Organic fertilizer for tobacco based on mild fermentation method using rape stalks as raw materials and preparation method thereof

Abstract

The present invention belongs to the technical field of organic fertilizers and microbial fertilizers. The present invention provides a method for preparing tobacco organic fertilizer based on a mild fermentation method using rapeseed stalks as raw materials, which comprises the following steps: chopping the rapeseed stalks and steaming them in a steam atmosphere, and then placing them in a steam explosion tank for steam explosion treatment; removing moisture from the surface of the rapeseed stalks after the steam explosion treatment, mashing the stems of Dipsacus asper and placing them together in a fermentation liquid for 2.5 to 3.5 hours; then inoculating fermentation bacteria, fermenting for 6 to 8 hours, and collecting the fermentation product. The tobacco organic fertilizer provided by the present invention has a short production cycle, with the fermentation stage taking less than 10 hours, and can be efficiently connected with other production links, and has good prospects for mechanized production. At the same time, the tobacco organic fertilizer obtained by the present invention can significantly improve the yield and quality of tobacco leaves.

Description

Technical Field

[0001] The invention belongs to the technical field of organic fertilizers and microbial fertilizers, and particularly relates to an organic fertilizer for tobacco produced by a mild fermentation method using rapeseed stalks as a raw material and a preparation method thereof. Background Art

[0002] Bio-organic fertilizer is a new type of fertilizer with the advantages of low production cost and rich nutrients. It has been widely used in the agricultural field. Specifically in tobacco cultivation, bio-organic fertilizer is also being used more and more widely. It has certain advantages in improving tobacco yield and quality, regulating soil microbial ecology, improving soil, increasing yield and quality, improving tobacco plant resistance, reducing the incidence of soil-borne diseases and increasing the yield of green tobacco. [1] .

[0003] However, with the large-scale production and use of bio-organic fertilizers, people have also found that they have some disadvantages, mainly reflected in: (1) the production time is long. The production cycle of bio-organic fertilizers prepared by fermentation is usually several days to dozens of days; (2) the environmental protection cost is high. Since microbial fermentation usually adopts methods such as composting that are easy to pollute the surrounding environment, the environmental protection cost of producing bio-organic fertilizers is high; (3) the effect on increasing tobacco yield, improving quality and resisting diseases is limited.

[0004] Numerous attempts have been made to shorten the fermentation cycle. For example, Wang Lin et al. of Hubei China Tobacco Industrial Co., Ltd., in Chinese Patent No. 102108031B, provide a method for rapidly fermenting tobacco cake fertilizer. This method requires only 15 to 25 days of fermentation to produce cake fertilizer for tobacco cultivation. Another example is Shi Zhentuo et al., in Chinese Patent No. 103449863B, provide a method for rapidly fermenting compost from traditional Chinese medicine residues to produce organic fertilizer. This method requires only 5 to 7 days of composting. These two patents, representative of the methods mentioned above, achieve this goal by adding a composting agent and selecting specific fermentation substrates and fermentation bacteria. In addition, research has been conducted on improving the physical environment of fermentation. For example, Yan Weiwei et al. proposed a method for rapid fermentation of olive fertilizer in Chinese Patent CN104892051B. This method involves digging a special fermentation tank and treating it with lime powder, utilizing the principle that lime generates heat when it comes into contact with water to accelerate the fermentation of fertilizer. Another example is Chi Jingliang et al., who proposed a method for rapidly fermenting livestock and poultry manure to produce organic fertilizer in a fermentation tunnel in Chinese Patent CN105503304B. This method, by setting up a special fermentation tunnel, can shorten the fermentation cycle to 4-5 days. Furthermore, researchers have also shortened the fermentation cycle by selecting special fermentation bacteria. For example, Li Liyan et al. proposed a synergistic bacterial agent for rapid fermentation of organic materials in Chinese Patent CN 114045232B. This agent has the characteristics of rapid compost fermentation start-up, rapid temperature rise, and long-term high temperature maintenance, which can quickly and efficiently degrade organic materials.

[0005] However, to the best of the inventor's knowledge, the production cycle of existing bio-organic fertilizers used in tobacco cultivation is still relatively long, and methods with shorter production cycles still require more than 4 to 5 days for fermentation, which has not effectively addressed environmental cost issues. Furthermore, since fermentation is a relatively complex biological activity affected by multiple factors, the cost of controlling process stability is also relatively high. Therefore, shortening the fermentation cycle can also reduce corresponding production control costs.

