Paenibacillus polymyxa type bio-organic fertilizer and application thereof

Abstract

The invention belongs to the field of fertilizers, and particularly relates to a paenibacillus polymyxa type bio-organic fertilizer which comprises the following components in parts by weight: 5-15 parts of rice straw, 10-20 parts of walnut green seedcase, 35-45 parts of biogas residue, 2-8 parts of chicken manure, 2-8 parts of sheep manure, 3-7 parts of zymophyte, 6-10 parts of paenibacillus polymyxa fermentation liquor, 4-9 parts of humic acid, 2-6 parts of zinc sulfate and 2-6 parts of borax. The biological organic fertilizer disclosed by the invention can be used for preventing and treating watermelon fusarium wilt and increasing the stem diameter and plant height of watermelons; and the method is environment-friendly.

Description

Technical Field

[0001] The invention belongs to the field of fertilizers, and in particular relates to a Paenibacillus polymyxa type biological organic fertilizer and application thereof. Background Art

[0002] In recent years, with the continuous planting of high value-added crops and the development of protected areas, soil-borne diseases have become more serious year by year. If not intervened in time, it will lead to crop failure in severe cases. However, in the prevention and control of diseases, the use of a single, especially targeted, agent is not only difficult to achieve the ideal prevention and control effect, but also causes the pathogen to develop drug resistance.

[0003] Biofertilizers are biological products containing specific active microorganisms, including microbial inoculants, compound biofertilizers, and bio-organic fertilizers. Bio-organic fertilizers are widely used because they contain specific functional microorganisms and rich nutrients. Unlike organic fertilizers, bio-organic fertilizers introduce special functional strains, such as microorganisms with growth-promoting and broad-spectrum antagonistic effects. These microorganisms can effectively decompose macromolecules in organic fertilizers and prevent the occurrence of plant diseases. The microorganisms used in biological control are mainly actinomycetes and bacteria, and a few fungi also have biocontrol potential. Among them, Bacillus has an inhibitory effect on the occurrence of many plant diseases and is an important group in crop biological control. Actinomycetes can inhibit the occurrence of diseases through antibiotics, competition, heavy parasitism, and induced resistance.

[0004] Paenibacillus polymyxa is a multifunctional Gram-positive bacterium that can produce endospores. It is widely found in soil, plant surfaces and rhizosphere, and is also distributed in marine sediments. Its spores have strong resistance to radiation, heat, dryness, extreme pH values ​​and toxic chemicals, and can remain active for a long time in a dormant state. The spores have the characteristics of fast reproduction, simple nutritional requirements and easy colonization in the rhizosphere of plants. Paenibacillus polymyxa can produce abundant biologically active metabolites, induce host plants to resist diseases, and prevent and control diseases. These active substances are mostly classified into polypeptides, polysaccharides, proteins, as well as nucleoside analogs, aldehyde acids and pyrazines according to their structures.

[0005] This study selected functional strains with growth-promoting and antagonistic effects, used different organic fertilizers as nutrient carriers of the functional strains, prepared biological organic fertilizers, and studied the effects of different carrier biological organic fertilizers on the incidence of watermelon wilt disease, providing certain support for the biological control of soil-borne diseases of watermelon and scientific planting management. Summary of the invention

[0006] The present invention provides a polymyxa-type bio-organic fertilizer and its application. The technical solution of the present invention is:

[0007] A Paenibacillus polymyxa type biological organic fertilizer comprises the following ingredients in parts by weight: 5-15 parts of rice straw, 10-20 parts of walnut green peel, 35-45 parts of biogas residue, 2-8 parts of chicken manure, 2-8 parts of sheep manure, 3-7 parts of fermentation bacteria, 6-10 parts of Paenibacillus polymyxa fermentation liquid, 4-9 parts of humic acid, 2-6 parts of zinc sulfate and 2-6 parts of borax; the fermentation bacteria are EM bacteria.

[0008] The polymyxa bacillus of the present invention can be any known strain in the prior art, preferably, polymyxa bacillus ZJU0901, which is deposited in the China Type Culture Collection (CCTCC) at Wuhan University, Luojia Mountain, Wuhan, on July 19, 2009, with a deposit number of CCTCC NO: M 209157. The strain is a known strain of CN101712941B in the existing literature, and the strain is used to complete the experiment of the present invention.

[0009] The EM bacteria of the present invention comes from Zhengzhou Baiyibao Biotechnology Co., Ltd.

