Preparation of peptide fertilizer for promoting long-acting growth of microorganisms and application of peptide fertilizer in phosphorus-reducing, calcium-reducing and smoke-increasing tobacco leaf planting

Abstract

The invention belongs to the technical field of tobacco planting, and mainly relates to preparation of a peptide fertilizer for promoting long-acting growth of microorganisms and application of the peptide fertilizer in phosphorus-reducing, calcium-reducing and smoke-increasing tobacco planting. Polypeptide, quick and long-acting carbon source-molasses, long-acting carbon source-paraffin and waste activated carbon containing amino acid are compounded to form the quick and long-acting multifunctional compound peptide fertilizer with a microbial porous living space, the peptide fertilizer is used for tobacco planting, the growth performance of tobacco can be improved, the growth effect of the peptide fertilizer exceeds that of a special fertilizer, use of phosphorus and calcium is reduced by 30-50%, and the cost is reduced. The method is beneficial to reducing tobacco leaf quality reduction and influence on river and lake non-point source pollution caused by too high available phosphorus content in tobacco planting soil, can promote long-acting growth of microorganisms, improves the number of soil microbial populations, ensures good growth of tobacco leaves, improves nutritional ingredients such as soluble sugar, amino acids and alcohols in the tobacco leaves, has obvious fragrance, and improves the quality of the tobacco leaves. Phosphorus and calcium reduction and tobacco leaf increase in tobacco leaf planting can be achieved, and obvious economic benefits and application values are achieved in the field of tobacco planting.

Description

Technical Field

[0001] The invention relates to the technical field of tobacco planting, and in particular to the development and application of a fertilizer suitable for tobacco planting that can reduce phosphorus, reduce calcium and enhance fragrance, that is, by compounding polypeptide fertilizers, regulating the number of soil microbial populations, increasing the release of soil phosphorus by microorganisms, and reducing the use of phosphorus fertilizer (superphosphate), reducing the conventional nitrogen, phosphorus and potassium ratio of 1:2:3 to 1:1:3, achieving the effect of reducing phosphorus and reducing calcium, achieving a reduction of phosphorus and calcium by 30-50%, and establishing a tobacco planting model, achieving the development and application of a complete set of technologies for reducing phosphorus and reducing calcium while increasing tobacco leaves and promoting soil microbial diversity. Background Art

[0002] The tobacco industry plays a very important role in the national economy. The poor quality of tobacco leaves seriously affects the scale and benefits of the tobacco industry. Therefore, exploring the production technology of high-quality tobacco leaves is important for expanding the production of high-quality tobacco leaves and improving the benefits of the tobacco processing industry and tobacco planting industry.

[0003] The high yield and high quality of tobacco leaves are closely related to the supply and absorption balance of soil nitrogen, phosphorus, potassium, calcium, etc. Nitrogen, phosphorus, potassium and calcium are all essential nutrients for tobacco growth, and their deficiency and excess will affect the growth and quality of tobacco. For example, excessive calcium will not only cause tobacco leaves to be rough and stiff, increase leaf hardness, and reduce their use value, but also cause imbalances in other metal elements, inhibit tobacco's absorption of potassium, iron, and manganese, thereby seriously affecting the quality of tobacco leaves. Studies have found that the exchangeable calcium content in the soil is extremely significantly positively correlated with the calcium, boron, and chlorine content of tobacco leaves, and is significantly or extremely significantly negatively correlated with the nitrogen, zinc, manganese, and potassium content of tobacco leaves. For another example, under high phosphorus levels, the root / crown ratio of tobacco plants decreases, nutrient absorption is unbalanced, and the stress resistance of tobacco plants decreases. Moreover, when there is too much phosphorus, the nitrogen / phosphorus and phosphorus / potassium ratios in the flue-cured tobacco will be unbalanced, which will lead to excessive nitrogen absorption and metabolic disorders in the tobacco plants, often causing the tobacco plants to be greedy for green and mature late, and to fall yellow poorly. After baking, the tobacco leaves lack oil and elasticity, are easy to break, have poor combustibility, and easily become too thick. In addition, excessive phosphorus will reduce the tobacco plants' absorption of zinc, iron, manganese, etc., and even induce a deficiency of these elements, which will adversely affect the quality and aroma of tobacco.

