CN111394281A - Bacillus belgii BV03 microbial agent and preparation method and application thereof - Google Patents
Bacillus belgii BV03 microbial agent and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of microorganism application, in particular to a Bacillus belgii BV03 microbial inoculum and a preparation method thereof and application thereof in growth of solanaceae plants and/or cucurbitaceae plants, wherein the Bacillus belgii BV03 is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation address is No. 3 of West Luo No.1 Hospital of sunward area in Beijing, the preservation number is CGMCC NO.18936, the preservation date is 11 months and 11 days in 2019, and the Bacillus belgii velezensis is named in classification; the preparation method comprises obtaining seed liquid, fermenting the seed liquid to obtain fermentation liquid pre-preparation, and preparing into microbial inoculum; the growth promoting effect of the microbial inoculum in industrial seedling raising reaches 30%, the strong seedling index rises by more than 45%, the yield increasing effect of the microbial inoculum during transplanting is 48% and 34%, and the maturing rate exceeds 85%.
Description
Technical Field
The invention relates to the technical field of microorganism application, in particular to a bacillus beiLeisi BV03 microbial inoculum and a preparation method and application thereof.
Background
Since the 21 st century, with rapid economic growth, population growth and growing concern about food health, agricultural production areas are continuously expanded, and yields are increasing day by day, with little success. However, various problems have been increasingly highlighted in the development process. In order to guarantee the increase of the yield of agricultural products, particularly vegetables and fruits, farmers pursue economic benefits and increase the yield per mu, and the phenomena of applying chemical fertilizers without standard or exceeding the use standard are frequently seen. The agricultural products are damaged by the behaviors, so that the agricultural production cost is increased, and the benefit value of the agricultural products is reduced; in addition, the pollution problem of the agricultural environment caused by the residue of the excessive application of the chemical fertilizer is increasingly prominent, which directly causes the soil structure to be damaged and the drug resistance of pests, and some harmful substances, such as toxic compounds or heavy metals, even possibly enter the human body along with the food chain, thereby threatening the health of human beings. The appearance of these phenomena becomes a barrier in the development of agriculture and forestry production.
A large amount of mineral elements accumulate in soil, can lead to soil structure to suffer destruction, influence the growth and development of plant, and the microorganism can be through certain physiology transform mineral element's state, turns into the state that can supply the plant direct absorption to utilize. Therefore, in order to solve this problem, a novel functional microorganism replacement or partial replacement has been soughtResearch on the use of chemical fertilizers for agricultural product planting is of great interest. For example, some microorganisms can convert insoluble phosphorus into soluble phosphorus for direct absorption and utilization by plants, and the growth-promoting bacteria have a phosphorus dissolving function; in addition, some microorganisms can dissolve organic phosphorus and inorganic phosphorus, so that phosphorus in soil is utilized to a greater extent, and the phosphorus is prevented from depositing in the soil. Bacillus belgii is a bacillus from various ecological environments such as plant rhizosphere, soil and the like. Most of the Bacillus decibelicus that have been isolated from the plant rhizosphere can colonize the plant root by producing siderophins, IAA (indoleacetic acid) and NH3Growth-promoting substances play an important role in promoting the growth of plants. In the aspect of promoting plant growth, different strains are mostly adopted for compounding at present, so that the advantages of the strains can be complemented, and different functions of each strain can be fully exerted. However, because different strains have different environmental adaptability, culture and/or storage conditions, the functions of the strains in the compound microbial inoculum are often limited, and the efficacy is weakened after the compound microbial inoculum is applied.
The invention provides a Bacillus beiLeisi BV03 microbial inoculum, a preparation method thereof and application in growth of solanaceae plants and/or cucurbitaceae plants, aiming at improving yield of solanaceae plants and/or cucurbitaceae plants and ensuring product quality of the solanaceae plants and/or cucurbitaceae plants.
