Organic Coupling Cultivation Method and Application of Facility Vegetables

Abstract

The present invention provides a method for organically coupling the cultivation of protected vegetables and its application. The method includes: digging square trenches 1 month before transplanting the seedlings of protected vegetables, and sequentially adding the following components: corn straw - Penicillium cyclopium 455-1 bacterial suspension - virus vaccine - peanut cake - wheat bran - farmyard manure - bacterial fertilizer - compound fertilizer - water, then covering it with soil and sealing it with a plastic film. After 1 month, punch holes on the surface of the plastic film to transplant the vegetable seedlings. The organically coupled cultivation technology of the fermentation broth of the 455-1 mutant strain of protected vegetables + corn straw + organic fertilizer provided by the present invention significantly improves the growth, root development, photosynthetic efficiency of protected tomatoes, peppers, and cucumbers, as well as the resistance to soil-borne oomycete diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of protected vegetable cultivation, and specifically, to a method for organically coupling the cultivation of protected vegetables and its application. Background Art

[0002] The oomycete diseases of protected vegetables severely restrict the safe production and benefits of protected vegetables. The oomycete diseases of protected vegetables mainly include airborne oomycete diseases and soil-borne oomycete diseases. The soil-borne oomycete diseases of protected vegetables are important soil-borne root diseases of protected vegetables, mainly including cucumber, tomato, pepper, eggplant blight, Pythium rot and damping-off, which seriously affect the healthy development of the protected vegetable industry. At present, the prevention and control technologies for soil-borne oomycete diseases of protected vegetables mainly rely on chemical pesticide control, such as single application, excessive application and random application, which are difficult to effectively prevent and control the occurrence and harm of soil-borne oomycete diseases of protected vegetables. In recent years, the cultivation technologies of coupling water and fertilizer, coupling substrate fertilization and drip irrigation under film for protected vegetables have played an important role in enhancing the growth of protected vegetables, improving the quality, increasing the yield of organic vegetables, enhancing the resistance to soil-borne diseases and reducing the application amount of chemical fertilizers and pesticides, and have gradually become new organic cultivation and disease control technologies.

[0003] Establishing a perfect new organic cultivation and disease control technology for protected vegetables, reducing the application amount of pesticides and chemical fertilizers, effectively controlling the occurrence and harm of soil-borne oomycete diseases, and ensuring the increase of the yield of green organic vegetables and safe production. The substrate coupling technology is the main organic coupling cultivation technology for protected vegetables. Researching, innovating and improving the organic coupling cultivation technology of protected vegetable substrates is expected to become a new organic cultivation technology that can effectively promote the growth of protected vegetables, enhance disease resistance and effectively control the occurrence and harm of soil-borne oomycete diseases of protected vegetables. The substrate coupling cultivation technology is to deeply mix and apply straw and organic fertilizer or microbial fertilizer in deep trenches about one month before vegetable transplanting. This technology has low cost, simple operation and is convenient for popularization and utilization. However, it mainly relies on the deep trench soil ecological environment to degrade the straw substrate, and it is difficult to effectively degrade and utilize the organic components of straw in a short time, which easily leads to a large amount of undecomposed straw substrate remaining in the soil substrate, reducing the technical performance and effect of the substrate coupling cultivation.

[0004] The biodegradation of lignocellulose in crop straw mainly depends on the cellulase, hemicellulase and ligninase secreted by the degradable microorganisms themselves and their activities. The degradation of carbohydrates such as cellulose and hemicellulose will produce sugars. The changes in the contents of total sugar and reducing sugar during the biodegradation process of crop straw reflect the degradation of carbohydrates. Therefore, during the biodegradation process of crop straw, the determination of various lignocellulase activities and the changes in the contents of total sugar and reducing sugar are important technical parameters reflecting the degree of straw degradation. Thus, it provides a reference for isolating and identifying microbial strains with the ability to degrade crop straw from crop straw and improving the organic coupling cultivation technology of protected vegetable straw-fertilizer. Summary of the Invention

[0005] The object of the present invention is to provide a method for organic coupled cultivation of protected vegetables and its application.

