Cigarette organic fertilizer based on mild fermentation method and using rape stalks as raw materials and preparation method thereof

Abstract

The invention belongs to the technical field of organic fertilizers and microbial fertilizers. The invention provides a preparation method of a tobacco organic fertilizer based on a mild fermentation method by taking rape stalks as a raw material, which comprises the following steps: cutting the rape stalks into pieces, cooking in a water vapor atmosphere, and then placing in a steam explosion tank for steam explosion treatment; removing moisture on the surfaces of the rape straws subjected to steam explosion treatment, mashing the teasel root and chrysanthemum stems, putting the teasel root and chrysanthemum stems into fermentation liquor, and treating for 2.5-3.5 hours; and inoculating zymophyte, fermenting for 6-8 hours, and collecting a fermentation product, thereby obtaining the product. The organic fertilizer for the cigarettes provided by the invention is short in production period, the time required in the fermentation stage is less than 10 hours, and the organic fertilizer can be efficiently linked with other production links and has a good mechanical production prospect. Meanwhile, the obtained organic fertilizer for the tobacco can remarkably improve the yield and quality of the tobacco leaves.

Description

Technical Field

[0001] The present invention belongs to the technical field of organic fertilizers and microbial fertilizers, and particularly relates to a tobacco organic fertilizer based on rape straw by a mild fermentation method and a preparation method thereof. Background Art

[0002] Bio-organic fertilizer is a new type of fertilizer, which has the advantages of low production cost and rich nutrient elements, and has been widely used in the agricultural field. Specifically in tobacco planting, bio-organic fertilizer is also being increasingly widely used, and it has certain advantages in improving the yield and quality of tobacco, regulating the soil microbial ecology, improving the soil, increasing the yield and quality, enhancing the stress resistance of tobacco plants, 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 relatively long, and the production cycle of bio-organic fertilizers prepared by the fermentation method is usually several days to dozens of days; (2) The environmental protection cost is relatively high. Since the composting method is usually used during microbial fermentation, which is likely to pollute the surrounding environment, the environmental protection cost during the production of bio-organic fertilizers is relatively high; (3) The effects on increasing the yield, improving the quality and disease resistance of tobacco leaves are limited.

[0004] In terms of shortening the fermentation cycle, a lot of exploration has been carried out. For example, Wang Lin et al. from China Tobacco Hubei Industrial Co., Ltd. provided a method for rapid fermentation of tobacco cake fertilizer in Chinese Patent CN 102108031B. This method only requires 15 - 25 days of fermentation to obtain the cake fertilizer for tobacco leaf planting. Another example is that Shi Zhentuo et al. provided a method for rapid fermentation of traditional Chinese medicine residue compost to produce organic fertilizer in Chinese Patent CN 103449863B. This method only requires 5 - 7 days of composting to obtain the organic fertilizer. The methods represented by the above two patents are achieved by adding decomposing agents and selecting specific fermentation substrates and fermentation bacteria. In addition, research has also been carried out on improving the fermentation physical environment. For example, Yan Weiwei et al. proposed a method for rapid fermentation of olive fertilizer in Chinese Patent CN104892051B. This method accelerates the fermentation of the fertilizer by digging a special fermentation pool and treating it with lime powder, utilizing the principle that lime generates heat when encountering water. Another example is that Chi Jingliang et al. proposed a method for rapid fermentation of livestock and poultry manure to produce organic fertilizer using a fermentation tunnel in Chinese Patent CN105503304B. This method can shorten the fermentation cycle to 4 - 5 days by setting up a special fermentation tunnel. In addition, people also shorten the fermentation cycle by selecting special fermentation bacteria. For example, Li Liyan et al. provided a rapid fermentation synergistic bactericide for organic materials in Chinese Patent CN 114045232B. This bactericide has the characteristics of enabling the compost fermentation to start quickly, having a fast temperature rise, and a long high-temperature duration, and can degrade organic materials quickly and efficiently.

[0005] However, as far as the inventor knows, the production cycle of the existing bio-organic fertilizer for tobacco planting is still relatively long. The methods with a shorter cycle still require more than 4 - 5 days for fermentation, so the problem of environmental protection cost has not been well solved. In addition, since fermentation is a relatively complex biological activity and is affected by various factors, the cost of controlling process stability is also relatively high. Therefore, shortening the fermentation cycle can also reduce the corresponding production control cost.