[0004] In short, the current amount of phosphorus fertilizer used in tobacco cultivation is large, and the phosphorus fertilizers used are mainly superphosphate, double superphosphate, calcium magnesium phosphate, phosphate rock powder and other phosphorus fertilizers with high calcium content. Therefore, excessive phosphorus also leads to excessive calcium. Excessive phosphorus and calcium not only affect the nutrient balance in the tobacco plant, but also affect the growth and metabolism of the tobacco plant, and never affect the quality of tobacco leaves, reduce the quality of tobacco leaves, and hinder the production of high-quality tobacco leaves. Therefore, it is necessary to reasonably control the content of nutrient elements in tobacco planting, not only to ensure strong growth in the early and middle stages and obtain a relatively large biomass, but also to ensure that the nitrogen fertilizer tobacco leaves fall yellow well and the quality of tobacco leaves is high at the end of the vigorous growth. Summary of the invention

[0005] In view of the above problems in the prior art, the present invention provides a peptide fertilizer that promotes the long-term growth of microorganisms, a preparation method thereof, and its application in tobacco planting. The present invention compounds a peptide, a quick-acting carbon source-molasses, a long-acting carbon source-paraffin, and waste activated carbon containing amino acids to produce a fast and long-acting multifunctional compound peptide fertilizer with a porous living space for microorganisms, referred to as peptide fertilizer, which is applied to tobacco planting to improve tobacco growth performance, and its growth effect exceeds that of special fertilizers, and reduces the use of phosphorus and calcium, achieving a 30-50% reduction in phosphorus and calcium, which is conducive to reducing the excessive effective phosphorus content in tobacco-planting soil, resulting in a decline in tobacco quality and affecting river and lake surface source pollution, and can also promote the long-term growth of microorganisms, increase the number of soil microbial populations, and ensure good growth of tobacco leaves, increase the nutritional components of tobacco leaves such as soluble sugars, amino acids, alcohols, and obvious aroma, and can achieve phosphorus and calcium reduction and increase tobacco leaves in tobacco planting, and has obvious economic benefits and application value in the field of tobacco planting.

[0006] To achieve the above object, the present invention is specifically implemented through the following technical solutions:

[0007] The first aspect of the present invention provides a method for preparing a peptide fertilizer that promotes the long-term growth of microorganisms, comprising the following steps: compounding polypeptides, potassium fertilizers and phosphorus fertilizers according to a nitrogen, phosphorus and potassium ratio of 1:1:3, and adding molasses, paraffin and waste activated carbon containing amino acids, mixing evenly, and making into powder or granules.

[0008] Furthermore, each ton of the peptide fertilizer consists of 900 kg of polypeptide, 40 kg of calcium superphosphate, 20 kg of potassium sulfate, 10 kg of molasses, 10 kg of paraffin and 20 kg of waste activated carbon containing amino acids.

[0009] A second aspect of the present invention provides a peptide fertilizer for promoting the long-term growth of microorganisms, which is prepared by the method for preparing a peptide fertilizer for promoting the long-term growth of microorganisms as described above.

[0010] The third aspect of the present invention provides the use of the peptide fertilizer for promoting the long-term growth of microorganisms as described above in tobacco planting.

[0011] Furthermore, the peptide fertilizer is used as base fertilizer or topdressing fertilizer.

[0012] Furthermore, 60 kg of the peptide fertilizer is applied per mu.

[0013] Furthermore, the application includes at least one of the following (1)-(6):

[0014] (1) Application in improving agronomic traits of tobacco, wherein the improved agronomic traits include increasing leaf length, leaf width, reducing aspect ratio and / or increasing leaf area;

[0015] (2) Application in increasing the content of soluble sugar, amino acid and / or alcohol in tobacco leaves;

[0016] (3) Application in reducing the activity of sucrose synthase in tobacco leaves;

[0017] (4) Application in improving soil microbial diversity;

[0018] (5) Application in improving the aroma of tobacco leaves;

[0019] (6) Application in reducing the amount of phosphorus fertilizer and calcium fertilizer in tobacco cultivation.