[0006] In the present invention, a Penicillium strain 2554-2 (i.e., strain MP-03) that can improve the cellulase activity of corn straw is isolated from corn straw, and it is identified as Penicillium cyclopium. ( Penicillium cyclopium ) Then, the nitrosoguanidine method is used to induce mutation of strain 2554-2, and a strong corn straw-degrading mutant strain 455-1 (i.e., strain ΔMP-03) is screened.

[0007] Penicillium cyclopium ( Penicillium cyclopium ) Strain 455-1 has been deposited in the China General Microbiological Culture Collection Center (CGMCC), No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with the postcode 100101. The deposit number is CGMCC NO.41541, and the deposit date is October 12, 2024. The proposed taxonomic name is Penicillium cyclopium. ( Penicillium cyclopium ) .

[0008] Strain 2554-2 has also been deposited in the China General Microbiological Culture Collection Center (CGMCC), No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with the postcode 100101. The deposit number is CGMCC NO.41542, and the deposit date is October 12, 2024. The proposed taxonomic name is Penicillium cyclopium. ( Penicillium cyclopium ) .

[0009] In the first aspect, the present invention provides a method for organic coupled cultivation of protected vegetables, and the method includes: digging a square trench 1 month before transplanting the protected vegetable seedlings, and sequentially adding the following components: corn straw - Penicillium cyclopium 455-1 bacterial suspension - virus vaccine - peanut cake - wheat bran - farmyard manure - microbial fertilizer - compound fertilizer - water, then covering it with soil and sealing it with a plastic film. After 1 month, punch holes on the surface of the plastic film to transplant the vegetable seedlings.

[0010] Further, by weight percentage, the addition ratios of the components are as follows: corn straw (crushed or uncrushed) 50%, peanut cake 8.2%, wheat bran 8.2%, farmyard manure 9.7% (the main components include various nutrient elements such as organic matter, nitrogen, phosphorus, potassium, etc., purchased from Shandong Lubao Biotechnology Co., Ltd.), microbial fertilizer 6.8% (Kingenta Pro-Soil No. 1), compound fertilizer 5.2% (the nitrogen, phosphorus and potassium content is 45%-55%, purchased from Anyang Zhongsheng Fertilizer Industry Co., Ltd.), water 6.8%, Penicillium cyclopium 455-1 bacterial suspension (200 million spores / mL) 5%, vegetable virus vaccine (weak strain vaccine or double-stranded small RNA vaccine, mainly used to prevent and control the occurrence and harm of late vegetable viruses, purchased from Nanjing Ruimat Biotech Co., Ltd.) 0.1%.

[0011] Cultivate statically at 30°C. After 7 days, measure the weight loss rate of the straw and the change in cellulase activity. The non-mutated Penicillium cyclopium bacterial suspension with an inoculation amount of 5% (200 million spores / mL) is used as a control.

[0012] In the present invention, the vegetables include but are not limited to tomatoes, peppers and cucumbers.

[0013] In the second aspect, the method of the present invention is applied to the prevention and control of soil-borne phytophthora diseases of tomatoes, peppers and cucumbers.

[0014] The phytophthora diseases include but are not limited to tomato late blight caused by Phytophthora infestans, cucumber phytophthora blight caused by Phytophthora drechsleri, and pepper phytophthora blight caused by Phytophthora capsici.

[0015] The organic coupling cultivation technology of the fermented liquid of the 455-1 mutant strain of protected vegetables + corn straw + organic fertilizer provided by the present invention significantly improves the growth, root development, photosynthetic efficiency of protected tomatoes, peppers and cucumbers, and the resistance to soil-borne oomycete diseases. Description of the Drawings

[0016] Figure 1 This is the promotion of tomato stem height growth by ΔMP-03 organic coupling cultivation in a preferred embodiment of the present invention.

