Broad-spectrum tobacco epidemic disease resistant bio-organic fertilizer and preparation method thereof
By fermenting a mixture of straw, alcohol extract of Cyperus rotundus, water extract of Herba Cyperi and water extract of Ferns with mixed fermentation strains, a broad-spectrum bio-organic fertilizer with resistance to tobacco diseases was prepared, which solved the problem that the existing technology could only prevent and control a single tobacco disease and achieved efficient prevention and control effects on multiple diseases.
Patent Information
- Application Number
- CN202510901421.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-01
- Publication Date
- 2025-09-23
AI Technical Summary
Existing bio-organic fertilizers can only effectively prevent and control a certain disease in the prevention and control of tobacco diseases, and it is difficult to achieve effective prevention and control effects on multiple diseases at the same time, which limits its scope of application.
A mixture of straw, Cyperus rotundus alcohol extract residue, Herba schizonepetae water extract residue, and Pteris fasciata water extract residue was fermented using mixed fermentation strains Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 to prepare a broad-spectrum bio-organic fertilizer with resistance to tobacco blight. The specific steps included optimizing the strain ratio, extraction method, and fermentation conditions.
The prepared bio-organic fertilizer has a highly effective control effect on tobacco bacterial wilt, black shank disease and brown spot disease, with a control effect of more than 85%. It overcomes the single control defect of traditional bio-organic fertilizer and has good industrial application prospects.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of bio-organic fertilizers, and particularly relates to a bio-organic fertilizer with broad-spectrum resistance to tobacco diseases and a preparation method thereof. Background Art
[0002] As a globally important cash crop, tobacco's yield and quality directly impact farmers' income and the sustainable development of the tobacco industry. Under traditional cultivation methods, excessive use of chemical fertilizers leads to soil degradation, imbalanced microbial communities, and reduced tobacco leaf quality. The rational application of organic fertilizers is considered an important way to alleviate these issues.
[0003] Multiple field trials have shown that combining organic and chemical fertilizers can significantly improve tobacco yield and quality. For example, Chen et al. found in tobacco-growing areas in Yunnan that when organic fertilizer replaced 30% of chemical fertilizer, nicotine and total sugar content in tobacco leaves became more balanced, and sensory quality scores increased by 12.5%. Furthermore, organic fertilizers can promote root development, increase the thickness of mid- and upper-leaf leaves, and improve the appearance of flue-cured tobacco leaves.
[0004] Organic fertilizers also play a role in preventing and controlling tobacco diseases. They reduce disease incidence by enhancing plant disease resistance and soil antibacterial capacity. Studies have shown that applying well-rotted sheep manure can reduce the incidence of tobacco black shank by 28% to 35%. This mechanism may be related to the colonization of antagonistic microorganisms (such as Bacillus subtilis) in organic fertilizers and the induction of systemic resistance (ISR).
[0005] However, to the best of the inventor's knowledge, existing bio-organic fertilizers developed for tobacco disease prevention and control are often only highly effective against a specific disease, and are unable to effectively prevent and control multiple diseases simultaneously. This limits the application of existing bio-organic fertilizers and makes their further promotion and application difficult. Summary of the Invention
[0006] In view of the shortcomings of the prior art, the object of the present invention is to provide a method for preparing a bio-organic fertilizer, which can simultaneously have excellent preventive and control effects on multiple serious diseases.
[0007] The present invention provides the following technical solutions:
[0008] A method for preparing a broad-spectrum tobacco disease-resistant bio-organic fertilizer, comprising the following steps:
[0009] The straw, the alcohol extract residue of Cyperus rotundus, the water extract residue of Herba Cyperi, and the water extract residue of Pteris chinensis are mixed in a weight ratio of 3:1-2:3-5:1-2 to serve as a fermentation substrate;
[0010] Using a mixed bacteria of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 as fermentation bacteria, fermenting a fermentation substrate to obtain the bio-organic fertilizer;
[0011] The Bacillus subtilis B115 has a preservation number of CGMCC No. 1210; the Bacillus amyloliquefaciens ZH1 has a preservation number of CGMCC No. 8803.
[0012] Preferably, the weight ratio of the straw, the alcohol extract residue of Cyperus rotundus, the water extract residue of Herba Achyranthis Bidentatae, and the water extract residue of Pteris crenatae is 3:1:4:2.