[0006] Currently, 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. Such products mainly focus on types such as pastries, tea, meat products, and fruit juices. For example, Ge Zhenzhen et al. [2] studied the influence of micro-fermentation on the flavor of noodles and their quality characteristics during storage, and Wu Bin [3] et al. studied the influence 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 light fermentation conditions on the quality of black tea. Such methods, including the above research, only take a few minutes or hours to complete the required fermentation process. If the concept of light fermentation can be adopted in the preparation of bio-organic fertilizer, unnecessary fermentation time can be discarded, and the required fertilizer effect can be obtained, the defects of the aforementioned bio-organic fertilizer can be solved.

[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-growing 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 seabuckthorn 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 ‘Xiangbolu’ and ‘Qiuyeqi’ under light rolling and light fermentation conditions [J]. Tea Newsletter, 2024, 51(2): 202-209. Summary of the invention

[0012] In view of the shortcomings of the prior art, the present invention aims to provide an organic fertilizer for tobacco using rape stalks as raw materials based on a mild fermentation method and a preparation method thereof. The fermentation cycle required for the organic fertilizer is greatly reduced, and the organic fertilizer can significantly improve the yield and quality of tobacco leaves.

[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, the preparation method comprising the following steps:

[0015] (1) preparing rape straw and teasel stems in a weight ratio of 100:3-5; cutting the rape straw into pieces and steaming them 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 performing steam explosion treatment for 3-8 minutes;

[0017] (3) Remove the surface moisture of the steam-exploded rape straw, mash the Sonchus asper stems and place them together with the rape straw in the fermentation broth, and treat for 2.5 to 3.5 hours;

[0018] (4) Remove the surface moisture of the product obtained in step (3), inoculate the fermentation bacteria by spraying, ferment for 6 to 8 hours, and collect the fermentation product to obtain the product;

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

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

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

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

[0023] Preferably, when performing cooking, under normal pressure, cook in a water vapor atmosphere for 20 minutes.

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

[0025] Preferably, when placing the rape straw and the Sonchus asper stems in the fermentation broth, the solid-liquid weight ratio is 1 g:10 mL.

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

[0027] Preferably, in step (4), in 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 bacterium 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 a tobacco organic fertilizer prepared by the above preparation method.

[0031] Before the research of the present invention, the inventors realized that in the preparation of bio-organic fertilizers in the prior art, fermentation is often required to be carried out outdoors or in a dedicated composting production area, and it is difficult for the fermentation process to be efficiently linked with the mechanized treatment links such as the pretreatment of raw materials, resulting in a lack of continuity in the entire production line. At the same time, when preparing bio-organic fertilizers in the prior art, the required fermentation time is usually relatively long (from several days to dozens of days). During the long fermentation process, unpleasant odors and overflow of fermentation waste liquid are likely to occur, resulting in air pollution and soil pollution, and the corresponding environmental protection work costs are relatively high. To solve this problem, researchers in this field have conducted a lot of research on fermentation equipment systems and adjustment of fermentation processes. Although there have been results, the problem of too long fermentation time still cannot be fundamentally solved.

[0032] The food industry is an industry that has basically completed highly mechanized production. Although many products require fermentation, the fermentation cycle is very short, and its fermentation process is generally referred to as mild fermentation or micro-fermentation. Due to the short fermentation cycle, continuous production of related food products can be achieved, and the degree of mechanized production is relatively high. Inspired by this, the inventors considered whether it is possible to use mild fermentation to complete the production of products with corresponding fertilizer effects when producing bio-organic fertilizers.

[0033] Different from the production of related fermented foods, organic fertilizers usually need to be fermented to make the substrate mature, so as to facilitate crops to absorb the required nutrients; while the fermentation process required for related foods is only to improve the flavor of the food, and there are subsequent cooked food processing (such as steaming, frying, etc.) processes. Therefore, the production purposes of the two in the fermentation process are essentially different.

[0034] Bio-organic fertilizers are essentially to process bio-source materials (such as plant straws, animal excreta) into small molecular substances that are more conducive to crop absorption. Generally, it is impossible to achieve the above-mentioned purpose only by fermenting for several hours. Therefore, the inventors considered performing corresponding pretreatment on the fermentation substrate (bio-organic fertilizer raw material) to make the fermentation substrate a substance that is easy for microorganisms to utilize and metabolize, thereby shortening the entire fermentation cycle.