[0020] The advantages and positive effects of the present invention are:

[0021] 1. The peptide fertilizer provided by the present invention utilizes polypeptide to control nitrogen, and its nitrogen is organic nitrogen, which can be preserved and slowly released in the soil, avoiding the use of inorganic nitrogen fertilizer in conventional production, and rapidly releasing nitrogen into the soil to cause excessive local concentration and damage to plant growth and nitrogen loss. In addition, not only can the release of nitrogen be controlled, but also the release of potassium fertilizer and phosphate fertilizer can be controlled by complex adsorption with metal elements such as potassium and calcium, and the effectiveness of phosphorus can be controlled by regulating the soil environment, so that the release of nitrogen, phosphorus, potassium, calcium, etc. can be adjusted simultaneously. The tobacco leaf growth effect of the current nitrate nitrogen fertilizer can be achieved by using the peptide fertilizer of the present invention. It has obvious performance in promoting crop growth, and tobacco only needs 5-7 days to turn green after applying the peptide fertilizer of the present invention, and its speed is equivalent to urea, but it itself belongs to nitrogen as protein nitrogen, which is completely organic fertilizer, and overcomes the shortcomings of poor solubility, slow fertilizer efficiency, and easy loss of organic fertilizer and causing non-point source pollution.

[0022] 2. The peptide fertilizer of the present invention can quickly promote the growth of soil microorganisms. Previous studies have shown that protein polypeptides can increase the number of soil microorganisms faster than all current organic and inorganic fertilizers, especially better than urea (minimum), ammonium carbonate and other chemical fertilizers (ammonium nitrogen). At the same time, it has the function of microbial adsorption and binding to heavy metal ions, which can reduce the absorption of heavy metals by plants and establish a new technical foundation for the cultivation of high-quality tobacco leaves.

[0023] 3. The peptide fertilizer of the present invention is different from the characteristics of ammonium nitrogen such as urea and ammonium carbonate. The nitrogen it contains is all protein nitrogen. At the same time, the types of amino acids after conversion are complete, the proportion of metabolites formed is balanced, the crop growth ratio is coordinated, and the crop appearance is small and straight leaves, while urea and ammonium carbonate have large and long leaves, high water content, soft, yellow leaves, and thin leaves.

[0024] 4. The price of the peptide fertilizer developed by the present invention is comparable to that of conventional fertilizers, the yield of crops grown is comparable to or slightly higher than that of conventional fertilizers, and there is no negative cumulative effect when used for a long time.

[0025] 5. The yield of tobacco leaves grown with the peptide fertilizer of the present invention is currently the only organic fertilizer that can reach the yield of nitrate nitrogen tobacco-specific fertilizer, and the application quality improvement detection and evaluation have reached and partially exceeded the conventional planting.

[0026] 6. Activated carbon powder containing amino acids is added to the peptide fertilizer of the present invention, which has a porous matrix space peptide fertilizer that promotes the long-term growth of microorganisms, that is, the microorganisms have independent porous reproduction space, and the external environment has little interference with the living space of the microorganisms, which can achieve long-term and stable promotion of soil microbial proliferation and improve soil microbial diversity.

[0027] 7. The peptide fertilizer of the present invention can increase the flavor substances of tobacco leaves, such as alcohol, aldehyde, acid, fat, polysaccharide, vitamins and other substances, which are far richer than special nitrate nitrogen fertilizers, and is beneficial to improving the aroma components of tobacco leaves.