[0010] Preferably, the composition comprises the following parts by weight: 8-12 parts of rice straw, 14-16 parts of walnut green peel, 38-42 parts of biogas residue, 4-6 parts of chicken manure, 4-6 parts of sheep manure, 4-6 parts of fermentation bacteria, 7-9 parts of Paenibacillus polymyxa fermentation liquid, 6-8 parts of humic acid, 3-6 parts of zinc sulfate and 3-5 parts of borax.

[0011] Preferably, the composition comprises the following ingredients in parts by weight: 10 rice straw, 15 walnut green peel, 40 biogas residue, 5 chicken manure, 5 sheep manure, 5 fermentation bacteria, 8 Paenibacillus polymyxa fermentation liquid, 7 humic acid, 4 zinc sulfate, and 4 borax.

[0012] The polymyxa-type bio-organic fertilizer described in the present invention is prepared by solid-state aerobic fermentation. Specifically, the organic fertilizer carrier (rice straw, walnut green skin, biogas residue, chicken manure, sheep manure) is sterilized at 121°C for 1 hour, cooled to room temperature and then used for standby; conventional solid-state aerobic fermentation is used in the art, and after the fermentation is completed, the mixture is stirred evenly, and random samples are taken to detect whether the various technical indicators meet the standards of bio-organic fertilizer (NY 884-2012), and the bio-organic fertilizer that meets the standards is bagged and placed in a cool and dry place for storage.

[0013] The Paenibacillus polymyxa bio-organic fertilizer of the present invention is used in watermelon agricultural production, especially for preventing and treating watermelon wilt and increasing the stem thickness and plant height of watermelons.

[0014] By adopting the above technical solution, the present invention has the following beneficial effects:

[0015] The treatment of applying the bio-organic fertilizer of the present invention has a good prevention effect on watermelon wilt disease, and the application of the bio-organic fertilizer increases the stem thickness and plant height of the watermelon plants. DETAILED DESCRIPTION

[0016] The present invention is further explained below in conjunction with specific implementation modes.

[0017] Example 1: Preparation of Paenibacillus polymyxa type biological organic fertilizer

[0018] 1.1 Preparation of Paenibacillus polymyxa fermentation broth

[0019] The preparation method was described in the reference “The control effect of the fermentation liquid of Bacillus polymyxa KM2501-1 on tomato root-knot nematodes”.

[0020] Use an inoculation loop to take a small amount of bacterial culture preservation liquid and streak it on solid KMB medium (tryptone 2%, MgSO4·7H2O0.15%, K2HPO40.15%, glycerol 1%, pH7.2-7.4), and culture it at 28℃ for about 48 hours until a single colony grows. Use an inoculation loop to pick a single colony and inoculate it in a 250mL Erlenmeyer flask containing 50mL sterilized KMB liquid culture medium, and culture it at 28℃, 180r / min for about 24 hours until the logarithmic growth phase of the bacteria. Take 1.4mL of the above cultured bacterial liquid and inoculate it in a 500mL Erlenmeyer flask containing 140mL sterilized KMB liquid culture medium, and culture it at 28℃, 180r / min for about 12 hours until the logarithmic growth phase of the bacteria, as the activated seed liquid.

[0021] The activated seed liquid was added to the sterilized fermentation medium SBM medium (soybean meal 3%, soluble starch 3%, calcium chloride 0.01%, sodium chloride 0.25%, pH 8.5) for fermentation in a 15L mechanical stirring aeration fermenter. The fermentation process of the fermenter was as follows: the medium liquid volume was 7L, the fermentation temperature was 28°C, the rotation speed was 200r / min, the pH value of the sterilized medium was about 8.5, the inoculation amount was 2%, the defoamer was 5mL, the ventilation volume was 0.8m3 / h, and the tank pressure was 0.02-0.05MPa. The spore content in the fermentation liquid was determined by the dilution coating plate method, and the spore content was 1.5×10 10 cfu / mL.

[0022] 1.2 Formula of polymyxa-based bio-organic fertilizer

[0023] Formula 1: Contains the following ingredients in parts by weight: 10 parts of rice straw, 15 parts of walnut green peel, 40 parts of biogas residue, 5 parts of chicken manure, 5 parts of sheep manure, 5 parts of fermentation bacteria (EM bacteria), 8 parts of functional bacteria (Bacillus polymyxa fermentation liquid), 7 parts of humic acid, 4 parts of zinc sulfate, and 4 parts of borax.

[0024] Formula 2: Contains the following ingredients in parts by weight: 25 parts of rice straw, 40 parts of biogas residue, 5 parts of chicken manure, 5 parts of sheep manure, 5 parts of fermentation bacteria (EM bacteria), 8 parts of functional bacteria (Bacillus polymyxa fermentation liquid), 7 parts of humic acid, 4 parts of zinc sulfate, and 4 parts of borax.

