A whole-cycle planting method for improving quality and yield of tobacco

Abstract

The present application belongs to the technical field of tobacco planting, and provides a whole-cycle planting method for improving quality and increasing yield of tobacco, comprising the following steps: (1) seed treatment: after cleaning the seeds with water, the seeds are soaked in an elicitor composed of gibberellin with a concentration of 50-1000 mg / L and 10-15 mL / L of Bacillus polymyxa dgnkzx004 fungicide fermentation liquor, then subjected to light treatment, and washed and dried; (2) seedling raising: mixed bacteria cow manure fermentation products are added to the seedling raising substrate, and Bacillus polymyxa dgnkzx004 fermentation liquor is applied for seedling raising; (3) transplanting: the tobacco seedlings are transplanted, and the mixed bacteria cow manure fermentation products are used as fertilizer for fertilization. The present application promotes seed germination rate, improves seedling emergence rate and seedling survival rate, and can significantly improve the growth potential of tobacco root system and the yield of tobacco, and significantly reduce the arsenic content in tobacco leaves.

Description

Technical Field

[0001] This invention belongs to the field of tobacco planting technology, specifically relating to a full-cycle planting method for improving the quality and yield of tobacco. Background Technology

[0002] Yunnan Province is a major tobacco-producing area, accounting for approximately 40% of the national planting area, and is also an important research and development region for tobacco cultivation technology. Because different stages of tobacco cultivation present different technical challenges, current improvements to tobacco cultivation technology are mainly carried out in stages. For example, in seed treatment, research focuses on formulating different hormone compositions to investigate their effects on germination; in seedling substrates, research focuses on using livestock waste to replace traditional commercial seedling substrates; and in post-transplant fertilization, research focuses on improving inorganic fertilizers and bio-organic fertilizers. However, because the raw materials for treatment agents and fertilizers used at different stages vary, the utilization rate of raw materials is relatively low. Therefore, whether it is possible to prepare treatment agents and fertilizers suitable for different stages based on the same raw materials and effectively promote tobacco growth is one of the directions that needs further research in this field. Summary of the Invention

[0003] To address the shortcomings of existing technologies, the present invention aims to provide a formulation based on the same raw materials, applicable to all stages of tobacco cultivation, thereby achieving a cultivation technology that covers the entire tobacco cultivation cycle and improves tobacco quality and yield in multiple dimensions. To achieve the aforementioned objective, the present invention adopts the following solution:

[0004] A full-cycle planting method for improving the quality and increasing the yield of tobacco, the full-cycle planting method comprising the following steps:

[0005] (1) Seed treatment: After washing the seeds with water, soak the seeds in an initiator composed of gibberellin at a concentration of 50~1000mg / L and Bacillus polymyxa dgnkzx004 sterilized fermentation broth at a concentration of 10~15mL / L, then treat them with light, and wash and dry them.

[0006] (2) Seedling raising: Add mixed bacteria cow manure fermentation product to the seedling substrate and apply Bacillus polymyxa dgnkzx004 fermentation liquid for seedling raising;

[0007] (3) Transplanting: Transplant the tobacco seedlings and fertilize them with the fermented product of mixed bacteria and cow manure.

[0008] Among them, the sterilized fermentation broth of Bacillus polymyxa dgnkzx004 is obtained by culturing Bacillus polymyxa dgnkzx004, centrifuging to remove the bacterial cells and collecting the supernatant;

[0009] The mixed-bacterial cow manure fermentation product is obtained by composting cow manure as a fermentation substrate and using Bacillus polymyxa dgnkzx004 and Bacillus licheniformis ZJU12 as fermentation bacteria.

[0010] Preferably, the concentration of gibberellin in the initiator is 50-100 mg / L, and the amount of sterilized Bacillus polymyxa dgnkzx004 fermentation broth added is 10-15 mL / L.

[0011] Preferably, in step (1), the light intensity is 1200 LX and the light duration is 210 minutes during the light treatment.

[0012] Preferably, the amount of mixed bacteria and cow manure fermentation products added to the seedling substrate is 10%wt.

[0013] Preferably, the seedling substrate is composed of cow dung, peat moss, rice husks, perlite, and mixed microbial cow dung fermentation products in a weight ratio of 2:5:1:1:1.