[0013] Preferably, the preparation method of the Cyperus rotundus alcohol-extracted residue is: washing and chopping the Cyperus rotundus, adding it to a 70% v / v ethanol aqueous solution with a weight that is 6 times that of the Cyperus rotundus, extracting it at 60°C for 2 hours, and filtering the filtrate; adding the filter residue to a 70% v / v ethanol aqueous solution with a weight that is 4 times that of the filter residue, extracting it at 60°C for 2 hours, filtering the residue phase, washing away the residual ethanol with pure water, and controlling the water content at 10-12% w / w.
[0014] Preferably, the preparation method of the water extract residue of the Herba Camphoratae is as follows: wash and chop the Herba Camphoratae, add the Herba Camphoratae to water 5 times the weight of the Herba Camphoratae, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue to water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content at 10-12% w / w.
[0015] Preferably, the preparation method of the water extract of the fern is as follows: wash the fern, chop it, add it to water 5 times the weight of the fern, extract it at 80°C for 4 hours, and filter the filtrate; add the filter residue to water 3 times the weight of the filter residue, extract it at 80°C for 4 hours, filter the residue phase, and control the water content at 10-12% w / w.
[0016] Preferably, the cell number ratio of the mixed bacteria of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 is 1:1.
[0017] Preferably, the inoculation amount of the mixed bacteria is 5%.
[0018] Preferably, during fermentation, the starting temperature is 30° C. and the fermentation time is 72 hours.
[0019] Preferably, the water content of the fermentation substrate is 20% w / w.
[0020] The present invention also provides bio-organic fertilizer prepared by the above preparation method.
[0021] Beneficial effects of the present invention:
[0022] The bio-organic fertilizer obtained by the present invention has a highly effective control effect on tobacco bacterial wilt, black shank disease and brown spot disease, and the control effect can reach more than 85%. It overcomes the defect that traditional bio-organic fertilizers can only control one tobacco disease, and has good industrial application prospects. DETAILED DESCRIPTION
[0023] The present invention is described in detail below through examples. It is necessary to point out that the following examples are only used to further illustrate the present invention and are not to be construed as limiting the scope of protection of the present invention. Non-essential improvements and adjustments made by those skilled in the art based on the above-mentioned invention still fall within the scope of protection of the present invention.
[0024] Example 1
[0025] 1. Raw materials:
[0026] Bacillus subtilis B115, with a deposit number of CGMCC NO.1210, is a publicly available strain purchased from the China General Microbiological Culture Collection Center.
[0027] Bacillus amyloliquefaciens ZH1, with a preservation number of CGMCC No. 8803, is a published strain purchased from the China General Microbiological Culture Collection Center.
[0028] Water extraction residue of Pteris viridis: Wash and chop the Pteris viridis into 5-7 mm pieces, add the Pteris viridis into water 5 times the weight of the Pteris viridis, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content at 10-12% w / w.
[0029] Water extraction of the residue of the Herba Aspergids: wash the Herba Aspergids, chop it into 5-7mm pieces, add it into water 5 times the weight of the Herba Aspergids, extract it at 80°C for 4 hours, and filter the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract it at 80°C for 4 hours, filter the residue phase, and control the water content at 10-12% w / w.
[0030] Alcohol extraction of Cyperus rotundus residue: Wash and chop the Cyperus rotundus into 5-7 mm pieces, add it to a 70% v / v ethanol aqueous solution 6 times the weight of the Cyperus rotundus, extract at 60°C for 2 hours, and filter the filtrate; add the filter residue to a 70% v / v ethanol aqueous solution 4 times the weight of the filter residue, extract at 60°C for 2 hours, filter the residue phase, wash away the residual ethanol with pure water, and control the water content to 10-12% w / w.
[0031] Straw: collected from local corn fields.
[0032] 2. Preparation of organic fertilizer
[0033] Straw, Cyperus rotundus alcohol extract residue, Herba Cyperi water extract residue, and Pteris viridis water extract residue were mixed in a weight ratio of 3:1:4:2, and sprayed with water to a water content of 20% w / w to serve as a fermentation substrate.
[0034] The seed liquid of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 were mixed as fermentation bacteria at a bacterial cell ratio of 1:1, a fermentation substrate was inoculated with a 5% inoculum amount, and fermentation was carried out for 72 hours at a starting fermentation temperature of 30°C to obtain an organic fertilizer.