Disclosure of Invention
Aiming at the technical problems that the adaptability, the culture and/or storage conditions of different strains in the environment are different, the functions of all strains in a compound microbial inoculum are often limited, and the effect is weakened after application, the invention provides a Bacillus velezensis (Bacillus velezensis) BV03 microbial inoculum, a preparation method and application thereof.
In a first aspect, the invention provides a preparation method of a bacillus beilaisi BV03 microbial inoculum, and the preparation method comprises the following steps:
(1) inoculating the preserved Bacillus beilesiensis BV03 strain in liquid L B culture medium, culturing at 30 + -1 deg.C for 16h or to logarithmic growth phase to obtain strain for inoculationUsing a seed solution having a concentration of not less than 1.0 × 10 as a seed solution8cfu/mL；
Wherein the Bacillus belgii BV03 has been preserved in China general microbiological culture Collection center (CGMCC), the preservation address is No. 3 of Siro 1 of Beijing, Chaoyang, the preservation number is CGMCC NO.18936, the preservation date is 11 months and 11 days in 2019, and the Bacillus belgii Bacillus velezensis is classified and named;
(2) inoculating the seed liquid into a fermentation culture medium in an inoculation amount of 10-12% (volume percentage) to ensure that the initial concentration of thalli in a fermentation culture system is not lower than 1.0 × 107cfu/mL；
(3) Adding diatomite into a fermentation system according to the mass-to-volume ratio of 1: 4-1: 5, fully and uniformly mixing, and performing spray drying to obtain a fermentation liquid pre-preparation of Bacillus beiLensis BV 03;
(4) mixing the fermentation liquid pre-preparation, organosilicon BD-3077, sodium carboxymethylcellulose, chitosan, sodium lignosulfonate and coenzyme A, and pulverizing with jet mill to obtain Bacillus beijerinckii BV03 microbial inoculum with viable bacteria number not less than 2.0 × 1011cfu/g。
Further, the formula of the fermentation medium in the step (2) is as follows:
corn flour 2%, soybean meal 1%, yeast powder 0.3%, CaCl20.2％，MgSO4·7H2O 0.1％，K2HPO4·3H2O 0.15％，KH2PO40.2％，FeSO4·7H20.001 percent of O (mass volume ratio), the balance of water, pH 7.0-7.5 and 0.1MPa, sterilizing the mixture together with a fermentation tank at 115 ℃ for 60min, and naturally cooling the mixture to the normal temperature for use;
the culture conditions of the step (2) are as follows:
ventilating and maintaining the pressure of 0.05MPa at the temperature of 30 +/-1 ℃, keeping the dissolved oxygen content at 20-22%, rotating at the speed of 180-220 r/min, keeping the pH of the fermentation liquor at 7.0-7.5 in the fermentation process, measuring the total amount of the viable bacteria after culturing for 36 hours, wherein the total amount of the viable bacteria reaches 2.0 × 109cfu/m L, end of fermentation.
Further, the step (4) is:
fully mixing 92-95 parts of fermentation liquor prefabricated agent, 1-1.5 parts of organic silicon BD-3077, 1-1.5 parts of sodium carboxymethylcellulose, 1-1.5 parts of chitosan, 1-2 parts of sodium lignosulfonate and 1-1.5 parts of coenzyme A, and crushing the mixture to phi less than 45 mu m by using an air flow crusher to obtain the Bacillus belgii BV03 microbial inoculum, wherein the number of live bacteria in the microbial inoculum is not less than 2.0 × 1011cfu/g, the number of viable bacteria in the microbial inoculum is not less than 1.0 × 10 after half a year at room temperature11cfu/g。
In a second aspect, the invention provides a bacillus beilaisi BV03 microbial inoculum prepared by the preparation method.
Further, the dosage form of the bacillus beiLeisi BV03 microbial inoculum is wettable powder.
In a third aspect, the invention provides an application of the Bacillus beijerinckii BV03 microbial inoculum prepared by the preparation method in the growth of solanaceae plants and/or cucurbitaceae plants.