[0028] 8. The peptide fertilizer of the present invention can reduce phosphorus and calcium by 30-50%, that is, apply 30-50% less superphosphate, reduce the impact of excessive phosphorus and calcium content on tobacco quality and the pollution to surrounding river and lake water sources. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0030] Figure 1 The effect of peptide fertilizer on the soluble sugar content of flue-cured tobacco leaves in the present invention;

[0031] Figure 2 The effect of peptide fertilizer on the amino acid content of flue-cured tobacco leaves in the present invention;

[0032] Figure 3 The effect of peptide fertilizer on the activity of sucrose synthase in flue-cured tobacco leaves in the present invention;

[0033] Figure 4The Venn diagram of the effect of peptide fertilizer on rhizosphere soil microbial community in the embodiment of the present invention;

[0034] Figure 5 It is a bar graph at the microbial phylum level showing the effect of peptide fertilizer on rhizosphere soil microbial community according to an embodiment of the present invention;

[0035] Figure 6 A heat map at the microbial genus level showing the effect of the peptide fertilizer on the rhizosphere soil microbial community according to an embodiment of the present invention;

[0036] Figure 7 This is a PCA analysis diagram of the transcription level of the effect of peptide fertilizer on flue-cured tobacco leaf metabolites in the embodiment of the present invention;

[0037] Figure 8 This is a graph showing the difference in expression levels of the effects of peptide fertilizer on metabolites in flue-cured tobacco leaves according to an embodiment of the present invention;

[0038] Fig. 9 This is a one-way analysis of variance diagram of metabolites of the peptide fertilizer of the embodiment of the present invention on the metabolites of flue-cured tobacco leaves. DETAILED DESCRIPTION

[0039] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. The embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0040] Based on the information contained in the present invention, it is easy for those skilled in the art to make various changes to the precise description of the present invention without departing from the spirit and scope of the appended claims. It should be understood that the scope of the present invention is not limited to the defined processes, properties or components, because these embodiments and other descriptions are only for the purpose of illustrating specific aspects of the present invention. In fact, various changes that a person skilled in the art or related fields can obviously make to the embodiments of the present invention are covered within the scope of the appended claims.

[0041] In order to better understand the present invention rather than limit the scope of the present invention, all numerals and other numerical values ​​used in the present invention to represent dosage, percentage, etc., should be understood as modified by the word "about" in all cases. Therefore, unless otherwise specified, the numerical parameters listed in the specification and the appended claims are approximate values, which may be changed according to the different ideal properties attempted to be obtained. Each numerical parameter should at least be regarded as obtained according to the reported significant figures and by conventional rounding methods. In addition, it should be noted that the meaning of the terms "including", "comprising", "containing", "having" and the like is non-restrictive, and other steps and other ingredients that do not affect the results can be added.

[0042] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, specific embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0043] The embodiment of the present invention provides a method for preparing a peptide fertilizer for promoting the long-term growth of microorganisms, comprising the following steps:

[0044] According to the formula ratio requirements of nitrate nitrogen used in conventional planting - the conventional nitrogen, phosphorus and potassium ratio of 1:2:3, and the current situation of the problem of high effective phosphorus content in the soil, a new peptide fertilizer preparation method and raw material usage ratio for promoting long-term growth of microorganisms were determined. The nitrogen, phosphorus and potassium ratio is 1:1:3, and the application ratio of phosphorus and calcium-reducing phosphate fertilizer reaches 30-50%. Then, according to the determined application ratio of nitrogen fertilizer, potassium fertilizer and phosphate fertilizer, peptide fertilizer that meets the following requirements is formulated: each ton of peptide fertilizer consists of 900kg of polypeptide, 40kg of supercalcium phosphate, 20kg of potassium sulfate, 10kg of molasses, 10kg of paraffin and 20kg of waste activated carbon containing amino acids, and is mixed into powder or granulated into granular fertilizer.

[0045] During tobacco planting, peptide fertilizer is applied to the planting soil by spot-applying base fertilizer or used entirely as topdressing. 60 kg per mu can meet all fertilizer requirements for tobacco leaf growth.