[0006] At present, in the field of fermentation technology, there are some types of products with a short fermentation production cycle. These products only need to undergo mild fermentation (or micro-fermentation) to complete the production process. These products are mainly concentrated in the categories of pastries, tea, meat products and juice. For example, Ge Zhenzhen et al. [2] Wu Bin studied the effects of micro-fermentation on noodle flavor and quality characteristics during storage. [3] et al. studied the effect of mild fermentation on the volatile aroma components of sea buckthorn juice, and Huang Jing et al. [4]et al. studied the effects of light rolling and fermentation on black tea quality. Such methods, including those described above, only require minutes or hours to complete the required fermentation process. If the concept of light fermentation could be adopted in the production of bio-organic fertilizers, eliminating unnecessary fermentation time while still achieving the desired fertilizer effect, the aforementioned drawbacks of bio-organic fertilizers could be overcome.

[0007] As far as the inventor knows, there is no report in this regard yet.

[0008] [1] Li Jia, Li Lu, Yang Shengnan, He Pengpeng, Zhang Jishuai. Effects of bio-organic fertilizer on tobacco soil microorganisms and quality of flue-cured tobacco leaves[J]. Journal of Jiangxi Agricultural Sciences, 2019, 31(06): 63-67

[0009] [2] Ge Zhenzhen, Wang Weijing, Gao Shanshan, et al. Effects of micro-fermentation on noodle flavor and quality characteristics during storage[J]. Food Industry Science and Technology, 2021, 42(13): 8. DOI: 10.13386 / j.issn1002-0306.2020100025.

[0010] [3] Wu Bin, Jiang Lei, Hou Jichao, et al. Effect of mild fermentation on volatile aroma components of sea buckthorn juice [J]. Agricultural Products Processing (first half of the month), 2022(008):000.

[0011] [4] Huang Jing, Huang Huaisheng, Gong Le, et al. Quality analysis of black tea of ​​tea varieties ‘Xiangbo Green’ and ‘Qiuyeqi’ under light rolling and light fermentation conditions [J]. Tea Newsletter, 2024, 51(2): 202-209. Summary of the Invention

[0012] In response to the shortcomings of the prior art, the present invention aims to provide a tobacco organic fertilizer using rapeseed stalks as raw material through a mild fermentation process and a preparation method. This organic fertilizer significantly reduces the fermentation cycle required and can significantly increase tobacco yield and quality.

[0013] In order to achieve the above object, the present invention provides the following technical solutions:

[0014] A method for preparing tobacco organic fertilizer using rape stalks as raw materials based on a mild fermentation method comprises the following steps:

[0015] (1) preparing rape straw and teasel stems in a weight ratio of 100:3-5; chopping the rape straw and steaming it in a steam atmosphere for 15-25 minutes;

[0016] (2) removing moisture from the surface of the cooked rapeseed straw, placing it in a steam explosion tank, setting the pressure to 1.2-1.8 MPa, and steam explosion for 3-8 minutes;

[0017] (3) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid for 2.5 to 3.5 hours;

[0018] (4) removing the surface moisture of the product obtained in step (3), inoculating the fermentation bacteria by spraying, fermenting for 6 to 8 hours, and collecting the fermentation product;

[0019] The fermentation broth is composed of culture broth of Bacillus subtilis CH-001, Bacillus cereus DS1 and Pseudomonas sp. SG-1;

[0020] The fermentation bacteria is a mixed bacteria consisting of Bacillus subtilis CH-001, Bacillus cereus DS1 and Pseudomonas sp. SG-1;

[0021] The deposit number of the Bacillus subtilis CH-001 is CCTCC NO.M 2011111, the deposit number of the Bacillus cereus DS1 is CCTCC NO.M 2010145, and the deposit number of the Pseudomonas sp. SG-1 is CGMCC No.3556.

[0022] Preferably, the weight ratio of the rape straw to the Dipsacus asper stems is 100:5.

[0023] Preferably, the cooking is carried out at normal pressure in a steam atmosphere for 20 minutes.