[0017] Figure 2 This is the promotion of pepper stem height growth by ΔMP-03 organic coupling cultivation in a preferred embodiment of the present invention.

[0018] Figure 3 This is the promotion of cucumber stem height growth by ΔMP-03 organic coupling cultivation in a preferred embodiment of the present invention.

[0019] Figure 4 This is the promotion of tomato stem diameter growth by ΔMP-03 organic coupling cultivation in a preferred embodiment of the present invention.

[0020] Figure 5 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation promotes the stem diameter growth of cucumbers.

[0021] Figure 6 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation promotes the stem diameter growth of peppers.

[0022] Figure 7 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation promotes the root activity of tomatoes.

[0023] Figure 8 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation promotes the root activity of peppers.

[0024] Figure 9 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation promotes the root activity of cucumbers.

[0025] Figure 10 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the dry weight of tomato roots.

[0026] Figure 11 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the dry weight of cucumber roots.

[0027] Figure 12 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the dry weight of pepper roots.

[0028] Figure 13 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the leaf area of tomatoes.

[0029] Figure 14 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the leaf area of cucumbers.

[0030] Figure 15 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the leaf area of peppers.

[0031] Figure 16 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the chlorophyll content of tomato leaves.

[0032] Figure 17 In the preferred embodiment of the present invention, the effect of the ΔMP-03 organic coupling cultivation on the chlorophyll content of cucumber leaves.

[0033] Figure 18Effect of ΔMP-03 organic coupling cultivation on chlorophyll content in pepper leaves in the preferred embodiment of the present invention.

[0034] Figure 19 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation technology significantly reduces the disease index of tomato late blight.

[0035] Figure 20 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation technology significantly reduces the disease index of cucumber blight.

[0036] Figure 21 In the preferred embodiment of the present invention, the ΔMP-03 organic coupling cultivation technology significantly reduces the disease index of pepper blight. Specific embodiments

[0037] The following examples are used to illustrate the present invention, but are not used to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0038] In the following examples, the data processing method uses Excel of Office2003 software for data processing, table making and drawing. The PPSS18.0 statistical software is used for significant analysis of the data.

[0039] Example 1 Degradation of corn straw organic matrix coupling cultivation technology by Penicillium mutant strain 455-1

[0040] 1. Apply the suspension of Penicillium citrinum 455-1 with obvious ability to degrade corn straw to establish an improved organic matrix coupling cultivation technology for protected vegetables.

[0041] One month before transplanting the protected vegetable seedlings, dig square trenches with the following dimensions: the trench depth is 35 cm, the width is 60 cm, and the length is 8.2 m. About 30-35 square trenches are dug per mu.

[0042] The ratio of corn straw and other components added to a single trench:

[0043] By weight percentage, the addition ratios of each component are as follows: corn straw (crushed or uncrushed) 50%, peanut cake 8.2%, wheat bran 8.2%, farmyard manure 9.7% (the main components include various nutrient elements such as organic matter, nitrogen, phosphorus, potassium, etc., purchased from Shandong Lubao Biotechnology Co., Ltd.), microbial fertilizer 6.8% (Kingenta Pro-Soil No. 1), compound fertilizer 5.2% (nitrogen, phosphorus, and potassium content is 45%-55%, purchased from Anyang Zhongcheng Fertilizer Industry Co., Ltd.), water 6.8%, Penicillium cyclopium 455-1 bacterial suspension (200 million spores / mL) 5%, vegetable virus vaccine (weak strain vaccine or double-stranded small RNA vaccine, mainly used to prevent and control the occurrence and harm of late vegetable viruses, purchased from Nanjing Ruimat Biotechnology Co., Ltd.) 0.1%.