[0035] As shown by the relevant experimental results of the present invention, the inventors first tried to perform steam explosion treatment on rape straws, and then fermented for 2 days. The obtained product could not increase the yield of tobacco. After replacing different fermentation bacteria, the obtained experimental results were still not ideal. After continuous exploration, the inventors tried to soak the straws after steam explosion treatment in the fermentation bacteria culture solution and then ferment, and found that the obtained product had the effect of increasing the yield of tobacco leaves. Encouraged by this, the inventors continuously improved the treatment process and finally obtained a product with the required fertilizer effect.

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

[0037] As shown in the related examples and comparative examples of the present invention, the cooking process, steam explosion process, selection of common raw materials, selection of fermentation broth, and selection of fermentation bacteria all play crucial roles. When other fermentation broths are selected for soaking treatment, it has a greater impact on the tobacco leaf yield. The reason may be that the related substances in the fermentation broth have a greater 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 single fermentation bacteria fermentation broth or a mixture of three fermentation bacteria fermentation broths being inferior to the scheme of using a mixture of two fermentation bacteria fermentation broths. In addition, in the fermentation stage, it is very important to use three fermentation bacteria as inoculation bacteria at the same time. The possible reason is that these three fermentation bacteria have a synergistic effect in the initial stage of fermentation, accelerating the utilization and metabolism of the fermentation substrate.

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

[0039] In addition, it should be pointed out that among the raw materials used in the present invention, rape straw, as an agricultural waste, is generally discarded or simply recycled, while the present invention can use it as a raw material for producing high-quality tobacco organic fertilizer, realizing the high-value reuse of rape straw; at the same time, Sonchus asper, which is a common weed in Yunnan, has a low harvesting or acquisition cost, and the present invention can realize the high-value utilization of weeds. Therefore, the raw material cost of the present invention is low, and it has good industrial economy.

[0040] Advantages of the present invention:

[0041] The production cycle of the tobacco organic fertilizer provided by the present invention is short. The time required in the fermentation stage is less than 10 hours, and it can be efficiently connected with other production links, having 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. Specific embodiments

[0042] The present invention will be specifically described below through examples. It is necessary to point out here that the following examples are only used to further illustrate the present invention and cannot be understood as limiting the protection scope of the present invention. Some non-essential improvements and adjustments made by those skilled in the art according to the above invention content still belong to the protection scope of the present invention.

[0043] Example 1

[0044] 1. Raw materials

[0045] Rape straw: harvested locally in Yunnan; Sonchus asper (L.) Hill: fresh stem part, 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; The three strains were cultured in LB medium respectively to prepare seed solutions, and then mixed for fermentation inoculation. The cell concentration of each strain after mixing was 1×10 9 cfu / mL.

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

[0048] 2. Preparation of tobacco organic fertilizer

[0049] (1) Prepare rape straw and Sonchus asper (L.) Hill stems according to the weight ratio of 100:5;

[0050] (2) Cut the rape straw into pieces (length controlled within the range of 0.5 - 1.0 cm) and steam-cook it for 20 minutes in a steam atmosphere (atmospheric pressure);

[0051] (3) Remove the surface moisture of the steamed rape straw, place it in a steam explosion tank, set the pressure to 1.5 MPa, and perform steam explosion treatment for 5 minutes;

[0052] (4) Remove the surface moisture of the rape straw after steam explosion treatment, mash the Sonchus asper (L.) Hill stems and place them together with the rape straw in the fermentation broth, with a solid-liquid weight ratio of 1 g:10 mL, and treat at 35°C for 3 hours;

[0053] (5) Remove the surface moisture of the product obtained in step (4), inoculate the fermentation bacteria by spraying, with an inoculation amount of 8% v / v, ferment at 37°C for 8 hours, and collect the fermentation product.

[0054] Example 2

[0055] 1. Raw materials

[0056] Rape straw: harvested locally in Yunnan; Sonchus asper (L.) Hill: fresh stem part, 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), which were kindly provided by colleagues for research use; the three strains were cultured in LB medium respectively to prepare seed solutions, and after mixing, they were used for fermentation inoculation. The cell concentration of each strain after mixing was 1×10 9 cfu / mL.