[0024] Preferably, when performing the steam explosion treatment, the pressure is 1.5 MPa and the time is 5 minutes.

[0025] Preferably, when the rape straw and the teasel stem are placed in the fermentation liquid, the solid-liquid weight ratio is 1 g:10 mL.

[0026] Preferably, in step (3), the treatment temperature is 37° C. and the treatment time is 3 hours.

[0027] Preferably, in step (4), among the fermentation bacteria, the bacterial concentrations and volumes of Bacillus subtilis CH-001, Bacillus cereus DS1 and Pseudomonas sp. SG-1 are the same, and the inoculation amount of the fermentation bacteria is 8% v / v.

[0028] Preferably, the bacterial concentration of each bacteria in the fermentation bacteria is 1×10 9 cfu / mL.

[0029] Preferably, in step (4), the fermentation temperature is 37° C. and the fermentation time is 8 hours.

[0030] Another object of the present invention is to provide an organic fertilizer for tobacco prepared by the above preparation method.

[0031] Before the present invention is studied, the inventor realizes that the prior art, in terms of preparing bio-organic fertilizer, often needs to ferment in the open air or in a special composting production area, and the fermentation process is difficult to efficiently link with the mechanized processing links such as the pre-treatment of raw materials, so that the whole production line lacks continuity. Meanwhile, when the prior art is preparing bio-organic fertilizer, the required fermentation time is usually longer (several days to tens of days), and in the long fermentation process, it is easy to produce discomfort odor and fermentation waste liquid overflow phenomenon, thereby causing air pollution and soil pollution, making the corresponding environmental protection work cost higher. In order to solve this problem, researchers in this field have carried out many studies in terms of fermentation equipment system and adjustment fermentation process. Although effective, it is still impossible to fundamentally solve the problem that the fermentation time is too long.

[0032] The food industry is a highly mechanized industry. While many products require fermentation, the fermentation cycle is very short, a process generally referred to as mild fermentation or micro-fermentation. This short fermentation cycle allows for continuous production of related food products, resulting in a high degree of mechanization. Inspired by this, the inventors considered whether mild fermentation could be employed in the production of bio-organic fertilizers to achieve the desired fertilizer effect.

[0033] Unlike the production of fermented foods, organic fertilizers typically require fermentation to mature the substrate, facilitating the absorption of nutrients by crops. However, the fermentation process for these foods is solely for flavor enhancement, followed by subsequent cooked food processing (such as steaming, cooking, and frying). Therefore, the production objectives of the fermentation process are fundamentally different.

[0034] Bio-organic fertilizer is essentially that biological source material (such as plant straw, animal excrement) is carried out corresponding processing, becomes the small molecule substance that is more conducive to crop absorption.Generally speaking, only relying on the fermentation of a few hours, it is impossible to complete the aforementioned purpose.Therefore, the inventor considers that fermentation substrate (bio-organic fertilizer raw material) is carried out corresponding pre-treatment, makes fermentation substrate become the material that is easy to microorganism utilization and metabolism, thereby shortens whole fermentation cycle.

[0035] As shown in the experimental results related to the present invention, the inventors first tried steam-exploding rapeseed straw and then fermenting it for two days. The resulting product did not increase tobacco yield. They then used different fermentation bacteria, but the experimental results were still unsatisfactory. After continuous exploration, the inventors tried soaking the steam-exploded straw in a fermentation culture solution before fermenting it, and found that the resulting product did increase tobacco yield. Encouraged by this, the inventors continued to improve the treatment process and ultimately obtained a product with the desired fertilizer efficiency.

[0036] It is not difficult to know that among all the processes of the present invention, the physical treatment process accounts for a high proportion, the fermentation process is short in time and can be well connected with the pre- and post-treatment processes, has a high degree of mechanization, and can complete production work in a closed production line, avoiding unpleasant odors and the outflow of fermentation waste liquid.