[0046] The present invention utilizes polypeptide to control nitrogen, and its nitrogen is organic nitrogen, which can be preserved in the soil and slowly released, avoiding the use of inorganic nitrogen fertilizer in conventional production, and rapidly releasing nitrogen into the soil to cause excessive local concentration and damage to plant growth and nitrogen loss. The polypeptide can not only control the release of nitrogen, but also control the release of potash fertilizer and phosphate fertilizer by complexing and adsorbing with metal elements such as potassium and calcium, and can also regulate the soil environment to control the effectiveness of phosphorus, so that the release of nitrogen, phosphorus, potassium, calcium, etc. can be adjusted at the same time. Planting tobacco with the peptide fertilizer of the present invention can achieve the tobacco leaf growth effect of the current nitrate nitrogen fertilizer. It has obvious performance in promoting crop growth. After applying the peptide fertilizer of the present invention, tobacco only needs 5-7 days to turn green, and its speed is equivalent to urea, but it itself belongs to protein nitrogen, which is completely organic fertilizer, and overcomes the shortcomings of poor solubility, slow fertilizer efficiency, and easy loss of organic fertilizers that cause non-point source pollution. Moreover, it can quickly promote the growth of soil microorganisms. Previous studies have shown that protein polypeptides increase the number of soil microorganisms faster than all current organic and inorganic fertilizers, especially better than urea (minimum), carbon ammonium and other fertilizers (ammonium nitrogen). At the same time, it has microbial adsorption and binding functions for heavy metal ions, which can reduce the absorption of heavy metals by plants, and establish a new technical basis for high-quality tobacco planting. At the same time, the peptide fertilizer of the present invention adds activated carbon powder containing amino acids, which has a porous matrix space peptide fertilizer that promotes the long-term growth of microorganisms, that is, the microorganism has an independent porous breeding space, and the external environment interferes less with the living space of microorganisms, which can achieve long-term and stable promotion of soil microorganism proliferation and improve soil microbial diversity. These characteristics enable the peptide fertilizer of the present invention to improve the flavor substances of tobacco leaves, such as alcohol, aldehyde, acid, fat, polysaccharide, vitamins and other substances are far richer than special nitrate nitrogen fertilizers, which is conducive to improving the aroma components of tobacco leaves. In addition, the peptide fertilizer of the present invention can achieve 30-50% reduction in phosphorus and calcium, that is, 30-50% less superphosphate is applied, reducing the impact of excessive phosphorus and calcium content on tobacco quality and the pollution of surrounding river and lake water sources. In short, the present invention has obvious economic benefits and application value in the field of tobacco planting.

[0047] The polypeptide fertilizer described above is prepared by conventional techniques in the art, such as the method disclosed in the patent "Preparation of soil remediation agent using protein peptides and method for remediating soil (ZL201210040772.9)".

[0048] The present invention is further described below in conjunction with specific examples. The experimental methods in the following examples without specifying specific conditions are generally carried out according to the conditions recommended by the manufacturer.

[0049] In the following examples, the preparation method of the polypeptides is described in the patent "Preparation of soil remediation agent using protein peptides and method for remediating soil (ZL201210040772.9)". The waste activated carbon containing amino acids is derived from the decolorized waste of an amino acid production plant.

[0050] Example

[0051] The amount of nitrogen fertilizer is determined according to the nitrogen application amount of conventional fertilization, and the required amount of polypeptide is calculated by the nitrogen content of the polypeptide. A flue-cured tobacco field planting test was carried out with conventional tobacco-specific fertilizer (N-P2O5-K2O=8:16:24=1:2:3 by mass percentage (wt)) as a control. In the embodiment of the present invention, the nitrogen and potassium in the fertilization process are controlled to be consistent with the tobacco-specific fertilizer, and the phosphorus is controlled at 50% of the special fertilizer, that is, N-P2O5-K2O=1:1:3. Each ton of peptide fertilizer that promotes the long-term growth of microorganisms is composed of 900kg of polypeptide, 40kg of supercalcium hydrogen phosphate, 20kg of potassium sulfate, 10kg of molasses, 10kg of paraffin and 20kg of waste activated carbon containing amino acids. When planting tobacco, fertilizer is applied at 60kg per mu by point application, and 65kg of special fertilizer is applied.