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

[0059] 2. Preparation of organic fertilizer for tobacco

[0060] (1) Prepare rape straw and Sonchus asper stems according to the weight ratio of 100:3;

[0061] (2) Cut the rape straw into pieces (the length is controlled in the range of 0.5 - 1.0 cm) and steam-cook it for 15 minutes in a steam atmosphere (atmospheric pressure);

[0062] (3) Remove the surface moisture of the steamed rape straw, place it in a steam explosion tank, set the pressure at 1.2 MPa, and perform steam explosion treatment for 8 minutes;

[0063] (4) Remove the surface moisture of the rape straw after steam explosion treatment, mash the Sonchus asper stems and place them together with the rape straw in the fermentation broth. The solid-liquid weight ratio is 1 g:10 mL, and treat them at 35 °C for 3.5 hours;

[0064] (5) Remove the surface moisture of the product obtained in step (4), inoculate the fermentation bacteria in a spraying manner, the inoculation amount is 8% v / v, ferment at 37 °C for 6 hours, and collect the fermentation product.

[0065] Example 3

[0066] 1. Raw materials

[0067] Rape straw: harvested locally in Yunnan; Sonchus asper: fresh stem part, 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), which were kindly provided by colleagues for research use; the three strains were cultured in LB medium to prepare seed solutions respectively, and after mixing, they were used for fermentation inoculation. The cell concentration of each strain after mixing was 1×10 9 cfu / mL.

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

[0070] 2. Preparation of tobacco organic fertilizer

[0071] (1) Prepare rape straw and Sonchus asper stems according to a weight ratio of 100:4;

[0072] (2) Cut the rape straw into pieces (the length is controlled in the range of 0.5 - 1.0 cm) and steam-cook it for 25 minutes under normal pressure in a steam atmosphere;

[0073] (3) Remove the surface moisture of the steamed rape straw, place it in a steam explosion tank, set the pressure to 1.8 MPa, and perform steam explosion treatment for 3 minutes;

[0074] (4) Remove the surface moisture of the rape straw after steam explosion treatment, mash the Sonchus asper stems and place them together with the rape straw in the fermentation broth. The solid-liquid weight ratio is 1 g:10 mL, and treat at 35 °C for 2.5 hours;

[0075] (5) Remove the surface moisture of the product obtained in step (4), inoculate the fermentation bacteria by spraying, the inoculation amount is 8% v / v, ferment at 37 °C for 8 hours, and collect the fermentation product.

[0076] Comparative Example 1

[0077] Except for not using the steam explosion process, the rest is the same as in Example 1. The specific scheme is as follows:

[0078] 1. Raw materials

[0079] Rape straw: harvested locally in Yunnan; Sonchus asper: fresh stem part, 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), which were kindly provided by colleagues for research use; the three strains were cultured in LB medium respectively to prepare seed solutions, and after mixing, they were used for fermentation inoculation. The cell concentration of each strain after mixing was 1×10 9 cfu / mL.

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

[0082] 2. Preparation of tobacco organic fertilizer

[0083] (1) Rapeseed straw and Sonchus asper stems were prepared according to the weight ratio of 100:5.

[0084] (2) The rapeseed straw was cut into pieces (the length was controlled in the range of 0.5 - 1.0 cm) and steamed in a water vapor atmosphere (atmospheric pressure) for 20 minutes.

[0085] (3) The surface moisture of the rapeseed straw after steaming treatment was removed, the Sonchus asper stems were mashed and placed together with it in the fermentation broth, and the solid-liquid weight ratio was 1 g:10 mL, and it was treated at 35°C for 3 hours.

[0086] (4) The surface moisture of the product obtained in step (3) was removed, and the fermentation bacteria were inoculated by spraying. The inoculation amount was 8% v / v, and it was fermented at 37°C for 8 hours, and the fermentation product was collected.

[0087] Comparative Example 2

[0088] Except for not adopting the steaming process, the rest was the same as in Example 1. The specific scheme was as follows:

[0089] 1. Raw materials

[0090] Rapeseed straw: harvested locally in Yunnan; Sonchus asper: fresh stem part, 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), which were kindly provided by colleagues for research use; the three strains were cultured in LB medium to prepare seed solutions respectively, and then mixed for fermentation inoculation. The cell concentration of each strain after mixing was 1×10 9 cfu / mL.

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

[0093] 2. Preparation of tobacco organic fertilizer

[0094] (1) Rapeseed straw and Sonchus asper stems were prepared according to a weight ratio of 100:5.

[0095] (2) The rapeseed straw was cut into pieces (the length was controlled in the range of 0.5 - 1.0 cm), placed in a steam explosion tank, the pressure was set at 1.5 MPa, and the steam explosion treatment was carried out for 5 minutes.

[0096] (3) The surface moisture of the rapeseed straw after steam explosion treatment was removed, the Sonchus asper stems were mashed and placed together with it in the fermentation broth, the solid-liquid weight ratio was 1 g:10 mL, and it was treated at 35 °C for 3 hours.

