Method for preventing and treating pathogenic fungus infection after continuous cropping of tomatoes by combining calcium propionate with trichoderma harzianum

Abstract

The invention relates to the technical field of agricultural cultivation, in particular to a method for preventing and treating pathogenic fungus infection after continuous cropping of tomatoes by combining calcium propionate with trichoderma harzianum, and the method can enhance the disease resistance of the tomatoes after continuous cropping, improve the quality of the tomatoes and promote the growth of the tomatoes. It is found for the first time that combined use of calcium propionate and trichoderma harzianum can comprehensively prevent and treat common mixed fungus infection after tomato continuous cropping, and particularly, the composition has a comprehensive prevention and treatment effect on mixed fungus infection of Colletotrichum gloeosporioides, Colletotrichum oleifera, Fusarium graminearum and Amoerella almoka.

Description

Technical Field

[0001] The invention relates to the technical field of agricultural cultivation, and in particular to a method for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes by using calcium propionate in combination with Trichoderma harzianum. Background Art

[0002] The harm caused by pathogenic fungi after continuous cropping of tomatoes in my country is extremely serious. In particular, after continuous cropping of tomatoes, multiple pathogenic fungi often cause joint infection. It is difficult to prevent and control, and it is not easy to completely and effectively cure the phenomenon of simultaneous infection of several pathogenic fungi. The difficulty of combined use of drugs is relatively large, the selectivity is very wide, and the dosage screening is complicated. If you are not careful, the growth of tomatoes will be inhibited, that is, the infection problem is solved, but the growth of crops is hindered. Therefore, it is urgent to solve a method for preventing and controlling the mixed infection of multiple pathogenic fungi after continuous cropping of tomatoes, and at the same time promoting crop growth. Summary of the invention

[0003] The purpose of the present invention is to provide a method for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes by using calcium propionate in combination with Trichoderma harzianum, thereby enhancing the disease resistance of tomatoes, improving the quality of tomatoes, and promoting their growth. To achieve the above purpose, the present invention provides the following technical solutions.

[0004] A method for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes by using calcium propionate in combination with Trichoderma harzianum comprises the following steps:

[0005] (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside;

[0006] (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100-200 μM / m 2 / s, wavelength 400-700nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 CFU stops, and activated Trichoderma harzianum conidia liquid is obtained;

[0007] (3) After the tomatoes are transplanted, when pathogenic fungal infection occurs, the conidia solution of Trichoderma harzianum prepared in step (2) is applied together with irrigation water, once every three days, for a total of three times, with each application amount being 400-1600 kg per mu;

[0008] (4) On the second day and the fifth day after the irrigation in step (3), a mixed solution of equal volumes of gibberellin at a concentration of 20-50 mg / L and strigolactone at a concentration of 3-5 μM / L is evenly sprayed on the tomato leaves, once in the morning and once in the evening.

[0009] Furthermore, the tomato leaves are evenly sprayed with a mixed solution of equal volumes of gibberellin with a concentration of 20 mg / L and strigolactone with a concentration of 5 μM / L.

[0010] Further, in each 1g of the conidia of Trichoderma harzianum, the number of viable bacteria of Trichoderma harzianum is 10 9 ~10 10 CFU.

[0011] The present invention also provides a method for preparing a preparation for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes, comprising the following steps:

[0012] (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside;

[0013] (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100-200 μM / m 2 / s, wavelength 400-700nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 The CFU was stopped, and the activated conidia liquid of Trichoderma harzianum was obtained.

[0014] The preparation for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes obtained by the above method.

[0015] The above method is used for the application of the preparation for controlling pathogenic fungi infection after continuous cropping of tomatoes in any of the following items:

[0016] (1) Application in preventing and controlling any one or more fungal infections such as Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera and Aspergillus emericae;

[0017] (2) Application in the prevention and treatment of tomato wilt caused by Fusarium oxysporum f.sp. Lycopersici, Fol. after continuous tomato cropping;

[0018] (3) Application in promoting tomato growth and development;

[0019] (4) Application in improving the salt resistance of tomatoes.