Further, when the application is industrial seedling raising, 2.0 × 10 is added11The Bacillus beilis BV03 microbial inoculum of cfu/g is administered at the dosage of 10.0 g/L, 5.0 g/L, 2.0 g/L, 2.0g 2.0 × 1011The wettable powder of the Bacillus beilesensis BV03 microbial inoculum of cfu/g has the best application effect according to the dosage of 2.0 g/L.
Further, after the solanaceae plant and/or the cucurbitaceae plant are transplanted, 2.0 × 10 is added11The Bacillus belgii BV03 microbial inoculum of cfu/g is applied according to the dosage of 5.0 kg/mu, 2.0 kg/mu and 1.0 kg/mu, and 2.0 × 1011The application effect of cfu/g wettable powder of the bacillus beilesensis BV03 microbial inoculum is optimal according to the dosage of 2.0 kg/mu.
The beneficial effect of the invention is that,
compared with the chemical fertilizer in the market, the Bacillus belgii BV03 microbial inoculum provided by the invention has good effect of promoting plant growth, the growth promoting effect on solanaceae plants and/or cucurbitaceae plants in industrial seedling culture can reach about 30%, and the strong seedling index is increased by over 45%; the transplanted solanaceae plant and/or cucurbitaceae plant are additionally applied, the growth promoting effect is 48% and 34% respectively, and the maturing rate is over 85%;
the Bacillus belgii BV03 microbial inoculum provided by the invention is simple in preparation method, easy to operate and has great application value for cultivation of economic plants.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1 Bacillus belgii BV03 bacterial agent
A Bacillus belgii BV03 microbial inoculum is wettable powder containing Bacillus belgii BV03, the Bacillus belgii BV03 is stored in China general microbiological culture Collection center (CGMCC), the storage address is No. 3 of No.1 Hospital, West Luo, North Cheng, the sunny region in Beijing, the storage number is CGMCC NO.18936, the storage date is 11 months and 11 days in 2019, and the Bacillus belgii velezensis is named in classification;
the preparation method of the microbial inoculum comprises the following steps:
(1) inoculating the preserved Bacillus beilesiensis BV03 strain in liquid L B culture medium, culturing at 30 + -1 deg.C for 16h or to logarithmic growth phase to obtain seed liquid with concentration not lower than 1.0 × 108cfu/mL；
(2) Inoculating the seed liquid into a fermentation culture medium for culture in an inoculation amount of 10-12% (volume percentage), so that the initial concentration of thalli in a fermentation culture system is not lower than 1.0 × 107cfu/mL；
Wherein, the formula of the fermentation medium is as follows:
corn flour 2%, soybean meal 1%, yeast powder 0.3%, CaCl20.2％，MgSO4·7H2O 0.1％，K2HPO4·3H2O 0.15％，KH2PO40.2％，FeSO4·7H20.001 percent of O (mass volume ratio), the balance of water, pH 7.0-7.5 and 0.1MPa, sterilizing the mixture together with a fermentation tank at 115 ℃ for 60min, and naturally cooling the mixture to the normal temperature for use;
the culture conditions of the step (2) are as follows:
ventilating and maintaining the pressure of 0.05MPa at the temperature of 30 +/-1 ℃, keeping the dissolved oxygen content at 20-22%, rotating at the speed of 180-220 r/min, keeping the pH of the fermentation liquor at 7.0-7.5 in the fermentation process, measuring the total amount of the viable bacteria after culturing for 36 hours, wherein the total amount of the viable bacteria reaches 2.0 × 109cfu/m L, ending fermentation;