[0035] Example 2
[0036] Compared with Example 1, the difference is that the ratio of each component of the fermentation substrate is different. The specific scheme is as follows:
[0037] 1. Raw materials:
[0038] Bacillus subtilis B115, with a deposit number of CGMCC NO.1210, is a publicly available strain purchased from the China General Microbiological Culture Collection Center.
[0039] Bacillus amyloliquefaciens ZH1, with a preservation number of CGMCC No. 8803, is a published strain purchased from the China General Microbiological Culture Collection Center.
[0040] Water extraction residue of Pteris viridis: Wash and chop the Pteris viridis into 5-7 mm pieces, add the Pteris viridis into water 5 times the weight of the Pteris viridis, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content at 10-12% w / w.
[0041] Water extraction of the residue of the Herba Aspergids: wash the Herba Aspergids, chop it into 5-7mm pieces, add it into water 5 times the weight of the Herba Aspergids, extract it at 80°C for 4 hours, and filter the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract it at 80°C for 4 hours, filter the residue phase, and control the water content at 10-12% w / w.
[0042] Alcohol extraction of Cyperus rotundus residue: Wash and chop the Cyperus rotundus into 5-7 mm pieces, add it to a 70% v / v ethanol aqueous solution 6 times the weight of the Cyperus rotundus, extract at 60°C for 2 hours, and filter the filtrate; add the filter residue to a 70% v / v ethanol aqueous solution 4 times the weight of the filter residue, extract at 60°C for 2 hours, filter the residue phase, wash away the residual ethanol with pure water, and control the water content to 10-12% w / w.
[0043] Straw: collected from local corn fields.
[0044] 2. Preparation of organic fertilizer
[0045] Straw, Cyperus rotundus alcohol extract residue, Herba Cyperi water extract residue, and Pteris viridis water extract residue were mixed in a weight ratio of 3:2:3:1, and sprayed with water to a water content of 20% w / w to serve as a fermentation substrate.
[0046] The seed liquid of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 were mixed as fermentation bacteria at a bacterial cell ratio of 1:1, a fermentation substrate was inoculated with a 5% inoculum amount, and fermentation was carried out for 72 hours at a starting fermentation temperature of 30°C to obtain an organic fertilizer.
[0047] Example 3
[0048] Compared with Example 1, the difference is that the ratio of each component of the fermentation substrate is different. The specific scheme is as follows:
[0049] 1. Raw materials:
[0050] Bacillus subtilis B115, with a deposit number of CGMCC NO.1210, is a publicly available strain purchased from the China General Microbiological Culture Collection Center.
[0051] Bacillus amyloliquefaciens ZH1, with a preservation number of CGMCC No. 8803, is a published strain purchased from the China General Microbiological Culture Collection Center.
[0052] Water extraction residue of Pteris viridis: Wash and chop the Pteris viridis into 5-7 mm pieces, add the Pteris viridis into water 5 times the weight of the Pteris viridis, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content at 10-12% w / w.
[0053] Water extraction of the residue of the Herba Aspergids: wash the Herba Aspergids, chop it into 5-7mm pieces, add it into water 5 times the weight of the Herba Aspergids, extract it at 80°C for 4 hours, and filter the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract it at 80°C for 4 hours, filter the residue phase, and control the water content at 10-12% w / w.
[0054] Alcohol extraction of Cyperus rotundus residue: Wash and chop the Cyperus rotundus into 5-7 mm pieces, add it to a 70% v / v ethanol aqueous solution 6 times the weight of the Cyperus rotundus, extract at 60°C for 2 hours, and filter the filtrate; add the filter residue to a 70% v / v ethanol aqueous solution 4 times the weight of the filter residue, extract at 60°C for 2 hours, filter the residue phase, wash away the residual ethanol with pure water, and control the water content to 10-12% w / w.
[0055] Straw: collected from local corn fields.
[0056] 2. Preparation of organic fertilizer
[0057] Straw, Cyperus rotundus alcohol extract residue, Herba Cyperi water extract residue, and Pteris fasciata water extract residue were mixed in a weight ratio of 3:2:5:2, and sprayed with water to a water content of 20% w / w to serve as a fermentation substrate.