[0097] (4) The surface moisture of the product obtained in step (3) was removed, and the fermentation bacteria were inoculated by spraying, the inoculation amount was 8% v / v, and it was fermented at 37 °C for 8 hours, and the fermentation product was collected.

[0098] Comparative Example 3

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

[0100] 1. Raw materials

[0101] Rapeseed straw: harvested locally in Yunnan; Sonchus asper: fresh stem part, harvested locally;

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

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

[0104] 2. Preparation of tobacco organic fertilizer

[0105] (1) Prepare rape straw and Sonchus asper (L.) Hill stems according to the weight ratio of 100:5;

[0106] (2) Cut the rape straw into pieces (length controlled within the range of 0.5 - 1.0 cm) and steam-cook for 20 minutes in a steam atmosphere (atmospheric pressure);

[0107] (3) Remove the surface moisture of the steamed rape straw, place it in a steam explosion tank, set the pressure to 1.5 MPa, and perform steam explosion treatment for 5 minutes;

[0108] (4) Remove the surface moisture of the rape straw after steam explosion treatment, mash the Sonchus asper (L.) Hill stems and place them together in the fermentation broth, with a solid-liquid weight ratio of 1 g:10 mL, and treat at 35°C for 3 hours;

[0109] (5) Remove the surface moisture of the product obtained in step (4), inoculate the fermentation bacteria in a spraying manner, with an inoculation amount of 8% v / v, ferment at 37°C for 8 hours, and collect the fermentation product.

[0110] Comparative Example 4

[0111] Except that the fermentation broth was adjusted to a fermentation broth of one fermentation strain, the rest was the same as in Example 1. The specific scheme was as follows:

[0112] 1. Raw materials

[0113] Rape straw: Harvested locally in Yunnan; Sonchus asper (L.) Hill: Fresh stem part, harvested locally;

[0114] Fermentation strain: Bacillus subtilis CH-001 (preservation number: CCTCC NO.M 2011111) was kindly provided by colleagues for research use; The seed solution was prepared by culturing this strain in LB medium for fermentation inoculation, and the bacterial concentration of this strain was 3×10 9 cfu / mL.

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

[0116] 2. Preparation of tobacco organic fertilizer

[0117] (1) Prepare rape straw and Sonchus asper (L.) Hill stems according to the weight ratio of 100:5;

[0118] (2) Cut the rape straw into pieces (with the length controlled within the range of 0.5 - 1.0 cm) and cook it in a steam atmosphere (under normal pressure) for 20 minutes.

[0119] (3) Remove the surface moisture of the cooked rape straw, place it in a steam explosion tank, set the pressure to 1.5 MPa, and conduct steam explosion treatment for 5 minutes.

[0120] (4) Remove the surface moisture of the rape straw after steam explosion treatment, mash the Sonchus asper stems and place them together with the rape straw in a fermentation broth. The solid - liquid weight ratio is 1 g:10 mL, and treat at 35 °C for 3 hours.

[0121] (5) Remove the surface moisture of the product obtained in step (4), inoculate the fermentation bacteria in a spraying manner, with the inoculation amount being 8% v / v, ferment at 37 °C for 8 hours, and collect the fermentation product.

[0122] Comparative Example 5

[0123] Except that the fermentation bacteria are adjusted to a mixed bacteria composed of two kinds of bacteria, the rest are the same as in Example 1. The specific scheme is as follows:

[0124] 1. Raw materials

[0125] Rape straw: harvested locally in Yunnan; Sonchus asper: fresh stem part, harvested locally;

[0126] 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; Bacillus subtilis CH - 001 and Bacillus cereus DS1 are cultured with LB medium respectively to prepare seed solutions, and after mixing, they are used for fermentation inoculation. The cell concentration of each bacterium after mixing is 1.5×10 9 cfu / mL.

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

[0128] 2. Preparation of tobacco organic fertilizer

[0129] (1) Prepare rape straw and Sonchus asper stems according to the weight ratio of 100:5.

[0130] (2) Cut the rape straw into pieces (with the length controlled within the range of 0.5 - 1.0 cm) and cook it in a water vapor atmosphere (at normal pressure) for 20 minutes;

[0131] (3) Remove the surface moisture of the cooked rape straw, place it in a steam explosion tank, set the pressure at 1.5 MPa, and conduct steam explosion treatment for 5 minutes;

[0132] (4) Remove the surface moisture of the rape straw after steam explosion treatment, pound the Sonchus asper stems and place them together with the rape straw in a fermentation broth, with the solid-liquid weight ratio of 1 g:10 mL, and treat at 35 °C for 3 hours;

[0133] (5) Remove the surface moisture of the product obtained in step (4), inoculate the fermentation bacteria in a spraying manner, with the inoculation amount of 8% v / v, ferment at 37 °C for 8 hours, and collect the fermentation product.