[0014] Preferably, the seedling substrate is composed of corn stalk charcoal, peat moss, perlite, and mixed microbial cow manure fermentation products in a weight ratio of 5:2:2:1.

[0015] Preferably, when applying the fermentation broth of Bacillus polymyxa dgnkzx004, it is applied once when 70% of the seedlings have emerged, and then applied again after sealing the trays.

[0016] Preferably, one seed is sown per hole during seedling cultivation.

[0017] Preferably, when fertilizing after transplanting, fertilizer is applied on the day of transplanting and on the 20th day after transplanting.

[0018] Preferably, after transplanting, the amount of fertilizer applied each time is 30 kg / mu.

[0019] The beneficial effects of this invention are:

[0020] This invention is the first to discover that adding fermentation broth of Bacillus polymyxa dgnkzx004 to conventional fertilizer can promote tobacco growth. Furthermore, it was found that sterilized fermentation broth of Bacillus polymyxa dgnkzx004 can promote seed germination rate. It was also found that using fermentation products prepared by co-inoculating cow manure with Bacillus polymyxa dgnkzx004 and Bacillus licheniformis ZJU12 as a seedling substrate can significantly improve germination and seedling survival rates. Finally, it was found that adopting the full-cycle planting scheme of this invention can significantly improve tobacco root growth vigor and tobacco yield, and significantly reduce the arsenic content in tobacco leaves. Detailed Implementation

[0021] The embodiments described below are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the present invention are still within the protection scope of the present invention.

[0022] The sources of raw materials, formulations, and tobacco seedlings involved in the following embodiments, preliminary experimental examples, and experimental examples of this invention are as follows:

[0023] The commercial organic fertilizer (N:P2O5:K2O=2:1.1:1.9, organic matter ≥45%) was purchased from Anhui Lemjia Biotechnology Co., Ltd.

[0024] Polymyxobacterium dgnkzx004 (hereinafter referred to as dgnkzx004), with accession number CCTCC M 2014504, was purchased from the China Center for Type Culture Collection.

[0025] Bacillus licheniformis ZJU12 (hereinafter referred to as ZJU12), with accession number CCTCC M205072, was purchased from the China Center for Type Culture Collection.

[0026] Corn stalks: collected from local farmers;

[0027] Peat moss: Purchased from Fushun Xinmei Horticulture Co., Ltd.;

[0028] Perlite: Purchased from Lingshou County Haibin Mineral Products Trading Co., Ltd., capacity 100kg / m³ 3 Brand: 1145;

[0029] Rice husks: collected from local farmers;

[0030] Cow dung: collected from local farmers;

[0031] Cigarette variety: Yunyan 87.

[0032] The preparation methods of the articles involved in the following embodiments, preliminary experimental examples and experimental examples of the present invention are as follows:

[0033] Fermentation broth of dgnkzx004 bacteria: cultured in LB liquid medium until the plateau phase at a temperature of 32℃ for 20 hours;

[0034] dgnkzx004 bacterial powder: Centrifuge the dgnkzx004 bacterial fermentation broth, collect the bacterial precipitate, and dry it at room temperature to obtain the powder.

[0035] Sterilized fermentation broth of dgnkzx004 bacteria: Centrifuge the fermentation broth of dgnkzx004 bacteria and collect the supernatant.

[0036] ZJU12 bacterial fermentation broth: cultured in LB liquid medium until the plateau phase at a temperature of 30℃ for 34 hours;

[0037] ZJU12 bacteria cow dung fermentation product: using cow dung as the fermentation substrate, inoculating with ZJU12 bacteria at an inoculation rate of 4% and composting for 15 days to obtain the product;

[0038] Mixed microbial cow manure fermentation product: using cow manure as the fermentation substrate, inoculate dgnkzx004 bacteria and ZJU12 bacteria (bacterial number ratio of 1:1) at a total inoculation amount of 4% and compost for 15 days to obtain the product.

[0039] Corn stalk charcoal: Corn stalks are dried, crushed, and then carbonized at 750°C for 2 hours under a nitrogen atmosphere.