[0052] After the tobacco turned yellow, different indicators were measured to explore the effect of the peptide fertilizer of the present invention on tobacco growth and quality. The specific research results are as follows:

[0053] (1) Effects on agronomic traits of field tobacco

[0054] Special fertilizer (nitrogen, phosphorus and potassium ratio of 1:2:3) was used as the control group CK, a new peptide fertilizer that promotes the long-term growth of microorganisms (nitrogen, phosphorus and potassium ratio of 1:1:3) was used as the experimental group T2, and the same amount of amino acid fertilizer was used as the nitrogen source as the experimental reference group T1. The results of agronomic traits of field planting are shown in Table 1.

[0055] Table 1 Effects of different field treatments on agronomic traits

[0056]

[0057] The results show that tobacco fertilizer shows good agronomic indicators in the early stage of flue-cured tobacco growth, and has better effects on tobacco leaf growth than the other two treatments. Amino acid fertilizer shows its characteristics of long-term fertility in the middle and late stages of flue-cured tobacco growth, and has good agronomic effects. The peptide fertilizer for promoting long-term growth of microorganisms has the largest leaf area, better leaf opening, and high economic utilization value.

[0058] (2) Effect on soluble sugar content of field tobacco leaves

[0059] Effects of different treatments on soluble sugar content in tobacco leaves in field validation experiments Figure 1As shown in the results, at the cluster stage, the soluble sugar content of the T2 treatment was the highest at 45.13 mg / g, and the soluble sugar content of the T1 treatment was the lowest at 40.97 mg / g, with no significant difference among the three treatments; at the vigorous growth stage, the soluble sugar content showed that T2 and T1>CK, the soluble sugar content of the T2 treatment was the highest at 64.92 mg / g, and the soluble sugar content of the CK treatment was the lowest at 55.65 mg / g. The soluble sugar content of the T1 and T2 treatments was significantly increased by 7.90 mg / g and 9.28 mg / g respectively compared with the CK treatment. The results showed that the application of peptide fertilizer and amino acid fertilizer during the vigorous growth period promoted the production of soluble sugar.

[0060] (3) Effects on the amino acid content of field tobacco leaves

[0061] Effects of different treatments on the amino acid content of tobacco leaves in field validation experiments Figure 2 As shown, during the clumping stage, the amino acid content in T2 treatment was the highest at 75.33 μmol / g, and the amino acid content in CK treatment was the lowest at 67.10 μmol / g, and there was no significant difference among the three treatments; during the vigorous growth stage, the amino acid content showed that T2 and T1>CK, the amino acid content in T2 treatment was the highest at 102.31 μmol / g, and the amino acid content in CK treatment was the lowest at 70.36 μmol / g. The amino acid contents in T1 and T2 treatments were significantly increased by 16.08 μmol / g and 36.95 μmol / g, respectively, compared with those in CK treatment.

[0062] (4) Effects on the activity of sucrose synthase in field tobacco leaves

[0063] Effects of different treatments on sucrose synthase activity in tobacco leaves in field validation experiments Figure 3 As shown in the results, during the clustering stage, the sucrose synthase activity of each treatment was CK>T1>T2. The sucrose synthase (SS) activity of the CK treatment reached 694.66U / g, and the T1 and T2 treatments were significantly lower than the CK treatment, decreasing by 154.41U / g and 187.35U / g, respectively, and there was no significant difference between the T1 and T2 treatments. During the vigorous growth period, the overall trend of sucrose synthase in the clustering stage was the same as that of the treatments, showing CK>T1>T2, but the sucrose synthase activity was enhanced compared with the clustering stage, among which the sucrose synthase activity of the CK treatment was the highest at 871.64U / g, and the sucrose synthase activity of the T2 treatment was the lowest at 736.15U / g. The T1 and T2 treatments were significantly lower than the CK treatment by 95.86U / g and 135.48U / g. The results showed that tobacco-specific fertilizers promoted the activity of sucrose synthase and accelerated the decomposition of sucrose in tobacco leaves.