[0058] The seed liquid of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 were mixed as fermentation bacteria at a bacterial cell ratio of 1:1, a fermentation substrate was inoculated with a 5% inoculum amount, and fermentation was carried out for 72 hours at a starting fermentation temperature of 30°C to obtain an organic fertilizer.
[0059] Comparative Example 1
[0060] Compared with Example 1, the difference is that the fermentation bacteria is only Bacillus subtilis B115, and the rest is the same as Example 1. The specific scheme is as follows:
[0061] 1. Raw materials:
[0062] Bacillus subtilis B115, with a deposit number of CGMCC NO.1210, is a publicly available strain purchased from the China General Microbiological Culture Collection Center.
[0063] Water extraction residue of Pteris viridis: Wash and chop the Pteris viridis into 5-7 mm pieces, add the Pteris viridis into water 5 times the weight of the Pteris viridis, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content at 10-12% w / w.
[0064] Water extraction of the residue of the Herba Aspergids: wash the Herba Aspergids, chop it into 5-7mm pieces, add it into water 5 times the weight of the Herba Aspergids, extract it at 80°C for 4 hours, and filter the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract it at 80°C for 4 hours, filter the residue phase, and control the water content at 10-12% w / w.
[0065] Alcohol extraction of Cyperus rotundus residue: Wash and chop the Cyperus rotundus into 5-7 mm pieces, add it to a 70% v / v ethanol aqueous solution 6 times the weight of the Cyperus rotundus, extract at 60°C for 2 hours, and filter the filtrate; add the filter residue to a 70% v / v ethanol aqueous solution 4 times the weight of the filter residue, extract at 60°C for 2 hours, filter the residue phase, wash away the residual ethanol with pure water, and control the water content to 10-12% w / w.
[0066] Straw: collected from local corn fields.
[0067] 2. Preparation of organic fertilizer
[0068] Straw, Cyperus rotundus alcohol extract residue, Herba Cyperi water extract residue, and Pteris viridis water extract residue were mixed in a weight ratio of 3:1:4:2, and sprayed with water to a water content of 20% w / w to serve as a fermentation substrate.
[0069] Bacillus subtilis B115 was used as a fermentation bacterium, the fermentation substrate was inoculated at an inoculation rate of 5%, and the fermentation was carried out at a starting fermentation temperature of 30° C. for 72 hours to obtain an organic fertilizer.
[0070] Comparative Example 2
[0071] Compared with Example 1, the difference is that the fermentation substrate does not contain the residue of the water extract of the Herba Glechomae var. chinensis. The rest is the same as Example 1. The specific scheme is as follows:
[0072] 1. Raw materials:
[0073] Bacillus subtilis B115, with a deposit number of CGMCC NO.1210, is a publicly available strain purchased from the China General Microbiological Culture Collection Center.
[0074] Bacillus amyloliquefaciens ZH1, with a preservation number of CGMCC No. 8803, is a published strain purchased from the China General Microbiological Culture Collection Center.
[0075] Water extraction residue of Pteris viridis: Wash and chop the Pteris viridis into 5-7 mm pieces, add the Pteris viridis into water 5 times the weight of the Pteris viridis, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content at 10-12% w / w.
[0076] Alcohol extraction of Cyperus rotundus residue: Wash and chop the Cyperus rotundus into 5-7 mm pieces, add it to a 70% v / v ethanol aqueous solution 6 times the weight of the Cyperus rotundus, extract at 60°C for 2 hours, and filter the filtrate; add the filter residue to a 70% v / v ethanol aqueous solution 4 times the weight of the filter residue, extract at 60°C for 2 hours, filter the residue phase, wash away the residual ethanol with pure water, and control the water content to 10-12% w / w.
[0077] Straw: collected from local corn fields.
[0078] 2. Preparation of organic fertilizer
[0079] Straw, Cyperus rotundus alcohol extract residue and Pteris viridis water extract residue were mixed in a weight ratio of 3:1:2, and sprayed with water to a water content of 20% w / w to serve as a fermentation substrate.
[0080] The seed liquid of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 were mixed as fermentation bacteria at a bacterial cell ratio of 1:1, a fermentation substrate was inoculated with a 5% inoculum amount, and fermentation was carried out for 72 hours at a starting fermentation temperature of 30°C to obtain an organic fertilizer.