[0037] As shown in the relevant embodiments of the present invention and the comparative examples, the steaming process, the steam explosion process, the selection of common raw materials, the selection of fermentation liquid and the selection of fermentation bacteria all play a vital role. When other fermentation liquids are selected for soaking treatment, the impact on tobacco leaf yield is relatively large. The reason may be that the related substances in the fermentation liquid have a relatively large impact on the degradation effect of the raw materials. Among them, the related substances may have synergistic and antagonistic effects, resulting in the effect of using a fermentation liquid of one fermentation bacteria or a mixed solution of three fermentation liquids being not as good as the solution of using a mixed solution of two fermentation liquids. In addition, it is very important to use the three fermentation bacteria as inoculation bacteria at the same time during the fermentation stage. The possible reason is that these three fermentation bacteria have a synergistic effect in the early stage of fermentation, which accelerates the utilization and metabolism of the fermentation substrate.

[0038] On this basis, the inventors investigated the effects of some schemes on tobacco leaf quality. As shown in another experimental result of the present invention, the selection of fermentation liquid and fermentation bacteria not only affects the yield of tobacco leaves, but also has a great impact on the quality of tobacco leaves (especially the content of neutral aroma components).

[0039] Furthermore, it should be noted that among the raw materials used in the present invention, rapeseed straw, as agricultural waste, is generally discarded or simply recycled. However, the present invention can use it as a raw material for producing high-quality tobacco organic fertilizer, achieving high-value reuse of rapeseed straw. Furthermore, teasel is a common weed in Yunnan, and the cost of harvesting or purchasing it is relatively low. Therefore, the present invention can achieve high-value utilization of the weed. Therefore, the raw material cost of the present invention is low, and it has good industrial economic efficiency.

[0040] Beneficial effects of the present invention:

[0041] The tobacco organic fertilizer provided by the present invention has a short production cycle, requiring less than 10 hours during the fermentation stage, and can be efficiently integrated with other production processes, showing good prospects for mechanized production. Furthermore, the tobacco organic fertilizer obtained by the present invention can significantly increase tobacco leaf yield and quality. DETAILED DESCRIPTION

[0042] 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.

[0043] Example 1

[0044] 1. Raw materials

[0045] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0046] Fermentation strains: Bacillus subtilis CH-001 (Deposit Number: CCTCC NO.M 2011111), Bacillus cereus DS1 (Deposit Number: CCTCC NO.M 2010145), Pseudomonas sp. SG-1 (Deposit Number: CGMCC No.3556), kindly provided by colleagues for research use; seed solutions of the three strains were prepared using LB medium, and the mixture was used for fermentation inoculation. The bacterial concentration of each strain after mixing was 1×10 9 cfu / mL.

[0047] Fermentation broth: The three fermentation strains were cultured in LB liquid medium until the plateau phase (the bacterial concentration was 1×10 8 cfu / mL), and then mixed in equal volumes.

[0048] 2. Preparation of organic fertilizer for tobacco

[0049] (1) preparing rape straw and teasel stems in a weight ratio of 100:5;

[0050] (2) Chop the rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 20 minutes;

[0051] (3) After steaming, the rape straw was dehydrated and placed in a steam explosion tank with a pressure of 1.5 MPa for 5 minutes.

[0052] (4) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid with a solid-liquid weight ratio of 1 g:10 mL, and treating at 35°C for 3 hours;

[0053] (5) removing moisture from the surface of the product obtained in step (4), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0054] Example 2

[0055] 1. Raw materials

[0056] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0057] Fermentation strains: Bacillus subtilis CH-001 (Deposit Number: CCTCC NO.M 2011111), Bacillus cereus DS1 (Deposit Number: CCTCC NO.M 2010145), Pseudomonas sp. SG-1 (Deposit Number: CGMCC No.3556), kindly provided by colleagues for research use; seed solutions of the three strains were prepared using LB medium, and the mixture was used for fermentation inoculation. The bacterial concentration of each strain after mixing was 1×10 9 cfu / mL.

[0058] Fermentation broth: The three fermentation strains were cultured in LB liquid medium until the plateau phase (the bacterial concentration was 1×10 8 cfu / mL), and then mixed in equal volumes.

[0059] 2. Preparation of organic fertilizer for tobacco

[0060] (1) preparing rape straw and Dipsacus asper stems in a weight ratio of 100:3;

[0061] (2) Chop the rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 15 minutes;

[0062] (3) After steaming, the rape straw was dehydrated and placed in a steam explosion tank with a pressure of 1.2 MPa for 8 minutes.