[0020] The present application also provides a method for preventing pathogenic fungi from infecting tomatoes after continuous cropping, comprising the following steps:

[0021] (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside;

[0022] (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100 μM / m 2 / s, wavelength 400nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 CFU stops, and activated Trichoderma harzianum conidia liquid is obtained;

[0023] (3) Apply Trichoderma harzianum to the tomato plants when transplanting them. The dosage is 5 g of the conidia solution of Trichoderma harzianum prepared in step (2) per hole. Sprinkle the solution into the hole.

[0024] Beneficial effects of this application:

[0025] 1. The present invention first discovered that the combination of calcium propionate and Trichoderma harzianum can have a comprehensive control effect on common fungi such as Higgins Anthracnose, Camellia oleifera, Fusarium graminearum and Emmerichia subspora. In particular, it can solve the problem of mixed infection of multiple or more fungi after continuous cropping of tomatoes.

[0026] 2. The present invention is the first to study the solution for the continuous cropping pathogenic fungi of tomatoes, that is, when the continuous cropping pathogenic fungi appear in large areas and multiple combinations, by comparing the CK control group, the experimental group using calcium propionate alone, the experimental group using Trichoderma harzianum alone, and the experimental group combining calcium propionate + Trichoderma harzianum, it is found that the combination of Trichoderma harzianum and 0.25% CP has a good control effect on several mixed fungal infections: Fusarium graminearum, Higgins Anthracnose, Camellia oleifera and Amoris Sporangiae. When controlling the mixed fungi of Higgins Anthracnose, Camellia oleifera and Amoris Sporangiae, Trichoderma harzianum can be used alone, or Trichoderma harzianum combined with 0.25% CP can be used, both of which can achieve good technical effects.

[0027] 3. The present invention proposes for the first time to prevent and control tomato wilt caused by Fusarium oxysporum f. sp. Lycopersici (Fol), which is one of the most serious fungal diseases in tomato production and has caused huge economic losses to global tomato agricultural production. The combined use of calcium propionate and Trichoderma harzianum can effectively prevent and control Fol fungal infection.

[0028] 4. After adding calcium propionate and Trichoderma harzianum to the irrigation water, it can significantly prevent and control the damage of various fungal infections after continuous cropping of the soil. At the same time, it will cause soil salinization. The application of gibberellins and strigolactones can promote plant growth and enhance the salt tolerance of plants. The present application uniformly sprays a mixed solution of equal volumes of gibberellins at a concentration of 20 mg / L and strigolactones at a concentration of 5 μM / L on the leaves in combination with a prevention and control test for the joint infection of various fungi after continuous cropping of tomatoes. It is concluded that it has the best effect on promoting the growth of tomato plants. When the concentrations of gibberellins and strigolactones are increased accordingly, they will hinder the growth of plant leaves and the synthesis of chlorophyll, thereby hindering the growth of tomatoes.

[0029] 5. We found that when the conidia of Trichoderma harzianum and calcium propionate were used for the joint control of multiple continuous cropping pathogenic fungi, that is, applied with irrigation water after tomato transplanting, the control effect was optimal on the second day of irrigation and the fifth day after irrigation. Excessive content of calcium propionate significantly inhibited the biomass accumulation and plant development of tomatoes. With the increase of calcium propionate content, the fresh weight of the aboveground part decreased, the chlorophyll content also decreased significantly, and the disease index increased. This shows that excessive calcium propionate inhibited the growth of tomato plants. At the same time, the content of calcium propionate was low, it could not effectively prevent fungal infection, and the disease index also increased accordingly.

[0030] 6. The present applicant has made a lot of creative research and obtained the result that the activated conidia of Trichoderma harzianum has the highest antibacterial activity, that is, the light intensity is 100 μM / m 2 / s, wavelength 400nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 The antibacterial activity of the activated conidia of Trichoderma harzianum was the highest when the CFU was stopped. The increase of light intensity and wavelength led to a significant increase in the disease index. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Different fungal colony morphologies under PDA medium culture (A);

[0032] Figure 2 Diameters of different fungal colonies on PDA medium at 7 days (B);

[0033] Figure 3Growth rate of different fungal colonies on PDA medium (C);

[0034] Figure 4 Morphology of different fungal colonies on PDA medium (+0.25% CP) (A);