[0040] Example 1

[0041] Seed treatment: Immerse dormant tobacco seeds in clean water and stir with a magnetic stirrer for 10 minutes to make the seeds tumble. Then remove the seeds and rinse them with clean water until the water is clear. After that, immerse the rinsed seeds in an initiator at 28°C and irradiate them under 1200LX light for 210 minutes. Remove the seeds, rinse them with clean water, and air dry them. The initiator is an aqueous solution composed of 100mg / L gibberellin and 10mL / L dgnkzx004 bacteria sterilized fermentation broth.

[0042] Seedling raising: Mix corn stalk charcoal, peat moss, perlite, and mixed bacteria cow manure fermentation products in a weight ratio of 5:2:2:1 to form a seedling substrate in seedling trays. Sow one seed per hole. When 70% of the seedlings have emerged, apply 20 mL of dgnkzx004 bacteria fermentation liquid. After sealing the trays, apply another 20 mL of dgnkzx004 bacteria fermentation liquid.

[0043] Transplanting: After the seedlings have matured, transplant them. Use the fermented product of mixed bacteria and cow manure as fertilizer. Apply fertilizer on the day of transplanting and 20 days after transplanting, with each application amount being 30 kg / mu.

[0044] Example 2

[0045] Seed treatment: Immerse dormant tobacco seeds in clean water and stir with a magnetic stirrer for 10 minutes to make the seeds tumble. Then remove the seeds and rinse them with clean water until the water is clear. After that, immerse the rinsed seeds in an initiator at 28°C and irradiate them under 1200LX light for 210 minutes. Remove the seeds, rinse them with clean water, and air dry them. The initiator is an aqueous solution composed of 1000mg / L gibberellin and 10mL / L dgnkzx004 bacteria sterilized fermentation broth.

[0046] Seedling raising: Mix corn stalk charcoal, peat moss, perlite, and mixed bacteria cow manure fermentation products in a weight ratio of 5:2:2:1 to form a seedling substrate in seedling trays. Sow one seed per hole. When 70% of the seedlings have emerged, apply 20 mL of dgnkzx004 bacteria fermentation liquid. After sealing the trays, apply another 20 mL of dgnkzx004 bacteria fermentation liquid.

[0047] Transplanting: After the seedlings have matured, transplant them. Use the fermented product of mixed bacteria and cow manure as fertilizer. Apply fertilizer on the day of transplanting and 20 days after transplanting, with each application amount being 30 kg / mu.

[0048] Example 3

[0049] Seed treatment: Immerse dormant tobacco seeds in clean water and stir with a magnetic stirrer for 10 minutes to make the seeds tumble. Then remove the seeds and rinse them with clean water until the water is clear. After that, immerse the rinsed seeds in an initiator at 28°C and irradiate them under 1200LX light for 210 minutes. Remove the seeds, rinse them with clean water, and air dry them. The initiator is an aqueous solution composed of 50 mg / L gibberellin and 15 mL / L dgnkzx004 bacteria sterilized fermentation broth.

[0050] Seedling raising: Mix cow dung, peat moss, rice husks, perlite, and mixed bacteria cow dung fermentation products in a weight ratio of 2:5:1:1:1 to use as a seedling substrate in seedling trays, and sow one seed per hole; when 70% of the seedlings have emerged, apply 20mL of dgnkzx004 bacteria fermentation liquid, and after sealing the trays, apply another 20mL of dgnkzx004 bacteria fermentation liquid.

[0051] Transplanting: After the seedlings have matured, transplant them. Use the fermented product of mixed bacteria and cow manure as fertilizer. Apply fertilizer on the day of transplanting and 20 days after transplanting, with each application amount being 30 kg / mu.

[0052] Comparative Example 1

[0053] Compared to Example 1, when preparing fertilizer, the fermentation substrate was replaced with a fermentation substrate composed of cow dung and corn stalks in a weight ratio of 8:2, while the rest remained the same as in Example 1.

[0054] Comparative Example 2

[0055] Compared to Example 1, the fermentation substrate was changed from cow dung to sheep dung when preparing the fertilizer, while the rest remained the same as in Example 1.

[0056] Comparative Example 3

[0057] Compared to Example 1, during seedling cultivation, the fermentation liquid of dgnkzx004 bacteria was replaced with commercial organic fertilizer at an application rate of 20g, while the rest remained the same as in Example 1.