[0081] Comparative Example 3
[0082] Compared with Example 1, the difference is that the Cyperus rotundus alcohol extract residue is replaced by the Cyperus rotundus water extract residue, and the rest is the same as Example 1. The specific scheme is as follows:
[0083] 1. Raw materials:
[0084] Bacillus subtilis B115, with a deposit number of CGMCC NO.1210, is a publicly available strain purchased from the China General Microbiological Culture Collection Center.
[0085] Bacillus amyloliquefaciens ZH1, with a preservation number of CGMCC No. 8803, is a published strain purchased from the China General Microbiological Culture Collection Center.
[0086] Water extraction residue of Pteris viridis: Wash and chop the Pteris viridis into 5-7 mm pieces, add the Pteris viridis into water 5 times the weight of the Pteris viridis, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content at 10-12% w / w.
[0087] Water extraction of the residue of the Herba Aspergids: wash the Herba Aspergids, chop it into 5-7mm pieces, add it into water 5 times the weight of the Herba Aspergids, extract it at 80°C for 4 hours, and filter the filtrate; add the filter residue into water 3 times the weight of the filter residue, extract it at 80°C for 4 hours, filter the residue phase, and control the water content at 10-12% w / w.
[0088] Cyperus rotundus water extraction residue: wash and chop the Cyperus rotundus (5-7 mm pieces), add to water 5 times the weight of the Cyperus rotundus, extract at 80°C for 4 hours, and filter to obtain the filtrate; add the filter residue to water 3 times the weight of the filter residue, extract at 80°C for 4 hours, filter to obtain the residue phase, and control the water content to 10-12% w / w.
[0089] Straw: collected from local corn fields.
[0090] 2. Preparation of organic fertilizer
[0091] Straw, Cyperus rotundus water extract residue, Herba Cyperi serratae water extract residue, and Pteris fasciata water extract residue were mixed in a weight ratio of 3:1:4:2, and sprayed with water to a water content of 20% w / w to serve as a fermentation substrate.
[0092] The seed liquid of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 were mixed as fermentation bacteria at a bacterial cell ratio of 1:1, a fermentation substrate was inoculated with a 5% inoculum amount, and fermentation was carried out for 72 hours at a starting fermentation temperature of 30°C to obtain an organic fertilizer.
[0093] Comparative Example 4
[0094] 1. Raw materials:
[0095] Preparation of water extract from Pteris viridis
[0096] Wash the fern and chop it into 5-7 mm pieces. Add it into water 5 times the weight of the fern, extract it at 80°C for 4 hours, and filter the filtrate. Add the filter residue into water 3 times the weight of the filter residue, extract it at 80°C for 4 hours, filter the residue phase, and control the water content at 10-12% w / w.
[0097] King oyster mushroom residue: purchased from Huamo (Shandong) Biotechnology Co., Ltd.
[0098] Corn stalks: collected by local farmers.
[0099] The water extract residue of Pteris podioides, Pleurotus eryngii residue (purchased from Huamo (Shandong) Biotechnology Co., Ltd.) and corn stalks (collected from local farmers) were mixed in a weight ratio of 1:3:1, and the water content was controlled to 20% w / w by spraying water.
[0100] Prepare fermentation substrate, use Bacillus subtilis B115 as fermentation bacteria, inoculate the fermentation bacteria at an inoculum amount of 5%, start fermentation at 30° C., and control the fermentation time to be 72 hours.
[0101] Experimental Example 1
[0102] This experimental example investigates the effects of the organic fertilizers obtained from the examples and comparative examples on preventing and treating tobacco bacterial wilt.
[0103] Take soil with a bacterial wilt incidence rate of 100% as potted soil for the test, 10 kg of soil per pot, and transplant one tobacco seedling per pot. After transplanting the tobacco, the organic fertilizer obtained in Examples 1 to 3 and 1 to 4 was used as a base fertilizer, and 80 g was applied to each pot; then, on the 7th day after transplanting, a commercially available conventional water-soluble fertilizer (purchased from Guizhou Qiannan Jinfu Co., Ltd., N:P2O5:K2O=22:14:10, diluted with 100 times the weight of water) was sprayed once, with a spraying amount of 200 mL per pot, and then sprayed once every 10 days, with the same spraying amount as before.
[0104] The control group replaced the base fertilizer of each group with a special compound fertilizer for flue-cured tobacco (produced by Hubei Xiangqing Fertilizer Co., Ltd.). Other than that, the fertilizer application system was the same as that of each group.