[0134] Comparative Example 6

[0135] Except for not adding Sonchus asper stems as raw materials, the rest is the same as in Example 1.

[0136] 1. Raw materials

[0137] Rape 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; cultivate the three strains separately in LB medium to prepare seed solutions, mix them and use for fermentation inoculation, and the cell concentration of each strain after mixing is 1×10 9 cfu / mL.

[0139] Fermentation broth: Cultivate the three fermentation strains separately in LB liquid medium until the stationary phase (cell concentration is 1×10 8 cfu / mL), and then mix them in equal volumes.

[0140] 2. Preparation of tobacco organic fertilizer

[0141] (1) Cut the rape straw into pieces (with the length controlled within the range of 0.5 - 1.0 cm) and cook it in a water vapor atmosphere (at normal pressure) for 20 minutes;

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

[0143] (3) Remove the surface moisture of the steam-exploded rape straw and place it in the fermentation broth. The solid-liquid weight ratio is 1 g:10 mL, and treat it at 35 °C for 3 hours;

[0144] (4) Remove the surface moisture of the product obtained in step (3), inoculate the fermentation bacteria by spraying, the inoculation amount is 8% v / v, ferment at 37 °C for 8 hours, and collect the fermentation product.

[0145] Experimental Example

[0146] 1. Test soil

[0147] The previous crop of the soil used in the test is 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 Investigation of economic traits:

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

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

[0154] The volatile compounds in tobacco leaves were determined by static headspace-gas chromatography-mass spectrometry. Specifically: Weigh 0.5 g of tobacco leaf sample into a 20 mL headspace vial, immediately seal it with an aluminum foil cap and gently shake the headspace vial to make the sample evenly loose, and place it in the headspace sampler to balance before injecting into the GC-MS to collect data.

[0155] As shown in Table 2, the effects of the different products obtained in Examples 1-3 and Comparative Examples 1-6 on the tobacco leaf yield were investigated. Each group was applied before rotary tillage and ridging, and the application rate was 800 kg / ha; the control group was applied with the HK microbial organic fertilizer (N+P 2 O 5 +K 2 O = 8%) produced by Guangxi Dahua Fertilizer Industry Co., Ltd., and the application rate was 800 kg / ha.

[0156] Specifically, the fertilizer used in the control group in this experiment is a commercially available mature product with a certain growth-promoting effect. The inventor adjusted its application rate to 800 kg / ha and did not top-dress. Under this condition, its yield is 2167.2 kg / hm 2In Examples 1 to 3 as the experimental group and Comparative Examples 1 to 5, the products of Examples 1 - 3 have an obvious promoting effect on the tobacco leaf yield, and the yield is significantly higher than that of the control group. It can be seen that the obtained products can not only be used as biological organic fertilizers, but also have a better growth promoting effect 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 the fermentation broth for soaking treatment, the fermentation bacteria for mild fermentation, and the fermentation substrate has an obvious impact on the fertilizer efficiency of the obtained products; in addition, from Comparative Example 1 and Comparative Example 2, it can also be seen that whether to perform steam explosion treatment and whether to combine cooking and steam explosion treatment are also the keys of the present invention. At the same time, in the tobacco leaves obtained in Examples 1 - 3, the proportion of upper-middle tobacco leaves is also higher than that of the control group.

[0157] Table 2

[0158]

[0159] As shown in Table 3, the inventors investigated the effects of 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 have an obvious enhancing effect on the content of brown reactants.

[0160] Table 3

[0161]

Claims

1. A method for preparing tobacco organic fertilizer using rape stalks as raw materials based on a mild fermentation method, 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; cutting the rape straw into pieces and steaming them 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 performing steam explosion treatment for 3-8 minutes; (3) removing moisture from the surface of the rapeseed straw after the 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 for 6 to 8 hours, and collecting the fermentation product to obtain; 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 composed 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 under normal pressure and 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 1.5MPa 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 the rape straw and the stems of Dipsacus asper 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. 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), the fermentation temperature is 37° C. and the fermentation time is 8 hours.

10. An organic fertilizer for tobacco based on a mild fermentation method using rape 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 9.

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