(3) adding diatomite into a fermentation system according to the mass-to-volume ratio of 1: 4-1: 5, fully and uniformly mixing, and performing spray drying to obtain a fermentation liquid pre-preparation of Bacillus beiLensis BV 03;
(4) fully mixing 92-95 parts of fermentation liquor prefabricated agent, 1-1.5 parts of organic silicon BD-3077, 1-1.5 parts of sodium carboxymethylcellulose, 1-1.5 parts of chitosan, 1-2 parts of sodium lignosulfonate and 1-1.5 parts of coenzyme A, and crushing the mixture to phi less than 45 mu m by using an air flow crusher to obtain the Bacillus belgii BV03 microbial inoculum, wherein the number of live bacteria in the microbial inoculum is not less than 2.0 × 1011cfu/g, the number of viable bacteria in the microbial inoculum is not less than 1.0 × 10 after half a year at room temperature11cfu/g。
Example 2 growth promoting effect of Bacillus belgii BV03 microbial inoculum in industrialized seedling of Capsici fructus
The treatment group comprises adding Bacillus beilesensis BV03 prepared in example 1 into a seedling culture medium according to the weight of 10 g/L, 5 g/L and 2 g/L, mixing uniformly to obtain culture mediums I, II and III, sowing according to a conventional method, managing No. 6 Capsici fructus according to a conventional method,
the No. 6 Chinese prickly ash cultivation group directly using the seedling culture substrate as the cultivation substrate is used as a negative control,
the cultivation group of Zanthoxylum piperitum No. 6 with nitrogen phosphorus potassium compound fertilizer (nitrogen phosphorus potassium 15:15:15) added in the cultivation substrate is used as positive control (usage amount is 60 g/m)2)，
Each group was treated with 36 plants, which were repeated 3 times.
And observing and counting the plant height, stem thickness, the whole plant dry weight and the strong seedling index after 21 days, wherein the calculation formula of the strong seedling index is that the strong seedling index is stem thickness (cm)/plant height (cm) × the whole plant dry weight (g), and the test results are shown in table 1.
TABLE 1 growth promoting effect of Bacillus belgii BV03 on industrial seedling culture of Capsici fructus
| Negative control | Positive control | Cultivation substrate I | Culture medium II | Culture medium III | |
| Plant height (cm) | 15.43±0.67 | 17.12±1.01 | 18.76±1.89 | 18.21±2.17 | 17.14±1.38 |
| Stem diameter (mm) | 1.87±0.11 | 1.98±0.06 | 2.22±0.11 | 2.17±0.10 | 2.19±0.08 |
| Whole plant Dry weight (g) | 0.78±0.13 | 0.95±0.11 | 1.03±0.17 | 1.01±0.18 | 1.02±0.13 |
| Seedling strengthening index | 0.009±0.0011 | 0.011±0.0018 | 0.012±0.0012 | 0.012±0.0011 | 0.013±0.0015 |
The results in table 1 show that compared with negative control, the plant height, stem thickness, whole plant dry weight and strong seedling index of the medium pepper No. 6 which is industrially cultivated by using the culture substrates I, II and III of the Bacillus beleisi BV03 microbial inoculum are obviously increased, wherein the culture substrate III with the addition of 2 g/L has the most obvious effect, the biomass accumulation is increased by more than 30 percent (30.77 percent), and the strong seedling index is increased by nearly 45 percent (44.44 percent), and in addition, the growth promoting effect of the medium pepper No. 6 using the Bacillus beleisi BV03 microbial inoculum is obviously superior to or equal to that of a nitrogen-phosphorus-potassium compound fertilizer (nitrogen: phosphorus: potassium is 15:15), which shows that the Bacillus beleisi microbial inoculum has a good effect of promoting plant growth in the industrial cultivation of solanaceae plants.