[0064] (5) Effects of different treatments on root soil microorganisms

[0065] 1) Analysis of Alpha Diversity of Field Rhizosphere Soil Microorganisms

[0066] The species diversity index of field soil microbial community was obtained under the condition of 97% similarity level as shown in Table 2. The coverage of all treatments was greater than 0.97, indicating that the sequencing ability can truly reflect the characteristics of soil microbial community. The soil microbial sobs, ace, and shannon indexes under each treatment showed T2>T1>CK. Among them, the Sobs index of T1 and T2 treatments increased significantly by 176.67 and 202.67 compared with the CK treatment; the ACE index of T2 treatment increased significantly by 290.12 compared with the CK treatment; the Shannon index of T1 and T2 treatment groups increased significantly by 1.51 and 2.23 compared with the CK treatment. It can be seen that peptide fertilizer and amino acid fertilizer can effectively improve the richness and diversity of soil microorganisms.

[0067] Table 2 Alpha diversity index of rhizosphere soil microorganisms in the field

[0068]

[0069] 2) Effect of unique species composition of rhizosphere soil microorganisms in the field

[0070] like Figure 4 As shown in the Venn diagram of the rhizosphere soil microbial community in the field verification test, the three treatment combinations have a total of 2463 OTUs of the same microbial species, accounting for 28.77%. At the same time, there are the same microbial species between every two treatments, among which the same OTUs of the CK treatment and the T1 treatment are 443, accounting for 5.17%, the same OTUs of the T1 treatment and the T2 treatment are 471, accounting for 5.50%, and the same OUTs of the T2 treatment and the CK treatment are 529, accounting for 6.18%. The unique microbial OTU values ​​between different treatments show that T2>T1>CK, and the unique microbial OUTs of the T1 and T2 treatments are 424 (4.95%) and 533 (6.23%) higher than those of the CK treatment, respectively. The results show that compared with tobacco-specific fertilizers and amino acid fertilizers, the peptide fertilizer of the present invention can effectively increase the types of rhizosphere soil microorganisms.

[0071] 3) Bar chart analysis of field rhizosphere soil microbial community composition

[0072] The bar graph of the effects of different treatments on rhizosphere soil microbial communities in the field validation experiment is shown in Figure 2. Figure 5As shown in the figure, the top ten taxa in terms of average relative abundance at the phylum level are shown, namely Chloroflexi, Actinobacteriota, Proteobacteria, Acidobacteriota, Firmicutes, Planctomycetota, Verrucomicrobiota, Myxococcota, Patescibacteria and Cyanobacteria. Among them, Chloroflexi is absolutely dominant in all treatments, followed by Actinobacteria and Proteobacteria.

[0073] The relative abundance of Chloroflexi was generally CK>T1>T2. The relative abundance of Chloroflexi in CK treatment was 0.66% and 3.97% higher than those in T1 and T2 treatments, indicating that tobacco-specific fertilizer can increase the relative abundance of Chloroflexi. The relative abundance of Actinobacteria was generally T2>CK>T1. The relative abundance of Actinobacteria in T2 treatment was 11.29% and 10.29% higher than those in T1 and CK treatments, indicating that compound peptide fertilizer has a significant effect on the relative abundance of Actinobacteria.

[0074] 4) Heat map analysis of rhizosphere soil microbial community composition in the field

[0075] At the genus classification level, based on the species annotation and abundance information of the field demonstration test samples, the top 20 species in terms of rhizosphere soil microbial abundance were selected, and a heat map was drawn according to their abundance information. Figure 6 As shown in the figure, T1 and T2 treatments belong to the same branch, indicating that peptide fertilizer is the main reason affecting the structure of microbial communities. As shown in the box in the figure, the relative abundance of Gordonia and Mycobacterium was higher in the T2 treatment. Arthrobacter and Bacillus were greatly affected by peptide fertilizer, with higher relative abundance in the T2 treatment, which was significantly different from the other two treatments. The relative abundance of the genus GAL15 in the T1 treatment was higher than that in the other two treatments, indicating that peptide fertilizer may promote its growth and reproduction.