[0063] (4) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid with a solid-liquid weight ratio of 1 g:10 mL, and treating at 35°C for 3.5 hours;

[0064] (5) removing moisture from the surface of the product obtained in step (4), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 6 hours, and collecting the fermentation product.

[0065] Example 3

[0066] 1. Raw materials

[0067] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0068] Fermentation strains: Bacillus subtilis CH-001 (Deposit Number: CCTCC NO.M 2011111), Bacillus cereus DS1 (Deposit Number: CCTCC NO.M 2010145), Pseudomonas sp. SG-1 (Deposit Number: CGMCC No.3556), kindly provided by colleagues for research use; seed solutions of the three strains were prepared using LB medium, and the mixture was used for fermentation inoculation. The bacterial concentration of each strain after mixing was 1×10 9 cfu / mL.

[0069] Fermentation broth: The three fermentation strains were cultured in LB liquid medium until the plateau phase (the bacterial concentration was 1×10 8 cfu / mL) and then mixed in equal volumes.

[0070] 2. Preparation of organic fertilizer for tobacco

[0071] (1) preparing rape straw and teasel stems in a weight ratio of 100:4;

[0072] (2) Chop the rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 25 minutes;

[0073] (3) After steaming, the rape straw was dehydrated and placed in a steam explosion tank with a pressure of 1.8 MPa for 3 minutes.

[0074] (4) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid with a solid-liquid weight ratio of 1 g:10 mL, and treating at 35°C for 2.5 hours;

[0075] (5) removing moisture from the surface of the product obtained in step (4), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0076] Comparative Example 1

[0077] Except that the steam explosion process is not adopted, the rest is consistent with Example 1. The specific solution is as follows:

[0078] 1. Raw materials

[0079] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0080] Fermentation strains: Bacillus subtilis CH-001 (Deposit Number: CCTCC NO.M 2011111), Bacillus cereus DS1 (Deposit Number: CCTCC NO.M 2010145), Pseudomonas sp. SG-1 (Deposit Number: CGMCC No.3556), kindly provided by colleagues for research use; seed solutions of the three strains were prepared using LB medium, and the mixture was used for fermentation inoculation. The bacterial concentration of each strain after mixing was 1×10 9 cfu / mL.

[0081] Fermentation broth: The three fermentation strains were cultured in LB liquid medium until the plateau phase (the bacterial concentration was 1×10 8 cfu / mL) and then mixed in equal volumes.

[0082] 2. Preparation of organic fertilizer for tobacco

[0083] (1) preparing rape straw and teasel stems in a weight ratio of 100:5;

[0084] (2) Chop the rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 20 minutes;

[0085] (3) removing the surface moisture of the rape straw after cooking, mashing the stems of Dipsacus asper and placing them together in the fermentation liquid with a solid-liquid weight ratio of 1 g:10 mL, and treating at 35°C for 3 hours;

[0086] (4) removing moisture from the surface of the product obtained in step (3), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0087] Comparative Example 2

[0088] Except that the steaming process is not adopted, the rest is consistent with Example 1. The specific scheme is as follows:

[0089] 1. Raw materials

[0090] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0091] Fermentation strains: Bacillus subtilis CH-001 (Deposit Number: CCTCC NO.M 2011111), Bacillus cereus DS1 (Deposit Number: CCTCC NO.M 2010145), Pseudomonas sp. SG-1 (Deposit Number: CGMCC No.3556), kindly provided by colleagues for research use; seed solutions of the three strains were prepared using LB medium, and the mixture was used for fermentation inoculation. The bacterial concentration of each strain after mixing was 1×10 9 cfu / mL.

[0092] Fermentation broth: The three fermentation strains were cultured in LB liquid medium until the plateau phase (the bacterial concentration was 1×10 8 cfu / mL) and then mixed in equal volumes.