Example 3 influence of Bacillus belgii BV03 bacterial agent on content of available nitrogen, available phosphorus and available potassium in culture medium in industrial seedling raising of capsicum
Example 2 after sowing for 22 days, the culture mediums I, II and III of the treatment group and the culture mediums of the negative control group were subjected to detection of available nitrogen, available phosphorus and available potassium contents by referring to "soil agro-chemical analysis and environmental monitoring" (Yangxicai, Wangcheng, Renheng, China land Press, 2008) and detection of available nitrogen by alkaline hydrolysis diffusion methodThe available phosphorus detection adopts 0.5M sodium bicarbonate leaching-molybdenum-antimony colorimetric method and the available potassium detection adopts NH4OAc leaching-flame photometry. The test results are shown in Table 2.
TABLE 2 influence of Bacillus belgii BV03 inoculum on available nitrogen, available phosphorus and available potassium content in the cultivation substrate
| Available nitrogen (mg/kg) | Available phosphorus (mg/kg) | Effective potassium (mg/kg) | |
| Cultivation substrate I | 1041.27±3.01 | 239.57±2.98 | 832.44±2.71 |
| Culture medium II | 1024.17±2.87 | 232.01±1.74 | 817.11±2.13 |
| Culture medium III | 998.93±3.17 | 223.35±2.56 | 814.16±1.74 |
| Negative control | 148.92±5.57 | 106.57±7.78 | 110.41±5.32 |
The results in table 2 show that, compared with the negative control, the contents of available nitrogen, available phosphorus and available potassium of the culture substrates I, II and III using the Bacillus belief agent are all significantly improved, and the contents of the available nitrogen, the available phosphorus and the available potassium of the culture substrates I, II and III are similar and have no obvious difference. The content of available nitrogen, available phosphorus and available potassium of the culture medium I is respectively increased by 703.38%, 225.47% and 756.36%; the content of available nitrogen, available phosphorus and available potassium of the culture medium II is respectively increased by 691.90%, 218.88% and 742.73%; the content of available nitrogen, available phosphorus and available potassium of the culture medium III is respectively increased by 679.01%, 210.38% and 740.00%; this indicates that the Bacillus belgii fungicide realizes the effect of promoting the growth of solanaceae plants by increasing the contents of available nitrogen, available phosphorus and available potassium in the culture medium.
Example 4 growth promoting effect of Bacillus belgii BV03 microbial inoculum in factory seedling culture of cucumber
The treatment groups are that the Bacillus beilesensis BV03 microbial inoculum prepared in the example 1 is added into a seedling culture substrate according to the proportion of 10 g/L, 5 g/L and 2 g/L, culture substrates I, II and III are obtained after the mixture is fully mixed, the Yuanfengyuan No. 6 is sown and managed according to the conventional method, the Yuanfengyuan No. 6 culture group which directly uses the seedling culture substrate as the culture substrate is used as a negative control, and the Yuanfengyuan No. 6 culture group which adds nitrogen-phosphorus-potassium compound fertilizers (nitrogen: phosphorus: potassium: 1: 0.4: 1.6) into the culture substrate is used as a positive control (the usage amount is 60 g/m)2) Each group was treated with 36 plants, and the treatment was repeated 3 times.
After 15 days, observing and counting the plant height, stem thickness, the dry weight of the whole plant and the strong seedling index, wherein the calculation formula of the strong seedling index is that the strong seedling index is stem thickness (cm)/plant height (em) × dry weight (g), and the test results are shown in table 3.
TABLE 3 growth promoting effect of Bacillus belgii BV03 microbial inoculum in industrial seedling culture of cucumber
| Negative control | Positive control | Cultivation substrate I | Culture medium II | Culture medium III | |
| Plant height (cm) | 14.43±0.62 | 15.12+1.27 | 15.76±1.32 | 15.21±1.10 | 15.14±1.03 |
| Stem diameter (mm) | 1.72±0.11 | 1.95±0.13 | 2.01±0.10 | 2.07±0.14 | 2.16±0.07 |
| Whole plant Dry weight (g) | 0.92±0.11 | 1.30±0.15 | 1.36±0.14 | 1.34±0.13 | 1.33±0.11 |
| Seedling strengthening index | 0.011±0.0021 | 0.017±0.0018 | 0.017±0.0015 | 0.018±0.0010 | 0.019±0.0012 |
The results show that compared with negative control, the plant height, stem thickness, whole plant dry weight and seedling strengthening index of Yuanfengyuan No. 6 for industrial seedling raising by using the culture substrates I, II and III of the Bacillus belgii fungicide are remarkably increased, wherein the effect of the culture substrate III with the addition amount of 2 g/L is most remarkable, the biomass accumulation and the growth of the culture substrate III reach 44.56%, and the seedling strengthening index is increased by 72.72%.