[0035] Figure 5 Diameters of different fungal colonies on PDA medium (+0.25% CP) for 7 days (B);

[0036] Figure 6 Growth rate of different fungal colonies in PDA medium (+0.25% CP) (C);

[0037] Figure 7 The growth morphology of different fungal colonies and Trichoderma harzianum on PDA medium (A);

[0038] Figure 8 Diameters of growth of different fungal colonies confronting Trichoderma harzianum at 7 days (B);

[0039] Fig. 9 Growth rate of different fungal colonies against Trichoderma harzianum on PDA medium (C);

[0040] Fig.10 The growth morphology of different fungal colonies and Trichoderma harzianum on PDA medium (+0.25% CP) (A);

[0041] Fig.11 Diameters of growth of different fungal colonies confronting Trichoderma harzianum at 7 days (B);

[0042] Fig.12 Growth rate of different fungal colonies against Trichoderma harzianum on PDA medium (+0.25% CP) (C);

[0043] Fig.13 Comprehensive comparison of the growth diameters of different bacterial communities under four experimental conditions over 7 days;

[0044] Fig.14 Comprehensive comparison of the growth rates of different bacterial communities under four experimental conditions over 7 days;

[0045] Fig.15 ITS amplification bands of plant pathogenic fungi isolated from soil. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in 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.

[0047] Example 1

[0048] A method for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes by using calcium propionate in combination with Trichoderma harzianum comprises the following steps:

[0049] (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside;

[0050] (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100 μM / m 2 / s, wavelength 400nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 CFU stops, and activated Trichoderma harzianum conidia liquid is obtained;

[0051] (3) After the tomatoes are transplanted, when pathogenic fungal infection occurs, the conidia solution of Trichoderma harzianum of step (2) is applied together with irrigation water, once every 3 days, for a total of 3 times, with each application amount being 400 kg of the conidia solution of Trichoderma harzianum of step (2) per mu;

[0052] (4) On the second day and the fifth day after the irrigation in step (3), a mixed solution of equal volumes of 20 mg / L gibberellin and 5 μM / L strigolactone was evenly sprayed on the tomato leaves, once in the morning and once in the evening.

[0053] Example 2

[0054] A method for preventing pathogenic fungi from infecting tomatoes after continuous cropping, comprising the following steps:

[0055] (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside;

[0056] (2) Preparation of Trichoderma harzianum conidia liquid: The Trichoderma harzianum strain was inoculated into the above culture medium and cultured under the irradiation of a white cold fluorescent lamp with a light intensity of 100 μM / m2 / s and a wavelength of 400 nm at 25°C until the number of viable bacteria per 1 g of Trichoderma harzianum conidia liquid reached 10. 9 ~10 10 CFU stops, and activated Trichoderma harzianum conidia liquid is obtained;

[0057] (3) Apply Trichoderma harzianum to the tomato plants when transplanting them. The dosage is 5 g of the conidia solution of Trichoderma harzianum prepared in step (2) per hole. Sprinkle the solution into the hole.

[0058] Example 3

[0059] A method for preparing a preparation for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes, comprising the following steps:

[0060] (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside;

[0061] (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100 μM / m 2 / s, wavelength 400nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 The CFU was stopped, and the activated conidia liquid of Trichoderma harzianum was obtained.

[0062] Experimental methods:

[0063] The strains used in the following experiments: Trichoderma harzianum, Fusarium graminearum, Colletotrichum higginsianum, Colletotrichum phomoides, Didymella americana, Colletotrichum boninense, Fusarium oxysporum f. sp. Lycopersici, Fol are all known strains and can be purchased on the market. The materials for this application were purchased from Mingzhou Biotechnology and Huizao Biotechnology.

[0064] Culture medium: All fungi were cultured on PDA medium (potato dextrose ager). Wash, peel and cut fresh potatoes into pieces, weigh 200g, add appropriate amount of distilled water and boil for about 15 minutes until the potatoes become soft, filter with multi-layer gauze, dissolve 20g of glucose in the filtrate (if it is treated with 0.25% calcium propionate, add 2.5g of calcium propionate powder), and make the volume of the solution to 1L and dispense it into conical bottles with 1.5% agar powder, seal them, sterilize them at 121℃ for 20 minutes, and then pour them into plates for use.