[0025] Formula 3: Contains the following ingredients in parts by weight: 30 parts of rice straw, 35 parts of walnut green peel, 5 parts of chicken manure, 5 parts of sheep manure, 5 parts of fermentation bacteria (EM bacteria), 8 parts of functional bacteria (Bacillus polymyxa fermentation liquid), 7 parts of humic acid, 4 parts of zinc sulfate, and 4 parts of borax.

[0026] Formula 4: Contains the following ingredients in parts by weight: 10 parts rice straw, 15 parts walnut green peel, 40 parts biogas residue, 10 parts sheep manure, 5 parts fermentation bacteria (EM bacteria), 8 parts functional bacteria (Bacillus polymyxa fermentation liquid), 7 parts humic acid, 4 parts zinc sulfate, and 4 parts borax.

[0027] Formula 5: Contains the following ingredients in parts by weight: 10 parts of rice straw, 15 parts of walnut green peel, 40 parts of biogas residue, 5 parts of chicken manure, 5 parts of sheep manure, 5 parts of fermentation bacteria (EM bacteria), 5 parts of functional bacteria (Bacillus polymyxa fermentation liquid), and 15 parts of humic acid.

[0028] Formula 6: Contains the following ingredients in parts by weight: 10 parts rice straw, 15 parts walnut green peel, 40 parts biogas residue, 5 parts chicken manure, 5 parts sheep manure, 10 parts fermentation bacteria (EM bacteria), 7 parts humic acid, 4 parts zinc sulfate, and 4 parts borax.

[0029] The preparation method of the Paenibacillus polymyxa type biological organic fertilizer formula is as follows: the organic fertilizer carrier (rice straw, walnut green peel, biogas residue, chicken manure, sheep manure) is sterilized at 121° C. for 1 hour, cooled to room temperature and then used for later use; conventional solid aerobic fermentation is adopted in the art, and after the fermentation is completed, the mixture is stirred evenly, and random samples are taken to detect whether various technical indicators meet the standards of biological organic fertilizer (NY 884-2012), and the biological organic fertilizer that meets the standards is bagged and placed in a cool and dry place for storage for later use.

[0030] Example 2: Biological control of soil-borne diseases of watermelon by Paenibacillus polymyxa-based bio-organic fertilizer.

[0031] Test crop: Watermelon

[0032] Test soil: The test soil used for potting was the soil from continuous watermelon planting for three consecutive years. Non-soil components were removed before potting, and sterilized soil was obtained by steam moist heat sterilization at 121℃ for 1h.

[0033] Test pathogen: Fusarium oxysporum suspension

[0034] Experimental Design:

[0035] A: Sterilized soil + pathogens;

[0036] B: Sterilized soil + pathogens + organic fertilizer (Formula 6)

[0037] C1: Sterilized soil + pathogens + Paenibacillus polymyxa bio-organic fertilizer (Formula 1)

[0038] C2: Sterilized soil + pathogens + Paenibacillus polymyxa bio-organic fertilizer (Formula 2)

[0039] C3: Sterilized soil + pathogens + Paenibacillus polymyxa bio-organic fertilizer (Formula 3)

[0040] C4: Sterilized soil + pathogens + Paenibacillus polymyxa bio-organic fertilizer (Formula 4)

[0041] C5: Sterilized soil + pathogens + Paenibacillus polymyxa bio-organic fertilizer (Formula 5)

[0042] Bio-organic fertilizer and organic fertilizer were mixed with soil at an application rate of 1%. Healthy watermelon seedlings with good growth and uniform size were selected and transplanted into pots after the seedlings grew 3-4 true leaves. Ten days after transplanting, a suspension of watermelon wilt bacteria was inoculated into each pot, with a spore concentration of 10 5 CFU / mL. In the sterilized soil pot test, 10 plants were set up for each treatment, repeated 4 times, and conventional water and fertilizer management was used.

[0043] Six watermelon plants were randomly selected for each treatment, and the plant height and stem thickness of the watermelons were counted on the 30th and 60th days after transplanting. During the experiment, the number of diseased watermelon plants and the disease grade were counted, and the incidence rate, disease index and control rate were calculated.

[0044] Condition Investigation

[0045] On the 45th day after transplanting, the number of diseased watermelon plants and the disease grade were counted, and the incidence rate, disease index and control effect were calculated.