[0093] 2. Preparation of organic fertilizer for tobacco

[0094] (1) preparing rape straw and teasel stems in a weight ratio of 100:5;

[0095] (2) Cut the rape straw into pieces (length controlled within the range of 0.5-1.0 cm), place them in a steam explosion tank, set the pressure to 1.5 MPa, and steam explode for 5 minutes;

[0096] (3) After steam explosion, the surface moisture of rapeseed straw was removed, and the stems of Dipsacus asper were crushed and placed together with the stems in the fermentation liquid at a solid-liquid weight ratio of 1 g:10 mL, and the fermentation was carried out at 35°C for 3 hours;

[0097] (4) removing moisture from the surface of the product obtained in step (3), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0098] Comparative Example 3

[0099] Except that the fermentation broth was adjusted to fermentation broth of two fermentation bacteria, the rest was consistent with Example 1. The specific scheme is as follows:

[0100] 1. Raw materials

[0101] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0102] Fermentation strains: Bacillus subtilis CH-001 (Deposit Number: CCTCC NO.M 2011111) and Pseudomonas sp. SG-1 (Deposit Number: CGMCC No.3556), were kindly provided by colleagues for research use. The two strains were cultured in LB medium to prepare seed solutions, which were mixed for fermentation inoculation. The bacterial concentration of each strain after mixing was 1.5×10 9cfu / mL.

[0103] Fermentation broth: The two fermentation strains were cultured separately in LB liquid medium until the plateau phase (the bacterial concentration was 1×10 8 cfu / mL) and then mixed in equal volumes.

[0104] 2. Preparation of organic fertilizer for tobacco

[0105] (1) preparing rape straw and teasel stems in a weight ratio of 100:5;

[0106] (2) Chop the rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 20 minutes;

[0107] (3) After steaming, the rape straw was dehydrated and placed in a steam explosion tank with a pressure of 1.5 MPa for 5 minutes.

[0108] (4) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid with a solid-liquid weight ratio of 1 g:10 mL, and treating at 35°C for 3 hours;

[0109] (5) removing moisture from the surface of the product obtained in step (4), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0110] Comparative Example 4

[0111] Except that the fermentation liquid was adjusted to a fermentation liquid of a fermentation bacteria, the rest was consistent with Example 1. The specific scheme is as follows:

[0112] 1. Raw materials

[0113] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0114] Fermentation strain: Bacillus subtilis CH-001 (Deposit No.: CCTCC NO.M 2011111) was kindly donated by colleagues for research use; the bacteria were cultured in LB medium to prepare seed solution for fermentation inoculation. The bacterial concentration was 3×10 9 cfu / mL.

[0115] Fermentation broth: The fermentation strain was cultured in LB liquid medium until the plateau phase (bacterial concentration was 1×10 8 cfu / mL).

[0116] 2. Preparation of organic fertilizer for tobacco

[0117] (1) preparing rape straw and teasel stems in a weight ratio of 100:5;

[0118] (2) Chop the rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 20 minutes;

[0119] (3) After steaming, the rape straw was dehydrated and placed in a steam explosion tank with a pressure of 1.5 MPa for 5 minutes.

[0120] (4) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid with a solid-liquid weight ratio of 1 g:10 mL, and treating at 35°C for 3 hours;

[0121] (5) removing moisture from the surface of the product obtained in step (4), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0122] Comparative Example 5

[0123] Except that the fermentation bacteria were adjusted to a mixed bacteria composed of two bacteria, the rest was consistent with Example 1. The specific solution is as follows:

[0124] 1. Raw materials

[0125] Rapeseed straw: harvested locally in Yunnan; Dipsacus asper: fresh stems, harvested locally;

[0126] Fermentation strains: Bacillus subtilis CH-001 (Deposit No.: CCTCC NO.M 2011111), Bacillus cereus DS1 (Deposit No.: CCTCC NO.M 2010145), Pseudomonas sp. SG-1 (Deposit No.: CGMCC No.3556), were kindly provided by colleagues for research use; Bacillus subtilis CH-001 and Bacillus cereus DS1 were cultured in LB medium, and seed liquids were prepared separately. The mixed cultures were used for fermentation inoculation. The bacterial concentration of each strain after mixing was 1.5×10 9 cfu / mL.

[0127] Fermentation broth: Bacillus subtilis CH-001, Bacillus cereus DS1 and Pseudomonas sp. SG-1 were cultured in LB liquid medium to the stationary phase (the bacterial concentration was 1×10 8 cfu / mL) and then mixed in equal volumes.

[0128] 2. Preparation of organic fertilizer for tobacco

[0129] (1) preparing rape straw and teasel stems in a weight ratio of 100:5;

[0130] (2) Chop the rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 20 minutes;

[0131] (3) After steaming, the rape straw was dehydrated and placed in a steam explosion tank with a pressure of 1.5 MPa for 5 minutes.