[0058] Preliminary Experiment Example 1

[0059] This preliminary experimental example is used to record that, prior to conducting the research of this invention, the inventors discovered that dgnkzx004 bacteria has a certain promoting effect on tobacco yield.

[0060] Tobacco seedlings using the same floating seedling raising scheme (see Group 1 in Preliminary Experiment Example 3) were divided into two groups for transplanting. A tobacco experimental park in Yunnan Province was used as the transplanting site. The soil physicochemical parameters were: pH=6.84, organic matter content of 11.1 g / kg, and cation exchange capacity of 8.67 cmol / kg. Fertilizer was applied on the day of transplanting and 20 days after transplanting, with an application rate of 30 kg / mu each time. The composition of the applied fertilizer is as follows:

[0061] Experimental group: commercial organic fertilizer + dgnkzx004 bacteria fermentation liquid (weight ratio 10:1);

[0062] Control group: Commercial organic fertilizer.

[0063] Table 1

[0064]

[0065] As shown in Table 1, adding the fermentation broth of *Dgnkzx004* bacteria to commercial organic fertilizer resulted in a certain increase in tobacco yield, suggesting that *Dgnkzx004* bacteria itself or its fermentation metabolites have a certain promoting effect on tobacco growth. To further investigate the effect of *Dgnkzx004* bacteria on tobacco growth, the inventors added *Dgnkzx004* bacteria powder and *Dgnkzx004* bacteria de-sterilized fermentation broth as additives to commercial organic fertilizer and conducted experiments. As shown in Table 2, adding only *Dgnkzx004* bacteria powder had no significant effect on tobacco yield, indicating that the metabolites of *Dgnkzx004* bacteria have a positive effect on promoting tobacco growth.

[0066] Table 2

[0067]

[0068] Based on the aforementioned research findings, the inventors considered using *dgnkzx004* bacteria as the fermentation inoculum and inexpensive livestock and poultry manure as the substrate to prepare microbial fermentation products to replace commercial organic fertilizer. However, as shown in Table 3, using *dgnkzx004* bacteria as the fermentation inoculum (4% inoculum) failed to initiate the fermentation process when using cow manure as the fermentation substrate; after 3 days of fermentation, the temperature of the cow manure did not increase significantly. Through experimentation, the inventors discovered that combining *dgnkzx004* bacteria with *ZJU12* bacteria (total inoculum 4%) successfully initiated the fermentation process. Simultaneously, as shown in Table 3, inoculating cow manure solely with *ZJU12* bacteria (4% inoculum) also initiated fermentation. However, as shown in Table 4, neither adding the *ZJU12* bacteria fermentation liquid to commercial organic fertilizer nor replacing commercial organic fertilizer with fermented cow manure inoculated with *ZJU12* bacteria produced a promoting effect on tobacco growth. In addition, as shown in Table 4, the fermented product prepared by using cow dung as the fermentation substrate and dgnkzx004 and ZJU12 bacteria as fermentation bacteria has a more significant promoting effect on tobacco growth than commercial organic fertilizer.

[0069] Therefore, the addition of ZJU12 bacteria facilitates the smooth initiation of fermentation by dgnkzx004 bacteria, producing metabolic products, thereby enabling the fermented product, which uses cow dung as a fermentation substrate, to promote tobacco growth.

[0070] Table 3

[0071]

[0072] Table 4

[0073]

[0074] Preliminary Experiment Example 2

[0075] Based on Preliminary Experiment Example 1, this preliminary experiment example is used to record the inventor's experiment on using dgnkzx004 bacteria for seed treatment.

[0076] The experimental procedure is as follows:

[0077] Submerge dormant tobacco seeds in clean water and stir with a magnetic stirrer for 10 minutes to agitate the seeds. Then remove the seeds and rinse them with clean water until the water is clear. Next, immerse the rinsed seeds in an initiator at 28°C and irradiate them under 1200 LX light for 210 minutes. After irradiating, remove the seeds, rinse them with clean water, and air dry them.