[0065] Culture conditions: All strains were cultured under white cold fluorescent light (light intensity up to 100 μM / m 2 / s; wavelength 400nm) and cultured at 25°C.

[0066] Experiment 1: Measurement of growth rate and colony diameter of different fungi

[0067] Experimental method: Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera, Aspergillus emoricalis and Trichoderma harzianum grown on PDA medium for 3 days were plated along the young mycelium of the colony edge and inoculated into the center of PDA medium without 0.25% calcium propionate. The plates were marked at 1d, 2d and 7d of growth. The distance between the two marks was the growth rate of the fungus. Colony morphology (A), colony diameter (B) and colony growth rate (C) were directly measured after 7d of growth, and repeated 3 times. Different letters represent significant differences, P < 0.01, error value ± SDs (n = 3).

[0068] Experimental results: Figure 1 and Figure 4 By comparison, it can be clearly seen that the inhibition zones of Higgins Anthrax and Camellia oleifera in the experimental group of PDA medium (+0.25% calcium propionate) were significantly smaller than those in the control group without calcium propionate. Figure 2 It can be seen that after 7 days of colony growth, the diameters of different fungi are significantly different, among which Trichoderma harzianum has the largest diameter, followed by Colletotrichum oleifera. Figure 3 According to the statistical results of the growth rates of different fungal colonies, it can be seen that the growth rate of Trichoderma harzianum is the fastest, followed by Colletotrichum oleifera, and the results are consistent with the trend of the fungal colony diameter (B).

[0069] Experiment 2: Effect of adding 0.25% CP to the culture medium on the growth rate and colony diameter of different fungi

[0070] Experimental method: Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera, Aspergillus emoricalis and Trichoderma harzianum grown on PDA medium for 3 days were plated along the edge of the colony and inoculated into the center of PDA medium supplemented with 0.25% calcium propionate (CP). The plates were marked at 1d, 2d and 7d of growth. The distance between the two marks was the growth rate of the fungus. This was repeated 3 times. The colony morphology (A), colony diameter (B) and colony growth rate (C) were directly measured after 7d of growth. The statistical results are shown in Table 1. Different letters represent significant differences, P < 0.01, error value ± SDs (n = 3).

[0071] Experimental results: Figure 4 According to the results of different fungal colony morphology on PDA medium (plus 0.25% CP), the growth of Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera, and Subsporia emerica was obviously inhibited, while Trichoderma harzianum was not affected.

[0072] Figure 5 According to the results of the measurement of the diameters of different fungal colonies on PDA medium, which are consistent with the results of colony morphology observation, the diameters of Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera, and Aspergillus emericae were significantly lower than the results without adding 0.25% CP in Experiment 1. However, unlike other fungi, the addition of 0.25% CP promoted the growth of Trichoderma harzianum.

[0073] Figure 6 According to the statistical results of the growth rates of different fungal colonies, it can be seen that the trend is consistent with the results of the fungal colony diameter (B), that is, the growth rate of Trichoderma harzianum is the fastest, and the growth rates of other fungi are inhibited by 0.25% CP.

[0074] Experiment 3: Colony morphology of different plant pathogenic fungi growing against Trichoderma harzianum

[0075] Experimental method: Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera and Emmerikaia subspora were grown on PDA plates without 0.25% calcium propionate and against Trichoderma harzianum. The colony morphology (A), colony diameter (B) and colony growth rate (C) were recorded after 7 days. The results were repeated 3 times. Different letters represent significant differences, P < 0.01, error value ± SDs (n = 3).

[0076] Experimental results: Combination Figure 7 and Figure 8 The growth morphology (A) and diameter (B) of different fungal colonies confronting Trichoderma harzianum. The results show that Trichoderma harzianum has the greatest inhibitory effect on Aspergillus emericae, followed by Colletotrichum higgins. Trichoderma harzianum has the least inhibitory effect on Fusarium graminearum. Fig. 9 According to the statistical results of the growth rates of different fungal colonies confronting Trichoderma harzianum, it can be seen that the trend is consistent with the results of the confrontation diameter (B) of the fungal colonies, that is, the use of Trichoderma harzianum alone has the greatest inhibitory effect on Acanthopanax emerica.