[0132] (4) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid with a solid-liquid weight ratio of 1 g:10 mL, and treating at 35°C for 3 hours;

[0133] (5) removing moisture from the surface of the product obtained in step (4), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0134] Comparative Example 6

[0135] Except that the stems of Dipsacus asper were not added as raw materials, the rest were consistent with those in Example 1.

[0136] 1. Raw materials

[0137] Rapeseed straw: harvested locally in Yunnan;

[0138] Fermentation strains: Bacillus subtilis CH-001 (Deposit Number: CCTCC NO.M 2011111), Bacillus cereus DS1 (Deposit Number: CCTCC NO.M 2010145), Pseudomonas sp. SG-1 (Deposit Number: CGMCC No.3556), kindly provided by colleagues for research use; seed solutions of the three strains were prepared using LB medium, and the mixture was used for fermentation inoculation. The bacterial concentration of each strain after mixing was 1×10 9 cfu / mL.

[0139] Fermentation broth: The three fermentation strains were cultured in LB liquid medium until the plateau phase (the bacterial concentration was 1×10 8 cfu / mL) and then mixed in equal volumes.

[0140] 2. Preparation of organic fertilizer for tobacco

[0141] (1) Chop rape straw into small pieces (length controlled within the range of 0.5-1.0 cm) and steam them under a steam atmosphere (normal pressure) for 20 minutes;

[0142] (2) Remove the surface moisture of the cooked rapeseed straw, place it in a steam explosion tank, set the pressure to 1.5 MPa, and steam explode for 5 minutes;

[0143] (3) After steam explosion, the rape straw was dehydrated and placed in a fermentation solution with a solid-liquid weight ratio of 1 g:10 mL at 35 °C for 3 h.

[0144] (4) removing moisture from the surface of the product obtained in step (3), inoculating the fermentation bacteria by spraying, with an inoculum amount of 8% v / v, fermenting at 37° C. for 8 hours, and collecting the fermentation product.

[0145] Experimental example

[0146] 1. Test Soil

[0147] The previous crop of the soil used in the experiment was flue-cured tobacco, and the physical and chemical properties of the soil are shown in Table 1.

[0148] Table 1

[0149]

[0150] 2. Test items and test methods

[0151] 2.1 Economic characteristics survey:

[0152] Each plot is harvested, rod-woven, baked, graded and yield calculated separately, and graded according to the national standard GB2635-92, and the yield and the proportion of upper and middle tobacco are calculated separately.

[0153] 2.2 Content of neutral aroma components in tobacco leaves:

[0154] Static headspace gas chromatography-mass spectrometry (GC-MS) was used to determine the volatile compounds in tobacco leaves. Specifically, 0.5 g of cut tobacco sample was weighed into a 20 mL headspace vial, immediately sealed with aluminum foil, and gently shaken to loosen the sample. The vial was then placed in a headspace sampler for equilibration and then loaded into the GC-MS for data acquisition.

[0155] As shown in Table 2, the effects of different products obtained in Examples 1 to 3 and Comparative Examples 1 to 6 on tobacco leaf yield were investigated. Each group broadcasted the fertilizer before rotary tillage and ridging, with the application rate of 800 kg / hectare; the control group broadcasted HK microbial organic fertilizer (N+P2O5+K2O=8%) produced by Guangxi Dahua Fertilizer Co., Ltd. before rotary tillage and ridging, with the application rate of 800 kg / hectare.

[0156] Specifically, the fertilizer used in the control group of this experiment was a mature commercial product with a certain growth-promoting effect. The inventor adjusted its application rate to 800 kg / hectare without topdressing. Under this condition, its yield was 2167.2 kg / hm 2. In Examples 1 to 3 and Comparative Examples 1 to 5 serving as the experimental groups, the products of Examples 1-3 have a significant promoting effect on tobacco leaf yield, and the yield is significantly higher than that of the control group. It can be seen that the resulting products can not only be used as bio-organic fertilizers, but also have a growth-promoting effect that is better than general commercially available products; however, the products obtained in Comparative Examples 3-6 are all inferior to the control group to varying degrees, indicating that the selection of fermentation liquid used for soaking treatment, fermentation bacteria used for mild fermentation, and fermentation substrates has a significant impact on the fertilizer efficiency of the resulting products; in addition, it can be seen from Comparative Examples 1 and 2 that whether to perform steam explosion treatment and whether to combine steaming and steam explosion treatment are also key to the present invention. At the same time, in the tobacco leaves obtained in Examples 1-3, the proportion of upper and middle tobacco leaves is also higher than that in the control group.

