent for preventing and treating cucumber fusarium wilt.

[0054] Experimental Example 4

[0055] This experimental example is used to verify the inhibitory effect of Irpex lacteus LL210 on Botrytis cinerea Pers, Alternaria alternata and Fusarium oxysporum. The specific experimental method is as follows: a 0.8 cm diameter Irpex lacteus cake is placed on one side of a 9 cm diameter PDA medium plate, and a pathogenic fungus cake of the same size is placed on the other side. The two cakes are separated by 4.5 cm and placed in a 28°C constant temperature incubator for culture. The growth conditions are counted on the 1st, 2nd, 3rd, 4th, 5th and 6th days of culture, and the radius of the pathogenic fungus cake is recorded as the experimental group colony radius. At the same time, control group 1 and control group 2 are set up. Control group 1 has no Irpex lacteus cake on one side of the PDA medium plate, but only has a pathogenic fungus cake on the other side. The radius of the pathogenic fungus cake is recorded on the 1st, 2nd, 3rd, 4th, 5th and 6th days of culture as the control group 1 colony radius. Control group 2 has an Irpex lacteus cake on one side of the PDA medium plate, but no pathogenic fungus cake on the other side. The radius of the pathogenic fungus cake is recorded on the 1st, 2nd, 3rd, 4th, 5th and 6th days of culture as the control group 2 colony radius. After the experiment, the inhibition rate is calculated. The inhibition rate on the Xth day = [(control group colony radius on the Xth day - control group colony radius on the X-1th day) - (experimental group colony radius on the Xth day - experimental group colony radius on the X-1th day)] x 100% / (control group colony radius on the Xth day - control group colony radius on the X-1th day). The control group in the inhibition rate calculation formula refers to control group 1. The inhibition rate calculation result is rounded to two decimal places. When X = 1, the experimental group colony radius on the X-1th day and the control group colony radius on the X-1th day are both 0.4 cm. The growth conditions of different mycelia are shown in Figures 1-7 Figure 1 is the mycelial growth condition picture when only Irpex lacteus is contained in the plate, Figure 2 is the mycelial growth condition picture when only Botrytis cinerea is contained in the plate, Figure 3 is the mycelial growth condition picture when both Irpex lacteus and Botrytis cinerea are contained in the plate, Figure 4 is the mycelial growth condition picture when only Alternaria alternata is contained in the plate, Figure 5 is the mycelial growth condition picture when both Irpex lacteus and Alternaria alternata are contained in the plate, Figure 6 is the mycelial growth condition picture when only Fusarium oxysporum is contained in the plate, Figure 7 ​Figures of mycelium growth of tomato gray mold in the presence of white-cyst Inonotus in the plate at different times, bc represents the mycelium growth curve of tomato gray mold in the presence of white-cyst Inonotus in the plate, bcck represents the mycelium growth curve of tomato gray mold in the absence of white-cyst Inonotus in the plate. Figure 1 In the figures, "positive" means taking a photo from the front of the culture dish, and "negative" means taking a photo from the back of the culture dish. Figure 3 In the figures, "positive" means taking a photo from the front of the culture dish, and "negative" means taking a photo from the back of the culture dish.

[0056] Then, the relationship between the radius of different mycelium and time was made into a table and a picture, and the results are shown in Table 2 and Fig. 2. Figures 8-11 Figure 8 Figures of mycelium growth of tomato gray mold in the presence of white-cyst Inonotus in the plate at different times, bc represents the mycelium growth curve of tomato gray mold in the presence of white-cyst Inonotus in the plate, bcck represents the mycelium growth curve of tomato gray mold in the absence of white-cyst Inonotus in the plate. Figure 9 Figures of mycelium growth of tomato gray mold in the presence of white-cyst Inonotus in the plate at different times, bc represents the mycelium growth curve of tomato gray mold in the presence of white-cyst Inonotus in the plate, bcck represents the mycelium growth curve of tomato gray mold in the absence of white-cyst Inonotus in the plate. Figure 10 Figures of mycelium growth of tomato gray mold in the presence of white-cyst Inonotus in the plate at different times, bc represents the mycelium growth curve of tomato gray mold in the presence of white-cyst Inonotus in the plate, bcck represents the mycelium growth curve of tomato gray mold in the absence of white-cyst Inonotus in the plate. Figure 11 Figures of mycelium growth of tomato gray mold in the presence of white-cyst Inonotus in the plate at different times, bc represents the mycelium growth curve of tomato gray mold in the presence of white-cyst Inonotus in the plate, bcck represents the mycelium growth curve of tomato gray mold in the absence of white-cyst Inonotus in the plate.

[0057] Table 4 Inhibition rate of white-cyst Inonotus on tomato gray mold, pepper alternaria leaf spot and cucumber fusarium wilt

[0058]

[0059] ​The results showed that Lrpex lacteus LLC210 had antagonistic effect on Botrytis cinerea Pers, Alternaria alternata and Fusarium oxysporum.

Claims

1. Application of biological antimicrobial agents in antagonizing plant pathogenic fungi, characterized in that: The biological antibacterial agent is white cyst or the volatile metabolites of white cyst, and the plant pathogenic fungus is pepper alternatifida; the preservation number of the white cyst is CGMCC NO.21057.

2. Application of biological antimicrobial agents in preventing and controlling plant diseases, characterized in that: The biological antibacterial agent is white sac fungus or a volatile metabolite of white sac fungus, and the plant disease is pepper brown spot disease; the preservation number of the white sac fungus is CGMCC NO.21057.

3. The use of a biological antimicrobial agent in the preparation of a plant disease control agent, characterized in that: The biological antibacterial agent is white sac rhizoctonia solani or a volatile metabolite of white sac rhizoctonia solani. The plant disease control agent is used for controlling pepper brown spot disease. The preservation number of the white sac rhizoctonia solani is CGMCC NO.21057.

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