[0077] Experiment 4: Colony morphology of different plant pathogenic fungi confronting Trichoderma harzianum on plates supplemented with 0.25% CP for 7 days

[0078] Experimental method: Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera and Aspergillus emoricalis were grown on PDA plates supplemented with 0.25% CP for 7 days against Trichoderma harzianum. The colony morphology (A), colony diameter (B) and colony growth rate (C) were recorded and statistical results were repeated 3 times. Different letters represent significant differences, P < 0.01, error value ± SDs (n = 3).

[0079] Experimental results: Fig.10 Fig.11 Combining the growth morphology (A) and diameter (B) results of different fungal colonies confronting Trichoderma harzianum, it can be seen that after adding 0.25% CP, the inhibitory effect of Trichoderma harzianum on different fungal colonies was significantly increased, among which the inhibitory effect on Higgins Anthracnose was the greatest, followed by Fusarium graminearum. Fig.12 According to the statistical results of the growth rates of different fungal colonies and Trichoderma harzianum, it can be seen that the trend is consistent with the results of the fungal colony confrontation diameter (B), that is, Trichoderma harzianum has the greatest inhibitory effect on Higgins Anthrax, followed by Fusarium graminearum.

[0080] Fig.13 Fig.14The CK control group, the experimental group using calcium propionate alone, the experimental group using Trichoderma harzianum alone, and the experimental group using calcium propionate + Trichoderma harzianum were summarized. The comparison results show that the treatment method of using Trichoderma harzianum in combination with 0.25% CP (CP0.25% + Trichoderma harzianum) has a more obvious inhibitory effect on each bacterial group, especially on Fusarium graminearum. The method of using Trichoderma harzianum in combination with 0.25% CP has a particularly significant inhibitory effect. The method of using 0.25% CP alone has no significant effect on the four bacterial groups, especially on Fusarium graminearum and Emorikkaya. The confrontation method of using Trichoderma harzianum alone also has a good inhibitory effect on Higgins Anthrax, Camellia Anthrax and Emorikkaya Anthrax, but the effect on Fusarium graminearum is not significant. Therefore, for the prevention and control of Fusarium graminearum, the method of using Trichoderma harzianum in combination with 0.25% CP should be adopted; and for the prevention and control of Higgins Anthracnose, Camellia oleifera and Emmerichia erythrocytes, the confrontation method of Trichoderma harzianum can be adopted, or the method of using Trichoderma harzianum in combination with 0.25% CP can be used to achieve better results.

[0081] Experiment 5 Effects of Gibberellic Acid + Strigolactone on Salt Resistance and Growth of Tomato

[0082] Experimental methods: Fig.15 The three plant pathogenic fungi bands isolated from the soil of tomato continuous cropping for 5 years were amplified using fungal ITS sequences. The results of the ITS sequence comparison of plant pathogenic fungi isolated from the soil are as follows: Colletotrichum oleifera, Fusarium oxysporum tomato-specific species Fol, and Aspergillus emorykaya, indicating that the above pathogens are common species in the soil of tomato continuous cropping; and then the experimental field with 5 years of continuous cropping and tomato seedlings were used as tomato research materials. At the beginning of the experiment, Fusarium graminearum (Fusarium oxysporum tomato-specific species Fol), Colletotrichum higgins, Colletotrichum oleifera, Colletotrichum oleifera, Colletotrichum oleifera and Aspergillus emorykaya were added with irrigation water. The groups were completely randomized with 3 replicates per treatment and 5 strains per replicate. Specifically, the method of Example 1 was used to treat tomato crops, and gibberellin and strigolactone were sprayed on the leaves of tomato transplanted seedlings, once in the morning and once in the evening. The spraying concentrations are shown in Table 1; among them, Test Examples 1-5, Control Examples 1-2 and the blank group were all sprayed with equal volumes of mixed solutions of gibberellin and strigolactone on the tomato leaves on the second day of irrigation and the fifth day, respectively, and sprayed once in the morning and once in the evening, Test Example 6 was sprayed on the day of irrigation and the fifth day; Test Example 7 was sprayed on the second day of irrigation and the third day; Control Examples 3-4 only changed the concentration of calcium propionate in Example 1 to 0.2% and 0.3%, respectively, and Control Example 5 only increased the light intensity of Example 1 to 200 μM / m 2 / s, wavelength 400nm; Comparative Example 6 only increases the light intensity of Example 1 to 100μM / m 2 / s, wavelength 700nm; 20 days after the above-mentioned treatment of spraying the growth regulator, the above-ground part of the whole plant was collected to determine the fresh weight, disease index, total chlorophyll content, and MDA content.