[0078] Remove the seeds treated as described above and place them in a glass petri dish with moistened filter paper. Germinate the seeds at 26°C with a light intensity of 1200 LX and a light duration of 12h / 24h. Record the number of normal seedlings on day 14 and calculate the germination rate. Germination rate = (Number of normally germinated seeds on day 14 / Total number of seeds) × 100%.

[0079] The different initiator formulations and germination rates are shown in Table 5.

[0080] Table 5

[0081]

[0082] As shown in Table 5, the conventional method of soaking seeds in gibberellin can achieve a germination rate of nearly 90%, but requires a large amount of gibberellin. However, combining the sterilized fermentation broth of *dgnkzx004* bacteria with gibberellin significantly improves the germination rate, reaching over 95%. Notably, even when the gibberellin concentration is reduced to 100 mg / L, the germination rate does not decrease significantly; furthermore, even when the gibberellin concentration is reduced to 50 mL / L, increasing the amount of sterilized fermentation broth of *dgnkzx004* bacteria to 15 mL / L still yields a germination rate of 95.1%.

[0083] Preliminary Experiment Example 3

[0084] Based on Preliminary Experiment Examples 1 and 2, this preliminary experimental example is used to record the inventor's experimental situation in preparing floating seedling substrate using dgnkzx004 bacteria.

[0085] The experimental procedure is as follows:

[0086] After disinfecting the seedling trays, different materials as shown in Table 6 were used as seedling substrates, with one seed sown per cell. Commercial organic fertilizer was used as fertilizer, applied once when 70% of the seedlings had emerged, and again after sealing the trays, at a rate of 20g per seedling tray each time. The germination rate and seedling survival rate were then assessed.

[0087] As shown in Table 6, adding mixed bacteria cow manure fermentation products (hereinafter referred to as fermentation products) to commonly used seedling substrates can improve the germination rate and seedling survival rate.

[0088] Table 6

[0089]

[0090] Experimental Example

[0091] This experiment examines the effects of Examples 1-3 and Comparative Examples 1-3 on tobacco yield, tobacco root growth potential, and arsenic content in tobacco leaves.

[0092] Experimental scheme 1: A tobacco experimental park in Yunnan Province was used as the tobacco transplanting site. No arsenic was detected in the experimental soil of the park. The soil physicochemical parameters were: pH=6.84, organic matter content was 11.1 g / kg, and cation exchange capacity was 8.67 cmol / kg.

[0093] Planting was carried out using the schemes of Examples 1-3, Comparative Examples 1-3, and the control group. Yield, single-plant fresh root weight, and longest root length were investigated. The control group, compared to Example 1, was fertilized with commercial organic fertilizer after transplanting. The experimental results are shown in Table 7.

[0094] Table 7

[0095]

[0096] As shown in Table 7, the present invention can significantly improve the root growth vigor of tobacco, with a marked increase in both fresh root weight and root length. This improved root growth vigor is beneficial to tobacco growth, and consequently, the yield of the present invention is significantly higher than that of the methods used in Comparative Examples 1-3.

[0097] Encouraged by the aforementioned positive results of this invention, the inventors considered adding some agricultural and forestry waste to the fermentation substrate to diversify the raw materials. However, according to the experimental results of Comparative Example 1, adding corn stalks to the fermentation substrate had a significant antagonistic effect on the fermentation products' ability to promote tobacco growth. This indicates that the fermenting bacteria produced metabolites detrimental to tobacco growth during the fermentation of the stalks. Based on this, the inventors replaced the fermentation substrate with sheep manure. As shown in the experimental results of Comparative Example 2, although the root growth potential and product yield were improved compared to commercial organic fertilizer, the improvement was still relatively low. This indicates that the fermenting bacteria produced fewer substances beneficial to tobacco growth during the fermentation of sheep manure.

[0098] As shown in the experimental results of Comparative Example 3, when the fermentation liquid of dgnkzx004 bacteria was replaced with commercial organic fertilizer during the seedling stage, even after transplanting, the root growth vigor and yield were still low. This suggests that replacing the fertilizer with one that does not contain the metabolites of dgnkzx004 bacteria during the transplanting process has a significant impact on the growth of tobacco.