[0046] The disease levels of plants are divided into 5 levels:

[0047] Level 0: No symptoms of infection;

[0048] Level 1: 1 leaf to 1 / 4 of the leaves of the whole plant show symptoms of infection;

[0049] Level 2: 1 / 4 to 1 / 2 of the leaves of the whole plant show symptoms of infection;

[0050] Level 3: More than 1 / 2 to 3 / 4 of the leaves of the whole plant show symptoms of infection;

[0051] Level 4: More than 3 / 4 of the leaves show symptoms of infection or the plant has died.

[0052] The calculation formula is as follows:

[0053] (1) Incidence rate = (number of diseased plants treated / total number of plants treated) × 100%;

[0054] (2) Disease index D = [∑ (number of diseased plants at each level × corresponding level) / (total number of plants × highest disease level)] × 100%;

[0055] (3) Control effect = [(disease index of control area - disease index of treated area) / disease index of control area] × 100%.

[0056] Results and analysis:

[0057] The control effects of each treatment on watermelon wilt in the sterilized soil test are shown in Table 1. Compared with treatment A, the incidence of treatments B and C1-C5 was reduced to varying degrees. Compared with treatment B, the incidence of treatment C1-5 was significantly reduced, and the control effect was significantly improved. The results showed that the treatment with bio-organic fertilizer had a better control effect on watermelon wilt; and different components in bio-organic fertilizer will have different degrees of influence on the control effect. Compared with treatment C2-5, bio-organic fertilizer with C1 formula can effectively reduce the incidence of watermelon wilt and improve the control effect.

[0058] Table 1: Control effect of bio-organic fertilizer on watermelon wilt

[0059] deal with Incidence % Control effect% A 65.0％ - B 55.0％ 22.4％ C1 27.5％ 68.5％ C2 40.0％ 50.8％ C3 45.0％ 36.4％ C4 42.5％ 45.0％ C5 42.5％ 42.8％

[0060] The stem diameter and plant height of each treatment in the sterilized soil test are shown in Table 2. The results showed that the application of biological organic fertilizer increased the stem diameter and plant height of watermelon plants.

[0061] Table 2: Effects of bio-organic fertilizer on watermelon growth characteristics

[0062]

[0063] Finally, it should be noted that the above examples are only some specific embodiments of the present invention. Obviously, the present invention is not limited to the above examples, and there are many variations. All variations that can be directly derived or associated with the content disclosed by a person skilled in the art are considered to be within the scope of protection of the present invention.

Claims

1. A polymyxa-type bio-organic fertilizer, characterized in that: The invention comprises the following ingredients in parts by weight: 5-15 parts of rice straw, 10-20 parts of walnut green peel, 35-45 parts of biogas residue, 2-8 parts of chicken manure, 2-8 parts of sheep manure, 3-7 parts of fermentation bacteria, 6-10 parts of Paenibacillus polymyxa fermentation liquid, 4-9 parts of humic acid, 2-6 parts of zinc sulfate and 2-6 parts of borax; the fermentation bacteria are EM bacteria.

2. A kind of Paenibacillus polymyxa type biological organic fertilizer according to claim 1, characterized in that, The invention comprises the following ingredients in parts by weight: 8-12 parts of rice straw, 14-16 parts of walnut green peel, 38-42 parts of biogas residue, 4-6 parts of chicken manure, 4-6 parts of sheep manure, 4-6 parts of fermentation bacteria, 7-9 parts of Paenibacillus polymyxa fermentation liquid, 6-8 parts of humic acid, 3-6 parts of zinc sulfate and 3-5 parts of borax.

3. A kind of Paenibacillus polymyxa type biological organic fertilizer according to claim 2, characterized in that, The invention comprises the following ingredients in parts by weight: 10 parts of rice straw, 15 parts of walnut green peel, 40 parts of biogas residue, 5 parts of chicken manure, 5 parts of sheep manure, 5 parts of fermentation bacteria, 8 parts of Paenibacillus polymyxa fermentation liquid, 7 parts of humic acid, 4 parts of zinc sulfate and 4 parts of borax.

4. A kind of polymyxa type biological organic fertilizer according to any one of claims 1-3, characterized in that, Prepared by solid-state aerobic fermentation.

5. A method of using a Paenibacillus polymyxa type bio-organic fertilizer according to any one of claims 1 to 3 for watermelon agricultural production.

6. Use of a Paenibacillus polymyxa type bio-organic fertilizer according to claim 5 for preventing and treating watermelon wilt.

7. Use of the Paenibacillus polymyxa type bio-organic fertilizer according to claim 5 for increasing the stem thickness and plant height of watermelon.

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