[0083] Table 1 Proportions of plant regulators at different concentrations

[0084]

[0085]

[0086] Table 2 Effects of different treatments on salt tolerance and growth of tomatoes

[0087]

[0088] Experimental results:

[0089] Biomass is the most intuitive data reflection of seedling growth. Compared with the control 1-2, which used gibberellins or strigolactones alone, and the blank group, the leaves of the present application were evenly sprayed with a mixed solution of equal volumes of gibberellins at a concentration of 20 mg / L and strigolactones at a concentration of 5 μM / L, and sprayed once in the morning and evening. The effect on tomato leaf growth (fresh weight of the aboveground part) was significant. After adding calcium propionate and Trichoderma harzianum to the irrigation water, various fungal infections after continuous cropping of the soil can be significantly prevented and treated. At the same time, the soil will be salinized. The application of gibberellins and strigolactones can promote plant growth and enhance the salt tolerance of the plants.

[0090] In addition, we compared several regulators commonly used in salt resistance of crops such as tomatoes, such as salicylic acid, melatonin, and brassinolide, other than gibberellins and strigolactones, and concluded that the application of gibberellins and strigolactones had the best effect, which could significantly improve the stress caused by spraying calcium propionate after pathogenic fungi infection. In addition, the total chlorophyll content also showed that gibberellins + strigolactones had the technical effect of coordinated synergism. After adding calcium propionate and Trichoderma harzianum for prevention and treatment, the test example 1 of this application could also effectively improve the effect of total chlorophyll content of tomatoes. After the treatment of gibberellins and strigolactones, the accumulation of MDA in tomato cells was reduced in the treatment groups, which alleviated and reduced the damage of salt to tomato seedlings.

[0091] The present application's uniform foliar spraying of a mixed solution of equal volumes of 20 mg / L gibberellin and 5 μM / L strigolactone combined with a prevention and control test for co-infection of multiple fungi after continuous cropping of tomatoes showed the best growth-promoting effect on tomato plants. Compared with Test Examples 2-5, when the concentrations of gibberellin and strigolactone were increased accordingly, they would hinder the growth of plant leaves and the synthesis of chlorophyll, thereby hindering the growth of tomatoes.

[0092] In addition, we found that when the conidia solution of Trichoderma harzianum and calcium propionate was used for joint control of multiple continuously cropped pathogenic fungi, that is, applied with irrigation water after tomato transplanting, the control effect was best on the second day of irrigation and the fifth day after. Compared with Test Examples 6-7, the disease index of spraying on the day of irrigation and the fifth day, and spraying on the second day of irrigation and the third day were higher than that of spraying on the second day of irrigation and the fifth day after Experimental Example 1.

[0093] We also found that excessive calcium propionate content will significantly inhibit the biomass accumulation and plant development of tomatoes. With the increase of calcium propionate content, the fresh weight of the aboveground part decreased, the chlorophyll content also decreased significantly, and the disease index increased. This shows that excessive calcium propionate inhibits the growth of tomato plants. At the same time, low calcium propionate content cannot effectively prevent fungal infection, and the disease index also increases accordingly.

[0094] Light intensity 100μM / m 2 / s, wavelength 400nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 The CFU stopped, and the antibacterial activity of the activated Trichoderma harzianum conidia liquid was the highest. The increase in light intensity and wavelength was not conducive to the comprehensive prevention and control effect of the Trichoderma harzianum conidia liquid combined with calcium propionate of the present application, resulting in a significant increase in its disease index.