[0105] The incidence rate, disease index and control effect of tobacco bacterial wilt were investigated. The experimental results are shown in Table 1.
[0106] Incidence rate = number of diseased plants / total number of surveyed plants × 100%.
[0107] The disease index is investigated according to the tobacco disease classification and investigation method of the tobacco industry standard of the People's Republic of China:
[0108] Level 0: All plants are disease-free;
[0109] Level 1: There are occasional chlorotic spots on the stem, or a few leaves wither on the side with streaks;
[0110] Level 2: There are black streaks on the stem but they have not yet reached the top, or more than half of the leaves on the diseased side are withered;
[0111] Level 3: Black streaks on the stem reach the top of the plant, or more than 2 / 3 of the leaves on the diseased side wilt; Level 4: The diseased plant is basically dead.
[0112] Disease index = [∑(disease level × number of strains at this level) / (highest level × total number of strains)] × 100.
[0113] Control effect = (control disease index - treatment disease index) / control disease index × 100%.
[0114] Table 1
[0115]
[0116]
[0117] Experimental Example 2
[0118] This experimental example investigates the effects of the organic fertilizers obtained from the examples and comparative examples on preventing and treating tobacco black shank disease.
[0119] Take the soil with 100% black shank disease incidence as potted soil for the test, 10 kg of soil per pot, and transplant one tobacco seedling per pot. After transplanting the tobacco, the organic fertilizer obtained in Examples 1 to 3 and 1 to 4 was used as a base fertilizer, and 80 g was applied to each pot; then, on the 7th day after transplanting, a commercially available conventional water-soluble fertilizer (purchased from Guizhou Qiannan Jinfu Co., Ltd., N: P2O5: K2O = 22:14:10, diluted with 100 times the weight of water) was sprayed once, and the spraying amount was 200 mL per pot. Thereafter, the fertilizer was sprayed once every 10 days, and the spraying amount was the same as before.
[0120] The control group replaced the base fertilizer of each group with a special compound fertilizer for flue-cured tobacco (produced by Hubei Xiangqing Fertilizer Co., Ltd.). Other than that, the fertilizer application system was the same as that of each group.
[0121] The incidence, disease index and control effect of tobacco black shank were investigated. The experimental results are shown in Table 2.
[0122] Incidence rate = number of diseased plants / total number of surveyed plants × 100%.
[0123] The disease index is investigated according to the tobacco disease classification and investigation method of the tobacco industry standard of the People's Republic of China:
[0124] Level 0: disease-free, no lesions at the base of the stem.
[0125] Level 1: Slight discoloration or water-soaked spots at the base of the stem, with the length of the spots ≤ 1 / 3 of the stem circumference.
[0126] Level 2: The lesions extend to 1 / 3 to 2 / 3 of the stem circumference, and the plant wilts slightly.
[0127] Level 3: The lesions cover more than 2 / 3 of the stem circumference, the plant wilts obviously or some leaves die.
[0128] Level 4: The whole plant wilts and dies, and the base of the stem is completely rotten.
[0129] Disease index = [∑(disease level × number of strains at this level) / (highest level × total number of strains)] × 100.
[0130] Control effect = (control disease index - treatment disease index) / control disease index × 100%.
[0131] Table 2
[0132]
[0133] Experimental Example 3
[0134] This experimental example investigates the effects of the organic fertilizers obtained from the examples and comparative examples on preventing and treating tobacco brown spot disease.
[0135] Take soil with 100% brown spot disease incidence as potted soil for the test, 10 kg of soil per pot, and transplant one tobacco seedling per pot. After transplanting the tobacco, the organic fertilizer obtained in Examples 1 to 3 and 1 to 4 was used as base fertilizer, and 80 g was applied to each pot; then, on the 7th day after transplanting, a commercially available conventional water-soluble fertilizer (purchased from Guizhou Qiannan Jinfu Co., Ltd., N:P2O5:K2O=22:14:10, diluted with 100 times the weight of water) was sprayed once, with a spraying amount of 200 mL per pot, and then sprayed once every 10 days, with the same spraying amount as before.
[0136] The control group replaced the base fertilizer of each group with a special compound fertilizer for flue-cured tobacco (produced by Hubei Xiangqing Fertilizer Co., Ltd.). Other than that, the fertilizer application system was the same as that of each group.