Example 5 application of Bacillus belgii BV03 microbial inoculum in post-pepper-transplant field production
The test object is that the seedlings of the Chinese prickly ash No. 6 treated by the culture medium III containing the Bacillus belgii BV03 microbial inoculum (2 g/L) in the example 2 are taken as seedlings, and the growth vigor of the seedlings is consistent;
test site: a facility land of Dongchun in Jinan City of Shandong province;
test time/climate: in 2019, in 5 months, the average temperature is 25 ℃ in the test period, and the climate is dry;
and (3) test treatment: before facility land soil is planted, weeds are thoroughly removed, land is prepared, fertilizer is applied to the decomposed organic fertilizer by 5000 kg/mu, and 5 treatments including Bacillus belgii BV03 treatment I (wettable powder is 5.0 kg/mu), Bacillus belgii BV03 treatment II (wettable powder is 2.0 kg/mu), Bacillus belgii BV03 treatment III (wettable powder is 1.0 kg/mu), a special compound fertilizer group for hot pepper (calcium superphosphate is 50.0 kg/mu, ammonium bicarbonate is 50.0 kg/mu, potassium sulfate is 25.0 kg/mu and calcium fertilizer is 50.0 kg/mu) and clear water are respectively arranged;
the test method comprises the steps of ploughing 30-40 cm after uniform spreading, curing soil, leveling land, making furrows (connecting ditches) according to 150cm, processing 0.06 mu each time, repeating for 3 times, keeping the area of 15 cells to be 0.9 mu each time, reserving 2 rows of protection lines for each cell, after slow seedling planting, carrying out field cultivation management according to local optimal measures, fixing 5 points for sampling in each cell by adopting a double diagonal line in a growth period, investigating 35 plants each time, recording the growth condition of an investigation object, after fructification and harvesting, counting the fruit setting rate, counting the yield and converting the yield into the yield per mu, and obtaining the fruit setting rate (fruit number/percentage) × 100% according to a fruit setting rate/flowering technical regulation method and requirements specified in the national standards of the people's republic of China (NY/T5105-2002) in the field application method of the compound fertilizer.
TABLE 4 influence and Effect of Bacillus belgii BV03 inoculum on post-pepper-transplant field production
As can be seen from Table 4, compared with the clear water control, the field growth of the peppers using the Bacillus belgii BV03 fungicide is good, the fruit setting rate and the yield per mu are improved, the pepper field growth is obviously superior to that of a special compound fertilizer group for the peppers, and the yield is respectively increased by 46.58%, 48.27% and 35.21%; the fruit setting rate and the yield per mu of the Bacillus beilis BV03 processed II are better than those of the Bacillus beilis BV03 processed I and the Bacillus beilis BV03 processed III, the fruit setting rate reaches 85 percent, and the yield increase reaches 48 percent. The result shows that the Bacillus belgii BV03 microbial inoculum has the function of promoting the growth and the yield increase of the peppers, and the best effect is achieved in the promotion of the pepper production when the dosage of the Bacillus belgii BV03 microbial inoculum is 2.0 kg/mu of wettable powder.