[0044] Cultivate statically at 30°C. After 7 days, measure the weight loss rate of the straw and the change in cellulase activity. The non-mutated Penicillium cyclopium bacterial suspension with an inoculation amount of 5% (200 million spores / mL) is used as a control.

[0045] One month before transplanting the seedlings of protected vegetables, the addition order of each component in the dug square trench is as follows: evenly spread the corn straw - mutated or non-mutated Penicillium cyclopium bacterial suspension - virus vaccine - peanut cake - wheat bran - farmyard manure - microbial fertilizer - compound fertilizer - water are added in sequence (the liquid components are evenly sprayed, and the solid or powder components are evenly spread), then cover it with soil and seal it with plastic film. After one month, punch holes on the surface of the plastic film to transplant the vegetable seedlings.

[0046] 2. Determination methods for various technical indexes of adding 455-1 bacterial suspension to degrade corn straw and organically coupled cultivation of tomatoes, peppers, and cucumbers

[0047] 2.1 Determination of plant height and stem diameter of tomatoes, peppers, and cucumbers in the improved organically coupled cultivation of protected vegetables

[0048] 1) Plant height: Measure the plant height of tomatoes, peppers, and cucumbers at the seedling stage, flowering stage, fruiting stage, and full fruiting stage. Use the 5-point sampling method to measure. At each point, measure the plant height of 50 plants of tomatoes, peppers, and cucumbers. For each single plant, take the highest measured height as the standard and calculate the average value.

[0049] 2) Stem diameter: Measure the stem diameter of tomatoes, peppers, and cucumbers at the seedling stage, flowering stage, fruiting stage, and full fruiting stage. Use a vernier caliper to measure. Use the 5-point sampling method to measure. At each point, measure the stem diameter of 50 plants of tomatoes, peppers, and cucumbers. For each single plant, take the thickest part at the base of the stem as the standard and calculate the average value.

[0050] 2.2 Determination of root growth of tomatoes, peppers, and cucumbers in the improved organically coupled cultivation technology of protected vegetables

[0051] During the seedling stage, flowering stage, fruiting stage, and full fruiting stage after transplanting of greenhouse tomatoes, peppers, and cucumbers, the dry weight and root activity of the roots of tomato, pepper, and cucumber plants were measured. The root activity was measured using the TTC method (Li Hesheng, 2002), and the dry weight of the roots was measured using a one-ten-thousandth balance after drying.

[0052] 2.3 Determination of Chlorophyll Content and Leaf Area of Tomato, Pepper, and Cucumber in the Improved Organic Coupling Cultivation Technology for Protected Vegetables

[0053] During the seedling stage, initial flowering stage, fruiting stage, and full fruiting stage of tomatoes, peppers, and cucumbers, the chlorophyll content and leaf area of the leaves were measured.

[0054] 1) Chlorophyll content: Acetone-ethanol mixture method.

[0055] 2) Leaf area: The length × width of the leaves of tomatoes, peppers, and cucumbers were measured respectively during the seedling stage, flowering stage, fruiting stage, and full fruiting stage. Five-point sampling was used for measurement. The leaf sizes of 50 tomato, pepper, and cucumber plants were measured at each point. For each plant, the length × width of 25 middle leaves of tomato, pepper, and cucumber plants were measured, and the average value was taken.

[0056] 3. Results and Analysis

[0057] 3.1 Effects of ΔMP-03 Organic Coupling Cultivation Technology on Promoting the Stem Height Growth of Greenhouse Tomatoes, Peppers, and Cucumbers