[0099] Experimental Scheme 2: A tobacco experimental park in Yunnan Province was used as the tobacco transplanting site. The soil in the park had an arsenic content of 0.71 mg / kg, pH=6.85, organic matter content of 11.3 g / kg, and cation exchange capacity of 8.72 cmol / kg. The park was divided into 7 experimental areas, and the planting was carried out using the schemes of Examples 1-3, Comparative Examples 1-3, and the control group, respectively. Compared to Example 1, the control group was fertilized with commercial organic fertilizer after transplanting.

[0100] After the tobacco leaves mature, the upper leaves are harvested, dried, ground, and passed through a 200-mesh sieve. They are then digested with concentrated nitric acid, and the arsenic content of the resulting digestate is determined by ICP-MS. As shown in Table 8, the method of this invention can effectively reduce the arsenic content in the upper leaves of tobacco, which is beneficial for planting in arsenic-contaminated soil using the planting method of this invention.

[0101] Table 8

[0102]

Claims

1. A full-cycle planting method for improving the quality and increasing the yield of tobacco, characterized in that, The full-cycle planting method includes the following steps: (1) Seed treatment: After washing the seeds with water, soak the seeds in an initiator composed of gibberellin at a concentration of 50~1000mg / L and Bacillus polymyxa dgnkzx004 sterilized fermentation broth at a concentration of 10~15mL / L, then treat them with light, and wash and dry them. (2) Seedling raising: Add mixed bacteria cow manure fermentation product to the seedling substrate and apply Bacillus polymyxa dgnkzx004 fermentation liquid for seedling raising; (3) Transplanting: Transplant the tobacco seedlings and fertilize them with the fermented product of mixed bacteria and cow manure. Among them, the sterilized fermentation broth of Bacillus polymyxa dgnkzx004 is obtained by culturing Bacillus polymyxa dgnkzx004, centrifuging to remove the bacterial cells and collecting the supernatant; The mixed-bacterial cow manure fermentation product is obtained by composting cow manure as a fermentation substrate and using Bacillus polymyxa dgnkzx004 and Bacillus licheniformis ZJU12 as fermentation bacteria. The fermentation broth of the polymyxa bacillus dgnkzx004 was prepared by culturing it in LB liquid medium until the plateau phase at a temperature of 32°C for 20 hours.

2. The full-cycle planting method for improving tobacco quality and increasing yield according to claim 1, characterized in that, In the initiator, the concentration of gibberellin is 50-100 mg / L, and the amount of *Bacillus polymyxa* dgnkzx004 sterilized fermentation broth added is 10-15 mL / L.

3. A full-cycle planting method for improving tobacco quality and increasing yield according to claim 1 or 2, characterized in that, In step (1), the light intensity is 1200 LX and the light duration is 210 minutes.

4. The full-cycle planting method for improving tobacco quality and increasing yield according to claim 1, characterized in that, The amount of mixed bacteria and cow manure fermentation products added to the seedling substrate is 10%wt.

5. The full-cycle planting method for improving tobacco quality and increasing yield according to claim 4, characterized in that, The seedling substrate is composed of cow dung, peat moss, rice husks, perlite, and mixed bacteria fermentation products of cow dung in a weight ratio of 2:5:1:1:

1.

6. The full-cycle planting method for improving tobacco quality and increasing yield according to claim 4, characterized in that, The seedling substrate is composed of corn stalk charcoal, peat moss, perlite, and mixed bacteria cow manure fermentation products in a weight ratio of 5:2:2:

1.

7. A full-cycle planting method for improving tobacco quality and increasing yield according to any one of claims 4 to 6, characterized in that, When applying the fermentation broth of Bacillus polymyxa dgnkzx004, apply it once when 70% of the seedlings have emerged, and then apply it again after sealing the trays.

8. The full-cycle planting method for improving tobacco quality and increasing yield according to claim 7, characterized in that, When raising seedlings, sow one seed in each hole.

9. The full-cycle planting method for improving tobacco quality and increasing yield according to claim 1, characterized in that, When fertilizing after transplanting, apply fertilizer on the day of transplanting and on the 20th day after transplanting.

10. A full-cycle planting method for improving tobacco quality and increasing yield according to claim 9, characterized in that, After transplanting, apply fertilizer at a rate of 30 kg / mu each time.

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