Example 6 application of Bacillus belgii BV03 microbial inoculum in field production after cucumber transplantation
The test object is that the Yuanfengyuan No. 6 seedling treated by the culture medium III containing the Bacillus belgii BV03 microbial inoculum (2 g/L) in the example 4 serves as a seedling, and the growth vigor of the seedling is consistent;
test site: a green house for cucumber of age 2 in Dongcun city of Jinan, Shandong province;
test time/climate: 8 months in 2019, the average temperature is 28 ℃ and the soil humidity is 60-90% in the test period;
and (3) test treatment: before facility land soil is planted, weeds are thoroughly removed, land is prepared, 6000 kg/mu of decomposed organic fertilizer is fertilized, 5 treatments are respectively set for treating I (5.0 kg/mu of wettable powder) by using Bacillus beilesensis BV03, II (2.0 kg/mu of wettable powder) by using Bacillus beilesensis BV03, III (1.0 kg/mu of wettable powder) by using Bacillus beilesensis BV03, a special compound fertilizer group for cucumber (30.0 kg/mu of calcium superphosphate, 60.0 kg/mu of urea and 2.0 kg/mu of potassium sulfate) and clear water;
the test method comprises the steps of ploughing 30-40 cm after uniform spreading, curing soil, ploughing the furrows after land leveling, planting the furrows with a dark water method, wherein the width of each furrow is 1.2m, the height of each furrow is more than 15cm, the area of each furrow is 0.75 mu according to small row spacing of 40cm, large row spacing of 80cm and plant spacing of 25-30 cm, the furrow is processed by 3 times, 2 rows of protection rows are reserved in each furrow, after seedling revival and field planting, field cultivation management is carried out according to local optimal measures, in a growth period, each furrow is fixed by double diagonal lines for 5 points for sampling, 35 plants are planted at each point, the growth condition of an investigation object is recorded, after fruiting and harvesting, the average fruit weight, the fruit setting rate and the per mu statistical yield are counted and converted into the per mu yield, the fruit setting rate calculation formula is that the fruit setting rate (%) (real number per fruit number) is × 100%, the application method refers to the national and national standards (NY/T5-2002) of Chinese people and the field technical requirements and the production rules of the production method.
TABLE 5 influence and Effect of Bacillus belgii BV03 inoculum on post-cucumber-transplant field production
As can be seen from Table 5, compared with the clear water control, the cucumber field using Bacillus belgii BV03 microbial inoculum has good growth vigor, the fruit setting rate and the yield per mu are both obviously improved, the effect is superior to or close to that of the special compound fertilizer group, and the yields per mu are respectively increased by 32.84%, 34.35% and 27.14%; particularly, the promotion effect of the Bacillus beiLensis BV03 treatment II on the field production of cucumbers is better than that of the Bacillus beiLensis BV03 treatment I and that of the Bacillus beiLensis BV03 treatment III, the average fruit weight reaches 230g, the fruit setting rate is over 86 percent, and the yield per mu is increased by 34 percent. The result shows that the Bacillus belgii BV03 microbial inoculum has the functions of promoting the growth and increasing the yield of cucumbers, and the best effect is achieved in promoting the cucumber production when the dosage of the Bacillus belgii BV03 microbial inoculum is 2.0 kg/mu of wettable powder.
Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. The preparation method of the Bacillus belgii BV03 microbial inoculum is characterized by comprising the following steps:
(1) inoculating the preserved Bacillus beilesiensis BV03 strain in liquid L B culture medium, culturing at 30 + -1 deg.C for 16h or to logarithmic growth phase to obtain seed liquid with concentration not lower than 1.0 × 108cfu/mL；
Wherein the Bacillus belgii BV03 has been preserved in China general microbiological culture Collection center, the preservation address is No. 3 of West Lu No.1 of the morning-Yang district, Beijing, the preservation number is CGMCC NO.18936, the preservation date is 11 months and 11 days in 2019, and the Bacillus belgii Bacillus velezensis is classified and named;
(2) inoculating the seed liquid to a fermentation culture medium in an inoculation amount of 10-12%, so that bacteria in a fermentation culture system are culturedInitial concentration of the product is not less than 1.0 × 107cfu/mL；
(3) Adding diatomite into a fermentation system according to the mass-to-volume ratio of 1: 4-1: 5, fully and uniformly mixing, and performing spray drying to obtain a fermentation liquid pre-preparation of Bacillus beiLensis BV 03;
(4) mixing the fermentation liquid pre-preparation, organosilicon BD-3077, sodium carboxymethylcellulose, chitosan, sodium lignosulfonate and coenzyme A, and pulverizing with jet mill to obtain Bacillus beijerinckii BV03 microbial inoculum with viable bacteria number not less than 2.0 × 1011cfu/g。
2. The method of claim 1, wherein the fermentation medium of step (2) has a formulation of:
corn flour 2%, soybean meal 1%, yeast powder 0.3%, CaCl20.2％，MgSO4·7H2O 0.1％，K2HPO4·3H2O0.15％，KH2PO40.2％，FeSO4·7H20.001% of O and the balance of water, the pH value is 7.0-7.5, the pressure is 0.1MPa, the materials and a fermentation tank are sterilized at 115 ℃ for 60min, and the materials are naturally cooled to be used under the condition of normal temperature;
the culture conditions of the step (2) are as follows:
ventilating and maintaining the pressure of 0.05MPa at the temperature of 30 +/-1 ℃, keeping the dissolved oxygen content at 20-22%, rotating at the speed of 180-220 r/min, keeping the pH of the fermentation liquor at 7.0-7.5 in the fermentation process, measuring the total amount of the viable bacteria after culturing for 36 hours, wherein the total amount of the viable bacteria reaches 2.0 × 109cfu/m L, end of fermentation.
3. The method of claim 1, wherein the step (4) is:
fully mixing 92-95 parts of fermentation liquor prefabricated agent, 1-1.5 parts of organic silicon BD-3077, 1-1.5 parts of sodium carboxymethylcellulose, 1-1.5 parts of chitosan, 1-2 parts of sodium lignosulfonate and 1-1.5 parts of coenzyme A, and crushing the mixture to phi less than 45 mu m by using an air flow crusher to obtain the Bacillus belgii BV03 microbial inoculum, wherein the number of live bacteria in the microbial inoculum is not less than 2.0 × 1011cfu/g, half year after strain at room temperatureThe number of viable bacteria in the preparation is not less than 1.0 × 1011cfu/g。
4. A Bacillus belgii BV03 bacterial preparation prepared by the process according to claim 1.
5. The microbial inoculum according to claim 4, in the form of wettable powder.
6. Use of the bacterial agent of claim 4 for the growth of solanaceae and/or cucurbitaceae plants.
7. The application of claim 6, wherein the application is 2.0 × 10 in industrial seedling raising11The Bacillus belgii BV03 microbial inoculum of cfu/g is applied according to the dosage of 10.0 g/L, 5.0 g/L and 2.0 g/L.
8. The application of claim 7, wherein the application is 2.0 × 10 in industrial seedling raising11cfu/g of Bacillus belgii BV03 microbial inoculum is applied according to the dosage of 2.0 g/L, and the dosage form of the microbial inoculum is wettable powder.
9. The use according to claim 6, wherein 2.0 × 10 is added after transplanting solanaceae and/or cucurbitaceae plants11The Bacillus belgii BV03 microbial inoculum of cfu/g is applied according to the dosage of 5.0 kg/mu, 2.0 kg/mu and 1.0 kg/mu.
10. The use according to claim 9, wherein 2.0 × 10 is added after transplanting solanaceae and/or cucurbitaceae plants11The bacterial agent of the Bacillus beilesensis BV03 of cfu/g is applied according to the dosage of 2.0 kg/mu, and the dosage form of the bacterial agent is wettable powder.
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| CN113897316B (en) * | 2021-10-29 | 2023-09-22 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Bacillus licheniformis BLc06, functional melon and fruit seedling biological matrix prepared from same and application |
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