[0095] The materials used in this experiment are proposed to prevent and control tomato wilt caused by Fusarium oxysporum tomato-specific type (Fol) and Colletotrichum oleraceus. Fol is one of the most serious fungal diseases in tomato production, causing huge economic losses to global tomato agricultural production. Experiments show that the method of using calcium propionate + Trichoderma harzianum in combination with the present application can effectively prevent and control Fol and Colletotrichum oleraceus fungal infection, and the method of using calcium propionate + Trichoderma harzianum in combination with the present application can also effectively prevent and control Fusarium graminearum (Fusarium oxysporum tomato-specific type Fol) after continuous cropping of tomatoes, Higgins Anthracnose, Camellia oleifera, Colletotrichum oleraceus and Emericae subspora co-infection.

[0096] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes by using calcium propionate in combination with Trichoderma harzianum, characterized in that: The steps include: (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside; (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100-200 μM / m 2 / s, wavelength 400-700nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 CFU stops, and activated Trichoderma harzianum conidia liquid is obtained; (3) After the tomatoes are transplanted, when pathogenic fungal infection occurs, the conidia solution of Trichoderma harzianum prepared in step (2) is applied together with irrigation water, once every three days, for a total of three times, with each application amount being 400 kg per mu; (4) On the second day and the fifth day after the irrigation in step (3), a mixed solution of equal volumes of gibberellin at a concentration of 20-50 mg / L and strigolactone at a concentration of 3-5 μM / L is evenly sprayed on the tomato leaves, once in the morning and once in the evening.

2. The method according to claim 1, characterized in that The tomato leaves are evenly sprayed with a mixed solution of equal volumes of 20 mg / L gibberellin and 5 μM / L strigolactone.

3. The method according to claim 1, characterized in that The light intensity is 100 μM / m 2 / s, wavelength 400nm.

4. A method for preparing a preparation for preventing and controlling pathogenic fungi infection after continuous tomato cropping, characterized in that: The steps include: (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside; (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100-200 μM / m 2 / s, wavelength 400-700nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 The CFU was stopped, and the activated conidia liquid of Trichoderma harzianum was obtained.

5. The method according to claim 4, characterized in that The light intensity is 100 μM / m 2 / s, wavelength 400nm.

6. A preparation for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes obtained by the method of claim 1 or 4.

7. Use of the preparation for preventing and controlling pathogenic fungi infection after continuous cropping of tomatoes as claimed in claim 6 in any of the following: (1) Application in preventing and controlling any one or more fungal infections such as Fusarium graminearum, Colletotrichum higginsii, Colletotrichum oleifera and Aspergillus emericae; (2) Application in the prevention and treatment of one or more fungal infections caused by Fusarium oxysporum f.sp. Lycopersici, Fol., Higgins Anthracnose, Camellia oleifera, Colletotrichum oleifera and Aspergillus emericae after continuous cropping of tomatoes; (3) Application in promoting tomato growth and development; (4) Application in improving the salt resistance of tomatoes.

8. A method for preventing pathogenic fungi from infecting tomatoes after continuous cropping, characterized in that: The steps include: (1) Preparation of culture medium: wash, peel and cut fresh potatoes into pieces, weigh 200 g, add an appropriate amount of distilled water and boil for about 15 min until the potatoes become soft, filter using multi-layer gauze, dissolve 20 g of glucose and 2.5 g of calcium propionate powder in the filtrate, and adjust the volume of the solution to 1 L and dispense into conical bottles with 1.5% agar powder, seal the bottles, sterilize at 121° C. for 20 min, and set aside; (2) Preparation of Trichoderma harzianum conidia solution: The Trichoderma harzianum strain was inoculated into the above culture medium and irradiated with a white cold fluorescent lamp at a light intensity of 100 μM / m 2 / s, wavelength 400nm, shake culture at 25℃ until the number of viable bacteria per 1g of Trichoderma harzianum conidia liquid is 10 9 ~10 10 CFU stops, and activated Trichoderma harzianum conidia liquid is obtained; (3) Apply Trichoderma harzianum to the tomato plants when transplanting them. The dosage is 5 g of the conidia solution of Trichoderma harzianum prepared in step (2) per hole. Sprinkle the solution into the hole.

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