[0137] The incidence, disease index, and control effect of tobacco brown spot disease were investigated. The experimental results are shown in Table 3.
[0138] Incidence rate = number of diseased plants / total number of surveyed plants × 100%.
[0139] The disease index is investigated according to the tobacco disease classification and investigation method of the tobacco industry standard of the People's Republic of China:
[0140] Level 0: No lesions.
[0141] Level 1: The lesion area accounts for ≤5% of the leaf area, or the number of lesions on a single leaf is ≤5.
[0142] Level 2: The lesion area accounts for 6% to 15% of the leaf area, or the number of lesions on a single leaf is 6 to 10.
[0143] Level 3: The lesion area accounts for 16% to 25% of the leaf area, or the number of lesions on a single leaf is 11 to 20.
[0144] Level 4: The area of lesions accounts for >25% of the leaf area, or the number of lesions on a single leaf is >20, and the leaves are severely wrinkled or partially dead.
[0145] Disease index = [∑(disease level × number of strains at this level) / (highest level × total number of strains)] × 100.
[0146] Control effect = (control disease index - treatment disease index) / control disease index × 100%.
[0147] Table 3
[0148]
Claims
1. A method for preparing a broad-spectrum tobacco disease-resistant bio-organic fertilizer, characterized in that: The preparation method comprises the following steps: The straw, the alcohol extract residue of Cyperus rotundus, the water extract residue of Herba Cyperi, and the water extract residue of Pteris chinensis are mixed in a weight ratio of 3:1-2:3-5:1-2 to serve as a fermentation substrate; Using a mixed bacteria of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 as fermentation bacteria, fermenting a fermentation substrate to obtain the bio-organic fertilizer; The Bacillus subtilis B115 has a preservation number of CGMCC No. 1210; the Bacillus amyloliquefaciens ZH1 has a preservation number of CGMCC No. 8803.
2. The preparation method according to claim 1, characterized in that The weight ratio of the straw, the alcohol extract residue of Cyperus rotundus, the water extract residue of Herba Cyperi, and the water extract residue of Pteris viridis is 3:1:4:
2.
3. The preparation method according to claim 1, characterized in that The preparation method of the Cyperus rotundus alcohol-extracted residue comprises the following steps: washing and chopping the Cyperus rotundus, adding the Cyperus rotundus to a 70% v / v ethanol aqueous solution whose weight is 6 times that of the Cyperus rotundus, extracting at 60°C for 2 hours, and filtering to obtain the filtrate; adding the filter residue to a 70% v / v ethanol aqueous solution whose weight is 4 times that of the filter residue, extracting at 60°C for 2 hours, filtering to obtain the residue phase, washing away the residual ethanol with pure water, and controlling the water content to be between 10% and 12% w / w.
4. The preparation method according to claim 1, characterized in that The preparation method of the aqueous extract of the Chinese yarrow stalk comprises the following steps: washing and chopping the Chinese yarrow stalk, adding the extract to water 5 times the weight of the Chinese yarrow stalk, extracting at 80°C for 4 hours, and filtering to obtain the filtrate; adding the filter residue to water 3 times the weight of the filter residue, extracting at 80°C for 4 hours, filtering to obtain the residue phase, and controlling the water content to be 10-12% w / w.
5. The preparation method according to claim 1, characterized in that The preparation method of the water extract residue of the fern is as follows: washing and chopping the fern, adding the fern to water 5 times the weight of the fern, extracting at 80°C for 4 hours, and filtering to obtain the filtrate; adding the filter residue to water 3 times the weight of the filter residue, extracting at 80°C for 4 hours, filtering to obtain the residue phase, and controlling the water content to be 10-12% w / w.
6. The preparation method according to claim 1, characterized in that The cell number ratio of the mixed bacteria of Bacillus subtilis B115 and Bacillus amyloliquefaciens ZH1 was 1:
1.
7. The preparation method according to claim 1 or 6, characterized in that The inoculation amount of the mixed bacteria is 5%.
8. The preparation method according to claim 7, characterized in that During fermentation, the starting temperature is 30°C and the fermentation time is 72 hours.
9. The preparation method according to claim 8, characterized in that The water content of the fermentation substrate is 20% w / w.
10. A broad-spectrum bio-organic fertilizer resistant to tobacco diseases, characterized in that: The bio-organic fertilizer is prepared by the preparation method according to any one of claims 1 to 9.