[0058] Figure 1 、 Figure 2 and Figure 3 The results showed that compared with MP-03 and WT (without adding bacterial suspension), the organic coupling cultivation technology of adding ΔMP-03 bacterial suspension to degrade corn straw was beneficial to increasing the plant height growth of tomatoes, peppers, and cucumbers in the later stage after seedling transplanting. The measurement data showed that this technology made the plant heights of tomatoes, peppers, and cucumbers show a gradually increasing trend from the initial flowering stage to the full fruiting stage, and the plant heights reached the highest at the full fruiting stage. In contrast, the plant height growth of pepper plants was relatively low from the initial flowering stage to the full fruiting stage, probably because the natural plants of peppers are shorter. Therefore, the organic coupling cultivation technology of adding ΔMP-03 bacterial suspension to degrade corn straw significantly accelerated the plant height growth rate of greenhouse tomatoes, peppers, and cucumbers in the later stage after seedling transplanting, which was beneficial to the growth and development of protected vegetables. Figure 1 、 Figure 2 and Figure 3 The different asterisks in

[0059] 3.2 Effects of ΔMP-03 Organic Coupling Cultivation Technology on Promoting the Stem Diameter Growth of Greenhouse Tomatoes, Cucumbers, and Peppers

[0060] Figure 4 、Figure 5 and Figure 6 The results showed that, compared with MP-03 and WT (without adding bacterial suspension), the organic coupling cultivation technology of adding ΔMP-03 bacterial suspension to degrade corn straw significantly increased the growth of the basal stem thickness of tomato, cucumber and pepper at the initial flowering stage, fruiting stage and full fruiting stage. The basal stem thickness of the three vegetables showed a gradually increasing trend, and the basal stem thickness of tomato, cucumber and pepper increased most significantly at the full fruiting stage. Compared with tomato and cucumber, due to the variety characteristics of pepper, the growth of the basal stem thickness was relatively less from the initial flowering stage to the full fruiting stage. Based on the relevant data, the organic coupling cultivation technology of adding ΔMP-03 bacterial suspension to degrade corn straw significantly accelerated the growth rate of the basal stem thickness of greenhouse tomato, cucumber and pepper from the initial flowering stage to the full fruiting stage, which was beneficial to the growth and development of greenhouse tomato, cucumber and pepper and improved the resistance to soil-borne diseases. Figure 4 、 Figure 5 and Figure 6 The different asterisks marked on the column chart represent the significant difference degrees of the basal stem thickness of the three vegetables, **P<0.01, ***P<0.001.

[0061] 3.3 Effects of ΔMP-03 organic coupling cultivation technology on the root activity of greenhouse tomato, pepper and cucumber

[0062] Figure 7 、 Figure 8 and Figure 9 The results showed that, compared with MP-03 and WT (without adding bacterial suspension), the organic coupling cultivation technology of adding ΔMP-03 bacterial suspension to degrade corn straw significantly improved the root activity of tomato, pepper and cucumber at the initial flowering stage, fruiting stage and full fruiting stage. The root activity of the three tested vegetables showed an increasing trend from the initial flowering stage to the full fruiting stage, and reached the highest at the full fruiting stage. Based on the relevant data, the organic coupling cultivation technology of adding ΔMP-03 bacterial suspension to degrade corn straw was beneficial to enhancing the root growth and development of greenhouse tomato, cucumber and pepper, significantly enhancing the growth of vegetables and improving the disease resistance to soil-borne fungi and oomycete diseases.

[0063] Figure 7 、 Figure 8 and Figure 9 The different asterisks marked on the column chart represent the significant difference degrees of root activity, *P<0.05, **P<0.01, ***P<0.001.

[0064] 3.4 Effects of ΔMP-03 organic coupling cultivation technology on the dry weight of roots of greenhouse tomato, pepper and tomato

[0065] Figure 10 、 Figure 11 and Figure 12The results showed that, compared with MP-03 and WT (no bacterial suspension added), the organic-coupled cultivation technique of corn straw degradation with the addition of ΔMP-03 bacterial suspension significantly increased the root dry weight of tomatoes, peppers, and cucumbers from the initial flowering stage to the peak fruiting stage. The root dry weight of all three vegetables showed a gradual increase from the initial flowering stage, with the most significant increase in root dry weight during the peak fruiting stage for tomatoes, peppers, and cucumbers. Based on the relevant data, the organic-coupled cultivation technique of corn straw degradation with the addition of ΔMP-03 bacterial suspension is beneficial for increasing root growth and development of greenhouse tomatoes, cucumbers, and peppers, significantly enhancing the growth of greenhouse vegetables.