[0076] (6) Metabolomics analysis

[0077] The changes in transcription and metabolites in different treatment groups were determined. The results of PCA principal component analysis at the transcriptomic level are shown in Figure 7 The results showed that there were significant differences in the transcription levels between the experimental group and the verification special fertilizer control group of different peptide fertilizer groups. The expression level differences were further analyzed at the transcription level. Figure 8 The results showed that compared with the control group, the levels of multiple transcriptomes in the peptide fertilizer group were significantly upregulated.

[0078] Metabolomics analysis of metabolites ( Fig. 9 ), the results showed that the peptide fertilizer group significantly increased the content of ornithine, sucrose, proline, glycine nicotine, octolose, apiglycoside, cinnamyl glycoside, methyl acetate glucoside, broccoli glycoside, hydroxycaproic acid, oxidized sulfonic acid and other sugars, amino acids and organic acids, and significantly reduced the content of glutamine and nicotine (nicotine)

[0079] (7) Inhalation test

[0080] The tobacco leaves grown with peptide fertilizer were baked, processed and made into finished cigarette products, and then manually tasted. The quality of the tobacco leaves was tasted by many experts, and the results (Table 3) are as follows: The results show that many indicators of tobacco leaves fertilized with peptide fertilizer are similar to those of special fertilizers, and some are better than special fertilizers, which also shows the advancement and feasibility of the technology.

[0081] Table 3 Peptide fertilizer tobacco leaf smoking test results

[0082]

[0083] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention. The purpose of the invention is to develop a new full-nutrition organic fertilizer - compound peptide fertilizer under the background of reducing phosphorus and calcium by 30-50%, so as to establish a technical foundation for growing high-quality tobacco leaves.

Claims

1. A method for preparing a peptide fertilizer for promoting long-term growth of microorganisms, characterized in that: The following steps are involved: According to the nitrogen, phosphorus and potassium ratio of 1:1:3, polypeptides, potassium fertilizers and phosphorus fertilizers are compounded, and molasses, paraffin and waste activated carbon containing amino acids are added. Mix well and make into powder or granules.

2. The method for preparing a peptide fertilizer for promoting long-term growth of microorganisms according to claim 1, characterized in that: Each ton of the peptide fertilizer is composed of 900 kg of polypeptide, 40 kg of calcium superphosphate, 20 kg of potassium sulfate, 10 kg of molasses, 10 kg of paraffin and 20 kg of waste activated carbon containing amino acids.

3. A peptide fertilizer for promoting long-term growth of microorganisms, characterized in that: The peptide fertilizer is prepared by the preparation method for promoting the long-term growth of microorganisms as described in any one of claims 1-2.

4. Application of the peptide fertilizer for promoting long-term growth of microorganisms as claimed in claim 3 in tobacco planting.

5. The use of the peptide fertilizer for promoting long-term growth of microorganisms in tobacco planting according to claim 4, characterized in that: The peptide fertilizer is used as base fertilizer or topdressing fertilizer.

6. The use of the peptide fertilizer for promoting long-term growth of microorganisms in tobacco planting according to claim 4, characterized in that: 60 kg of the peptide fertilizer was applied per mu.

7. The use of the peptide fertilizer for promoting long-term growth of microorganisms in tobacco planting according to claim 4, characterized in that: The application includes at least one of the following (1)-(6): (1) Application in improving agronomic traits of tobacco, wherein the improved agronomic traits include increasing leaf length, leaf width, reducing aspect ratio and / or increasing leaf area; (2) Application in increasing the content of soluble sugar, amino acid and / or alcohol in tobacco leaves; (3) Application in reducing the activity of sucrose synthase in tobacco leaves; (4) Application in improving soil microbial diversity; (5) Application in improving the aroma of tobacco leaves; (6) Application in reducing the amount of phosphorus fertilizer and calcium fertilizer in tobacco cultivation.

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