[0157] Table 2

[0158]

[0159] As shown in Table 3, the inventors investigated the effects of the different products obtained in Examples 1 to 3 and Comparative Examples 1 to 5 on the neutral aroma components (brown reactants) of tobacco leaves. The products obtained in Examples 1 to 3 significantly increased the content of brown reactants.

[0160] Table 3

[0161]

Claims

1. A method for preparing tobacco organic fertilizer based on a mild fermentation method using rapeseed stalks as raw materials, characterized in that: The preparation method comprises the following steps: (1) preparing rape straw and teasel stems in a weight ratio of 100:3-5; chopping the rape straw and steaming it in a steam atmosphere for 15-25 minutes; (2) removing moisture from the surface of the cooked rapeseed straw, placing it in a steam explosion tank, setting the pressure to 1.2-1.8 MPa, and steam explosion for 3-8 minutes; (3) removing moisture from the surface of the rapeseed straw after steam explosion treatment, crushing the stems of Dipsacus asper and placing them together in the fermentation liquid for 2.5 to 3.5 hours; (4) removing the surface moisture of the product obtained in step (3), inoculating the fermentation bacteria by spraying, fermenting at 37° C. for 8 hours, and collecting the fermentation product; The fermentation broth is composed of culture broth of Bacillus subtilis CH-001, Bacillus cereus DS1 and Pseudomonas sp. SG-1; The fermentation bacteria is a mixed bacteria consisting of Bacillus subtilis CH-001, Bacillus cereus DS1 and Pseudomonas sp. SG-1; The deposit number of the Bacillus subtilis CH-001 is CCTCC NO.M 2011111, the deposit number of the Bacillus cereus DS1 is CCTCC NO.M 2010145, and the deposit number of the Pseudomonas sp. SG-1 is CGMCC No.3556.

2. The method for preparing tobacco organic fertilizer based on a mild fermentation method using rape stalks as raw materials according to claim 1, characterized in that: The weight ratio of the rape straw to the teasel stem is 100:

5.

3. The method for preparing tobacco organic fertilizer based on a mild fermentation method using rape stalks as raw materials according to claim 1, characterized in that: When steaming, the mixture is steamed for 20 minutes at normal pressure in a steam atmosphere.

4. The method for preparing tobacco organic fertilizer based on a mild fermentation method using rape stalks as raw materials according to claim 1, characterized in that: When performing steam explosion treatment, the pressure is 1.5 MPa and the time is 5 minutes.

5. The method for preparing tobacco organic fertilizer based on a mild fermentation method using rape stalks as raw materials according to claim 1, characterized in that: When rape straw and Dipsacus asper stems are placed in the fermentation liquid, the solid-liquid weight ratio is 1 g:10 mL.

6. The method for preparing tobacco organic fertilizer based on a mild fermentation method using rape stalks as raw materials according to claim 5, characterized in that: In step (3), the treatment temperature is 37°C and the treatment time is 3 hours.

7. The method for preparing tobacco organic fertilizer based on a mild fermentation method using rape stalks as raw materials according to claim 1, characterized in that: In step (4), among the fermentation bacteria, the bacterial concentrations and volumes of Bacillus subtilis CH-001, Bacillus cereus DS1 and Pseudomonas sp. SG-1 are the same, and the inoculation amount of the fermentation bacteria is 8% v / v.

8. The method for preparing tobacco organic fertilizer based on a mild fermentation method using rape stalks as raw materials according to claim 7, characterized in that: The bacterial concentration of each bacteria in the fermentation bacteria is 1×10 9 cfu / mL.

9. An organic fertilizer for tobacco produced by a mild fermentation method using rapeseed stalks as raw materials, characterized in that: The tobacco organic fertilizer is prepared by the preparation method according to any one of claims 1 to 8.

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