[0066] Figure 10 , Figure 11 and Figure 12 Each experiment was sampled at 5 points and the data were analyzed three times. The dry weight of at least 60 roots of each vegetable was measured each time. Different asterisks on the bar graph represent the significant differences in the dry weight of roots of the three vegetables, *P<0.05, **P<0.01, ***P<0.001.

[0067] 3.5 Effects of ΔMP-03 organic coupled cultivation technology on leaf area changes of greenhouse tomatoes, peppers, and cucumbers

[0068] Figure 13 , Figure 14 and Figure 15 The results showed that compared with MP-03 and WT (no bacterial suspension added), the addition of ΔMP-03 suspension to corn straw-degrading coupled cultivation significantly increased leaf area during the initial flowering, fruiting, and peak fruiting stages of greenhouse tomatoes, cucumbers, and peppers. This trend gradually increased, with leaf area reaching its maximum during the peak fruiting stage. Based on the relevant data, the addition of ΔMP-03 suspension to corn straw-degrading coupled cultivation significantly increased leaf area in greenhouse tomatoes, cucumbers, and peppers, contributing to improved photosynthetic efficiency and light energy utilization rate of tomato, cucumber, and pepper leaves.

[0069] Figure 13 , Figure 14 and Figure 15 Sampling was carried out at 5 points in each experiment, and at least 50 leaves were collected from each vegetable at each point. The area measurement was repeated 3 times. Different asterisks marked on the bar graph represent the significant difference in the change of leaf area of each vegetable, *P<0.05, **P<0.01, ***P<0.001.

[0070] Effects of ΔMP-03 organic coupled cultivation technology on chlorophyll content in greenhouse tomato, pepper, and cucumber leaves

[0071] Figure 16 , Figure 17 and Figure 18The results showed that, compared with MP-03 and WT (no bacterial suspension added), the organic-coupled cultivation technique using the ΔMP-03 bacterial suspension to degrade corn straw significantly increased chlorophyll content in tomato leaves during the initial flowering, fruiting, and peak fruiting stages. Chlorophyll content in tomato leaves was highest during the fruiting stage and began to decline during the peak fruiting stage. Chlorophyll content in pepper and cucumber leaves showed a gradual increase from the initial flowering, fruiting, and peak fruiting stages, with peak content during the peak fruiting stage. Based on these data, the organic-coupled cultivation technique using the ΔMP-03 bacterial suspension to degrade corn straw significantly increased chlorophyll content in greenhouse tomatoes, cucumbers, and peppers, contributing to improved photosynthetic efficiency in these leaves.

[0072] Figure 16 , Figure 17 and Figure 18 Each experiment involved sampling at 5 points and three replicates. At least 50 leaves of each vegetable were measured each time. The different asterisks on the bar graph represent the significant differences in the chlorophyll content of each vegetable leaf, *P<0.05, **P<0.01, ***P<0.001.

[0073] Example 2 Application of the Improved Facility Vegetable Organic Coupling Cultivation Technology in Preventing and Controlling Soil-Borne Diseases of Tomatoes, Peppers, and Cucumbers

[0074] 1. Investigation method of the damage caused by improved organic coupling cultivation technology of facility vegetables to tomato, pepper and cucumber root and stem blight. The tomato, pepper and cucumber greenhouses treated with improved organic coupling cultivation technology of facility vegetables and one control greenhouse were investigated starting from the sporadic outbreak of soil-borne blight of tomatoes, peppers and cucumbers. The investigation was conducted once every 7 days, and samples were taken at 5 points on the diagonal each time. 50 plants were investigated at each point. The investigation was conducted 3 times in a row and the experiment was repeated 3 times.

[0075] 2. Improved organic coupled cultivation technology for greenhouse vegetables: Tomato, pepper, and cucumber disease classification standards and disease index calculation

[0076] Level 0: No lesions on leaves or fruits;

[0077] Level 1: The lesion area accounts for less than 5% of the leaf area. The fruit lesions are watery and light gray, accounting for 10-20% of the fruit area. There are basically sporadic lesions on the stem.

[0078] Level 2: The lesion area accounts for 6%-10% of the leaf area, the fruit lesions are dark gray, accounting for 21%-30% of the fruit area, there are sporadic mold layers, and the number of lesions at the base of the stem increases;

[0079] Level 3: The lesion area accounts for 11%-20% of the leaf area, the fruit lesions are dark gray, accounting for 31%-40% of the fruit area, the mold layer accounts for 30%-40% of the lesions, and the lesions at the base of the stem are continuous, accounting for more than 20%-40% of the basic height of the stem;

[0080] Level 4: The lesion area accounts for 21%-50% of the leaf area. The fruit lesions are dark gray, accounting for 41%-50% of the fruit area. The mold layer accounts for 41%-50% of the lesion area. The lesion at the base of the stem accounts for 41%-50% of the basic height of the stem.

[0081] Level 5: The lesion area accounts for more than 50% of the leaf area. The mold layer accounts for more than 60% of the lesion area. The fruit is rotten. The lesion at the base of the stem accounts for more than 60% of the basic height of the stem.

[0082] Disease index = 100×Σ(number of diseased leaves at each level × representative value at each level) / (total number of leaves surveyed × representative value of the highest level).

[0083] 3. Results and Analysis

[0084] The effects of the ΔMP-03 organic coupling cultivation technology on the changes in the disease index of soil-borne blight of greenhouse tomatoes, peppers, and cucumbers are as follows:

[0085] Figure 19 、 Figure 20 and Figure 21 The results showed that compared with MP-03 and WT (without adding bacterial suspension), the organic coupling cultivation technology of adding ΔMP-03 bacterial suspension to degrade corn straw significantly reduced the incidence and disease index of late blight of greenhouse tomatoes (caused by Phytophthora infestans), Phytophthora blight of cucumbers (caused by Phytophthora drechsleri), and Phytophthora blight of peppers (caused by Phytophthora capsici) from the seedling stage to the full fruit stage. Compared with the treatment with ΔMP-03 suspension, the organic coupling cultivation technology of MP-03 and WT suspensions to degrade corn straw showed a gradually increasing trend in the disease index of soil-borne blight at the seedling stage, early flowering stage, fruiting stage, and full fruit stage of the three vegetables, and the disease was the most severe at the full fruit stage. However, the organic coupling cultivation technology of ΔMP-03 suspension to degrade corn straw significantly reduced the incidence and disease index of soil-borne blight at different development stages of the three greenhouse vegetables. Therefore, the organic coupling cultivation technology of ΔMP-03 suspension to degrade corn straw significantly inhibited the incidence of soil-borne blight of greenhouse tomatoes, cucumbers, and peppers, and significantly reduced the occurrence and damage of soil-borne blight of tomatoes, cucumbers, and peppers.

[0086] Figure 19 、 Figure 20 and Figure 21 Different asterisks marked on each bar graph represent the significant difference degree of the disease index of soil-borne blight of each vegetable, *P<0.05, **P<0.01, ***P<0.001.

[0087] Comprehensive comparative analysis of the above results showed that, compared with MP-03 and WT (without adding bacterial suspension), in the conventional organic coupling cultivation technology, the technology of adding ΔMP-03 bacterial suspension to degrade corn straw in organic coupling cultivation: 1) significantly increased the degradation rate of corn straw, resulting in a significant decrease in the weight of corn straw and a significant increase in the degradation rates of the three major components of lignocellulose; 2) determined that at a temperature of 30 °C, an initial pH of 7.0, and adding farmyard manure or bacterial fertilizer, ΔMP-03 had a higher degradation effect on corn straw than MP-03; 3) clarified that the ΔMP-03 bacterial suspension had a stronger performance in activating the activities of endo-cellulase and exo-cellulase of corn straw than MP-03, significantly accelerating the production of cellobiose from corn straw and the decomposition and utilization rate of corn straw; 4) the technology of adding ΔMP-03 bacterial suspension to degrade corn straw in organic coupling cultivation increased the growth rates of plant height and stem diameter of facility tomato, pepper, and cucumber seedlings in the later stage of transplanting; 5) the technology of adding ΔMP-03 bacterial suspension to degrade corn straw in organic coupling cultivation enhanced the root growth and development of facility tomato, cucumber, and pepper seedlings in the later stage of transplanting, which was beneficial to increasing the growth potential and disease resistance of facility tomato, cucumber, and pepper; 6) the technology of adding ΔMP-03 bacterial suspension to degrade corn straw in organic coupling cultivation significantly increased the leaf area, chlorophyll content, photosynthetic efficiency, and light energy utilization rate of facility tomato, cucumber, and pepper; 7) the organic coupling cultivation technology with ΔMP-03 suspension significantly inhibited the incidence of soil-borne diseases of facility tomato, cucumber, and pepper, significantly improved the prevention and control effect, and reduced the occurrence and harm of soil-borne diseases of tomato, cucumber, and pepper.

[0088] Although the present invention has been described in detail with general descriptions and specific embodiments above, based on the present invention, some modifications or improvements can be made, which are obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of the present invention claimed.

Claims

1. An organic coupling cultivation method for protected vegetables, characterized in that, The method includes: digging square trenches 1 month before transplanting the seedlings of protected vegetables, and sequentially adding the following components: corn straw - Penicillium cyclopium ([ Penicillium cyclopium Penicillium cyclopium ) 455-1 bacterial suspension - vegetable virus vaccine - peanut cake - wheat bran - farmyard manure - microbial fertilizer - compound fertilizer - water, then covering with soil and sealing with plastic film. After 1 month, punch holes on the surface of the film-covered area to transplant the vegetable seedlings; Among them, Penicillium cyclopium 455-1 has the ability to degrade corn straw, and its preservation number is CGMCC No. 41541.

2. The method according to claim 1, characterized in that By weight percentage, the addition ratios of each component are as follows: corn straw 50%, peanut cake 8.2%, wheat bran 8.2%, farmyard manure 9.7%, microbial fertilizer 6.8%, compound fertilizer 5.2%, water 6.8%, 5% of Penicillium cyclopium 455-1 bacterial suspension with a concentration of 200 million spores / mL, and vegetable virus vaccine 0.1%. The main components of the farmyard manure are organic matter, nitrogen, phosphorus, and potassium. The microbial fertilizer is Kingenta Pro-Soil 1. The main components of the compound fertilizer are nitrogen, phosphorus, and potassium.

3. The method according to claim 2, wherein The vegetable virus vaccine is a weak strain vaccine or a double-stranded small RNA vaccine, purchased from Nanjing Remate Biotechnology Co., Ltd.

4. The method according to claim 1, wherein The vegetables include tomatoes, peppers, and cucumbers.

5. Use of the method according to any one of claims 1-4 in the prevention and control of soil-borne diseases of tomatoes, peppers, and cucumbers.

6. The application according to claim 5, characterized in that The diseases include late blight of tomato caused by Phytophthora infestans ( Phytophthora infestans ), Phytophthora blight of cucumber caused by Phytophthora drechsleri ( Phytophthora drechsleri ), and Phytophthora blight of pepper caused by Phytophthora capsici ( Phytophthora capsici ).

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